Contents
• Organizing Committee ·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·4
• Floor Plan ·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·5
• Schedule ·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·6
• Program ·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·7
• 발표연사자료 ·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·11
Session 1_Drug Discovery & Pharma ·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·13
Session 2_Plant Metabolomics ·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·23
Session 3_Microbial Metabolomics ·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·33
Session 4_Data Analysis & Bioinformatics ·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·41
Session 7_Human Diseases ·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·51
Session 8_Environmental Toxicity ·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·61
Session 9_Food & Nutrition ·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·71
• Young Scientists ·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·81
Young Scientists 1 ·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·83
Young Scientists 2 ·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·93
• 포스터 초록 ·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·103
Poster 1. Human and Animal Metabolomics ·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·105
Poster 2. Plant Metabolomics ·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·141
Poster 3. Food and Microbial Metabolomics ·
·
·
·
·
·
·
·
·
·
·
·
·
·
·161
Poster 4. Environmental Metabolomics ·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·
·173
Organizing Committee
￭ ￭ ￭ 임원명단
￭ ￭ ￭ 운영위원명단
회 장
운 영 위 원 장
정봉철
한국과학기술연구원
부회장
한국식품연구원
고 문
이정애
한국과학기술연구원
황금숙
한국기초과학지원연구원
김경헌
고려대학교
김현진
경상대학교
최형균
중앙대학교
윤영란
경북대학교
류광현
경북대학교
재 무 위 원 장
유장렬
한국생명공학연구원
감 사
학 술 위 원 장
안여환
영인프런티어㈜
이 사
|
건국대학교
총 무 위 원 장
권대영
2
이충환
김경헌
고려대학교
김석원
한국생명공학연구원
김영석
이화여자대학교
김재광
농촌진흥청
김현진
경상대학교
류광현
경북대학교
백남인
경희대학교
윤영란
경북대학교
이동호
고려대학교
이충환
건국대학교
이백석
CJ 제일제당 바이오연구소
이정애
한국과학기술연구원
장인진
서울대학교
최형균
중앙대학교
황금숙
한국기초과학지원연구원
국제협력위원장
홍보조직위원장
기 획 위 원 장
Floor Plan
2F
2013. 4. 4 (목) - 5 (금)
The-K 서울호텔 2층 (가야금홀)
|
3
Schedule
April 4 [Thur]
시 간
가 야 금 홀 (2층 )
거 문 고 홀 (3층 )
08:00-09:00
등 록 (2층 등 록 대 )
09:05-09:15
개 회 사 (가 야 금 홀 )
09:15-10:35
Session 1 (Drug Discovery & Pharma) - 가 야 금 홀
Coffee Break
10:35-10:50
10:50-12:10
Session 2 (Plant Metabolomics) - 가 야 금 홀
12:20-12:50
Luncheon Seminar 2 : Sponsored by Scinco & Thermo Fisher Scienific - 가 야 금 홀
13:40-14:40
Session 3 (Microbial Metabolomics) - 가 야 금 홀
14:40-15:00
General Board Meeting - 가 야 금 홀
15:00-15:40
Poster Presentation : (with Coffee Break) - 가 야금 홀
15:40-17:00
Session 4 (Data Analysis & Bioinformatics) - 가 야 금 홀
17:00-18:00
Session 5 (Young Scientists) - 가 야 금 홀
18:00-20:00
Welcome Reception - 거 문 고 홀
April 5 [Fri]
시 간
등 록 (2층 등 록 대 )
09:30-10:30
Session 6 (Young Scientists) - 가 야 금 홀
Coffee Break - 가 야 금홀
10:50-12:10
Session 7 (Human Diseases) - 가 야 금 홀
12:20-12:50
Luncheon Seminar 2 : (Sponsored by AB SCIEX Korea) -가 야 금 홀
13:15-13:30
|
거 문 고 홀 (3층 )
08:00-09:30
10:30-10:50
4
가 야 금 홀 (2층 )
Coffee Break - 가 야 금홀
13:30-14:50
Session 8 (Environmental Toxicity) - 가 야 금 홀
15:00-15:30
Tea Time Seminar : (Sponsored by 영 인 프 런 티 어 ) - 가 야 금 홀
15:40-17:00
Session 9 (Food & Nutrition) - 가 야 금 홀
17:00-17:30
Award Ceremony and Closing Remarks - 가 야 금 홀
후원기업 감사장 / 우수 포스터 시상 / 행운권 추첨
Program
￭ ￭ ￭ April 4 (Thur)
S1
Drug Discovery & Pharma S1-1
09:15 - 09:35
09:15-10:35
Chair: 김동현 (인제대학교) / 장인진 (서울대학교)
Metabolism of botanical drug: metabolic pathways of iridoid glucosides
이혜숙 (가톨릭대학교)
S1-2
09:35 - 09:55
Lipidomic profiling after statin drug treatment
정병화 (한국과학기술연구원)
S1-3
09:55 - 10:15
ER stress and sphingolipid metabolism
박태식 (가천대학교)
S1-4
10:15 - 10:35
Metabolic analysis of heavy metal toxicity using NMR techniques
김석만 (부산대학교)
S2
Plant Metabolomics
S2-1
10:50 - 11:10
10:50-12:10
Chair: 김영식 (서울대학교) / 최형균 (중앙대학교)
Biochemical genetics of brassinosteroid metabolic pathways
최성화 (서울대학교)
S2-2
11:10 - 11:30
DART-MS Snapshot on natural products and plant metabolomics
장영표 (경희대학교)
S2-3
11:30 - 11:50
A simple and rapid prediction system by metabolic fingerprinting of Fourier transform
infrared spectroscopy
김석원 (한국생명공학연구원)
S2-4
11:50 - 12:10
Prediction of bioactivity and active constituents for botanical drugs
성상현 (서울대학교)
|
5
S3
Microbial Metabolomics
S3-1
13:40 - 14:00
13:40-14:40
Chair: 성문희 (국민대학교) / 임용현 (한국표준과학연구원)
Global optimization of microbial systems and process for biochemical production
박용철 (국민대학교)
S3-2
14:00 - 14:20
System response of metabolic networks in Chlamydomonas reinhardtii to temporal
and degree-wise nutritional challenge
이도엽 (국민대학교)
S3-3
14:20 - 14:40
Metabolic engineering of industrial microorganisms based on metabolomics
이백석 (CJ 바이오 연구소)
S4
Data Analysis & Bioinformatics
S4-1
15:40 - 16:00
15:40-17:00
Chair: 서진호 (서울대학교) / 김경헌 (고려대학교)
Exploring the human diseasome
고광일 (고려대학교)
S4-2
16:00 - 16:20
Model-based genome design for the development of viral vaccine platforms
임광일 (숙명여자대학교)
S4-3
16:20 - 16:40
Application of systems metabolomics to cell factories
이동엽 (Nat. Univ. of Singapore)
S4-4
16:40 - 17:00
Adaptive laboratory evolution of Escherichia coli - harnessing the power of
genome-scale sciences for metabolic network evolution
이대희 (한국생명공학연구원)
S5
Young Scientists
S5-1
17:00 - 17:15
17:00-18:00
Chair: 오세량 (한국생명공학연구원)
Yeast metabolome sampling methodology for metabolomics and metabolic engineering
김수아 (고려대학교)
S5-2
17:15 - 17:30
Classification of black raspberry fruits at different ripening stages by NMR and multivariate
statistical analysis and effect of antiinflammatory and antioxidant activities
양승옥 (농촌진흥청)
6
|
S5-3
17:30 - 17:45
NMR-based metabolite profiling for early detection and non-invasive diagnosis of gastric
cancer
정지연 (한국기초과학지원연구원)
S5-4
17:45 - 18:00
Evaluation of CYP2D6 allele activity through metabolomic profile using liquid
chromatography-mass spectrometry
서정주 (경북대학교)
￭ ￭ ￭ April 5 (Fri)
S6
Young Scientists
S6-1
09:30 – 09:45
09:30-10:30
Chair: 김윤곤 (숭실대학교)
Evaluation of endogenous metabolic markers of CYP3A activity using metabolic profiling
and midazolam pharmacokinetics
신광희 (서울대학교)
S6-2
09:45 – 10:00
Patterns of gene and metabolite define the effects of extracellular osmolality on kidney
collecting duct
최효정 (경북대학교)
S6-3
10:00 – 10:15
Comprehensive analysis of central carbon metabolites and
metabolic flux analysis in yeast
13
C-isotope dynamics for
정준영 (고려대학교)
S6-4
10:15 – 10:30
LC-MS based metabolomics approach for natural products screening and identification
이종석 (경기바이오센터)
S7
Human Diseases
S7-1
10:50 - 11:10
10:50-12:10
Chair: 박민수 (세브란스병원) / 황금숙 (한국기초과학지원연구원)
Metabolomics in the kidney diseases
권태환 (경북대학교)
S7-2
11:10 - 11:30
Reverse of renal cell carcinoma by blocking autophagy
김수열 (국립암센터)
S7-3
11:30 - 11:50
Biochemical roles of endocrine and neurologic systems on the hormonal dependent diseases
최만호 (한국과학기술연구원)
|
7
S7-4
11:50 - 12:10
Application of imaging mass spectrometry of phospholipid metabolites for
biomarker identification
김광표 (건국대학교)
S8
Environmental Toxicity
S8-1
13:30 - 13:50
13:30-14:50
Chair: 장윤석 (포항공과대학교) / 표희수 (한국과학기술연구원)
Circulating serum dioxin-like compound-induced mitochondrial dysfunction and diabetes
pandemic
김영미 (경희대학교)
S8-2
13:50 - 14:10
Polybromodiphenyl ethers (PBDEs) profiling in dietary intake and pentabromodiphenyl
ether exposed rat
서정주 (한국기초과학지원연구원)
S8-3
14:10 - 14:30
Persistent organic pollutants and obesity-related metabolic dysfunction
이덕희 (경북대학교)
S8-4
14:30 - 14:50
Urinary bisphenol-A concentration and its association with estrogen metabolism
in Korean adults based on environmental metabolomics
이정애 (한국과학기술연구원)
S9
Food & Nutrition
S9-1
15:40 - 16:00
15:40-17:00
Chair: 권대영 (한국식품연구원) / 윤정한 (한림대학교)
Metabolomic profiling as a useful tool for diagnosis and treatment of chronic disease
김오연 (동아대학교)
S9-2
16:00 - 16:20
Determination of key volatile and non-volatile components related to beef flavor in
glutathione Maillard reaction products using metabolomic approach
김영석 (이화여자대학교)
S9-3
16:20 - 16:40
Metabolomic analysis of natural products: Age discrimination of Panax ginseng
이동호 (고려대학교)
S9-4
16:40 - 17:00
The role of microRNAs in metabolic diseases and the involvement of food components
안지윤 (한국식품연구원)
8
|
발표연사 자료
Session 1
Drug Discovery & Pharma
Chair: 김동현_인제대학교 / 장인진_서울대학교
|
1
이 혜 숙 _Hye Suk Lee, 가톨릭대학교 약학대학
Office: 02-2164-4061 / C.P.: 010-6435-6817
E-mail: sianalee@catholic.ac.kr
학력/경력
1980.3-1984.2
1984.3-1986.2
1986.3-1989.8
1986.3-1995.8
1990.6-1991.6
1995.9-2010.12
2003.9-2005.8
2007.1-2008.12
2011.1-현재
2012.6-2014.05
성균관대학교 약학 약학사
성균관대학교 분석약학 약학석사
성균관대학교 분석약학 약학박사
한국화학연구원 안전성센터 연구원/선임연구원
일본 Showa대학교 약학부 박사후연구원
원광대학교 약학대학 조교수/부교수/교수
원광대학교 약학대학 학장
원광대학교 산학협력단 부단장
가톨릭대학교 약학대학 교수
한국연구재단 기초연구본부 의약학단 전문위원
주요연구실적
• H.Y. Ji, H.R. Lee, S.R. Im, J.H. Kim, H.S. Lee, Effect of efavirenz on UDP-glucuronosyltransferase 1A1, 1A4,
1A6, and 1A9 activities in human liver microsomes. Molecules 17(1), 851-860
• H.Y. Ji, K.H. Liu, J.H. Jeong, D.Y. Lee, H.J. Shim, M. Son, H.S. Lee, Effect of a new prokinetic agent DA-9701
formulated with Corydalis Tuber and Pharbitidis Semen on cytochrome P450 and UDP-glucuronosyltransferase
enzyme activities in human liver microsomes. Evid Based Complement Alternat Med 2012:650718 (2012)
• H.Y. Ji, H.R. Lee, J.H. Kim, K.H. Kim, K.R. Lee, H.J. Shim, M. Son, H.S. Lee, In vitro metabolism of corydaline
in human liver microsomes and hepatocytes using liquid chromatography-ion trap mass spectrometry. J Sep
Sci 35(9), 1102-1109 (2012)
• Y.Y. Cho, M.H. Lee, C.J. Lee, K. Yao, H.S. Lee, A.M. Bode, Z. Dong, RSK2 as a key regulator in human
skin cancer. Carcinogenesis 33, 2529-2537 (2012) 외 SCI논문 145편
14
|
S1-1
Metabolism of botanical drug: metabolic pathways
of iridoid glucosides
Hye Suk Lee
Drug Metabolism & Bioanalysis Laboratory, College of Pharmacy, The Catholic University of Korea,
Bucheon 420-743, Korea
Email: sianalee@catholic.ac.kr
The market of botanical therapeutics is growing and the development has been increasingly received
attention. US FDA has approved an orally administered botanical, Fulyzag for use in treating HIV-related
diarrhea on 31, December, 2012 and a topical containing green tea (Veregen) as the new drug.
Pharmacokinetic studies of botanical drugs should address the followings in order to establish the
pharmacological basis for the efficacy of botanical drugs: (i) bioavailability to assess to what degree and
how fast compounds are absorbed after administration of botanical drugs, (ii) elucidation of metabolic
pathways of active compounds, (iii) assessment of elimination routes and their kinetics, and (iv) interactions
of botanical drugs with synthetically derived drug products. The bioavailability of herbal compounds is
associated with many presystemic processes such as solubility in gastrointestinal fluid, membrane
permeability, degradation in the gastrointestinal tract, transporter-mediated intestinal efflux, P-glycoprotein,
presystemic gut wall metabolism, and presystemic hepatic metabolism.
The metabolism study of botanical drug can (i) assessment of quantitative content of main components,
(ii) identification of components that can be absorbed from gastrointestinal (GI) tract, (iii) metabolic
stability, metabolic profile and characterization of metabolic enzymes in GI tract (intestinal bacteria, acid, or
intestinal metabolic enzymes) and liver (hepatic metabolic enzymes), (iv) effect of botanical drug
components and/or their metabolites on intestinal or hepatic metabolic enzymes.
Verproside, a catalpol derivative iridoid glycoside isolated from Pseudolysimachion rotundum
var.subintegrum, is a biologically active compound with anti-inflammatory, antinociceptic, antioxidant, and
anti-asthmatic activities. Twenty-one metabolites were identified in bile and urine samples obtained after
intravenous administration of verproside in rats using liquid chromatography-quadrupole Orbitrap mass
spectrometry. Verproside was metabolized by O-methylation,glucuronidation,sulfation, and hydrolysis to
verproside glucuronides (M1 and M2), verproside sulfates (M3 and M4), picroside II (M5), M5 glucuronide
(M7), M5 sulfate (M9), isovanilloylcatalpol (M6), M6 glucuronide (M8), M6 sulfate (M10),
3,4-dihydroxybenzoic acid (M11), M11 glucuronide (M12), M11 sulfates (M13 and M14),
3-methyoxy-4-hydroxybenzoic acid (M15), M15 glucuronides (M17 and M18), M15 sulfate (M20),
3-hydroxy-4-methoxybenzoic acid (M16), M16 glucuronide (M19), and M16 sulfate (M21). Incubation of
verproside with rat hepatocytes resulted in thirteen metabolites (M1-M11, M13, and M14). We characterized
human UDP-glucuronosyltransferase and sulfotransferase enzymes responsible for verproside sulfates and
glucuronides, respectively.
|
15
정 병 화 _Byung Hwa Jung, 한국과학기술연구원 분자인식연구센터 / 책임연구원, 센터장
Office: 02-958-5062 / C.P.: 010-4629-8603
E-mail: jbhluck@kist.re.kr
학력/경력
1988
1990-2000
2004.05-2005.05
2006.02- 현재
2010.03- 현재
2011.04- 현재
이화여대 약학 학사
서울대 약제학 석사/박사
University of North Carolina at Chapel Hill 약학 Post-doc.
과학기술연합대 겸임부교수
한국과학기술연구원 책임연구원
한국과학기술연구원, 분자인식연구센터 센터장
주요연구실적
• Kumar BS, Chung BC, Kwon OS, Jung BH. Discovery of common urinary biomarkers for hepatotoxicity induced
by carbon tetrachloride, acetaminophen and methotrexate by mass spectrometry-based metabolomics. J. Appl.
Toxicol. 2012 32(7), 505-20 (2012).
• Lee SH, An JH, Park HM, Jung BH. Investigation of endogenous metabolic changes in the urine of pseudo
germ-free rats using a metabolomic approach. J. Chromatogr. B. 887–888, 8-18 (2012).
• Kumar BS, Chung BC, Lee YJ, Yi HJ, Lee BH, Jung BH. GC-MS-based simultaneous quantitative analytical
method for urinary oxysterols and bile acids in rat. Anal. Biochem. 408, 242-252 (2011)
• Kumar BS, Lee YJ, Yi HJ, Chung BC, Jung BH. “Discovery of safety biomarkers for atorvastatin in rat urine
using mass spectrometry based metabolomics combined with global and targeted approach.” Anal. Chim. Acta,
19, 661, 47-59 (2010)
• 스타틴의 약효 및 간 독성 측정용 마커 및 측정방법:
등록번호- 10-1099615, 등록일-2011. 12. 21. 등 논문 60여편, 특허등록 20여건
16
|
S1-2
Lipidomic profiling after statin drug treatment
1
1
Byung Hwa Jung , Jong Min Choi , Joo-Youn Cho
2
1
2
Molecular Recognition Research Center, KIST, Seoul, 136-791, Korea
Department of Pharmacology and Clinical Therapeutics, Seoul National University College of Medicine and Hospital
E-mail: jbhluck@kist.re.kr
Lipid is very important biological component since it plays multiple roles in cells and organisms. It
organizes materials of membranes, has functions in the energy production and storage, regulate cell
signaling, ligands or mediators of cell-cell interactions and protein-protein interaction. Therefore, lipids are
associated with various diseases, such as diabetes, atherosclerosis, obesity, and cardiovascular disease. Those
diseases are much related to hyperlipidemia.
Statins are the first-line therapy for the treatment of cholesterol induced hypercholesterolemia. Statins
inhibit the synthesis of cholesterol by competitively blocking the enzyme HMG-CoA reductase, which
converts HMG-CoA to mevalonate, the precursor of cholesterol.
Statin has a function in the production of cholesterol and its metabolites, so overall lipid metabolism
could be changed by the administration of statin. It also affect to the toxicity in the drug treatment and
body functions related to the lipid metabolism. Therefore it is important and meaningful to check overall
change of lipid metabolism in the treatment of statin drug.
Lipidomics, which is a critical part of metabolomics, can be defined as the system-wide characterization
of lipids and their interaction with cells and other biochemical compounds. As mentioned above, lipidomics
has expected to assume a notable role in systems biology and has attained more interest these day since
dysregulated lipid metabolism is related to human diseases,
In this presentation, the research on the administration of two representative statin drug, atorvastatin and
rosuvastatin will be given. In the case of atorvastatin study, lipid profiling by quantitative analysis of target
lipid and relations between liver toxicity and change of lipid profiling in the treatment of atorvastatin in rat
will be shown. In another study of rosuvastatin, the non targeted lipidomic research after the administration
in human will be described.
|
17
박 태 식 _Tae Sik Park, 가천대학교 생명과학과
Office: 031-750-8824 / C.P.: 010-2757-5903
E-mail: pts9918@gmail.com
학력/경력
1994
2001
2002
2005
2007
2011
고려대학교 농화학과 학사
Rutgers 대학교 식품과학과 박사
Pfizer (Ann Arbor, MI, USA) 심혈관연구부 Post-doc.
Columbia 의대 분자의학과 Associate Research Scientist
가천의과학대학교 분자의학과 조교수
가천대학교 생명과학과 부교수
주요연구실적
• 비만과 고지혈증으로 인한 지방산의 증가로 인한 지질대사체의 축적이 질병의 원인이라는 가설을 기초로 지질독성과
만성대사질환 (심혈관질환/당뇨병) 상관관계를 연구 중임.
• Lee SY, Kim JR, Ha MY, Shim SM, Park TS (2013). Measurement of diacylglycerols in plasma and tissues
by liquid chromatography-tandem mass spectrometry. Lipids. 48(3):287-96
• Park TS, Goldberg IJ (2012). Sphingolipids, lipotoxic cardiomyopathy and cardiac failure. Heart Failure Clinics,
8(4):633-41.
• Lee SY, Kim JR, Hu Y, Khan R, Kim SJ, Bharadwaj KG, Davidson MM, Choi CS, Shin KO, Lee YM, Park
WJ, Park IS, Jiang XC, Goldberg IJ, Park TS (2012). Cardiomyocyte specific deficiency of serine palmitoyltransferase
subunit 2 reduces ceramide but leads to cardiac dysfunction. J Biol Chem. 25;287(22):18429-39.
• Han MS, Lim YM, Quan W, Kim JR, Chung KW, Kang M, Kim S, Park SY, Han JS, Park SY, Cheon HG,
Rhee SD, *Park TS, Lee MS (2011). Lysophosphatidylcholine as an effector of fatty acid-induced insulin resistance.
J Lipid Res. 52(6):1234-46. *Co-corresponding author.
• Chang ZQ, Lee SY, Kim HJ, Kim JR, Kim SJ, Hong IK, Oh BC, Choi CS, Goldberg IJ, Park TS (2011).
Endotoxin activates de novo sphingolipid biosynthesis via nuclear factor kappa B-mediated upregulation of Sptlc2.
Prostaglandins Other Lipid Mediat. 94(1-2):44-52.
18
|
S1-3
Differential regulation of sphingolipid metabolism by ER stress
Tae-Sik Park
Departmetnt of Life Science, Gachon University, Sungnam 461-701, Korea
E-mail: pts9918@gmail.com
The endoplasmic reticulum is the principal organelle in the cell for protein folding and trafficking, lipid
synthesis and cellular calcium homeostasis. Perturbation of ER function results in activation of the unfolded
protein response (UPR). Chronic ER stress is reported to have an important role in abnormal lipid
biosynthesis and development of insulin resistance. The present study reports that transcription of
sphingosine kinase 2 (Sphk2) is differentially regulated by ER stress-mediated UPR pathways. Expression of
Sphk2, a major isotype of sphingosine kinase in the liver, was upregulated by tunicamycin and sphingosine
1-phosphate (S1P) was elevated in primary mouse hepatocytes. In contrast, chronic ER stress by high fat
diet suppressed Sphk2 expression. Overexpression of the activating transcription factor 4 (ATF4) upregulated
Sphk2 expression, whereas the spliced form of X-box binding protein 1 (sXBP1) downregulated Sphk2 as
demonstrated by Sphk2 promoter assays and western blot analyses. Adenoviral Sphk2 overexpression
activated pAKT with no alteration of IRS1 phosphorylation. In addition, cellular S1P levels were elevated
by Sphk2 overexpression while ceramide and sphingomyelin were not altered. Hepatic overexpression of
Sphk2 improved glucose tolerance in the mice fed a high fat diet and decreased hepatic accumulation of
lipid droplet by upregulating the genes in fatty acid oxidation. These results demonstrated that Sphk2 is
differentially regulated by ER stress-induced UPR pathways would contribute to the amelioration of hepatic
insulin resistance and steatosis via S1P production.
|
19
김 석 만 _Suhkmann Kim, 부산대학교 화학과
Office: 051-510-2240 / C.P.: 010-2599-7738
E-mail: suhkmann@pusan.ac.kr
학력/경력
1984.03-1992.02
1992.03-1994.02
1995.10-1998.12
1999.02-2000.10
2000.11-2002.10
2002.11-2006.08
2006.09-현재
2013.01-현재
부산대학교 화학과 학사
부산대학교 화학과 석사
일본 요코하마국립대학교 물질공학 박사
미국 아리조나대학 화학과 Post-doc.
미국 City of Hope, Cancer center Post-doc.
부산대학교 유전체물성연구소 연구교수
부산대학교 화학과 부교수
단백질체 생물물리 연구센터 센터장
주요연구실적
• 1H NMR spectroscopic identification of a glue sniffing biomarker, Forensic Science International (2011) 120–125
• Characterization of the Effects of Silver Nanoparticles on Liver Cell Using HR-MAS NMR Spectroscopy, Bulletin
of the Korean Chemical Society (2011) 32-6
• Study of metabolic profiling changes in colorectal cancer tissues using 1D 1H HR-MAS NMR spectroscopy,
Bulletin of the Korean Chemical Society (2013) 34-5
20
|
S1-4
Metabolic analysis of heavy metal toxicity using NMR techniques
Suhkmann Kim*
Department of Chemistry, Pusan National University, Busan, 609-735, Korea
Email: suhkmann@pusan.ac.kr
Heavy metals are common air and water pollutants, mainly as a result of various industrial activities,
which contamination has been a threat to our environment and human health. It is environmentally
ubiquitous, easily dissolved in and transported by water and accumulated in organism. Among the heavy
metals, mercury and cadmium are a hazardous contaminant in the marine environment, also induced the
many disease and carcinogenesis. [1] In addition, mercury cause oxidative stresses and DNA damages by
producing production of reaction oxygen species (ROS). [2] Also, cadmium involved in nervous system
damage and lung and prostate cancer. [3] Recently, Hsu etal. [4] reported that cadmium induced oxidative
stress down-regulates the gene expression of DNA mismatch recognition proteins MSH2 and MSH6 in
zebrafish (Danio rerio) embryos.
1
In this study, H NMR based metabolomics was applied to the adult zebrafish (Brachydanio rerio) to
1
investigate metabolic changes in whole body as a response to heavy metal exposure. We performed H
NMR experiments to confirm the difference of metabolites between control and heavy metal exposed
1
groups. Multivariate analysis was achieved using SIMCA-P+. Assignment of H NMR spectrum and
quantification of metabolites were accomplished by Chenomx.
References
[1] A. Takaki et al. Toxicology 2004, 203, 145-154
[2] X. Liu et al. Ecotoxicology 2011, 20, 177-186
[3] V. Verougstraete et al. Journal of Toxicology and Environmental Health, Part B: Critical Reviews
2003, 6, 227-255
[4] T. Hsu et al. Aquatic Toxicology 2013, 126, 9-16
|
21
22
|
Session 2
Plant Metabolomics
Chair: 김영식_서울대학교 / 최형균_중앙대학교
|
23
최 성 화 _Sunghwa Choe, 서울대학교 자연과학대학 생명과학부
Office: 02-880-6691 / C.P.: 010-9222-6691
E-mail: shchoe@snu.ac.kr
학력/경력
1984-1989
1989-1991
1994-1997
1997-2000
2000-2001
2008-2009
2012-현재
2001-현재
서울대학교 식물학 이학사
서울대학교 식물학 이학사
University of Arizona 식물과학 이학박사
University of Arizona 식물과학 연구교수
Ceres, Inc. 선임연구원
University of Wisconsin Madison 객원교수 Biotech center
서울대학교 차세대융합기술연구원 부원장
서울대학교 생명과학부 조교수, 부교수, 교수
주요연구실적
박사 학위과정부터 현재까지 일관되게 식물의 스테로이드성 생장호르몬인 브라시노스테로이드의 대사 경로 해석
연구를 수행하고 있다. 현재 총 40여편의 관련 논문을 Plant Cell, Plant Journal, Plant Physiology 저널 등에 발표하였다.
브라시노스테로이드 대사 경로에 관여하는 효소를 유전학적으로 발굴하여 기능을 분석하는 방법을 주로 사용하고
있으며, 그 결과로 현재는 20여 단계의 합성 및 분해 경로를 거의 완벽하게 이해하게 되었다. 또한, 최근에는 이
대사경로를 조절하는 신호전달 기작에 관한 연구를 수행하여 브라시노스테로이드 합성이 또 다른 생장호르몬인
옥신에 의해 촉진됨을 밝힌 바 있다.
24
|
S2-1
Biochemical genetics of brassinosteroid metabolic pathways
Yuhee Chung and Sunghwa Choe
School of biological sciences, College of natural sciences, Seoul National University, Seoul 151-747, Korea
Advanced institute of convergence technologies, Suwon-si, Gyeonggi-do 443-270, Korea
Email: shchoe@snu.ac.kr
Brassinosteroids (BRs) are plant steroid hormones that regulate diverse aspects of growth and
development throughout the life cycle of plants. The physiological processes that are regulated by BRs
include vascular system differentiation, pollen maturation, cell elongation, cell division, and others. Mutants
being defective in BR biosynthesis or signaling display characteristic dwarf phenotypes, confirming their
roles as growth-promoting hormones. BRs are synthesized using sterols including campesterol as a
precursor. Therefore, mutants that have lesions in sterol biosynthesis, dwarf1, dwarf5, and dwarf7, as well
as downstream BR biosynthetic step such as dwarf3/cpd, dwarf4, det2/dwf6, and BR6ox exhibit dwarf
phenotypes. In spite of their pivotal roles in plant physiology, a signal that turns on biosynthesis of the
bioactive BR compounds has been elusive. Recently, we have found that auxin stimulates the expression of
DWARF4 which encodes rate-determining step enzyme in the BR pathways. Treating Arabidopsis seedlings
with auxin resulted in elevation of BR biosynthesis especially in the roots. Furthermore, chromatin
immunoprecipitation assays showed that auxin inhibits the binding of the BR-specific transcriptional
repressor, BZR1, to the DWARF4 promoter. Transcriptome analysis after treatment with auxin alone or
auxin plus brassinazole (Brz, a BR biosynthetic inhibitor) revealed that genes previously characterized as
being auxin responsive are not properly regulated when BR biosynthesis is disrupted by Brz. Therefore, our
results support the ideas that auxin regulates BR biosynthesis and auxin thus relies on synthesized BRs for
some of its physiological effects in Arabidopsis.
|
25
장 영 표 _Young Pyo Jang, 경희대학교 약학대학
Office: 02-961-9421 / C.P.: 010-9989-9924
E-mail: ypjang@khu.ac.kr
학력/경력
1992
1994
2001
2005
2007
2007
서울대학교 약학대학 약학과 학사
서울대학교 약학대학 약학과 석사
서울대학교 약학대학 약학과 박사
Columbia University, NY, USA Post. Doc. Fellow
Columbia University, NY, USA Researcher
경희대학교 약학대학
주요연구실적
• Chemometric Classification of Morphologically Similar Umbelliferae Medicinal Herbs by DART-TOF-MS
Fingerprint. Phytochemical Analysis
• Rapid purification method for vitamin A-derived aging pigments A2E and iso-A2E using cation exchange resin.
Journal of Chromatography A
• A rapid, simple method for the genetic discrimination of intact Arabidopsis thaliana mutant seeds using metabolic
profiling by direct analysis in real-time mass spectrometry. Plant Methods
• Identification of ambiguous cubeb fruit by DART-MS-based fingerprinting combined with principal component
analysis. Food Chemistry
• Quantitative analysis of major dibenzocyclooctane lignans in schisandrae fructus by online TLC-DART-MS.
Phytochemical Analysis
26
|
S2-2
DART-MS Snapshot on Natural Products Analysis and Plant
Metabolomics
Young Pyo Jang
Division of Pharmacognosy, College of Pharmacy, Kyung Hee University, Seoul, Korea
Email: ypjang@khu.ac.kr
The ultimate goal of metabolomics would be analyze all the metabolomes from the specified biological
samples both in qualitative and quantitative manner. So far, unfortunately, no instrument can afford this
kind of analysis capacity. Therefore, multi-disciplinary and multi-instrumental collaboration is critical to get
reasonable and significant answers for various biological questions we raise. Direct analysis in real time
(DART) is relatively new ionization technique that provides rapid ionization of various types of sample
under atmospheric pressure. The ionization mechanism involves the reaction of excited-state helium with
water molecule in the air to produce protonated water clusters followed by proton transfer to the analytes.
Since M+H molecular ions are mostly observed by DART-TOF-MS for most compounds, DART-MS
produces relatively simple and clear mass snapshots on crude raw materials containing complex components.
The absence of sample preparation step in DART-MS measurement makes it very attractive instrument
when large numbers of samples are ready to be analyzed in metabolomics study. In this presentation, a few
examples of DART-MS analysis on natural products and crude herbal drugs and its application on plant
metabolomics will be introduced.
|
27
김 석 원 _Suk Weon Kim, 한국생명공학연구원
Office: 042-860-4646 / C.P.: 010-4448-9369
E-mail: kimsw@kribb.re.kr
학력/경력
2004
1994-현재
2008-현재
1991-현재
2011-현재
한국과학기술원 식물분자생물학 박사
한국생명공학연구원 선임/책임연구원
과학기술연합대학원 기능유전체학/조교수
한국식물생명공학회 정회원/평의원
한국대사체학회 이사
주요연구실적
• 식물조직배양 및 분자육종
• 식물대사체 및 다변량통계분석 기반 활용 기술개발(두과작물, 십자화과 작물)
• 원형질체 비대칭융합 기술을 이용한 사이브리드 잡종 식물체 개발
28
|
S2-3
Simple and rapid prediction system by metabolic fingerprinting of
Fourier transform infrared spectroscopy
Seung Yeob Song1, Myungsuk Ahn1 Jong Hyun Kim1, Jang R. Liu1 and Suk Weon Kim2*
1
Green Bio Research Center
Biological Resource Center, Korea ResearchI nstituteof Bioscience and Biotechnology
(KRIBB),125 Gwahak-ro, Yuseong-gu, Daejeon, 305-806, Korea.
Email: kimsw@kribb.re.kr
2
Plant metabolomics is a research field for identifying all of the metabolites found in a certain plant cell,
tissue, organ, or whole plant in a given time and conditions and for studying changes in metabolic
profiling as time goes or conditions change. Metabolomics is one of the most recently developed omics for
holistic approach to biology. Metabolomics or metabolite fingerprinting techniques usually involves collecting
spectra of crude solvent extracts without purification and separation of pure compounds or not in
standardized conditions.Therefore, that requires a high degree of reproducibility, which can be achieved by
using a standardized method for sample preparation and data acquisition and analysis. In plant biology,
metabolomics is applied for various research fields including rapid discrimination and screening of plant
species, cultivar and GM plants, metabolic evaluation of commercial food stocks and medicinal herbs,
understanding various physiological responses, and determination of gene functions. Recently, plant
metabolomics is applied for characterization of gene function often in combination with transcriptomics by
analyzing tagged mutants of the model plants of Arabidopsis and rice.The use of plant metabolomics
combined by other omics data in plant phenomics will be the challenge for the coming year. This study
attempted to introduce applications and prospects of plant metabolomics research.
|
29
성 상 현 _Sang Hyun Sung, 서울대학교 약학대학
Office: 02-880-7859 / C.P.: 010-6220-0746
E-mail: shsung@snu.ac.kr
학력/경력
1990
1992
1998
2000
2003
2005
2008
2012
서울대학교 약학과 약학 학사
서울대학교 대학원 약학과 생약학 석사
서울대학교 대학원 약학과 생약학 박사
서울산업대학교 식품공학과 겸임교수
㈜ 엘컴사이언스 R&D 이사
일본 카나자와 대학 연구원
서울대학교 약학과 조교수
서울대학교 약학과 부교수
주요연구실적
• Jeong EJ, Lee HK, Lee KY, Jeon BJ, Kim DH, Park JH, Song JH, Huh J, Lee JH, Sung SH. The effects
of lignan-riched of Shisandra chinensis on amyloid-β- induced cognitive impairment and neurotoxicity in the
cortex and hippocampus of mouse, J Ethnopharmacol, 146(1):347-54 (2013)
• Yang H, Yoo G, Kim HS, Kim JY, Kim SO, Yoo YH, Sung SH. Implication of the stereoisomers of ginsenoside
derivatives in the antiproliferative effect of HSC-T6 cells, J Agric Food Chem, 60(47):11759-64 (2012).
• Lee DY, Kim SH, Kim YC, Kim HJ, Sung SH, Discrimination of Scrophulariae Radix according to geographical
origin and determination of active constituents by near infrared spectroscopy (NIRS), Microchem J, 99(2):213-217
(2011)
• Yang H, Cho YW, Kim SH, Kim YC, Sung SH. Triterpenoidal saponins of Pulsatilla koreana roots, Phytochemistry,
71(16):1892-1899 (2010)
• Kim SH, Kim DH, Park JH, Choi EJ, Park SH, Lee KY, Jeon MJ, Kim YC, Sung SH, Discrimination of
Scrophularia spp. according to geographic origin with HPLC-DAD combined with multivariate analysis, Microchem
J, 94(2):118-124 (2010)
30
|
S2-4
Prediction of Bioactivity and Active Constituents for Botanical Drugs
Sang Hyun Sung
College of Pharmacy, Seoul National University, Seoul 151-742, Korea
E-mail: shsung@snu.ac.kr
Statistical techniques have been playing more and more important roles in many scientific fields.
Combined with analytical techniques, statistical techniques have formed a characteristic research field, called
metabolomics. Applied in natural product researches, metabolomics has allowed various advances including
dereplication and fingerprint-based quality control. However, advances in analytical techniques allow us to
acquire more information from natural products, and advances in data mining technology allow us to handle
more sophisticated data sets.
One of advanced application of metabolomics techniques in natural products would be a computational
approach to discover correlation between compositional and pharmacological data sets. This approach, called
a quantitative composition-activity relationship (QCAR), has been attempted for two major purposes. At
first, Biological potency could be predicted from the chemical profiles of natural products applying the
discovered correlation. Chemical analyses can be performed more simply than biological assays, so the
prediction of biological activity could be an effective method for an indirect biological evidence based
quality control. In addition, we would estimate which constituents played important roles in the bioactivity
of natural products. This correlational approach is expected to be an effective and holistic method for
discovering active compounds from natural products.
Two correlational approach models have been created, and their predictive abilities were considered. At
first, 42 different extracts were obtained from 6 samples of Morus alba root bark under 7 different
extraction solvent conditions. HPLC-DAD chromatograms and mushroom tyrosinase inhibitory activities of
these extract samples were obtained, and the partial least squares (PLS) regression model between the
chromatograms and the bioactivities of samples was created. The cross validations were performed, and the
PLS models showed excellent performance in bioactivity prediction. Additionally, an estimation of the
bioactive component was made from the regression coefficient, and the result of the estimation was similar
to the experimental results of reference papers.
Secondly, a non-linear model was also attempted, applying UPLC-qTOF/MS data and protective effects
for H2O2-induced AGS cells of 41 Artemisia princeps samples. This study applied bioactivity data of
cellular assay, so we supposed that non-linear algorithms should be used for discover complex relations
between constituents and treated cells. Non-linear data mining algorithms including artificial neural network
and random forest were applied to the correlational models, and validations for predictive abilities of the
model were also performed for each model.
|
31
32
|
Session 3
Microbial Metabolomics
Chair: 성문희_국민대학교 / 임용현_한국표준과학연구원
|
33
박 용 철 _Yong-Cheol Park, 국민대학교
Office: 02-910-5462 / C.P.: 010-2082-9965
E-mail: ycpark@kookmin.ac.kr
학력/경력
1994
1996
2003
2004-2006
2006-2009
2009
서울대학교 식품공학과 농학사
서울대학교 생물화학공학전공 공학석사
서울대학교 생물화학공학전공 공학박사
미국 Rice University 생화학과/박사후연구원
서울대학교 농업생물신소재연구소 연구교수
국민대학교 발효융합학과 조교수, 부교수
주요연구실적
• 미생물 대사공학 및 발효공정최적화 기술을 이용한 바이오소재 및 바이오연 생산기술 개발
- 섬유소 바이오매스를 이용한 바이오에탄올 생산기술 개발
- 고에탄올 생산용 발효공정 최적화
- 해조류 바이오매스 기반 바이오소재 생산용 미생물 플랫폼 기술 개발
34
|
S3-1
Global optimization of microbial systems and process
for biochemical production
Yong-Cheol Park
Department of Advanced Fermentation Fusion Science and Technology, Kookmin University, Seoul 136-702, Korea
Email: ycpark@kookmin.ac.kr
For sustainable development, many research groups have made an effort to change the currently available
crude oil to the renewable biomass as resource. To overcome the general problems of corn- and
sugar-based biomass, non-food biomass such as cellulosic biomass (tree, straw, agricultural residue et al.)
has been concerned as alternative biomass for production of biochemicals. Cellulosic biomass is composed
of cellulose, hemicellulose and lignin. By its decomposition, various mono-sugars are released. Among them,
most abundant sugars of glucose and xylose should be converted to biochemicals for gaining their price
competitiveness against petroleum-oriented chemicals. To develop a commercially available bioprocess,
meanwhile, engineering of genetic and microbial systems should be complementary to bioprocess
engineering by feed-forward and feed-back cycles. Global Optimization of Systems and Process (GoSysPro)
armed with genetic, microbial and metabolic engineering, –omic technology and fermentation optimization is
a promising technology able to meet the system and process complementation, and applicable for rapid
development of biochemical production process. In this presentation, microbial systems were engineered by
the GoSysPro technology for mass production of two value-added chemicals, 2,3-butanediol and D-ribose,
sources of synthetic rubber and riboflavin, respectively. The engineered microorganisms of Klebsiellaoxytoca
and Bacillussubtilis could produce the target chemicals with high conversion yields and production rates.
And high titers of 2,3-butanediol and D-ribose were able to be obtained by fermentation optimization.
|
35
이 도 엽 _Do Yup Lee, 국민대학교
Office: 02-910-5733 / C.P.: 010-9918-9338
E-mail: rome73@kookmin.ac.kr
학력/경력
1998
2003
2009
2010
2012
연세대학교 생명공학과 학사
서울대학교 식품공학과 석사
UC Davis Food Sci. & Tech. 박사
UC Davis 대사체/단백질체학 포스닥
Lawrence Berkeley National Laboratory 대사체학 포스닥
주요연구실적
• Low-Dose Ionizing Radiation-Induced Blood Plasma Metabolic Response in a Diverse Genetic Mouse Population.
Do Yup Lee, Benjamin P. Bowen, David H. Nguyen, Sara Parsa, Yurong Huang, Jian-Hua Mao and Trent
R. Northen. Radiation Research, 2012;178(6)
• System response of metabolic networks in Chlamydomonas reinhardtii to total available ammonium. Do Yup
Lee, Jeong-Jin Park, Dinesh Kumar, Oliver Fiehn. Molecular & Cellular Proteomics, 2012;11(10):973-988
• Resolving brain regions using nanostructure initiator mass spectrometry imaging of phospholipids. Do Yup Lee*,
Virginia Platt*, Ben Bowen, Katherine Louie, Christie Canaria, Cynthia T. McMurray and Trent Northen. Integrative
Biology, 2012;4(6):693-699
*These authors contributed equally to this work.
• Pharmacogenetics meets Metabolomics: Metabolomics is useful tool for screening of new endogenous OCT2
substrate related to metformin disposition. Im-Sook Song*, Do Yup Lee*, Min-Hye Shin, Hyunmi Kim, Yun
Gyong Ahn, Inmyoung Park, Kyoung Heon Kim, Tobias Kind, Oliver Fiehn, Jae-Gook Shin, and Kwang-Hyeon
Liu. PLoS ONE, 2012;7(5):e36637
*These authors contributed equally to this work.
• Retinoic acid induces a metabolic switch in SH-SY5Y cells from glycolysis to oxidative Phosphorylation. Xun
Z., Do Yup Lee, Lim J., Canaria C., Barnebey A., Yanonne S., McMurray CT. Mechanisms of Ageing and
Development, 2012;133:176–185
36
|
S3-2
System response of metabolic networks in Chlamydomonas reinhardtii
to temporal and degree-wisen utritionalc hallenge
Do Yup Lee
Department of Advanced Fermentation Fusion Science and Technology, Kookmin University, Seoul 136-702, Korea
Email:rome73@kookmin.ac.kr
Drastic alterations in macronutrients are known to cause large changes in biochemistry and gene
expression in the photosynthetic alga Chlamydomonas reinhardtii. However, metabolomic and proteomic
responses to gradual challenge in macronutrients availability have not yet been systematically studied.
Responses of 145 identified metabolites were quantified using gas chromatography-time of flight mass
spectrometry; 495 proteins (including 187 enzymes) were monitored using liquid chromatography-ion trap
mass spectrometry with label-free spectral counting approach.
Stress response and carbon assimilation processes (Calvin cycle, acetate uptake and chlorophyll
biosynthesis) were modified first, in addition to enrichment in enzyme contents for lipid biosynthesis and
accumulation of short chain free fatty acids. Nitrogen/carbon metabolism was re-modulated only under
chronic conditions, for example in TCA cycle tightly linked to amino acid metabolism. In conclusion,
Chlamydomonas metabolic network sensitively and precisely responds to total nitrogen availability with
transient increases in short-chain free fatty acids and turnover of internal proteins, long before nitrogen
resources are depleted.
|
37
이 백 석 _Baek-Seok Lee, CJ Cheil Jedang Co. Ltd
Office: 82-2-3660-0537
E-mail: bslee@cj.net
학력/경력
2003
1999
1997
Inha University Ph.D
Inha University M.S.
Inha University B.S.
주요연구실적
2008-present
2005-2008
2003-2005
38
|
CJ Research Institute of Biotechnology, CJ Cheil Jedang Co. Ltd.
Research Associate, Institute for Advanced Biosciences, Keio University, Japan
Postdoctoral Fellow, Department of Bioengineering, University of California, San Diego, CA,
USA
S3-3
Metabolic engineering of industrial microorganisms based
on metabolomics
Baek-Seok Lee
CJ Research Institute of Biotechnology, CJ Cheil Jedang Co. Ltd.
E-mail: bslee@cj.net
Industrial fermentation processes are continuously being improved using a combination of strain
improvement and bioprocess optimization. Currently target selection, i.e. the identification of bottle-necks
and bioprocess parameters that affect production yield and productivity, is the rate-limiting step in
bioprocess optimization.
With the recently introduced metabolomics technology it is now possible to compare wild-type's
metabolite profile with overproduction strain's and the metabolic profile of low production phase with high
production phase. We have successfully applied this metabolomics platform for selecting targets for
improvement of product yield and productivity based on different metabolite profile.
As the biochemical level of the metabolome is closest to that of the phenotype (i.e. productivity, yield),
metabolomics is the most relevant functional genomics tool for understanding biological production. By
calculating the correlation between the large numbers of metabolites measured and the production outcome
under different conditions, metabolomics enables the identification of bottlenecks in the production strain.
After biological interpretation of the results, the genetic and bioprocess targets are identified. CJ has
successfully applied this approach for identifying targets in metabolite production in bacteria. The targets
have been validated and large improvements in yield and productivity of amino acids and nucleotides were
achieved in these improved strains and fermentation optimization.
|
39
40
|
Session 4
Data Analysis & Bioinformatics
Chair: 서진호_서울대학교 / 김경헌_고려대학교
|
41
고 광 일 _Kwang-Il Goh, 고려대학교
Office: 02-3290-3115 / C.P.: 011-1736-3641
E-mail: kgoh@korea.ac.kr
학력/경력
2004
2004-5
2005-6
2005-6
2007-
서울대학교 물리학과 통계물리학 이학박사
서울대학교 이론물리학연구센터 박사후연구원
University of Notre Dame 박사후연구원
Dana-Farber Cancer Institute 방문연구원
고려대학교 물리학과 조교수, 부교수
주요연구실적
• The human disease network, Proc. Natl. Acad. Sci. USA 104, 8685-8690 (2007) 외 연구논문 50여편.
42
|
S4-1
Exploring the human diseasome
Kwang-Il Goh
Departmetnt of Physics, Korea University, Seoul 136-713, Korea
Email: kgoh@korea.ac.kr
Advances in genome-scale molecular biology and molecular genetics have greatly elevated our knowledge
on the basic components of human biology and diseases. At the same time, the importance of cellular
networks between those biological components is increasingly appreciated. Built upon these recent
technological and conceptual advances, a new discipline called the network medicine, an approach to
understand human diseases from a network point-of-view, is about to emerge. In this review article, we
will survey some recent endeavors along this direction, centered on the concept and applications of the
human diseasome and the human disease network. Questions, and partial answers thereof, such as how the
connectivity between molecular parts translates into the relationships between the related disorders on a
global scale and how central the disease-causing genetic components are in the cellular network, will be
discussed. The use of the diseasome in combination with various interactome networks and other
disease-related factors is also reviewed.
|
43
임 광 일 _Kwang-il Lim, 숙명여자대학교 의약과학과
Office: 02-2077-7627 / C.P.: 010-3052-3548
E-mail: klim@sm.ac.kr
학력/경력
1989-1993
1993-1995
1994-2000
2000-2005
2005-2011
2011-현재
포스텍 화학공학 학사
포스텍 화학공학 석사
한화에너지 (현 SK에너지) 공정연구원
University of Wisconsin-Madison, Dept. of Chemical and Biological Engineering 시스템생물학 박사
University of California at Berkeley, Dept. of Chemical and Biomolecular Engineering 바이러스공학/줄기
세포공학 박사후연구원
숙명여자대학교 의약과학과/조교수
주요연구실적
• R. S. Ashton, A. Conway, C. Pangarkar, J. Bergen, K. Lim, P. Shaw, M. Bissell and D. V. Schaffer. 2012.
EphrinB2 signaling regulates adult hippocampal neurogenesis. Nature Neuroscience. 15(10): 1399-1406.
• K. Lim , R. Klimczak, J. H. Yu and D. V. Schaffer. 2010. Specific insertions of zinc finger domains into
Gag-Pol yield engineered retroviral vectors with selective integration properties. PNAS. 107(28): 12475-12480.
• J. C. Burnett, K. Lim, A. Calafi, J. J. Rossi, D. V. Schaffer and A. P. Arkin. 2010. Combinatorial latency
reactivation for HIV-1 subtypes and variants. Journal of Virology. 84(12):5958-5974
• K. Lim and J. Yin. 2009. Computational fitness landscape for all gene-order permutations of an RNA virus.
PLoS Computational Biology. Feb 6;5(2):e1000283.
• D. V. Schaffer*, J. T. Koerber and K. Lim. 2008. Molecular engineering of viral gene delivery vehicles. Annual
Review of Biomedical Engineering. 10:169-194.
• K. Lim, T. Lang, V. Lam and J. Yin. 2006. Model-based design of growth-attenuated viruses. PLoS Computational
Biology. Sep 1;2(9):e116.
44
|
S4-2
Model-based genome design for the development
of viral vaccine platforms
Kwang-il Lim
Dept. of Medical and Pharmaceutical Sciences
Sookmyung Women’s University
Email: klim@sm.ac.kr
Although many viruses are linked to diseases that adversely impact the health of their human, animal,
and plant hosts, viruses could help promote wellness and treat disease if their ‘‘good traits’’ could be
harnessed. The engineering of viral genomes provides ways not only to explore viral regulatory mechanisms
at a genomic level, but also to produce recombinant viruses that may serve as vaccines, gene delivery
vectors, and oncolytic (tumor-killing) agents. However, the complexity of interactions among viral and
cellular components involved in the life cycle of a virus can make it challenging to anticipate how altering
viral components will influence the overall behavior of the virus. We have developed a system-level
computer model that begins to mechanistically account for key virus–cell interactions in viral intracellular
development. In this presentation, I will show how the model predictions captured the experimentally
observed effects of some gene rearrangements of viral genomes on the levels and timing of viral protein
expression and viral progeny production, aspects that mainly determine the immunogenicity and safety of
live-virus vaccines, respectively. Further, our model was applied to quantitatively analyze the growth of all
possible gene-rearranged variants of a prototype RNA virus covering its complete gene-order design space.
Refinement and extension of our approach to current and other virus systems has the potential to advance
the application of viruses as therapeutic agents by providing a virus genome design tool.
|
45
이 동 엽 _Dong-Yup Lee, 싱가포르국립대
Office: +65 6516-6907 / C.P.: +65 9793-8059
E-mail: cheld@nus.edu.sg
학력/경력
1998
2000
2004
2004-2005
2005-현재
2005-현재
연세대학교 화학공학과 학사
한국과학기술원 생명화학공학과 석사
한국과학기술원 생명화학공학과 박사
한국과학기술원 생물정보연구센터 선임연구원
싱가포르국립대 조교수
Bioprocessing Technology Institute, Bioinformatics group leader
주요연구실적
• 모델링과 오믹스 데이터의 융합 연구를 통해 시스템 생명공학 및 생물정보학 기법을 새롭게 개발.
• 미생물, 동물세포, 식물세포등 다양한 생물시스템의 가상세포를 개발하여 균주개량 및 바이오 리파이너리 연구에
활용
• 인공유전자 합성을 위한 합성생물학 기법 개발
• 현재까지 약 60여개의 논문발표와 10개 이상의 국제특허출원
46
|
S4-3
Application of Systems Metabolomics to Cell Factories
Dong-Yup Lee
Department of Chemical & Biomolecular Engineering, National University of Singapore,
4 Engineering Drive 4, Singapore 117576, Singapore.
Bioprocessing Technology Institute, A*STAR (Agency for Science, Technology and Research),
20 Biopolis Way, #06-01, Singapore 138668, Singapore,
Email: cheld@nus.edu.sg
Metabolomics is a rapidly emerging field involving the measurement and study of small molecules in
biological systems. These small molecules, known as metabolites, are the end products of cellular processes,
and thus their levels can closely reflect the phenotypic state of a biological system. This makes
metabolomics a valuable tool within the systems biology framework for investigating cellular responses to
perturbations, with the ultimate aim of understanding complex systems which include microbes, plant,
human tissues and cell, as well as other mammalian systems. In this talk, I will present our recent efforts
in establishing LC-MS-based systems metabolomics platform with application to mammalian cell factories,
e.g., Chinese hamster ovary cells, and metabolite-centric model-driven analysis for characterizing the cell
physiology. In addition, future perspectives on “Systems Bioinformatics” are also discussed to suggest new
opportunities and challenges in this field.
[This work was supported by a grant from the Next-Generation BioGreen 21 Program (No. PJ009520),
Rural Development Administration, Republic of Korea].
|
47
이 대 희 _Daehee Lee, 한국생명공학연구원
Office: 042-879-8225 / C.P.: 010-3493-3215
E-mail: dhlee@kribb.re.kr
학력/경력
1996-2000
2000-2003
2003-2007
2007-2010
2010-현재
서울대학교 식품공학 학사
서울대학교 식품생명공학 석사
서울대학교 식품생명공학 박사
University of California, San Diego Dept. of Bioengineering 박사후연구원
한국생명공학연구원 바이오합성연구센터 전임연구원
주요연구실적
• Haythem Latif, Joshua A. Lerman, Vasiliy A. Portnoy, Yekaterina Tarasova, Harish Nagarajan, Alexandra C.
Schrimpe-Rutledge, Richard D. Smith, Joshua N. Adkins, Dae-Hee Lee, Yu Qiu, Bernhard O. Palsson, and
Karsten Zengler, 2013, PLoS Genetics (Accepted) The genome organization of Thermotoga maritima reflects
its lifestyle
• Hojung Nam, Nathan E. Lewis, Joshua A. Lerman, Dae-Hee Lee, Roger L. Chang, Donghyuk Kim, and Bernhard
O. Palsson, 2012, Science, 337(6098): 1101-1104, Network context and selection in the evolution to enzyme
specificity.
• Sung Ho Yoon, Mee-Jung Han, HaeyoungJeong, ChoongHoon Lee, Xiao-Xia Xia, Dae-Hee Lee, JiHoon Shim,
Sang Yup Lee, Tae Kwang Oh, and Jihyun F. Kim, 2012, Genome Biology 13:R37, Comparative multi-omics
systems analysis of Escherichia coli strains of B and K-12
• Dae-Hee Lee, Adam M. Feist, Christian L. Barrett, and Bernhard O. Palsson, 2011, PLoS One 6(10): e26172,
Cumulative number of cell divisions as a meaningful timescale for adaptive laboratory evolution of Escherichia
coli
• Dae-Hee Lee and Bernhard O. Palsson, 2010, Applied and Environmental Microbiology 76(3): 4158-4168, Adaptive
evolution of Escherichia coli K-12 MG1655 on a non-native carbon source, L-1,2-propanediol
48
|
S4-4
Adaptive laboratory evolution of Escherichia coli - harnessing the
power of genome-scale sciences for metabolic network evolution
Dae-Hee Lee
Systems and Synthetic Biology Research Center, Korea Research Institute of Bioscience
and Biotechnology, Daejeon 305-806, Korea
Email: dhlee@kribb.re.kr
Adaptive laboratory evolution (ALE) studies can provide key information to address a wide range of
issues in evolutionary biology through direct observation of the evolution process. They not only enable
testing of evolutionary theory and principles, but also have applications to metabolic engineering and human
health. In this study, the genetic and biochemical basis for bacterial adaptation was determined to gain
understating of the plasticity of bacterial genomes. Using next-generation sequencing technology, we
identified all accumulated mutations that appear during ALE of an Escherichia coli K-12 strain from no
growth at all initially to maximal growth on an unnatural carbon source, L-1,2-propanediol (L-1,2-PDO).
We obtained proof that the observed spontaneous mutations were responsible for improved fitness by
creating multiple defined site-directed mutants that had growth rates matching those of the evolved strains.
To elucidate the mechanisms underlying the adaptation, we assessed metabolic changes using global
transcriptome, proteome, and metabolite profiling in conjunction with a genome-scale in silico model
(iAF1260) of E. coli metabolism. Following adaptation to growth on L-1,2-PDO, growth rates of the
evolved strains are consistent with the computationally-predicted optimal growth rates using iAF1260. In
addition, ALE tunes the transcriptional program of evolved E. coli by changing expression for pathways
needed for efficient metabolism of the non-native substrate. This optimization of expression for growth on
L-1,2-PDO is further validated by metabolite profiling when the L-1,2-PDO-evolved strain growing on
glycerol is shifted back to L-1,2-PDO. ALE studies on a non-native carbon source can provide a detailed
understanding into how metabolism and its regulation change at the genome-scale to form an optimal
growth phenotype, and that the integration of ‘omics’ data and genome-scale in silico models enable deep
insight into mechanisms of bacterial adaptation.
|
49
50
|
Session 7
Human Diseases
Chair: 박민수_세브란스병원 / 황금숙_한국기초과학지원연구원
|
51
권 태 환 _Tae-Hwan Kwon, 경북대학교 의학전문대학원
Office: 053-420-4825 / C.P.: 010-9033-0904
E-mail: thkwon@knu.ac.kr
학력/경력
2004-현재
2000-2003
1997-2000
2005.02
2001.02
1988-1992
1982-1988
경북대학교 의학전문대학원 생화학세포생물학교실 분자세포생리학/신장내과학/교수
동국대학교 의과대학 생리학교실 분자세포생리학/신장내과학/조교수
University of Aarhus, Denmark 분자세포생리학/조교수
University of Aarhus, Denmark 분자세포생리학 의학박사 (dr.med.: Habilitation degree)
경북대학교 대학원 내과학 의학박사 (PhD)
경북대학교 병원 내과 전문의
경북대학교 의과대학 의학과 졸업 의사
주요연구실적
• 연구분야 및 논문
분야: 1) 분자세포생리학; 수분통로 및 전해질 운반체 조절기전
2) 수분대사질환: 신성 요붕증의 병태생리, 고혈압의 병태생리
3) 산-염기대사질환: 산-염기 운반체 이상
4) 신장내과학: 급,만성 신부전 병태생리, 당뇨병성 신증 병태생리
논문: SCI 논문 150 편 이상 (Nature, PNAS, JCI, Am J Physiol Renal 등)
Book Chapter 11 편 (Brenner, Schrier, Hebert 등 국외 Kidney Textbook)
52
|
S7-1
Metabolomics in the kidney diseases
Tae-Hwan Kwon
Department of Biochemistry and Cell Biology,
School of Medicine, Kyungpook National University
Email:thkwon@knu.ac.kr
There are a number of metabolites in urine, plasma, cells or tissues, and altered metabolic profiling could
provide important information which indicates organ dysfunction. Metabolites are endogenous and exogenous
molecules which play a role in the regulatory and biological systems of the cell. Since metabolites are
regulated through a number of metabolic pathways, investigation of the whole feature of metabolites rather
than several selected metabolites enables us to understand the underlying physiological and pathophysiological
status more comprehensively. Accordingly, a relatively new metabolic profiling approach based on 1H NMR
spectroscopy coupled with multivariate pattern recognition was exploited to understand the metabolic
response of the kidney. Kidney is an organ which metabolizes a large number of substrates. These
substrates can be changed into metabolites by the kidney cells (e.g., tubular epithelial cells, interstitial cells,
vascular endothelial cells, and cells composing of glomerulus) which are heterogenous in structure, function,
and metabolism. These metabolites are secreted into tubular lumen (i.e., urine), and hence metabonomics
(i.e., a metabolic profiling analysis) of both kidney tissues and urine could provide important biological
information regarding the ongoing status of kidney function via quantitative determination of the
intermediary metabolites. Recently, we exploited metabonomics on lithium-induced nephrogenic diabetes
insipidus, and patients with chronic kidney disease and kidney transplantation. Moreover, an integrated
analysis of metabolome and transcriptome was applied to the kidney inner medullary collecting duct cells
exposed to different extracellular osmolalities for comprehensive understanding of the response of the kidney
collecting duct cells to the changes of extracellular environment. The successful application of 1H NMR
spectroscopy of tissues and biofluids for studying metabolic diseases and toxic process has now been
established and this method provides novel diagnostic markers for a disease state or biomarkers for drug
response phenotypes, and mechanistic information on cellular perturbations and pathways.
|
53
김 수 열 _Soo-Youl Kim, 국립암센터
Office: 031-920-2221 / C.P.: 010-4657-4369
E-mail: kimsooyoul@gmail.com
학력/경력
1985
1991
1994
2001
2005
연세대학교 생화학과 생화학 학사
서울대학교 의학과 생화학 박사
National Institutes of Health Post-doc.
National Institutes of Health Visiting Scientist
Cornell 의대 조교수 현재 국립암센터 수석연구원/부장
주요연구실적
• Transglutaminase Inhibitor: A New Anti-Inflammatory Approach in Allergic Conjunctivitis. J. Clin. Invest. 111,
121-8, 2003.
• Transglutaminase 2 induces NF-kB activation via a novel pathway in BV-2 microglia. J. Biol. Chem. 279,
53725-53735, 2004
• Reversal of drug resistance in breast cancer cells by transglutaminase 2 inhibition and nuclear factor-kappaB
inactivation. Cancer Res. 66, 10936-43, 2006
• A New Regulatory Mechanism of NF-kB Activation by I-kBain Cancer Cells. J Mol Biol. 384, 756-65, 2008
• Transglutaminase 2 as a cisplatin resistance marker in non-small cell lung cancer. J Cancer Res Clin Oncol.
136, 493-502, 2010
• Anti-inflammatory effects of the R2 peptide, an inhibitor of transglutaminase 2, in a mouse model of allergic
asthma, induced by ovalbumin. Br J Pharmacol. 162(1):210-25. 2011
• Cancer Cell Promotes Survival through Depletion of the von Hippel-Lindau Tumor Suppressor by Protein
Cross-linking. Oncogene 30:4780-90, 2011
• TANK-binding kinase 1(TBK1) controls cell survival through PAI-2/serpinBe and transglutaminase 2. PNAS
109(4)177-186. January 24 2012.
54
|
S7-2
Reverse of rena cell carcinoma by blocking autophagy
Soo-Youl Kim
Head, Cancer Biology Division
National Cancer Center
Email:kimsooyoul@gmail.com
Radiation and conventional cytotoxic chemotherapies are ineffective in treating renal cancer. RCC is
characterized by alterations in the metabolic phenotype related to glucose and energy metabolism also
referred to as aerobic glycolysis or Warburg effect. However, it does not reflect typical RCC phenotype to
induce resistance to cancer therapy. Recently we found that RCC adopted highly increased autophagy for
survival. Interestingly we found a target X to be highly expressed commonly in RCC cell lines and RCC
tumors. Target X is known for playing in many physiologically important roles for survival and defense.
One of them was autophagy processing. Target X physically interacts with p62 and recruited into
autophagosomes for degradation. Therefore targeting molecule of target X can be a victim of autophagy.
We found that treatment of target X siRNA induced apoptosis in RCC cells. Furthermore, target X directly
modifies DNA binding domain of p53, leading to p53 depletion via autophagy in RCC. This implies that
induced expression of target X promotes tumor cell survival through p53 depletion in RCC. The novel
small molecule inhibiting target X was developed. A single treatment of target X inhibitor nearly abrogated
tumor growth by stabilizing p53 in the ACHN and CAKI-1 preclinical xenograft tumor models. The
combination of target X inhibitor with sorafenib showed efficacy in RCC and was driven by target X
inhibition, suggesting that target X inhibitor may overcome the resistance to tyrosine kinase inhibitors and
represent a new therapeutic approach to RCC.
(Target X is a novel enzyme in autophagy process.)
|
55
최 만 호 _Man Ho Choi, 한국과학기술연구원
Office: 02-958-5081 / C.P.: 010-7741-5081
E-mail: mh_choi@kist.re.kr
학력/경력
2002.2
2004.9
현재
성균관대학교/약학대학 임상분석 박사
MIT Post-doc.
한국과학기술연구원 책임연구원
주요연구실적
∙ 생체 내 호르몬 분석 연구: SCI 논문 70여편 발표
∙ 머리카락에 의한 심혈관 및 치매 예측 연구: 기술이전 완료
56
|
S7-3
Biochemical roles of endocrine and neurologic systems
on the hormonal dependent diseases
Man Ho Choi1, Hong Seog Seo2 and Bong Chul Chung1
1
Future Convergence Research Division, Korea Institute of Science and Technology, Seoul 136-791, Korea;
2
Cardiovsacular Center, Korea University College of Medicine, Seoul 152-703, Korea
E-mail: mh_choi@kist.re.kr
Abnormalities in steroid hormones synthesized from cholesterol in the adrenal cortex, ovaries, and testes
are responsible for development and prevention of many age- and hormonal dependent diseases including
cardiovascular events. Due to their biochemical roles in endocrine and neurologic systems, the quantitative
metabolite profiling is needed to elucidate altered expression of endogenous hormones.
In contrast to classical bioanalyses that mainly focus on single metabolites or defined sets of linked
reactions and cycles, metabolite profiling involves the collection of quantitative signatures on a broad series
of metabolites to understand metabolic dynamics associated with biological conditions of interest. In
particular, mass spectrometry (MS)–based metabolomics has been considered in a wide range of applications,
and it provides good specificity and reproducibility. It is mainly coupled to sample pretreatment and
chromatographic separation steps to reduce the biological complexity. Biological samples (e.g., urine, blood,
hair) are collected, pretreated for metabolite extraction, and analyzed with either gas chromatography–mass
spectrometry (GC-MS) or liquid chromatography–mass spectrometry (LC-MS).
This presentation will be introduced that the quantitative metabolite profiles of endogenous hormones
expressions between healthy and disease states may allow the causes of disease, stages of progression, and
diagnostic biomarkers to be identified.
|
57
김 광 표 _Kwang Pyo Kim, 건국대
Office: 02-458-7682 / C.P.: 010-9574-8880
E-mail: kpkim@konkuk.ac.kr
학력/경력
2013
2004-2012
2002-2004
1997-2002
1992-1997
1990-1992
1986-1990
건국대학교 분자생명공학과/교수
건국대학교 분자생명공학과/조교수
Harvard Medical School Post-doc. Fellow
Univ. of Illinois at Chicago Ph.D
CJ제일제당 선임연구원
서울대학교 화학과/석사
서울대학교 화학과/학사
주요연구실적
• Shanta SR, Kim TY, Hong JH, Lee JH, Shin CY, Kim KH, Kim YH, Kim SK, Kim KP. A new combination
MALDI matrix for small molecule analysis: application to imaging mass spectrometry for drugs and metabolites.
Analyst. 2012 Dec 21;137(24):5757-62.
• Lee GK, Lee HS, Park YS, Lee JH, Lee SC, Lee JH, Lee SJ, Shanta SR, Park HM, Kim HR, Kim IH, Kim
YH, Zo JI, Kim KP, Kim HK. Lipid MALDI profile classifies non-small cell lung cancers according to the
histologic type. Lung Cancer. 2012 May;76(2):197-203.
• Shanta SR, Choi CS, Lee JH, Shin CY, Kim YJ, Kim KH, Kim KP. Global changes in phospholipids identified
by MALDI MS in rats with focal cerebral ischemia. J Lipid Res. 2012 Sep;53(9):1823-31.
• Shanta SR, Zhou LH, Park YS, Kim YH, Kim Y, Kim KP. Binary matrix for MALDI imaging mass spectrometry
of phospholipids in both ion modes. Anal Chem. 2011 Feb 15;83(4):1252-9.
• Kim Y, Shanta SR, Zhou LH, Kim KP. Mass spectrometry based cellular phosphoinositides profiling and phospholipid
analysis: a brief review. Exp Mol Med. 2010 Jan 31;42(1):1-11.
58
|
S7-4
Application of imaging mass spectrometry of phospholipid metabolites
for biomarker identification
Kwang Pyo Kim
Department of Molecular Biotechnology, Konkuk University, Seoul 143-701
E-mail: kpkim@konkuk.ac.kr
Recent developments in imaging mass spectrometry (IMS) have allowed complete mapping of the
biological molecules including phospholipids (PLs) that are the major building block molecules of cellular
membranes. The IMS technique can detect different classes of PLs as well as their location information
directly from tissue sections. PL head groups carry either positive and/or negative charges; therefore, IMS
experiments must be conducted in both positive- and negative-ion mode to detect all types of phospholipids.
Recently, we developed an optimized matrix preparation for IMS experiments in both ion modes that
maximize PL identification from a single brain tissue section. The optimized matrix showed improved
stability and consistency during both ion mode experiments and successfully identified >100 peaks of PLs
determined by parent ion m/z value. Further tandem mass spectrometric analysis (MS/MS) was performed to
those PLs that are anatomically important according to their distribution on rat brain tissue section.
|
59
60
|
Session 8
Environmental Toxicity
Chair: 장윤석_포항공과대학교 / 표희수_한국과학기술연구원
|
61
김 영 미 _Youngmi Kim Pak, 경희대학교 의과대학 생리학교실
Office: 02-961-0908 / C.P.: 010-9965-0958
E-mail: ykpak@khu.ac.kr
학력/경력
1983/1985
1991
1991-1995
1996-2002
2002-2007
2007-현재
서울대학교 약학과 학사/석사
미국 퍼듀대학교 생화학 박사
미국 스탠포드대학 의과대학 Post-doc., Res. Associate
국립보건원 대사질환과 보건연구관/과장
울산의대 아산생명과학연구소 부교수
경희대 의과대학 생리학교실 교수
주요연구실적
Out of total 84 SCI papers since 1988
• Kim MS*, Pak YK*, Jang PG, Namkung C, Choi YS, Jeon MJ, Han SM, Kim SW, Kim HS, Park JY, Kim
YB, and Lee KU (2006) Role of forkhead transcription factor FOXO1 in the regulation of food intake and
energy homeostasis, Nature Neuroscience, 9, 901-906 (* co-first author)
• Chang E-J, Ha J, Oerlemans F, Lee YJ, Lee SW, Ryu J, Kim HJ, Lee Y, Kim H-M, Choi J-Y, Kim J Y,
Shin CS, Pak YK, Tanaka S, Wieringa B, Lee ZH, and Kim HH (2008) Brain-type creatine kinase has a
crucial role in osteoclast-mediated bone resorption, Nature Medicine, 14(9):966-972
• Lim S, Ahn SY, Song IC, Chung MH, Jang HC, Park KS, Lee KU, Pak YK*, and Lee HK* (2009) Chronic
exposure to the herbicide, atrazine, causes mitochondrial dysfunction and insulin resistance, PLoS One 4(4):
e5186- (* Co-corresponding authors)
• Ahn SY, Choi YS, Koo HJ, Jeong JH, Park WH, Kim M, Piao Y, and Pak YK (2010) Mitochondrial dysfunction
enhances the migration of vascular smooth muscles cells via suppression of Akt phosphorylation, BBA-General
Subjects, 1800: 275-281
• Choi YS*, Jeong JH*, Min H-K, Jung H-J, Hwang D, Lee S-W#,and Pak YK# (2011) Shot-gun proteomic
analysis of mitochondrial D-loop DNA binding proteins: Identification of mitochondrial histones, Molecular
BioSystems, 7: 1523-1536 6
• Jeon J, Jeong J-H, Baek J-H, Koo H-J, Park W-H, Yang J-S, Yu M-H, Kim S, Pak YK (2011) Network clustering
revealed the systemic alterations of mitochondrial protein expression, PLoS Computational Biol. 7(6): e1002093
(IF=5.759)
• Piao Y, Kim H-G, Oh MS and Pak YK (2012) Overexpression of TFAM, NRF-1 and myr-AKT protects the
MPP+-induced mitochondrial dysfunctions in neuronal cells, BBA-General Subjects, 1820: 577-585
• Koo H-J, Piao Y, and Pak YK (2012) ER stress impairs insulin signaling through mitochondrial damage in
SH-SY5Y cells, Neuro-Signals, 20: 265-280
• Park W-H, Jun DW, Kim JT, Park H-K, Chang Y-S, Park KS, Lee HK, and Pak YK (2013) Novel cell-based
assay reveals associations of circulating serum AhR-ligands with metabolic syndrome and mitochondrial dysfunction.
BioFactors, in press
62
|
S8-1
Circulating serum dioxin-like compound-induced mitochondrial
dysfunction and diabetes pandemic
Youngmi Kim Pak
Department of Physiology, College of Medicine, Kyung Hee University, Seoul 130-701, Korea
Email:ykpak@khu.ac.kr
Serum concentrations of environmental pollutants have been positively correlated with diabetes and
metabolic syndrome in epidemiologic studies. In turn, abnormal mitochondrial functions have been associated
with the diseases. The relationships between these variables, however, have not been studied. We analyzed
whether low-dose circulating dioxins in human serum are associated with parameters of metabolic syndrome
and mitochondrial functions. We developed the newly designed cell-based aryl hydrocarbon receptor (AhR)
agonist bioassay system. Serum AhR agonist activities were measured as serum 2,3,7,8-tetrachlorodibenzo-p-dioxin
equivalent (sTCDDeq) in pM using 10 μl human sera from 97 Korean participants (47 with glucose
intolerance and 50 matched controls). sTCDDeq levels were higher in participants with glucose intolerance
than normal controls and were positively associated (p<0.01) with obesity, blood pressure, serum
triglyceride, and fasting glucose, but not with HDL-cholesterol. Body mass index was in a positive linear
relationship with sTCDDeq in healthy participants. When myoblast cells were incubated with human sera,
ATP generating power of mitochondria became impaired in a dioxin concentration-dependent manner. Our
results support that circulating dioxin-like substances may directly reduce mitochondrial functions in tissues,
leading to weight gain, glucose intolerance, and metabolic syndrome. Our rapid cell-based assay using
minute volume of human serum may provide one of the best monitoring systems for circulating dioxins,
good clinical biomarkers for the progress of disease and therapeutic efficacy.
|
63
서 정 주 _Jungju Seo, 한국기초과학지원연구원
Office: 02-6943-7686 / C.P.: 010-9570-7686
E-mail: jjseo@kbsi.re.kr
학력/경력
1987
1997
2013
고려대학교 화학 이학사
고려대학교 물리화학 이학박사
한국기초과학지원연구원 책임연구원 분석연구부장
주요연구실적
• Jeoung Hwa Shin, Hee Ock Bo, Eunjung Bang, Shela Gorinstein, Jungju Seo*, Development of a
cleanup method for polybrominated diphenyl ether (PBDE) in fish by freezing-lipid filtration, EUROPEAN
FOOD RESEARCH AND TECHNOLOGY, 235(2), 295-301, 2012
• Jinsung An, Ki-Hyun Kim, Hye-On Yoon, Jungju Seo*, Application of the wavelength dispersive
X-ray fluorescence technique to determine soil fluorine with consideration of iron content in the matrix,
SPECTROCHIMICA ACTA PART B-ATOMIC SPECTROSCOPY, 69(1) 38-43, 2012
• Seung In Hong, Na Ma, Jungju Seo, In-Hwan Kim, Lipase-catalyzed synthesis of capsiate analog
using vanillyl alcohol and conjugated linoleic acid under vacuum system, Process Biochemistry, 47,
2317-2322, 2012
• Yong-Kook Kwon, Young Sang Jung, Jong-Chul Park, Jungju Seo, Man-Sik Choi, Geum-Sook Hwang,
Characterizing the effect of heavy metal contamination on marine mussels using metabolomics, MARINE
POLLUTION BULLETIN 64(9), 1874-1879, 2012
64
|
S8-2
Polybromodiphenyl ethers (PBDEs) profiling in dietary intake and
penta bromodiphenyl ether exposed rat
Jungju Seo
Seoul Center, Korea Basic Science Institute, Seoul 136-713
Email:jjseo@kbsi.re.kr
Polybrominated diphenyl ethers (PBDEs) are a group of brominated flame retardants, which have been
manufactured in large quantities and widely used in a variety of consumer goods. PBDEs have come under
increased scrutiny because of their potential to impact upon the environment and human health.
A market basket study is a useful method for estimating the average intake level of PBDEs in regions,
based on a model of the average total diet. It is possible to provide information for the daily intake of
food groups. These monitoring provide current estimates of the exposure of human to PBDEs and are a
valuable tool to improve risk assessments and to develop the appropriate strategies to manage risks that
may be associated with these contaminant.
PBDEs may leach or volatilize from products and have been shown to bioaccumulate in the environment.
Bioaccumulation of PBDEs has been extensively studied several decades ago. But little information is
available about the metabolic characterization of PBDEs in animal models. In this work, we investigate the
metabolic changes in rat force-fed with pentabromodiphenyl ether (PeBDE) by NMR-based metabolomics.
1H-NMR spectra obtained from rat urine and liver tissue samples.
In order to examine the bioavailability and bioaccumulation of PBDEs at low exposure levels, we have
performed a mass balance study in rats fed a low dose of a penta-BDE for 7 days. Suitable extraction and
analytical methods were developed to achieve the chromatographic separation of pentabromodiphenyl ether
metabolites formed in vivo and to study their structure.
The identification of the OH-PBDE metabolites was also supported by full scan electron ionization mass
spectra and MS/MS spectra of LC-orbitrap mass spectrometer.
Six biomarkers were candidate from NMR analysis of urine metabolites isolated from rat dosed with
BDE119.
This study provided the valuable information of metabolic difference between treatment and control group
by using multivariate analysis and could be applied for understanding the toxicity of PBDEs.
|
65
이 덕 희 _Duk-Hee Lee, 경북대학교 의학전문대학원
Office: 053-420-4866 / C.P.: 010-8510-9774
E-mail: lee_dh@knu.ac.kr
학력/경력
1989.02
1992.02
1995.02
1989.03-1990.02
1990.03-1993.02
1993.03-2001.05
2001.05-2002.04
2002.05-2003.08
2003.09-현재
경북대학교 의과대학 졸업
경북대학교 예방의학(역학) 석사
경북대학교 예방의학(역학) 박사
경북대학교 의과대학 부속병원 인턴
경북대학교 의과대학 예방의학교실 전공의
고신대학교 의학부 예방의학교실 전임강사, 조교수
하버드대학교 보건대학원 방문교수
미네소타대학교 보건대학원 Reseach associate
경북대학교 의학전문대학원 예방의학교실 조교수, 부교수, 교수
주요연구실적
• Lee DH, Steffes MW, Sjödin A, Jones RS, Needham LL, Jacobs DR Jr. Low dose of some persistent organic
pollutants predicts type 2 diabetes: a nested case-control study. Environ Health Perspect. 2010;118(9):1235-42.
• Lee DH, Lee IK, Song K, Steffes M, Toscano W, Baker BA, Jacobs DR Jr. A strong dose-response relation
between serum concentrations of persistent organic pollutants and diabetes: results from the National Health
and Examination Survey 1999-2002. Diabetes Care. 2006;29(7):1638-44.
• Lee DH, Lind PM, Jacobs DR Jr, Salihovic S, van Bavel B, Lind L. Polychlorinated Biphenyls and Organochlorine
Pesticides in Plasma Predict Development of Type 2 Diabetes in the Elderly: The Prospective Investigation
of the Vasculature in Uppsala Seniors (PIVUS) study. Diabetes Care. 2011;34(8):1778-84.
• Lee HS, Lee JC, Lee IK, Moon HB, Chang YS, Jacobs DR Jr, Lee DH. Associations among organochlorine
pesticides, Methanobacteriales, and obesity in Korean women. PLoS One. 2011;6(11):e27773.
• Vandenberg LN, Colborn T, Hayes TB, Heindel JJ, Jacobs DR Jr, Lee DH, Shioda T, Soto AM, vom Saal
FS, Welshons WV, Zoeller RT, Myers JP. Hormones and endocrine-disrupting chemicals: low-dose effects and
nonmonotonic dose responses. Endocr Rev. 2012 Jun;33(3):378-455
66
|
S8-3
Persistent organic pollutants and Obesity-related Metabolic
Dysfunctions
Duk-Hee Lee
Department of Preventative Medicine, School of Medicine,
Kyungpook National University, Daegu, Korea
Email:lee_dh@knu.ac.kr
As one of the most important health-related issues worldwide, obesity is closely linked to many chronic
diseases. Among them, type 2 diabetes is a typical example of well-known obesity-related metabolic
dysfunctions. In the current paradigm for type 2 diabetes, obesity results from energy imbalance, insulin
resistance from obesity, and exhaustion of pancreatic beta cells from overproduction of insulin to
compensate for insulin resistance, which ultimately progresses to diabetes. However, there is growing
evidence that low dose exposure to Persistent Organic Pollutants (POPs) is involved in all these steps of
pathogenesis of type 2 diabetes.
POPs are various chemicals which share characteristics of high lipophilicity, the ability to accumulate in
fat, and resistance to biodegradation. Although most chlorinated POPs such as polychlorinated biphenyls
(PCBs) and organochlorine (OC) pesticides were banned several decades ago and the emission of dioxins
are strictly regulated in most developed countries, the exposure to these chemicals in the general population
still occurs because they have widely contaminated our food chain. Also, POPs that have accumulated in
human adipose tissue due to previous high exposure have become a continuous source of internal exposure
as POPs are slowly but continuously released from adipose tissue to the circulation and critical organs.
Several epidemiological studies showed associations between type 2 diabetes and a variety of POPs
including PCBs, dioxins, and OC pesticides such as DDT or Chlordane. A cross-sectional study in the U.S.
general population observed that obesity was not associated with type 2 diabetes among persons with very
low levels of POPs, which would suggest that the POPs accumulated in adipose tissue may play a critical
role in the pathogenesis of type 2 diabetes. Also, serum concentrations of some POPs were strongly related
to components of metabolic syndrome and insulin resistance among non-diabetes.
Following these cross-sectional findings, several recent prospective studies have observed that some OC
pesticides and PCBs predicted the future risk of type 2 diabetes in the general population although the
specific kinds of POPs predicting type 2 diabetes and the shapes of the dose-response curves varied across
studies. In addition, some OC pesticides and PCBs predicted future adiposity, dyslipidemia, and insulin
resistance among participants without diabetes. Recent animal experimental studies demonstrated that the
treatment of mixed POPs developed abdominal obesity, hepatosteatosis, and insulin resistance, which are
pre-diabetic conditions. Taken together with both epidemiological and experimental findings on POPs, the
background exposure to POPs may help to explain the recent epidemic of obesity-related metabolic
dysfunctions like metabolic syndrome and type 2 diabetes.
|
67
이 정 애 _Jeongae Lee, 한국과학기술연구원 (분자인식연구센터)
Office: 02-958-6632 / C.P.: 010-3263-5800
E-mail: frans@kist.re.kr
학력/경력
1985.2
1988.8
1999.2
1985.2-현재
2001.6-2002-12
단국대학교/화학과 화학 학사
단국대학교/대학원 물리분석화학 석사
고려대학교/대학원 분석화학 박사
한국과학기술연구원/분자인식연구센터 선임연구원
UC Berkeley Dept. Nutritional Sciences and Toxicology Post-doc. fellow
주요연구실적
논문
- 송나래, 온지원, 이정애, 박정덕, 권호장, 윤혜정, 표희수 (2013) Biomonitoring of urinary di(2-ethylhexyl) phthalate
metabolites of mother and child pairs in South Korea, Environment International 54:65-73
- Mina Ha, Hojang Kwon, Hae-Kwan Cheong, Sinye Lim, Seung Jin Yoo, Eun-Jung Kim, Seok Gun Park, Jeongae
Lee, Bong Chul Chung (2012) Urinary metabolites before and after cleanup and subjective symptoms in volunteer
participants in cleanup of the Hebei Spirit oil spill, Sci Tot Env 429:167-173
- Jeongae Lee, Min-hwa Kim, Mina Ha, Bong Chul Chung (2010) Biomed. Chromatogr. 24(5):562-568
특허
- 함정엽, 김태정, 권학철, 정봉철, 이정애, 정규혁, 아릴 나프탈렌 리그난 구조의 저스티시딘B 유도체 및 그의 제조방법
- 정혜선, 이정애, 장유라, 임종현, 정봉철, 수단 IV 를 이용한 유지의 정량 방법, 10-2012-0137304, 2012.11.29.
- 표희수, 이정애, 김승기, 정선아, 동위원소희석-기체크로마토그래피-이중질량분석기를 이용한 산성제초제의 분석,
10-2012-0103203, 2012.9.18.
- 이정애, 이수현, 정봉철, 표희수, 생체 시료 내 페놀 화합물 동시 분석 방법, 10-2012-0093684, 2012.8.27.
- 함정엽, 김태정, 이정애, 송중호, 정봉철, 신규한 포타슘 오가노-1Ｈ-1,2,3-트리아졸-4-일트리플루오로보레이트 유도체
및 그 제조방법, 10-2012-0058960, 2012.6.1.
- 표희수, 이정애, 박경수, 김미옥, 김승기, 생체 시료 중 프탈레이트 대사체 분석 방법, 10-2012-0060835, 2012.6.7.
- 이정애, 표희수, 정봉철, 생체 시료 내 다환 방향족 탄화수소 대사체 분석 방법, 10-2012-0018879, 2012.2.24.
- 정혜선, 양현옥, 이정애, 권학철, 정준영, 송영호, 박주영, 김지연, 생체적합성 용매의 용해도가 향상된 녹인 강황
추출물 및 커큐민, 10-2011-0127052, 2011.11.30.
68
|
S8-4
Urinary bisphenol-A concentration and its association with estrogen
metabolism in Korean adults based on environmental matabolomics
Eun Jee Kim1,2, Dongho Lee2, Bong Chul Chung1, Heesoo Pyo1, and Jeongae Lee1,*
1
Molecular Recognition Research Center, Korea Institute of Science and Technology, Hwarangno 14-gil 5,
2
Seongbuk-gu, Seoul 136-791, Korea; Department of Biotechnology, Korea University, 145,
Anam-ro, Seongbuk-gu, Seoul 136-701, Korea
E-mail contact: frans@kist.re.kr
Bisphenol-A (BPA), a monomer used to make polycarbonate, is estrogenic properties both in vitro and in
vivo known as one of the major environmental endocrine disrupting chemicals. Humans are cumulative or
long-term exposure to BPA. BPA was detected in 97.3% of 1904 urine specimens from Korean adults. We
investigated the urinary concentration of estrogens in low and high concentration of BPA and its possible
association with estrogen metabolism. Urine sample were selected higher BPA concentration group (BPA-H,
n=100, 11.05 ± 20.47 µg/g creatinine) and lower BPA concentration group (BPA-L, n=100, 0.70 ± 0.22
µg/g creatinine) from Biomonitoring of Hazardous Materials 2009-2010 survey years. Urinary estrogens was
hydrolyzed using enzymatic deconjugated, extracted, and then derivatized and measured using gas
chromatography-mass spectrometry. Urinary estrogens in BPA-H were higher than in BPA-L. The
concentrations of estrone, estradiol, and their hydroxylated metabolites in both gender were higher in
BPA-H than in BPA-L group with significantly (p<0.04). There were positive correlations between urinary
BPA and the estrogens. Furthermore, estrogen metabolism in BPA-H to 4-hydroxy estrogen was higher than
to 2-hydroxy estrogen. These results suggest that exposure to BPA in life-time may alter characteristic
differences concentrations of estrogens in BPA-L and BPA-H. Because human health effects are most likely
associated with long-term and low dose exposure, the relevance of single measurements of urinary BPA
concentrations has been questioned. Although single spot urine samples were limited measures of long-term
exposure to BPA, we found that prolonged exposure to BPA can adversely affect the estrogen metabolism
in circulating steroid hormone levels.
Keywords: Urinary bisphenol-A, Steroid hormone, Estrogen metabolism, Hydroxylase
|
69
70
|
Session 9
Food & Nutrition
Chair: 권대영_한국식품연구원 / 윤정한_한림대학교
|
71
김 오 연 _Oh Yoen Kim, 동아대학교 식품영양학과
Office: 051-200-7326 / C.P.: 010-3272-4515
E-mail: oykim@dau.ac.kr
학력/경력
1997.02
1999.02
2002.02
2012.03-현재
2009.02-2012.02
2006.12-2009.02
연세대학교 식품영양학과 식품영양학 학사
연세대학교 식품영양학과 영양학 석사
연세대학교 식품영양학과 임상영양학 박사
동아대학교 식품영양학과 조교수
연세대학교 노화과학연구소 연구교수
l’Institut Pasteur, Paris, FRANCE Unit 'Cytokines & Inflammation' Dept. of 'Infection & Epidemiology'
Post-doctor/Research Fellow
2002.03-2006.12 연세대학교 노화과학연구소 연구교수/박사후연구원 2013.01-현재 한국지질동맥경화학회 부총무, 학술/
교육위원
2010.11-현재
Journal “Frontiers in Cellular Endocrinology”, Frontiers Research Foundation, Switzerland Review Editor
주요연구실적
주요 연구분야: 관상동맥경화증, 당뇨병, 대사증후군 및 비만관련 질병의 위험요소와 영양요소를 포함한
환경요인의 상관관계를 산화스트레스 및 염증, 면역반응관련 지표의 비교분석, 질환위험군과 질환군을
대상으로 임상영양중재를 실시하여 위험개선효과를 관찰 연구. 이중 대사체 분석과 관련한 최근 논문 편은
다음과 같음.
1. Kim OY, Lee JH, Sweeney G. Metabolomic profiling as a useful tool for diagnosis and treatment
of chronic disease: focus on obesity, diabetes and cardiovascular diseases. Expert Rev Cardiovasc
Ther. 2013 Jan;11(1):61-8.
2. Kim JY, Kim OY, Paik JK, Kwon DY, Kim HJ, Lee JH. Association of age-related changes in
circulating intermediary lipid metabolites, inflammatory and oxidative stress markers, and arterial
stiffness in middle-aged men. Age (Dordr). 2012 Jul 18. [Epub]
3. C.Y. Ha, J.Y. Kim, J.K. Paik, Y.H.Paik, E.J.Lee, J.H.Lee. The association of specific metabolites of
lipid metabolism with markers of oxidative stress, inflammation and arterial stiffness in men with
newly diagnosed type 2 diabetes. 76;674–682, 2012.05
72
|
S9-1
Metabolomic profiling as a useful tool for diagnosis and treatment of
chronic disease: focus on obesity, diabetes and cardiovascular diseases
Oh Yoen Kim and Jong Ho Lee
Department of Food Science and Nutrition, Dong-A University, Busan Korea; Department of Food and Nutrition,
Yonsei University, Seoul, Korea
E-mail: oykim@dau.ac.kr
There have been considerable improvements in therapeutics for chronic diseases. Currently, there is a
deep appreciation and strong interest in personalized medicine as a more effective means of disease
management. However, the maximum benefit of these or other options are hard to achieve in practice,
partly due to the difficulties associated with determining optimal targets for such interventions. Recent
developments have suggested that understanding changes in metabolite profiles will confer a high degree of
predictive accuracy in terms of understanding the fundamental mechanisms resulting in perturbations of the
metabolic state. Metabolomics involves the establishment of relationships between phenotype and a
metabolic signature, which are key aspects of biological function. Metabolite profiling for the diagnosis of
disease pathogenesis is rapidly emerging and holds great promise for successful clinical implementation.
These approaches have been applied to the identification of serum/plasma metabolic markers involved in
obesity, diabetes and vascular disease using animal models or in humans. Different kinds of metabolite
profiling techniques using nuclear magnetic resonance spectroscopy, mass spectrometry, ultraperformance
liquid chromatography and so on are currently employed to generate global metabolic profiles. Scientific
information derived from these techniques can be applied to provide accurate and clinically useful
prognostic/diagnostic capability for the management of major chronic diseases. One current consideration
limiting the widespread use of metabolomic profiling is the analysis of its cost-effectiveness. In summary, it
is hoped that the information derived from metabolite profiling will make it possible to suggest
individualized therapies that more effectively treat disease
Supported by grants from Korean Research Foundation (2012-0001851).
|
73
김 영 석 _Young-Suk Kim, 이화여자대학교
Office: 02-3277-3091 / C.P.: 010-8884-8253
E-mail: yskim10@ewha.ac.kr
학력/경력
1986-1990
1990-1992
1993-1997
1997-1999
1999-2000
2000-현재
서울대학교 식품공학과 학사
서울대학교 식품공학과 석사
Rutgers University 식품공학과 박사
University of Minnesota 식품공학과/Post-doc.
Griffith Laboratories Company Principal Scientist
이화여자대학교 식품공학과/조교수, 부교수, 교수
주요연구실적
Differences in volatile compositions of ginseng species (Panax sp.), Journal of Agricultural and Food Chemistry,
60, 76176, (2012)
Metabolomic approach for determination of key volatile compounds related to beef flavor in glutathione-Maillard
reaction products, Analytica Chimica Acta, 703(2), 204 (2011)
Metabolic analysis of guava (Psidium guajava L.) fruits at different ripening stages using different data-processing
approaches, Journal of Chromatography B, 878, 2983 (2010)
74
|
S9-2
Determination of key volatile and non-volatile components related to beef
flavor in glutathione Maillard reaction products using metabolomic approach
Sang Mi Lee1, Nahyun Kim2, Dongho Lee2, Kwang-Ok Kim1 and Young-Suk Kim1
1
Department of Food Science and Engineering, Ewha Womans University, Seoul 120-750, Korea
2
School of Life Sciences and Biotechnology, Korea University, Seoul 136-713, Korea
E-mail: yskim10@ewha.ac.kr
Flavor perception depends on the combined responses of our senses and the cognitive processing of those
inputs. In particular, olfaction, taste and somatosenses can affect our brain, resulting in our flavor
experience. The multimodal sensation occurred by divers components can be efficiently studied on the base
of metabolomic approach which allows a fast and unbiased comparative multivariate analysis of the whole
small molecules related to the sensory inputs. In this study, the non-target analysis was performed using
gas chromatography-time-of-flight mass spectrometry (GC-TOFMS) and ultra performance liquid
chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOFMS) combined by sensory analysis
to investigate some novel volatile and non-volatile components related to sensory attributes of
glutathione-Maillard reaction products (GSH-MRPs). The following 18 descriptors were developed to
describe the sensory attributes of the beef stock samples containing 12 different types of GSH-MRP: 6
odors (beef, chicken, sulfur, potato, soy sauce, and chestnut), 5 tastes (salty, sour, sweet, bitter, and MSG),
6 flavors (beef, chicken, sulfur, potato, soy sauce, and chestnut), and 1 mouthfeel attribute (mouthcoating).
Then, the PLS regression method of multivariate analysis was employed to interpret and visualize
correlations between the results obtained by sensory analysis and other types of variables, such as volatile
and non-volatile components in GSH-MRPs. We demonstrated that the unbiased non-targeted analysis based
on metabolomic approach allowed the determination of key volatile and non-volatile components related to
the beef flavor attribute in GSH-MRPs.
|
75
이 동 호 _Dongho Lee, 고려대학교 생명과학대학 생명공학부
Office: 02-3290-3017 / C.P.: 010-5438-1748
E-mail: dongholee@korea.ac.kr
학력/경력
1997-2001
2002-2003
2003-2006
2006-현재
University of Illinois at Chicago 약학 약학박사
Research Triangle Institute Postdoctoral chemist
한국생명공학연구원 선임연구원
고려대학교 생명과학대학 부교수
주요연구실적
천연물로부터 의약품등 기능성 소재개발
약용식물의 표준화
76
|
S9-3
Metabolomic Analysis of Natural Products: Age Discrimination
of Panax ginseng
Dongho Lee
College of Life Sciences and Biotechnology, Korea University, Seoul 136-713, Korea
E-mail: dongholee@korea.ac.kr
Metabolomics, a fine combination of analytical and statistical techniques, provides major insights into the
similarities and differences of samples in various environmental conditions by quantitatively and qualitatively
measuring the dynamic range of metabolites in organisms. Metabolomics has been applied to research on
natural products in various ways, especially for quality control of medicinal plants. The diverse metabolome
data obtained by using this approach enables comparison among samples based on multivariate statistical
methods. By applying various metabolite selection methods to the data set, the optimal number of
metabolites, which are possible biomarkers, is suggested to derive the best result for discriminating sample
groups.
Here, the ultraperformance liquid chromatography-quadrupole time-of-flight mass spectrometry was applied
to discriminate the cultivation ages of Panax ginseng. Metabolite profiling of P. ginseng samples aged from
1 to 6 years has been performed, and various metabolite selection methods, including random forest,
prediction analysis of microarray, and partial least squares-discriminant analysis, have been applied to
improve the interpretability of age discrimination data. From the results on the clear discrimination of
ginseng cultivation ages, 10 possible age-dependent key constituents have been proposed. Their structures
were identified using multi-stage mass spectrometric analyses, such as tandem mass and accurate mass, by
comparison with an in-house ginsenoside library and with literature data.
Although long-term and more comprehensive studies are required to build the classification system and
validate the biomarker candidates, this metabolomic approach is worthy of further exploration as a method
for assessing the quality of diverse medicinal plants.
|
77
안 지 윤 _Jiyun Ahn, 한국식품연구원
Office: 031-780-9079 / C.P.: 010-2236-9079
E-mail: jyan@kfri.re.kr
학력/경력
1999
2001
2006
2001
2007
2008
건국대학교 수의학 학사
건국대학교 수의생리학 석사
건국대학교 수의생리학 박사
한국식품연구원 연구원
한국식품연구원 선임연구원
과학기술연합대학원대학교 조교수
주요연구실적
○ 관심 연구분야 : 비만, 당뇨, 인슐린 저항성, 식품소재의 기능성 연구
○ 주요 연구실적 :
• J Ahn, MY Um, H Lee, CH Jung, SH Heo, T Ha. (2013) Eleutheroside E, an active component of Eleutherococcus
senticosus, ameliorates insulin resistance in type 2 diabetic db/db mice. Evid Based Complement Alternat
Med (accepted) 외 3건
• J Ahn, MY Um, H Lee, CH Jung, SH Heo, T Ha. (2012) Lycopene inhibits hepatic steatosis via
microRNA-21-induced downregulation of fatty acid-binding protein 7 in mice fed a high-fat diet. Mol Nutr
Food Res 56: 1665-1674 외 9건
• J Ahn, H Lee, CH Chung, T Ha. (2011) High fat diet induced downregulation of microRNA-467b increased
lipoprotein. BBRC 414: 664-669 7건
• J Ahn, H Lee, S Kim, T Ha. (2010) Curcumin-induced suppression of adipogenic differentiation is accompanied
by activation of Wnt/β-catenin signaling. Am J Physiol Cell Physiol 298: C1510-C1516 외 3건
• J Ahn, H Lee, S Kim, D Kwon, T Ha. (2008) Dietary resveratrol alters lipid metabolism-related gene expression
of mice on an atherogenic diet. J Hepatol 49: 1019-1028 외 6건
• J Ahn, H Lee, S Kim, T Ha. (2007) Resveratrol inhibits TNF-a-induced changes of adipokines in 3T3-L1
adipocytes. BBRC 364: 972-977 외 1건
78
|
S9-4
The role of microRNAs in metabolic disorders and the involvement
of food components
Jiyun Ahn and Tae Youl Ha
Metabolism and Nutrition Research Group, Korea Food Research Institute, Seongnam, Korea
E-mail: jyan@kfri.re.kr
MicroRNAs (miRNAs) are highly conserved, small non-coding RNAs that regulate gene expression at the
post-transcriptional level by binding to complementary sites on target transcripts. Thus, miRNAs are
important modulators of developmental and physiological processes, including energy homeostasis, lipid
metabolism, pancreatic β-cell development, adipogenesis, and weight gain due to a high fat diet.
Dysregulation of miRNAs may contribute to metabolic abnormalities and this has led to studies of miRNA
that may potentially serve as therapeutic targets for ameliorating metabolic disorders.
Metabolic syndrome is a major public health challenge and is associated with obesity, type 2 diabetes,
and cardiovascular disease. Life style modification focusing on diet has been suggested as a successful
approach for managing metabolic disorders. Food contains various bioactive compounds including
phytochemicals and the health beneficial effects of food components through modulation of gene expressions
have been widely reported. However, there are relatively few studies on food components for the regulation
of miRNAs involved in various metabolic disorders.
Here, we will discuss our understanding of the roles of miRNAs in metabolic disorders, with specific
emphasis on the hepatic steatosis and obesity. In addition, the underlying mechanisms of food component
through the regulation of miRNA expressions are also be discussed.
Supported by grants from Korea Food Research Institute
|
79
80
|
YOUNG SCIENTISTS
Young Scientists 1
Chair: 오세량_한국생명공학연구원
|
1
김 수 아 _Sooah Kim, 고려대학교 생명공학과
Office: 02-3290-3471 / C.P.: 010-6327-7322
E-mail: tndklove@korea.ac.kr
학력/경력
2003.03.-2007.02 고려대학교 식품공학부 학사
2007.03.-2009.02 고려대학교 식품공학과 석사
2009.09-현재
고려대학교 생명공학과 박사과정 (지도교수 : 김경헌)
주요연구실적
• S Kim, DY Lee, G Wohlgemuth, HS Park, O Fiehn, KH Kim. Analytical Chemistry 2013, 85, 2169-2176
• S Kim, EJ Yun, MA Hossain, H Lee, KH Kim. Analytical and Bioanalytical Chemistry 2012, 404, 553-562
• S Kim, MH Shin, MA Hossain, EJ Yun, H Lee, KH Kim. Analytical and Bioanalytical Chemistry 2011, 399,
3519-3528
• S Kim, EJ Yun, JS Bak, H Lee, SJ Lee, CT Kim, J-H Lee, KH Kim. Food Chemistry 2010, 121, 521-526
• YH Jung, S Kim, TH Yang, HJ Lee, D Seung, Y-C Park, J-H Seo, I-G Choi, KH Kim. Bioprocess and Biosystems
Engineering 2012, 35, 1497-1503.
• SH Chu, H-N Noh, S Kim, KH Kim, S-W Hong, H Lee. Plant Molecular Biology 2010, 74, 493-502
• MA Hossain, S Kim, KH Kim, S-J Lee, H Lee. Hortscience 2009, 44, 1907-1913
• SR Kim, HT Kim, HJ Park, S Kim, HJ Choi, G-S Hwang, JH Yi, DH Ryu, KH Kim. International Journal
of Food Microbiology 2009, 134, 190-195
84
|
Y1-1
Yeast metabolome sampling methodology for metabolomics and
metabolic engineering
Sooah Kim and Kyoung Heon Kim
Food Bioengineering Laboratory, School of Life Sciences and Biotechnology
Korea University, Seoul 136-713, Korea
E-mail: tndklove@gmail.com
Metabolomics, which is the study of global changes of the entire set of metabolites from a living
organism, can be a powerful tool in the metabolic and physiological study. The rapid arrest of cell
metabolism and the reproducible, nonselective and efficient recovery of all existing metabolites are the
essential prerequisites to obtain reliable biological data that can closely show the actual events taking place
in the living system. Saccharomyces cerevisiae is one of the most popular industrial strains for production
of ethanol and have been used as a model microorganism in microbial metabolomics and metabolic
engineering. However, a few studies have been conducted to develop or systematically evaluate the sample
preparation methodology for metabolite analysis and profiling of S. cerevisiae. In this study, we have
studied the development and evaluation of the sampling and extraction methods for the global metabolite
profiling of S. cerevisiae using gas chromatography-time-of-flight mass spectrometry (GC/TOF MS). These
results can be used as a customized protocol for the yeast or an experimental strategy for microbial
metabolome analysis and microbial metabolic engineering.
This work was supported by the Pioneer Research Center Program (2011-0002327) and the Advanced
Biomass R&D Center of Korea for the Global Frontier Program (2011-0031353), both funded by Korean
Government (MEST).
|
85
양 승 옥 _Seung-Ok Yang, 농촌진흥청 인삼특작부 인삼특작이용팀
Office: 043-871-5684 / C.P.: 010-6339-6399
E-mail: yangso@wm.cau.ac.kr/ yangso@korea.kr
학력/경력
2005
2007
2011
2012-현재
서울산업대 식품공학 학사
서울산업대 식품공학/식품화학 석사
중앙대 약학대학 생약학/ 천연물 대사체학 박사
농촌진흥청 인삼특작부 인삼특작이용팀 대사체학을 이용한 인삼 및 특작물 품질관리 박사후 연구원
주요연구실적
과제 경력
• 2006~현재: 식물, 식품 및 생체시료를 소재로 대사체학을 이용한 9건의 프로젝트 수행
학위 논문
• 석사학위: Riboflavin과 chlorophyll b 광감제 첨가 광산화에 의한 daidzein과 genistein의 안정성 연구 (2007. 02. 23)
• 박사논문: 대사체학 기법을 이용한 인삼의 연근분리와 감초의 종 분리를 위한 예측모델 시스템 개발 (2011. 08. 26)
논문실적 (최근 5년)
• 2007~현재: 국, 내외 논문 21편 투고
대사체학 관련 논문 16편 투고
주요연구분야
• GC-MS, LC, NMR을 이용한 천연물, 식품 및 생체시료 대사체 연구
• MDGC를 이용한 정밀분석기기 연구
• GC, GC-MS를 이용한 지방산화 연구
• HPLC를 이용한 콩의 isoflavone 연구
• 식물세포배양
• in vitro 생리활성 assay
86
|
Y1-2
Classification of black raspberry (Rubus Occidentalis) fruits at
different ripening stages by NMR and multivariate statistical analysis
and effect of antiinflammatory and antioxidant activities
1
1
1
1
1
Seung-Ok Yang , Sang-Hyun Sohn , Sang-Won Lee , Young-Ock Kim , Hyung-Don Kim ,
1
2
1
Seung-Yu Kim , Hyung-Kyoon Choi , Yu Su Shin
1
Department of Herbal Crop Research, NIHHS, RDA, Eumsung, Chungbuk, 369-873, Republic of Korea
2
College of pharmacy, Chung-Ang University, Seoul 156-756, Republic of Korea
E-mail: yangso@wm.cau.ac.kr
The black raspberry (BR) is fruit of Rubus Occidentalis L. of the family Rosaceae. Rubus Occidentalis is
a species of Rubus native to eastern North America. Black raspberries have been used as a medicinal
plant, and exhibits anticancer, antioxidant, antiinflammatory, immune modulation activities. Different stage of
maturation of the Rubus occidentalis harvested in Gochang, South Korea (GPS N 35° 29′ 54.23, E 126°
39′ 14.53), was investigated through global metabolite profiling by NMR spectroscopy. NO (nitric oxide)
and DCF-DA (2',7'-dichlorofluorescein diacetate) assays were performed to evaluate anti-inflammatory effect
and ROS (reactive oxygen species) production by the treatment of the water and ethanol extracts of BR
fruits in animal cells.
The PCA and PLS-DA clearly distinguished BR according to the unripened, mid-ripened, and ripened
maturation stage. The metabolites in BR compounds were assigned as isoleucine, valine, threonine, alanine,
glutarate, acetate, succinate, methylamine, citrate, asparagines, 2-oxoglutarate, malonate, glucose, fructose,
xylose, tyrosine, and formate. The loading plot of 3 maturation stage BR fruits D2O direct extracts samples
showed that the unripened extract contained more isoleucine, valine, threonine, alanine, glutarate, acetate,
succinate, citrate, malonate, xylose, and tyrosine. The compounds of asparagines, 2-oxoglutarate in
mid-ripened fruits were relatively higher than unripened and ripened maturation stages, whereas those of
glucose, fructose, glycine were higher in ripened stage than the other stage. The unripened fruits of BR
were more effective than the ripened stage fruits in production of inflammatory mediator and hydrogen
peroxide (H2O2)induced-oxidative stress. Antiinflamatory and antioxidant effect was higher in unripened BR
fruits containing higher contents of amino acid, oganic acid and xylose.
This study was supported by 2013 Post Doctoral Course Program of Department of Medicinal Crop
Research, Rural Development Administration, Republic of Korea.
|
87
정 지 연 _Jeeyoun Jung, 한국기초과학지원연구원
Office: 010-9102-0918 / C.P.: 010-9102-0918
E-mail: jjy0918@kbsi.re.kr
학력/경력
2006
2010
2012
2007
2013
원광대학교 한의학과 한의학 학사
원광대학교 한의학과 한희학 석사
원광대학교 한의학과 한의학 박사
LG 생활건강 연구원
한국기초과학지원연구원 박사후연구원
주요연구실적
• 1H NMR based metabolomic study on resistance to diet-induced obesity in AHNAK knock-out mice, 2010,
Biochemical and Biophysical Research Communications, 403, 428-434
• 1H-NMR Based metabolomics study of cerebral infarction, 2011, Stroke, 42, 1282-1288
• 1H NMR based metabolic profiling of naproxen-induced toxicity in rats, 2011, Toxicology letters, 200, 1-7
• 1H NMR-based metabolite profiling of diet-induced obesity in a mouse mode, 2012, BMB Reports, 45, 419-424
• Serum metabolomics reveals pathways and biomarkers associated with asthma pathogenesis, Clinical & Experimental
Allergy, 2013
88
|
Y1-3
NMR-Based metabolite profiling for early detection and non-invasice
diagnosis of gastric cancer
Jeeyoun Jung, Ho-Sub Lee, Eun Jung Bang, Sung-il Cho, You-Jin Jang, Jung-Myun Kwak,
Do Hyun Ryu, Sung-Soo Park, Geum-Sook Hwang
Integrated Metabolomics Research Group, Seoul center, Korea Basic Science Institute, Seoul, 136-701, Republic Korea
Department of physiology, College of Oriental Medicine, Wonkwang University, Iksan, 540-549, Republic of Korea
E-mail: jjy0918@kbsi.re.kr
Diagnosis and monitoring of gastric cancer (GC) on the basis of non-invasive measurements is difficult
clinically. Thus, we used a metabolomics approach to investigate altered metabolic patterns in urine from
patients with gastric cancer and find urinary metabolic biomarkers for the non-invasive diagnosis of gastric
cancer, and compare the urinary and tissue metabolic profiling of GC patients.
In total, 154 urine samples from GC patients and healthy individuals, and 30 pairs of matched tumor
and normal stomach tissues were collected. Multivariate analysis was performed on the urinary and tissue
1
1
metabolic profiles, which were acquired using H-NMR and H HR-MAS spectroscopy, respectively.
Multivariate statistical analysis showed a significant separation between GC patients and healthy
individuals in both the urinary and tissue data. The metabolites perturbed in the urine of GC patients were
related to amino acid and lipid metabolism, consistent with the changes in metabolism of gastric cancer
tissue. Furthermore, the presence of gastric cancer in the external validation of the urine model was
predicted with high accuracy; patients with early gastric cancer were also predicted correctly.
These data illustrate that a urinary metabolomics approach can be effectively used for early diagnosis of
gastric cancer, and balanced metabolite profiles between systemic environment and tumor microenvironment
not only provide insights into the mechanisms associated with gastric cancer but also allow reliable
identification of potential markers for gastric cancer.
Supported by National Research Foundation of Korea Grant funded by the Korean Government
(2010-0024825, 2011-002827) and Korea Basic Science Institute (T33409).
|
89
서 정 주 _Jeong Ju Seo, 경북대학교
Office: 053-420-5443 / C.P.: 010-8948-1651
E-mail: honeysjj@hanmail.net
학력/경력
2005
2013
충북대학교 생명약학 석사
경북대학교 임상약리학 박사수료
주요연구실적
Research paper
Jeong Ju Seo, Jeonghyeon Park, Min Ho Bae, Mi-sun Lim, Sook Jin Seong, Joomi Lee, Sung Min Park, Hae
Won Lee, Young-Ran Yoon ‘Rapid determination of sumatriptan in human plasma by ultra performance liquid
chromatography-tandem mass spectrometry and its application to clinical pharmacokinetic study' J Chromatogr
B Analyt Technol Biomed Life Sci. 2013 Mar 1;919-920:38-42. doi: 10.1016/j.jchromb.2013.01.004. Epub 2013
Jan 18.
Oral presentation
Jeong Ju Seo, Jeonghyeon Park, Min Ho Bae, Sook Jin Seong, Mi-sun Lim, Hae Won Lee, Young-Ran Yoon
'Metabolic phenotype and its relationship with CYP2D6 genetic polymorphisms and pharmacokinetic variation'
(KSCPT) Nov../16/2012
90
|
Y1-4
Evaluation of CYP2D6 allele activity through metabolomic profile
using liquid chromatography-mass spectrometry
Jeong Ju Seo1,2, Mi-Ri Gwon1,2, Ji Yeon Hyun1,2, Yong-Chul Shin1,2,
1
1,2
Hae Won Lee , Young-Ran Yoon
1
Departmetnt of Biomedical Science, Kyungpook National University Graduate School, 680 Gukchaebosang-ro,
Jung-gu, Daegu 700-842, South Korea
2
Clinical Trial Center, Kyungpook National University Hospital, 130 Dongdeok-ro, Jung-gu, Daegu 700-721, South Korea
E-mail: honeysjj@hanmail.net
The CYP2D6*10B/*10B allele is a clinically important gene that is found in about 50% of Koreans.
This allele is responsible for the pharmacokinetic (PK) variability. Metabolomics is a science that
establishes the relationship between the metabolic signature, which is the key aspect of biological functions,
and the phenotype. Based on pharmacometabolomics, individual variations not only in PK but also
pharmacogenomics can be explained. In this study, the enzyme activity variations of the CYP2D6*10B/*10B
allele are evaluated through the metabolic profiling.
A total of 16 healthy Korean males were divided into the CYP2D6*1/*1 (wild) and CYP2D6*10B/*10B
(mutant) groups through the RT-PCR method before 100-mg of metoprolol tartrate was administered to
them. The blood samples for the measurement of the blood concentrations of the metoprolol and
α-hydroxymetoprolol, and for the calculation of the metabolic ratio (MR), were collected for 24h and then
analyzed through LC-MS. Metabolic profiling was conducted using the urine samples that were collected for
12h before and after the drug administration using LC-MS. The multivariate analysis of metabolic profiles
were performed by SIMCA software.
The PK parameters and MR were significantly different with wild and mutant groups. To explain the
enzyme activities in the mutant group that showed high degree individual variations, phenylethylamine,
phenylalanine, and estradiol were selected through PLS modeling. Using the selected metabolites, the
predicted MR from mutant group was well correlated to the actual MR. In conclusion, the study suggests
potential of clinical application for enzyme activity evaluation using metabolic profiling.
This research was supported by grants from the Korea Healthy 21 R&D Project, Ministry of Health &
Welfare, and the Republic of Korea (A070001) & the National Project for Personalized Genomic Medicine
(A111218-PG02), the Ministry of Health & Welfare, Republic of Korea, and the Basic Science Research
Program through the National Research Foundation of Korea (NRF) funded by the Minisry of Education
Science and Technology (2010-0022996), Republic of Korea.
|
91
92
|
Young Scientists 2
Chair: 김윤곤_숭실대학교
|
93
신 광 희 _Kwang-Hee Shin, 서울대학교 의과대학
Office: 02-740-8910 / C.P.: 010-9502-7745
E-mail: happyi69@snu.ac.kr
학력/경력
2010.03- 2012.08
2008.03- 2010.02
2003.03- 2007.02
2012.09- 현재
2012.08- 2012.12
2007.02- 2008.02
서울대학교 의과대학 의학과(임상약리학) 박사
서울대학교 의과대학 의학과(임상약리학) 석사
서울대학교 약학대학 약학과 학사
서울대학교 의과대학 의학연구원 선임연구원
덕성여자대학교 약학대학 시간 강사 약물대사학
서울대학교병원 임상약리학과 연구원
주요연구실적
• K.H. Shin et al, Pharmacokinetic and Pharmacodynamic Properties of a New Long-Acting Granulocyte
Colony-Stimulating Factor (HM10460A) in Healthy Volunteers, Biodrugs, (2013-Jan-29 epub ahead of print)
• K.H. Shin et al, A Positron Emission Tomography Microdosing Study with Sertraline in Healthy Volunteers,
International Journal of Clinical Pharmacology and Therapeutics, 2012:50(3);224-232
• K.H. Shin et al, The Effect of the Newly Developed Angiotensin Receptor II Antagonist Fimasartan on the
Pharmacokinetics of Atorvastatin in relation to OATP1B1 in Healthy Male Volunteers, Journal of cardiovascular
pharmacology, 2011:58(5);492-499
• K.H. Shin et al, Pharmacokinetic Comparison of a New Sustained-Release Formulation of Glimepiride/Metformin
1/500 mg Combination Tablet and a Sustained-Release Formulation of Glimepiride/Metformin 2/500 mg Combination
Tablet in Healthy Korean Male Volunteers: A Randomized, 2-Sequence, 2-Period, 2-Treatment Crossover Study,
Clinical Therapeutics, 2011:33(11);1809-1818
• K.H. Shin et al. Effect of ketoconazole on the pharmacokinetics of udenafil in healthy Korean subjects. British
Journal of Clinical Pharmacology. 2010; 69(3):307-310
• K.H. Shin et al. The Effects of Ketoconazole and Rifampicin on the Pharmacokinetics of Mirodenafil in Healthy
Korean Male Volunteers: An Open-Label, One-Sequence, Three-Period, Three-Treatment Crossover Study. Clinical
Therapeutics. 2009;31(12):3009-3020
• K.H. Shin et al. Comparison of the Pharmacokinetics between Udenafil 100 mg tablets and 200 mg tablets
in Healthy Volunteers. Korean Journal of Clinical Pharmacology & Therapeutics, 2008;16(1):45-52
94
|
Y2-1
Evaluation of endogenous metabolic markers of CYP3A activity using
metabolic profiling and midazolam pharmacokinetics
Kwang-Hee Shin1, Manho Choi2, In-Jin Jang1 and Joo-Youn Cho1
1
Departmetnt of Clinical Pharmacology and Therapeutics, Seoul National University College of Medicine and
Hospital, Seoul, 110-744, Korea
2
Future Convergence Research Division, Korean Institute of Science and Technology, Seoul, Korea
E-mail: happyi69@snu.ac.kr
Cytochrome P450(CYP)3A is a major drug-metabolizing enzyme and the activity marker for CYP3A has
clinically meaningful. The aim of this study was to evaluate endogenous metabolic markers of CYP3A
activity in healthy subjects using a metabolomics approach.
An open-label, 1-sequence, 3-period, 3-treatment, crossover study was conducted in 24 healthy Korean
male volunteers. Each subject received a midazolam in each of 3 study periods: midazolam 1 mg alone
(control phase); midazolam 1 mg after pretreatment with 400 mg ketoconazole once daily for 4 days
(CYP3A inhibited phase); and midazolam 2.5 mg after pretreatment with 600 mg rifampicin once daily for
10 days (CYP3A induced phase). Serial samples were collected for pharmacokinetic analysis for midazolam,
1’-hydroxy midazolam and 4-hydroxy midazolam. Pharmacokinetic parameters were obtained
noncomoopartmental analysis and compartmental analysis. The 12-hours interval urine samples were
collected for metabolomics analysis between 24 hours before and 24 hours after of midazolam
administration in each study period. Plasma and urine samples were determined using gas
chromatography-mass spectrometry and liquid chromatography-mass spectrometry. From the untargeted
metabolomics, isobutyryl-L-carnitine decreased in CYP3A inhibition state and glycochenodeoxycholate-sulftae
was increased in CYP3A induction state. From the targeted steroid profiling, the known CYP3A activity
markers, urine 6β-hydroxy-cortisol / cortisol and 6β-hydroxy-cortisone / cortisone, and plasma
4β-hydroxy-cholesterol, but also 7β-dehydroepiandrosteron (DHEA) / DHEA and 16α-hydroxy-DHEA /
DHEA were well correlated with midazolam clearance. From the correlation analysis between midazolam
clearance and metabolites changes, the equation for midazolam clearance was obtained; Ln(CL) = 2.1539 +
0.2724 * I + 0.0101 * DHEA + 0.0910 * R1 + 0.6502 * R2 (I = 1 if CYP3A5*1/*3, otherwise 0, R1:
7β-hydroxy DHEA / DHEA ratio, R2: 6β-hydroxy cortisone / cortisone). We showed that a combination of
the concentrations and ratios of several endogenous metabolites and the CYP3A5*3 genotype is a reliable
predictive marker of CYP3A activity during CYP3A induction and inhibition.
This work was supported by a grant from the National Research Foundation of Korea (NRF), which was
funded by the Korean government (MEST) (no. 2010-0009796)
|
95
최 효 정 _Hyo-Jung Choi, 경북대학교 의학전문대학원
Office: 053-420-4828 / C.P.: 010-5511-7867
E-mail: sky1339@nate.com
학력/경력
2011.03- present Kyungpook National University, Daegu, Korea Biomedical Science Ph.D. student
2007.03- 2009.02 Kyungpook National University, Daegu, Korea Biomedical Science M.S.
2003.03- 2007.02 Kyungpook National University, Daegu, Korea Life Sciences and Biotechnology B.S.
주요연구실적
주저자 논문
Choi HJ, Kwon TH. Fluorescence quenching to measure dDAVP-induced cell volume changes in kidney collecting
duct. The Korean Journal of Nephrology 2009;28:317～325 (제1저자, 연구재단등재)
Kim HY, Choi HJ, Lim JS, Park EJ, Jung HJ, Lee YJ, Kim SY, Kwon TH. Emerging role of Akt substrate
protein AS160 in the regulation of AQP2 translocation. Am J Physiol Renal Physiol. 2011 Jul;301(1):F151-61.
(공동 제1저자)
Choi HJ, Yoon YJ, Kwon YK, Lee YJ, Chae S, Hwang D, Hwang GS, Kwon TH. Patterns of gene and metabolite
define the effects of extracellular osmolality on kidney collecting duct. J Proteome Res. 2012 Jul 6;11(7):3816-28.
(제1저자)
공동저자 논문
Lee M, Lee SJ, Choi HJ, Jung YW, Frøkiaer J, Nielsen S, Kwon TH. Regulation of AQP4 protein expression
in rat brain astrocytes: role of P2X7 receptor activation. Brain Res. 2008 Feb 21;1195:1-11.
Lee YJ, Choi HJ, Lim JS, Earm JH, Lee BH, Kim IS, Frøkiaer J, Nielsen S, Kwon TH. A novel method of
ligand peptidomics to identify peptide ligands binding to AQP2-expressing plasma membranes and intracellular
vesicles of rat kidney. Am J Physiol Renal Physiol. 2008 Jul;295(1):F300-9.
Lee YJ, Lee JE, Choi HJ, Lim JS, Jung HJ, Baek MC, Frøkiær J, Nielsen S, Kwon TH. E3 ubiquitin-protein
ligases in rat kidney collecting duct: response to vasopressin stimulation and withdrawal. Am J Physiol Renal
Physiol. 2011 Oct;301(4):F883-96.
Jung HJ, Lim JS, Choi HJ, Lee MS, Kim JH, Kim SY, Kim S, Kim E, Kwon TH. Vasopressin V2R-targeting
peptide carrier mediates siRNA delivery into collecting duct cells. PLoS One. 2012;7(6):e40010.
96
|
Y2-2
Patterns of gene and metabolite define the effects of extracellular
osmolality on kidney collecting duct
Hyo-Jung Choi,†, Yu-Jeong Yoon,‡, Yong-Kook Kwon,‡,∥ Yu-Jung Lee,† Sehyun Chae,§
§
‡,∥
†
and Tae-Hwan Kwon
Daehee Hwang, Geum-Sook Hwang,
†
Department of Biochemistry and Cell Biology, School of Medicine, Kyungpook National University, Taegu, Korea
‡
Integrated Metabolomics Research Group, Seoul Center, Korea Basic Science Institute, Seoul, Korea
§
Department of Interdisciplinary Bioscience and Bioengineering, POSTECH, Pohang, Korea
∥
Graduate School of Analytic Science & Technology, Chungnam National University, Daejeon, Korea
Email: thkwon@knu.ac.kr
To investigate the effects of changes in extracellular osmolality on the function of kidney collecting duct
cells, particularly on water and sodium reabsorption in the conditions of diuresis and antidiuresis, we
generated transcriptome and metabolome profiles of primary cultured inner medullary collecting duct
(IMCD) cells. They were grown in hyperosmolar culture medium (640 mOsm) for 4 days and then exposed
to either reduced (300 mOsm) or same osmolality for 1 or 2 days more. Integrated analysis of the
transcriptome and metabolome revealed that decreased extracellular osmolality was associated with decreased
levels of organic osmolytes, glucose, intermediates of citric acid cycle, and branchedchain amino acids
(BCAA) in IMCD cells, along with significantly decreased gene expression and protein abundance of P-type
transporters (ATP1B1), ABC transporters (ABCC5 and ABCG1), and insulin signaling pathways (IRS2).
Quantitative real-time RT-PCR and semiquantitative immunoblotting confirmed the changes of transcript
levels of differentially expressed genes and protein levels. Taken together, integrated analysis of omics data
demonstrated that water and sodium reabsorption could be reduced by decreased extracellular osmolality per
se, through decreased levels of ABC transporters and IRS2, which play a potential role in the transport of
organic osmolytes, BCAA, glucose, and trafficking of epithelial sodium channel.
|
97
정 준 영 _Joon Young Jung, 고려대학교 화공생명공학과 시스템 생명공학 연구실
Office: 010-8931-6376 / C.P.: 010-8931-6376
E-mail: leonis3@korea.ac.kr
학력/경력
2006
2009
2009-현재
고려대학교 화공생명공학 학사
고려대학교 화공생명공학 석사
고려대학교 화공생명공학 박사
주요연구실적
•
•
•
•
•
98
|
S. cerevisiae 균주에서 글루코스와 글리세롤을 이용 대사공학적 접근을 통한 1,2-propanediol생산
E. coli 균주에서 lignin-hydrolysate compounds에 대한 전사체 연구
S. avermitilis 균주에서 statin 계열 물질 GC/MS분석
Metabolic profiling 기술을 이용하여 S. cerevisiae 균주에서 GCY1 유전자의 역할 규명
박테리아, 효모 균주에서 13C carbon source를 이용한 대사체 기반 대사흐름 측정 기술 개발
Y2-3
Comprehensive analysis of central carbon metabolites and
dynamics for metabolic flux analysis in yeast
13
C-isotope
Joon-Young Jung, Tae-Yeon Kim, Dae-Kyun Im and Min-Kyu Oh
Department of Chemical and Biological Engineering, Korea University 5-1 Anam-dong, Sungbuk-gu Seoul, South
Korea 136-71,
E-mail: mkoh@korea.ac.kr
Comprehensive analysis of carbon fluxes (metabolic reaction rates) in the central metabolic pathways
(glycolysis, pentose phosphate pathway and TCA cycle) has improved our understanding about microbial
physiology of metabolically engineered strains. For this purpose, accurate, reliable and reproducible
measurement of intracellular metabolites is prerequisite. The first critical step is the establishment of proper
quenching, extraction and sampling protocol, which ensures rapid arrest of all metabolic activity, effective
extraction of target metabolites from the cells, and verification of metabolites leakage via mass balance
analysis among different types of sample. The second step is the assessment of appropriate mass peaks
corresponding to the derivatized metabolites. However, the accurate measurement of intracellular metabolites
cannot be directly assessed to the metabolic fluxes. To measure metabolic flux, stable isotope tracers such
as 13C should be hired. Up to now, flux analysis using protein-bound amino acids combined with
computational calculation tracing site-specific 13C pattern has been widely used. But new technology,
metabolome-based 13C flux analysis, has been recently introduced, which directly measures isotope tracer
conversion rates of target metabolites for flux analysis
In this study, we performed qualitative and quantitative analysis of intermediate metabolites on glycolysis,
pentose phosphate pathway and TCA cycle with quadruple gas chromatography mass spectrometry (GC-MS).
First, extraction methods, quenching conditions, and leakage of metabolites were tested for central carbon
metabolites analysis from Saccharomyces cerevisiae and Kluyveromyces marxianus. Also, a bioreactor with
fast sampling device was designed. Then, different metabolite levels were detected when different carbon
substrates, such as glucose and xylose, were utilized in K. marxinus. Base on the metabolite profiling
method, isotope-based dynamic studies were performed with uniformly labeled 13C glucose. From time series
isotopic studies, conversion rates of glucose into the central metabolic pathway were well demonstrated,
which showed pattern of glucose utilization in the yeast strain.
This research is supported by grants from National Research Foundation of Korea (NRF) [Project No.
2012-0005359 and 2012M1A2A2026560]
|
99
이 종 석 _Jong Suk Lee, 경기과학기술진흥원
Office: 031-888-6930 / C.P.: 010-4816-0527
E-mail: jslee@gstep.re.kr
학력/경력
2001
2003
2003
2004-2010
2010
2010-
동국대학교 생화학 학사
부산대학교 미생물학 석사
서울대학교 BK21농생명공학사업단 연구원
한국생명공학연구원 연구원
서울대학교 농업생물공학 박사
경기과학기술진흥원 책임연구원
주요연구실적
• 논문: 국내 2편, SCI 27편, 특허 4건
• 주요 연구분야/공동연구 가능분야
(1) LC-MS기반 Metabolomics (대사체학): 의약품 및 천연물 구조분석, 다변량 통계분석
(2) 질량분석기기반 단백질의약품 특성분석
• 수행중인 연구과제
(1) 바이오시밀러등 재조합의약품의 특성분석법 고도화 연구(2012-2013, 식약청)
(2) 대사체학 분석기법을 이용한 유용생리활성소재 탐색 및 응용(2012-2014, 국립수산과학원)
(3) 미립화 유산균의 활성성분 분석(2012-2014, 바이오그린21 (주)RNA위탁사업)
(4) 초고속 기능성화합물은행 구축사업(2013, 경기과학기술진흥원)
100
|
Y2-4
LC-MS based metabolomics approach for natural product screening
and identification
Jong Suk Lee
Gyeonggi Biocenter, Gyeonggi Institute of Science & Technology Promotion, Suwon 443-270, Korea
E-mail: jslee@gstep.re.kr
A new strategy was proposed for platform technology development of efficient bioactive secondary
metabolite screening from natural products. This strategy contained LC-MS based metabolite profiling,
MS/MS spectral searching, and multivariate statistical analysis for biologically interesting target compound
discovery. In order to high-throughput metabolite identification, MS/MS spectral library has been developed
to automatically search tandem mass spectrometry (MSn) data against high quality experimental MS/MS data
from known metabolites. Searchable MS/MS spectral libraries, constructed using the results of liquid
chromatography-tandem mass spectrometry (LC-MS/MS) coupled with electrospray ionization (ESI), are here
presented with regard to the identification and confirmation of a variety of natural product samples. These
MS/MS spectra were then searched automatically against a total 5,188-substance mass spectral library (671
plant origin, 640 microbe origin, and 8,505 NIST library), which contained many previously acquired plant
and microbe origin natural product standards. Unsupervised data analysis by PCA was performed and
followed by principal component variable grouping to find correlated variables. This tool allows us to
distinguish of differences in the metabolite profile of individual bacteria. Using PCA, it was proved that SP
medium was more optimal for present of metabolite profile difference of myxobacteria strains. The HCA
dendrogram result, based on metabolite profiling, showed good correlation with 16s rRNA sequence based
myxobacteria taxonomy analysis. This result revealed that secondary metabolite profiling base
chemotaxonomy was represented of the each bacterial characteristic. By the factor analysis and LC-MS base
metabolite profiling, six target compounds were discovered from the four target myxobacteria strain, which
produce unique metabolites. All metabolite lists of target strain were then submitted to the constructed
MS/MS spectral library program which simultaneously searches several lists of potential unique metabolites
extracted from statistic analysis to propose known metabolite or novel metabolite candidates. The target
compounds were identified as four known compounds (myxochelin A, myxochelin B, myxalamide C, and
stigmatellin A) and two unknown compounds (m/z 419, m/z 388 of Cystobacter velatus DSM14718) by
MS/MS spectral library searching. This results how ed that MS/MS spectral library was very powerful tool
for high throughput structural analysis from complex sample. These results suggest that the proposed
strategy, LC-MS based metabolite profiling combined multivariate statistic analysis, was highly sophisticated
platform technology for bioactive secondary metabolite screening from microbial natural product, judgment
of relatedness between samples, and chemotaxonomy analysis.
|
101
102
|
포스터 초록
Poster 1.
Human and Animal Metabolomics
|
1
P1-1
Metabotyping of the C. elegans sir-2.1 mutant using in vivo labeling
13
and C-heteronuclear multidimensional NMR metabolomics
Yong Jin An1,2, Wen Jun Xu1, Xing Jin1, He Wen1, Hyesook Kim3, Junho Lee3 and Sunghyouk Park1,*
1
College of Pharmacy, Seoul National University, Sillim-dong, Gwanak-gu, Seoul, 151-724, Korea,
2
Department of Biochemistry, College of Medicine, Inha University, Incheon, 400-712, Korea,
3
WCU Department of Biophysics and Chemical Biology, Seoul National University, Sillim-dong, Gwanak-gu, Seoul,
151-724, Korea
E-mail: biochem.yong@gmail.com
The roles of sir-2.1 in C. elegans lifespan extension have been subjects of recent public and academic
debates. We applied an efficient workflow for in vivo 13C-labeling of C. elegans and 13C-heteronuclear
NMR metabolomics to characterizing the metabolic phenotypes of the sir-2.1 mutant. Our method delivered
two orders of magnitude higher sensitivity than unlabeled approach, enabling 2D and 3D NMR experiments.
Multivariate analysis of the NMR data showed distinct metabolic profiles of the mutant, represented by the
increases in glycolysis, nitrogen catabolism and initial lipolysis. The metabolomic analysis defined the
sir-2.1 mutant metabotype as the decoupling between enhanced catabolic pathways and ATP generation. We
also suggested the relationship between the metabotypes, especially the branched chain amino acids, and the
roles of sir-2.1 in the worm lifespan. Our results should contribute to solidifying the roles of sir-2.1, and
the described workflow can be applied to studying many other proteins in metabolic perspectives.
106
|
P1-2
In vitro identification of obovatol metabolites in human liver
microsomes using liquid chromatography-tandem mass spectrometry
Jeongmin Jooa, Doohyun Leea, Zhexue Wua, Jung-Hoon Shina, Hye Suk Leeb, Taeho Leea,
a
and Kwang-Hyeon Liu
a
College of Pharmacy and Research Institute of Pharmaceutical Sciences, Kyungpook National University, Daegu, Korea
College of Pharmacy and Integrated Research Institute of Pharmaceutical Sciences, Catholic University of Korea, Korea
E-mail: jjmin1422@naver.com
b
Obovatol is neolignan compound isolated from Magnolia obovata. It has inhibitory effects on arachidonic
acid-induced platelet aggregation and cancer cell growth through inhibition of NF-κB activity. This study
was performed to identify the metabolic pathway of obovatol in human liver microsomes. Human liver
microsomal incubation of obovatol in the presence of NADPH and/or UDPGA resulted in the formation of
six metabolites, M1-M6. M1 and M2 were identified as hydroxyobovatol, on the basis of liquid
chromatography/tandem mass spectrometric (LC-MS/MS) analysis. M1, M2 and obovatol were further
metabolized to their glucuronide conjugates, obovatol-glucuronide (M3), obovatol-diglucuronide (M4), and
hydroxyobovatol-glucuronide (M5 and M6).
|
107
P1-3
Profiling and characterization of skin ceramides using
nanoelectrosparay tandem mass spectrometry
Jung-Hoon Shin and Kwang-Hyeon Liu
College of Pharmacy and Research Institute of Pharmaceutical Sciences
Kyungpook National University, Daegu, Korea
E-mail:dstlkh@gmail.com
Lipids are important components in all biological tissues. They play important roles associated with the
proper function of the organism. Lipidomics, a branch of metabolomics, is a systems-based study of all
lipids and their function within the cell. Recently various techniques for global lipid profilling have been
developed to understand the function of lipids in a biological system and applied in drug and biomarker
development. Shotgun lipidomics which extracted lipids from cells, body fluids or tissues are directly
infused into a mass spectrometer, provide rapid semi-quantitative snapshots of the composition of complex
lipidomes in samples. In this study, we developed lipidomics methodology for skin lipid profiling and
identification using chip-based direct infusion nanoelectrospray tandem mass spectrometry. Skin ceramides
are produced by coupling fatty acid acyl chains onto sphingoid bases by an amide binding. Typical
sphingoid bases detected in skin comprise dihydrospingosine (DS), sphingosine (S), phytosphingosine (P),
and 6-hydroxysphingosine (H), and fatty acid acyl chains comprise non-hydroxy fatty acid (N), α-hydroxy
fatty acid (A), ω-hydroxy fatty acid (O), and estrified ω-hydroxy fatty acid (E). By the combination of
sphingoid bases and fatty acid acyl chains, 16 ceramide classes can be listed. The MS/MS fragmentation
pattern of these ceramides were characterized through the interpretation of product ion scan mass spectra of
them. Based on the MS fragmentation pattern of skin ceramides, an in silico MS/MS library for the
annotation and identification of unknown lipids from skin samples has generated. This library was applied
to identify the ceramide profiles of human and mouse skin.
108
|
P1-4
Lipidomic platform establishment for the identification of skin
ceramides having non-hydroxy acyl chain.
Jung-Hoon Shin1), Jong Cheol Shon1), Eun Sung Jung2) Hye Min Park2), Choong Hwan Lee2)
1)
and Kwang-Hyeon Liu
1)
College of Pharmacy and Research Institute of Pharmaceutical Sciences, Kyungpook National University, Daegu, Korea
2)
Department of Bioscience and Biotechnology, Konkuk University, Seoul, Korea
E-mail:dstlkh@gmail.com
The stratum corneum as the outermost layer of the skin has a barrier function and protects the organism
against environmental influences and transepidermal water loss. Its unique morphology consists of
keratin-enriched corneocytes embedded in a distinctive mixture of lipids that contains mainly ceramides, free
fatty acids, and cholesterol. Ceramides are sphingolipids consisting of sphingoid bases, which are
amide-linked to fatty acids. Typical sphingoid bases detected in skin comprise dihydrosphingosine (dS),
sphingosine (S), phytosphingosine (P), and 6-hydroxysphingosine (H), and fatty acid acyl chains comprise
non-hydroxy fatty acid (N), α-hydroxy fatty acid (A), ω-hydroxy fatty acid (O), and esterified ω-hydroxy
fatty acid (E). By the combination of sphingoid bases and fatty acid acyl chains, 16 ceramide classes can
be listed. MS/MS analysis of skin ceramides using tandem mass spectrometry provided the characteristic
fragmentation pattern of both acyl- and sphingoid-unit, which are informative for the structure identification
of ceramides. In this study, we developed lipidomic platform for the identification of skin ceramides having
non-hydroxy acyl chain (N-type ceramide; NS, NdS, NH, and NP). The MS/MS fragmentation pattern of
these ceramides were characterized and patternized through the interpretation of product ion scan mass
spectra of them. Based on the MS/MS fragmentation pattern of these ceramides, an in silico N-type
ceramide MS/MS library has generated.
|
109
P1-5
Skin ceramide profiling in stratum corneum obtained from Korean,
Malay, and Vietnamese using shotgun lipidomics.
Jung-Hoon Shin1), Kyohoon Lee2), Jong Cheol Shon1), Boram Lee1),Chang Seo Park2),
3)
2)
1)
Eung Ho Choi , Sunki Kim , and Kwang-Hyeon Liu
1)
College of Pharmacy and Research Institute of Pharmaceutical Sciences, Kyungpook National University, Daegu, Korea
2)
Dept. of Chemical and Biochemical Engineering, Dongguk University, Seoul, Korea
3)
Dept. of Dermatology, Yonsei University Wonju College of Medicine, Wonju, Korea
E-mail:dstlkh@gmail.com
In the stratum corneum of skin, ceramides represent the major constituent of the free extractable
intercellular lipids and play a significant role in maintaining the water permeability barrier of the skin.
Ceramides are sphingolipids consisting of sphingoid bases, which are amide-linked to fatty acids. Up to
now, 16 ceramide subclasses have been identified in human stratum corneum. In this study, we developed
the lipidomic platform for the skin lipid profiling and identification using chip-based direct infusion
nanoelectrospray tandem mass spectrometry. Using this platform, we identified 21 skin ceramides in human
skin stratum corneum, and compared ceramide composition among three ethnic groups (Korean, Malay, and
Vietnamese). However, there were no significant ethnic differences in skin ceramide profiles.
Fig. 1. The chemical structure of MGDG 16:3, 18:3 (A), DGDG 18:3, 18:3 (B), SQDG 16:0, 18:3 (C)
110
|
P1-6
Albendazole and fenbendazole hydroxylation is mainly mediated by
CYP2J2 and CYP2C19 in human liver microsomes
Zhexue Wu, Doohyun Lee, Jeongmin Joo, Jung-Hoon Shin, Taeho Lee and Kwang-Hyeon Liu
College of Pharmacy and Research Institute of Pharmaceutical Sciences, Kyungpook National University, Daegu, Korea
E-mail : wuzhexue527@163.com
Albendazole and fenbendazole are broad spectrum anthelmintics. Lee et al. recently reported that
albendazole hydroxylation is mediated by CYP2J2 isoform, however, there is relatively few reports in the
CYP2J2-mediated metabolism of benzimidazole drugs. We have investigated the hydroxylation of
albendazole and fenbendazole in human liver microsomes (HLM) and recombinant systems. In HLM,
ticlopidine and danazole inhibited hydroxyalbendazole and hydroxyfenbendazole formation, and
diethyldithiocarbamate and quinidine inhibited the hydroxylation of albendazole and fenbendazole,
respectively. Of the 10 cDNA-expressed P450s, CYP2C19, CYP2J2, and CYP2E1 are involved in the
albendazole hydroxylation, whereas CYP2C19, CYP2J2, and CYP2D6 are involved in the formation of
hydroxyfenbendazole. Correlation analysis between the known P450 enzyme activities and the rate of the
benzimidazole hydroxylation in the 15 HLMs showed that benzimidazole hydroxylation is correlated with
CYP2J2 and/or CYP2C19 activity.
In conclusion, our data suggested that both CYP2J2 and CYP2C19 play important roles in albendazole
and fenbendazole hydroxylation. The present data will be useful to understand the pharmacokinetics and
drug-interaction of albendazole and fenbendazole in vivo.
|
111
P1-7
Mass spectrometry-based skin metabolite profiling of ultraviolet
B-irradiated hairless mice
Hye Min Parka, Jeong Kee Kimb, Kwang-Hyeon Liuc, Choong Hwan Leea*
a
Departmetnt of Biosciense and Biotechnology, Konkuk University, Seoul 143-701, Korea
b
Food Research Institute, AmorePacific R&D Center, Yongin-si, Gyeonggi-do, Korea
c
College of Pharmacy and Research Institute of Pharmaceutical Sciences, Kyungpook National University,
Daegu 702-701, Korea
E-mail: ramgee@naver.com
Metabolite profiling related to ultraviolet (UV) radiation in skin has not yet been performed despite many
physiological evidence-based molecular mechanism studies of the relationship between the skin and UV
radiation. In this study, we observed the clinical and histological changes including the increased TEWL,
epidermal thickness and inflammatory cells and the changes of collagen fibre in chronic exposed mice skin
to UVB radiation for 12 week. Futhermore, we analyzed metabolite profiling of the mice skin in response
to UVB radiation for 6 and 12 wks, respectively, using ultra-performance liquid chromatography
(UPLC)-quadrupole time-of-flight (Q-TOF)-mass spectrometry (MS), and gas chromatography (GC)-TOF-MS
as well as multivariate statistical analysis. Amino acids, organic compounds, fatty acids, lipids, nucleosides,
carbohydrates, lysophosphatidylcholines (lysoPCs), lysophosphatidylethanolamines (lysoPEs) and other
metabolites were determined to be discriminators that characterized the differences between non-irradiated
and UVB-irradiated group. In addition, both of the changes in skin primary and secondary metabolite by
UVB exposure were generally higher in 12 wks than 6 wks. These results from primary and secondary
metabolite profiles explained the prolonged chronic exposure to UVB light may have a great influence on
skin by altering its metabolites, suggesting that each these changed metabolites possess potential as
biomarkers for skin diseases caused by UVB irradiation.
112
|
P1-8
Dietary fat and colon tumors induce distinctive changes in lysophospholipid
profiles in the sera and livers of obesity-resistant BALB/c mice
Hyang Yeon Kim1, Minhee Kim2, Hye Min Park1, Jiyoung Kim1, Eun Ji Kim3,
1
2,3
Choong Hwan Lee , Jung Han Yoon Park
1
Department of Bioscience and Biotechnology, Konkuk University, Seoul 143-701, Korea
Department of Food Science and Nutrition, Hallym University, Chuncheon 200-702, Korea
3
Center for Efficacy Assessment and Development of Functional Foods and Drugs, Hallym University, Chuncheon
200-702, Korea
E-mail: festivalkim@naver.com
2
Colorectal cancer is one of the most common cancers in the USA and its incidence appears to be rising
worldwide. Intake of a high-fat diet (HFD) often leads to weight gain and obesity in rodents and humans,
and is associated with a higher risk of colorectal cancer. Our previous study revealed that chronic
consumption of a high-fat diet (HFD) stimulates colon cancer progression in obesity-resistant BALB/c mice.
Recently, to better understand the biochemical mechanisms underlying obesity and the related dysfunction,
metabolomic profiles of body fluids have been investigated using ultra performance liquid
chromatography-quadrupole-time-of-flight-mass spectrometry (UPLC-Q-TOF-MS). The aim of the present
study was to investigate the significant alteration of metabolites caused by tumor progression and an HFD
in the serum and liver in the same mouse model.
Male BALB/c mice were fed either a control diet or an HFD for 20.5 weeks. The syngeneic CT26
colon carcinoma cells were injected into the right rear flank of mice after 16 weeks of feeding. Metabolites
in serum and liver samples were analyzed by UPLC-Q-TOF-MS-based metabolomics.
HFD feeding and tumor injection induced changes in the choline-containing phospholipids, namely,
phosphatidylcholines (PCs) and lysophosphatidylcholines (lysoPCs), as well as lysophosphatidylethanolamines
(lysoPEs) in the serum and liver. The majority of these metabolite changes were due to HFD feeding (11
in sera and 5 in livers) rather than tumors (3 in sera and 1 in livers). These data explain the HFD- and
tumor-related metabolite alterations of phospholipids, especially lysoPCs, in the liver and serum of
obesity-resistant mice, suggesting that the lysoPCs are biomarkers for the chronic consumption of HFD in
non-obese individuals.
|
113
P1-9
Predicting idiopathic toxicity of cisplatin by
a pharmacometabonomic approach
Hyuk Nam Kwon1, Mina Kim3, He Wen1, Sunmi Kang2,Hey ji Yang2, Myung Joo Choi3,
3
4
5
6
3
3
Hee Seung Lee , DalWoong Choi , In Suh Park , Young Ju Suh , Soon Sun Hong , Sunghyouk Park
1
College of Pharmact, Seoul National University, Seoul, Korea, 2Department of Biochemistry, 3Department of
Biomedical Sciaences, College of Medicine, Inha University, Incheon, Korea, 4Department of Environmental Health,
College of Health Sciences, Korea University, Seoul, Korea, 5Department of Pathology, 6Department of Biostatistics,
College of Medicine, Inha University, Incheon, Korea
E-mail: brian.biochem@gmail.com
Cisplatin has been one of the most widely used anticancer agents, but its nephrotoxicity remains a
dose-limiting complication. Here, we evaluated the idiopathic nature and the predose prediction of
cisplatin-induced nephrotoxicity using a nuclear magnetic resonance (NMR)-based pharmacometabonomic
approach. Cisplatin produced serious toxic responses in some animals (toxic group), but had little effect in
others (nontoxic group), as judged by hematological and histological results. The individual metabolic
profiles, assessed by urine NMR spectra, showed large differences between the post-administration profiles
of the two groups, indicating the relevance of the NMR approach. Importantly, multivariate analysis of the
NMR data showed that the toxic and nontoxic groups can be differentiated based on the pretreatment
metabolite profiles. Leave-one-out analysis, performed to evaluate the practical performance of our approach,
gave a 66% accuracy rate in predicting toxic responses based on the pretreatment metabolite profiles.
Hence, we provide a working model that can explain the idiopathic toxicity mechanism based on marker
metabolites found by NMR analysis consistent with tissue NADH measurements. Thus, a
pharmacometabonomic approach using pretreatment metabolite profiles may help expedite personalized
chemotherapy of anticancer drugs
114
|
P1-10
Lipidomic analysis of hypertriglyceridemic subjects based on ultra
performance liquid chromatography/Q-TOF mass spectrometry
Ju Yeon Park1, Jeeyoun Jung1, and Geum-Sook Hwang1,2
1
2
Integrated Metabolomics Research Group, Korea Basic Science Institute, Seoul 136-713, Korea
Graduate School of Analytical Science and Technology, Chungnam University, Daejeon 305-764, Korea
E-mail: gshwang@kbsi.re.kr
Hyperglyceridemia have been associated with an increased risk of cardiovascular disease, type 2 diabetes,
and metabolic syndrome. Lipidomic analysis offers a means of identifying markers that may be indicative
of pathways of hyperglyceidemia. Therefore we investigated the altered lipid pattern in hypertriglyceridemic
subjects and identified a relationship between lipid acyl chain content and hyperglyceridemia risk. The study
subjects were grouped into normal triglyceride (fasting triglyceride (TG) <150mg/dL, n=49) and
hypertriglyceridemia (TG ≥150mg/dL, n=26). Serum lipid analyzed ultra-performance liquid
chromatography/Q-TOF mass spectrometry, coupled with a multivariate statistical analysis. Principal
components analysis (PCA) showed separation pattern between hypertriglyceridemic subjects and normal
triglyceride subjects (R2X=89.0%, Q2=69.6%). There were significant differences in lyso-phosphatidyl
choline, phosphatidyl choline, sphingomyelin, diacylglycerol, and triacylglycerol between hypertriglyceridemic
subjects and normal triglyceride subjects. Additionally, we observed that lipid acyl chain content were
associated with hyperglyceridemia risk. Among these lipid metabolites, triacylglycerols level of lower carbon
number and double bond content were significantly increased in hyperglyceridemic subjects relative to
normal triglyceride subjects. Our study showed that lipidomic analysis may be useful for understanding the
alternation of lipid component in the stages of the pathologic process of hypertriglyderidemia.
|
115
P1-11
Metabolic profiling study in serum of patients with cardiovascular
disease using ultra-performance liquid chromatography/quadrupole
time-of-flight mass spectrometry
1,2
1
2
Jueun Lee , Youngae Jung , Do Hyun Ryu , and Geum-SookHwang
1
1,3
2
Korea Basic Science Institute, Seoul 136-713, Korea, Sungkyunkwan Univ. Natural Sciences Campus,
Cheoncheon-dong, Jangan-gu, Suwon-si, Gyeonggi-do 440-746 Korea, 3Graduate School of Analytical Science and
Technology, Chungnam University, Daejeon, 305-764
E-mail: gshwang@kbsi.re.kr; dhryu@skku.edu
Cardiovascular disease (CVD) refers any disease that affects the cardiovascular system such as Angina
pectoris and Myocardial infarction and is the most prevalent cause of death in developed nations. Although
there are a number of risk factors like age, gender, high blood pressure, high serum cholesterol levels,
obesity, and renal failure, the mechanisms underlying CVD are still not fully understood. Moreover, the
main obstacle to the treatment of CVD in clinical practice is the asymptomatic processes, that are
associated with plaque formation, develop causing silent yet progressive tissue damage. In this study, we
applied ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF
MS) to analyze metabolites in methanolic extracts of serum from Angina pectoris and Myocardial infarction
patients. The Partial least squares-Discriminant Analysis (PLS-DA) model was generated form metabolic
profiles and the score plot showed the significant difference between two disease models. The amino acids
and acyl-carnitine were up-regulated in Myocardial infarction compared to Angina pectoris, whereas the
levels of lysoPCs and lysoPEs were significantly decreased. These results suggest that metabolic profiling of
serum from Angina pectoris and Myocardial infarction patients using UPLC/Q-TOF MS could provide new
phenotypic biomarkers for CVD diagnosis and treatment.
116
|
P1-12
1
H NMR-based Metabolomic Profiling in Mice Infected
with Orientia Tsutsugamushi
Changhun Kim and Geum-Sook Hwang
Integrated Metabolomcis Research Group, Seoul Center, Korea Basic Science Institute, Seoul 136-701, Republic of Korea
E-mail: gshwang@kbsi.re.kr
Orientia tsutsugamushi is an obligate intracellular pathogen of scrub typhus, a chigger-borane zoonosis
that is a highly prevalent, hazardous illness of greatest public health in the Orient and part of the
Palearctic. In this study, we investigated metabolic profiling of multuple organs (spleen and liver) and
serum derived from Karp, serotype of O.tsutsugamushi infected models at 4 and 7 days, using 1H NMR
based metabolomics combined chemometrics analysis. Principle component analysis (PCA) and partial
least-squares discriminant analysis (PLS-DA) score plots of NMR data from tissue and serum showed clear
separations between control and O.tsutsugamushi-infected mice at 4 days and 7 days after infection.
Furthermore, two-way ANOVAs indicated that infection, duration time and their interaction (infection ×
duration time) affect the metabolite changes differently. Lactate, choline, and branched chain amin acids
(isoleucine, leucine, valine) were significantly lower after infection in liver tissue, while was higher in
spleen tissue. Succinate, acetate were lower in liver tissue at 7 days compared to 4 days and taurine was
lower in spleen tissue at the same condition. In addition, betaine, O-phosphocholine showed interaction
effect of time and infection in all tissue samples. This study suggests that NMR-based metabolite profiling
of multiple organ tissues and serum could provided insight into the metabolic changes in host infected with
virulent Orientia Tsutsugamushi and progress of the disease.
|
117
P1-13
Investigation on effect of gallic acid in a mouse model of
Alzheimer’s disease using NMR based metabolomics
Young-Sang Jung, Eosu Kim, and Geum-Sook Hwang
Integrated Metabolomcis Research Group, Seoul Center, Korea Basic Science Institutue, Seoul 136-701,
Republic of Korea
Geriatric Division, Yonsei University College of Medicine.
E-mail: gshwang@kbsi.re.kr
Alzheimer’s disease is a kind of dementia which causes troubles with memory, thinking and behavior.
Symptoms usually develop slowly and get worse over period of time. Up to date, the cause of Alzheimer’s
is still unknown and there are no ways to cure Alzheimer’s disease. Therefore, researchers have been
making great efforts to elucidate mechanism of onset and development and to find treatment to cure or to
stop development. In this study, we investigated on the effects of gallic acid in Alzheimer’s disease model
(n=10, by Aβ injection into brain), treated model (n=10, by Aβ injection and gallic acid), and control
(n=10) mice, using NMR based metabolomics. We measured and profiled NMR spectra of 30 serum
samples and analysed the profiles with multivariate statistical analysis. We found clear separation
Alzheimer’s mice model from the other mice models in the 2D scores plot of principle component analysis
(PCA) while the control and treated mice models were overlapped. This result indicates that the metabolic
profiles of Alzheimer’s mice model are significantly different with those of controls, and the metabolic
profiles of treated mice are similar to the controls’s, demonstrating that the gallic acid could improve the
Alzheimer’s disease. The major metabolites contributing in differentiation between Alzheimer’s and controls
were valine, lactate, alanine, pyruvate, citrate, creatine, dimethyl sulfone, glucose, tyrosine, and formate. In
conclusion, NMR based metabolomics study on the Alzheimer’s disease showed that gallic acid could be
potential treatment drug for Alzheimer’s disease and the significantly different metabolites might provide
clues for further understanding Alzheimer’s diseases.
118
|
P1-14
Hip2 influences cell survival following ultraviolet radiation
Sooyeon Kang and Seongman Kang
Department of Biotechnology, School of Life Sciences and Biotechnology, Korea University, Seoul 136-701, Korea
E-mail: sandrak@korea.ac.kr
Huntington interaction protein2 (Hip2) is a human ubiquitin conjugating enzyme (identical to bovine
E2-25K) that is known to associate with Huntington, the protein linked to Huntington’s disease (HD).
Huntington is crucial for normal development and may be regarded as a cell survival gene. Hip2 has
previously been shown to be involved in the suppression of apoptosis triggered in response to DNA
damage. Thus, the level of Hip2 expression in cells may significantly influence its survival following DNA
damage induced by ultraviolet (UV) radiation. In this study, human embryonic kidney (HEK293) cells were
exposed to ultraviolet radiation (UVP CL-1000 ultraviolet corsslinker) at different doses to induce DNA
damage. Cell viability and proliferation were analyzed and pro-/anti- apoptosis molecule antibodies such as
Bax, Bcl-2 and PARP were used. For cell cycle, 5mg/ml of PI (propidium iodide) was added and analyzed
with a FACSan flow cytometer (BD, science). The expression of Hip2 protein was increased as the dose of
UV was elevated. Cells transfected with additional Hip2 were found to be significantly more resistant to
cell death induced by UV radiation than the cells without Hip2. These results demonstrated that Hip2
appears to play an important role in cell survival by functioning as an inhibitor of apoptosis following
DNA damage. Therefore, the mechanism of Hip2 may contribute greatly to the development of more
effective cancer treatment strategies in the future.
|
119
P1-15
An NMR metabolomics approach for the diagnosis of leptomeningeal
carcinomatosis in an animal model
He Wenc,1, Hye Rim Choa,b,1, Young Jin Ryua, Yong Jin And, Hyo Cheol Kima,e, Woo Kyung
a,e
a,e
c,2
a,e,2
Moon , Moon Hee Han , Sunghyouk Park , Seung Hong Choi
a
b
Department of Radiology, Seoul National University College of Medicine, 110-744, Seoul, Korea,
Department of Radiation Applied Life Science, Seoul National University College of Medicine, 110-744, Seoul, Korea,
c
College of Pharmacy, Seoul National University, San 56-1 Sillim-dong, Gwanak-gu, 151-742, Seoul, Korea,
d
Department of Biochemistry, Center for Advanced Medical Education by BK21 project, College of Medicine,
Inha University, 400-712, Incheon, Korea,
e
College of Medicine, Seoul National University, 110-744, Seoul, Korea
E-mail: moonhyuk1223@gmail.com
Leptomeningeal carcinomatosis (LC), caused by malignant cells seeding the leptomeninges, is the third
most common metastatic complication of the central nervous system, but the performance of current
diagnostic modalities is not satisfactory. To develop a new metabolomic diagnostic approach, we
successfully established a rat LC model using F-98 glioma cells that stably express green fluorescent
protein. Cytological diagnosis gave 66.7% sensitivity for the 7-day LC group and 0% for the 3-day group,
whereas MR imaging suffered non-specific responses. Then, we applied NMR-based metabolomics on
cerebrospinal fluid (CSF), and detected metabolic difference between normal and LC groups. Predictions
based on the multivariate model gave sensitivities of 89 (7-day LC group) or 88% (3-day LC group), a
specificity of 89%, and an overall accuracy of 89% in the diagnosis of the LC. Further statistical analysis
identified lactate, acetate, and creatine as specific for the 7-day LC group and glucose as being specific for
the normal group (P < 0.006 for all). Overall, our metabolomics approach delivered both earlier and more
accurate diagnostic results than the currently-used cytology and MR imaging. As the first report on the
metabolomics diagnosis for LC, it may become a useful clinical protocol that can augment or replace
current diagnostic methods, if proven in human samples.
120
|
P1-16
Uch-L3 interacting partners in osteoblast cells
1,2
1
1,3
1,3,4
Eunsom Choi , Mi-Jung Choi , So Yeon Kim , Sang-ho Ahn
1,3
and Ji Young Kim *
Clinical Trial Center for Medical Devices1, Institute of Medical Science2, Institute of Biomedical Engineering3,
Department of Rehabilitation Medicine4, Yeungnam University, Daegu, Korea
E-mail: evernew100@gmail.com
Proteasomal ubiquitination system is an essential in various biological functions. Of important parts, the
process of deubiquitination is a cleavage of ubiquitin from ubiquitinated target protein. A deubiquitinating
enzyme, ubiquitin C-terminal hydrolase-L3 (Uch-L3) previously found the involvement of osteoblast
differentiation. Uch-L3 stabilizes Smad1 in BMP2-induced osteoblast differentiation and its knock-out mice
shows osteoporotic bone phenotype. Therefore, to investigate more detail function of Uch-L3, we tried to
find its interacting partner proteins in osteoblast cells. Retroviral transduction system was used to establish
over-expressed Uch-L3 cells. The cells were expanded and harvested, and then, the lysates were
immunoprecipitated against anti-FLAG antibody. After the elution, the eluent was re-immunoprecipitated
against anti-HA antibody. The final eluent was displayed in a SDS-PAGE gel and then the gel was
silver-stained and/or coomassie-stained for LC-MS/MS analysis. The identified proteins were confirmed by
immunoprecipitation. The pattern of ubiquitination was also examined by in vivo ubiquitination assay. The
results suggest that the identified proteins may have important functions in osteoblast maintenance and/or
differentiation.
Supported by a grant from the National Research Foundation of Korea (NRF) funded by the Ministry of
Education, Science and Technology (No. 2010-0011231).
|
121
P1-17
Lipidomic profiling as a potential sexual biomarker of gonadextomized
E
rat brain using a UPLC/Q-ToF MS
Jisun Yoo1, Man Ho Choi2, Young Hwan Kim1
1
Division of Mass Spectrometry Research, Korea Basic Science Institute, Ochang 368-883,
2
Korea Institute of Science and Technology, Seoul 136-791, South Korea
E-mail: yhkim@kbsi.re.kr
Lipidomics is a rapidly expanding research field in which multiple techniques are utilized to quantify the
precise chemical constituents in cell’s lipidome and identify their cellular organization. Numerous articles
have been reported that concerned with sex differences in lipid metabolism. As the lipid is related to sex
hormone, some disease is occur more frequently in men than women.
This study aimed to determine rat brain lipid species that could be used to sexual biomarker, we
investigated and compared the profiles of lipidomes in control rat brains with gonadectomized rat brains. 8
male and 8 female twenty week old rats were gonadectomized, 6 male and 6 female control group was
kept under regular condition. Sexual biomarker in rat brain sample were profiled by ultra performance
liquid chromatography coupled with quadrupole time of flight mass spectrometry (UPLC/Q-ToF MSE). The
acquired data was analyzed by principle component analysis (PCA) and orthogonal partial least squares
discriminant analysis (OPLS-DA) to identify potential sex
difference specific biomarkers.
122
|
P1-18
Obesity related metabolomic analysis of human subjects in black
soybean peptide (BSP) intervention study by ultra performance liquid
chromatography and quadrupole-time-of-flight mass spectrometry
1
1
1
2
3
Min Jung Kim , Hye Jeong Yang , Jin Hee Kim , Chang-Won Ahn , Jong Ho Lee ,
4
1
Kang Sung Kim and Dae Young Kwon
1
Department of Food Metabolism and Nutrition, Korea Food Research Institute, Kyongki-do, Korea
2
Research and Development Center, Nong Shim Co., Ltd., Seoul, Korea
3
Department of Food & Nutrition, Yonsei University, Seoul, Korea
4
Department of Food Science & Nutrition, Yongin University, Kyongki-do, Korea
E-mail: well@kfri.re.kr
The present study aimed to identify key metabolites related to weight reduction in humans by studying
the metabolic profiles of sera obtained from 34 participants who underwent dietary intervention with black
soybean peptides (BSP) for 12 weeks. This research is sequel to our previous work in which the effects of
BSP on BMI and blood composition of lipid was investigated [1]. Sera of the study were subjected to
ultra
performance
liquid
chromatography
and
quadrupole
time-of-flight
mass
spectrometry
(UPLC-Q-TOF-MS) and the data were analyzed using partial least-squares discriminate analysis (PLS-DA)
score plots. Body mass index and percent body fat of the test group were reduced. Levels of betaine,
benzoic acid, pyroglutamic acid, pipecolic acid, N-phenylacetamide, uric acid, l-aspartyl-l-phenylalanine, and
lysophosphatidyl cholines (lysoPCs) (C18:1, C18:2, C20:1, and C20:4), showed significant increases. Levels
of l-proline, valine, l-leucine/isoleucine, hypoxanthine, glutamine, l-methionine, phenylpyruvic acid, several
carnitine derivatives, and lysoPCs (C14:0, PC16:0, C15:0, C16:0, C17:1, C18:0, and C22:0) were
significantly decreased. In particular, lysoPC 16:0 with a VIP value of 12.02 is esteemed to be the most
important metabolite for evaluating the differences between the 2 serum samples. Our result confirmed
weight-lowering effects of BSP, accompanied by favorable changes in metabolites in the subjects’ blood.
Therefore, this research enables us to better understand obesity and increases the predictability of the
obesity-related risk by studying metabolites present in the blood.
|
123
P1-19
Metabolite profiling of aged-mice serum and tissue
1
1
1
2
2
Kim Seungwoo , Cheon Hyo-Soon , Song Jae-chun , Hwang Geum-Sook , Bang Eun Jung ,
Jeon Jae-pil1, Park Sang-Ick1 and Kim Young-Youl1
1
Division of Brain Diseases, Center for Biomedical Sciences, National Institute of Health, Korea Centers for Disease
Control and Prevention, Osong Health Technology administration Complex, Chungcheongbuk-do, Korea.
2
Division of Analytical Research, Korea Basic Science institute/Seoul Center, Seoul Korea
E-mail:elegans514@naver.com
Aging is known to be a risk factor for many diseases, such as diabetes, cardiovascular and
neurodegenerative disease. Most previous studies focused on age-related disease mechanism, but common
process of these diseases is uncovered. Although, DNA damage and oxidative stress contribute to aging, it
has been shown that metabolic dysfunction is a common hallmark of aging process. Therefore, we focused
on specific metabolic changes on aging process, to find biomarkers and therapeutic targets of age-related
diseases. The young (4-5month-old) and old (19-20month-old) C57BL/6 mice were used in this study. We
have characterized physical and mental function of aged-mice model using behavior test and metabolic
1
profiling compare these mice using H-NMR.
124
|
P1-20
Metabolic changes of steroids in precocious pubertal girls may not be
associated with urinary levels of bisphenol A
Su Hyeon Lee1,2, Se Mi Kang1,3, Man Ho Choi1, Jeongae Lee1, Mi Jung Park4,
4
2
3
1
Shin Hye Kim , Won-Yong Lee , Jongki Hong , Bong Chul Chung
1
Future Convergence Research Division, Korea Institute of Science and Technology, Seoul 136-791, Korea
2
Department of Chemistry, Yonsei University, Seoul 120-749, Korea
3
Department of Basic Pharmaceutical Science, Kyung Hee University, Seoul 130-701, Korea
4
Department of Pediatrics, Sanggye Paik Hospital, Seoul 139-707, Korea
E-mail: fomeandu@nate.com
Precocious puberty (PP) is responsible for either central activation of the hypothalamic-pituitary-gonadal
(HPG) axis (central-PP) or abnormal steroid production that does not result from sustained activation of the
HPG axis (peripheral-PP). Therefore, PP refers to the appearance of physical and hormonal signs of
pubertal development at an abnormal early age. Gas chromatography-mass spectrometry-based steroid
signatures was applied to urine samples obtained from 42 patients with central-PP and 40 patients with
peripheral-PP to evaluate metabolic changes against 32 age-matched healthy girls. The levels of androgens,
such
as
testosterone,
androstenedione,
androstenediol,
16α-hydroxy-dehydroepiandrosterone,
and
5α-androstanedione, tended to be high in both PP groups, and 17β-estradiol was higher in central-PP (P <
0.01) than in peripheral-PP and controls. Altered steroid metabolism was also associated with the urinary
BPA levels, and testosterone, 17β-estradiol, and pregnenolone were significantly increased in the high BPA
groups. In particular, a correlation of estrogen metabolism with BPA levels irrespective of the type of PPs
was observed. The result implies that BPA exposure may cause metabolic changes in steroidogenesis in
girls, but not in the early onset of PP.
|
125
P1-21
Quantification of serum cholesterol and oxysterols using
MALDI-MS/MS coupled to a hybrid-SPE
Seul Ki Been,1,2 Ju-Yeon Moon,1 Won-Yong Lee,2 Bong Chul Chung,1 and Man Ho Choi1
1
Future Convergence Research Division, Korea Institute of Science and Technology, Seoul 136-791
2
Department of Chemistry, Yonsei University, Seoul 120-749
E-mail: shmwsr@hanmail.net
Abnormal levels of cholesterol and its oxidation metabolites (oxysterols) in biological samples are
associated with many endocrine disorders including cardiovascular events. To achieve a high-throughput
screening method, the matrix-assisted lasers desorption ionization (MALDI) coupled to the linear ion-trap
tandem mass spectrometry (LTQ-MS/MS) was developed for analysis of cholesterol and oxysterols in serum
sample. Due to the background noise derived from both MALDI matrix and biological specimens, it has
been difficult to detect cholesterol and oxysterols using MALDI-MS. To improve the detectability, the
optimized dansylation was introduced with 200 µL solution containing 5 mg/mL dansyl chloride and 1.25
mg/mL DMAP in dichloromethane:acetone (70:30, v/v), and 10 µL triethylamine at 65 ˚C for 1 hr. For
quantitative MALDI-MS/MS analysis, precursor and product ions were selected as m/z 491 → m/z 369 for
cholesterol, m/z 489 → m/z 361 (4β-, 7α- and 7β-OH-cholesterol) for A/B-ring hydroxylated oxysterols,
m/z 507 → m/z 367 (4β-, 7α-,7β-, 19-, 20α-, 22R-, 22S-, 24S-, 25-OH-cholesterol) for total oxysterols, and
m/z 436 for 27-OH-cholesterol. A comparison between SPE methods with protein-precipitation Hybrid-SPE
and hydrophobic interaction Oasis HLB showed that the extraction efficiency of dansylated cholesterols was
significantly increased in Hybrid-SPE method. Matrix condition for deceasing interference of matrix in the
low mass range was optimized binary matrix mixture of 10 mg/mL CHCA and 5 mg/mL DHB in 100%
acetone, 0.2% TFA. The devised technique could be useful for a high-throughput analysis and improving
the detectability of cholesterol and oxysterols in serum samples.
126
|
P1-22
Metabolic alteration of ω-3 and ω-6 polyunsaturated fatty acids in
hypercholesterolemia model rabbits
In Hye Seo1,2, Su Hyeon Lee1,2, Hong Seog Seo3, Myeong Hee Moon2, Bong Chul Chung1,
1
Man Ho Choi
1
Future Convergence Research division, Korea Institute of Science and Technology; 2 Department of Chemistry,
Yonsei University; 3Department of Internal Medicine, Korea University College of Medicine
E-mail: inhye2277@naver.com
The ω-6 and ω-3 polyunsaturated fatty acids (PUFAs) have been associated with many physiological
disorders including cardiovascular events and inflammatory systems. Although there has been great interest
in the fatty acid classes, the individual fatty acids have been investigated in biochemical functions. Here,
we developed the analytical methods to measure the levels of ω-3 and ω-6 PUFAs from model rabbit with
hypercholesterolemia using liquid chromatography-mass spectrometry (LC-MS) and gas chromatography-mass
spectrometry (GC-MS). Including essential fatty acid precursor, linoleic acid and α-linolenic acid, 6 PUFAs
were conducted and quantitatively analyzed. In this study, PUFAs were analyzed combined with chemical
derivatization, such as silylation and methylation, to improve detection sensitivity and chemical stability.
This technique will be applied to evaluate metabolic alteration of ω-3 and ω-6 PUFAs from model rabbits
with hypercholesterolemia.
|
127
P1-23
Analysis of polybrominated diphenyl ethers in internal organs of
piglets according to diseases of piglet
Jeounghwa Shin
Korea Basic Science Institute, Seoul center, Seoul 136-701, Korea
E-mail: jhshin01@kbsi.re.kr
Polybrominated diphenyl ethers (PBDEs) are a group of brominated flame retardants (BFRs), which have
been manufactured in large quantities and widely used in a variety of consumer goods. Their chemical
structure similarities to polychlorinated dibenzo-p-dioxins and polychlorinated biphenyls (PCBs), their toxicity
has been studied. PBDEs are persistent and lipophilic, which results in their bioaccumulation in the fatty
tissues of organisms and enrichment throughout food chains. Polybrominated diphenyl ethers (PBDEs) are
found in a variety of foods. Ninety five per cent of human exposure to POPs comes from food. The major
food sources are fish and seafood and dairy products. A number of studies also reported high levels of
PBDEs in animals and public concern about PBDE levels in animals has been raised. The purpose of this
study was to evaluate interrelation between the levels of PBDEs in internal organs of piglets according to
diseases of piglet.
Salmonella spp. and Streptococcus is Bacillus nature diseases. PRRS is Virus diseases.
The concentrations of PBDEs in internal organs of piglet from Bacillus nature diseases were higher than
the concentrations from Virus diseases. The concentrations of PBDEs in piglet liver from Virus diseases
were higher than the concentrations from Bacillus nature diseases. BDE-66 was detected in all samples.
BDE-100 was detected in almost samples. BDE-138 and BDE-183 were detected more in the samples from
Bacillus nature diseases than from Virus diseases.
Supported by the grant of Creative Allied project from KRCF
128
|
P1-24
Metabolic Profiling of Gliomas and Detection of Onco-metabolites
by NMR and ESI-MS Analysis
Gregory Hyung Jin Park, Yun-Ju Lee, Eun-Hee Kim, Chaejoon Cheong, Seung-Ho Yang, and
Hyeon-Man Baek
Division of Magnetic Resonance Research, Korea Basic Science Institute, Cheongwon, 363-883, Korea
E-mail: ghjpark1@kbsi.re.kr
Gliomas are the most common and the most malignant forms of brain tumors. In some tumor cases,
mutations in isocitrate dehydrogenases (IDHs) are observed, which result in elevated levels of
2-hydroxyglutarate (2HG) as well as inhibition of reduction of isocitrate into α-ketoglutarate in TCA cycle.
In this study, we detected and quantified brain metabolites contained in human glioma tissue extracts by
NMR spectroscopy and ESI-MS, to determine metabolic pathways alterations between tumors with IDH
mutations such as IDH1-R132H and wild-type tumors. One-dimensional 1H NMR experiment of
IDH-mutated tumor tissue extracts samples showed peaks arising from 2HG at 1.84/1.99, 2.24/2.28, and
4.02 ppm as predicted, as well as peaks from other metabolites. Two-dimensional NMR techniques such as
1
H-13C HSQC experiments further confirmed the cross-peaks from 2HG, which also indirectly revealed three
of five 13C spectral peaks at 28.2, 30.6, and 69.2 ppm, respectively. In particular, the (δH, δC) = (4.02 ppm, 69.2 ppm) cross peak
from 2HG clearly distinguished itself from glutamate/glutamine peaks. In mass spectrometry, mass peaks at
m/z = 147.03 (ESI-negative mode) were confirmed to come from 2HG. Further quantification of related
metabolites will elucidate tumor-induced alterations in metabolic pathways more quantitatively. Moreover, the
sharp difference in NMR profiles between IDH-mutated gliomas and wild-type gliomas, particularly the
NMR-detectability of onco-metablites such as 2HG, can be utilized for novel diagnostic techniques such as
metabolite-specific chemical shift imaging (CSI) using clinical MRI equipments.
Supported by grants from KBSI-#T33416.
|
129
P1-25
Metabolomics view on obese mice liver metabolites modulation by
soy diet fermented with Bacillus amyloliquefaciens
Hye Ryun Kim, Jang-Eun Lee, So young Lee, Hyang Rin Kang and Byung Hak Ahn
Korean Alcoholic Beverage Research Center, Korea Food Research Institute, Seongnam 463-746, Republic of Korea
E-mail: hrkim@kfri.re.kr
Liver metabolites of obese and normal mice with various soy diets were analyzed through UPLC-Q-TOF
mass spectrometry and their metabolic profiles were compared to those different diet groups by multivariate
statistical analysis. We investigated the soy diet fermented by Bacillus amyloliquefaciens, which produce
high content of 1-deoxynojirimycin and have strong α-glucosidase inhibitory activity, showed remarkable
changes on liver metabolites compared with normal and commercial soy diets in obese mice group on
PLS-DA model. Otherwise there was no significant differentiation in control mice group, showing no
dependence of the each soy diet on liver metabolites. Whereas various lipidomic metabolites were strongly
related to normal diet obesity mice group, some metabolites were highly concerned in the soy diet
fermented by B. amyloliquefaciens M140 and B. amyloliquefaciens M26 demonstrating these soy diets were
directly effective in obesity control.
These comprehensive metabolic results can be used to better understand obesity control by a microbial
fermented soy food. Moreover the developed metabolomics approach can be applied to a wide range of
studies involving tissue metabolic profiling by UPLC-Q-TOF-MS as well as investigation of liver and
obesity research.
130
|
P1-26
A new biochemical insight of epitestosterone as a NF-kB regulator in
androgen dependent cell lines
Dong-Hyoung Lee, Su Hyeon Lee, Sung-Won Moon, Bong Chul Chung, and Man Ho Choi
Future Convergence Research Division, KIST, Seoul 136-791, Korea;
E-mail: d11018@kist.re.kr
Although the cross-talk between endogenous steroids and inflammatory activities in hormonal-dependent
diseases has been identified, epitestosterone is poorly elucidated. In this study, we introduce the
epitestosterone administration in company with dihydrotestosterone (DHT) attenuate and recover the reduced
androgen receptor (AR) expression via activated NF-κB expression which characterized by both
phosphorylation ratio and the alteration of NLS (Nuclear Localization Sequence) targeted p65 antibody in
androgen dependent prostate cancer cell line (LNCaP). In addition, the liver microsomal in-vitro assay
resulted in dose-dependent production of active metabolites including 5β-androstan-3β, 17α-diol and
5α-androstan-3β, 17β-diol by the epitestosterone treatment. These evidences suggest that the epitestosterone
may be a new androgenic stimulator allows control the level of NF-κB activation, which correlated with
prolonged inflammation status.
|
131
P1-27
Sex differences in human with vasospastic angina using
high-temperature GC-MS based serum cholesterol signatures
Hyun-Hwa Son1, Ju-Yeon Moon1, Hong Seog Seo2, Hyun Hee Kim2, Bong Chul Chung1,
1
and Man Ho Choi
1
Future Convergence Research Division, Korea Institute of Science and Technology, Seoul 136-791, Korea;
2
Cardiovsacular Center, Korea University College of Medicine, Seoul 152-703, Korea
E-mail: cat2010@swu.ac.kr
Abnormal cholesterol metabolism is associated with vasospastic angina, which is considered an early
manifestation of atherosclerotic vascular disease. Cholesterol signatures, simultaneous analysis of cholesterol,
4 cholesterol precursors, 9 oxysterols, and 7 cholesteryl ester (CEs) in serum was developed to
comprehensively evaluate cholesterol metabolism.
Methanol-based hybrid SPE minimized matrix effects, and HTGC-MS using a thermally stable MXT-1
stainless steel capillary column resulted in a good chromatographic resolution for the separation of lipophilic
compounds. In 20 μL serum, overall recoveries ranged 82.1–129.3%, except for CEs (26.1–64.0%). The
limit of quantification (LOQ) of cholesterol and CEs ranged from 0.2 to 10.0 μg/mL, except for cholesteryl
arachidonate (100 μg/mL), while OHCs and cholesterol precursors ranged from 0.01 to 0.10 μg/mL.
Linearity as the correlation coefficient was higher than 0.99 with the exception of cholesteryl laurate,
2
myristate, oleate, and linoleate (r > 0.98). The precision (% CV) and accuracy (% bias) ranged from 1.1%
to 10.9% and from 75.9% to 125.1%, respectively. Cholesterol signatures was applied to 154 patients with
vasospastic angina, the quantitative results were compared between 65 male and 89 female. It showed sex
differences in vasospastic angina, metabolic ratios of desmosterol to cholesterol (24-reductase; P < 0.0003)
and 7-dehydrocholesterol to cholesterol (7-reductases; P < 0.02) were higher in the female patients than the
male patients. This technique can be used to evaluate and compare metabolic changes in levels of
cholesterol and its metabolites as well as to better understand the pathophysiology of vasospastic angina.
132
|
P1-28
Toxicometabolomics study for prediction of hepatotoxicity by
troglitazone in rats
Ji Won Kim1, Sung Ha Ryu2, Siwon Kim3, Suhkmann Kim3, and Kyu-Bong Kim2
1
Department of Smart Food and Drug, Inje University, Obang-dong, Gimhae, Gyungnam 621-749, Republic of Korea
2
College of Pharmacy, Dankook University, 119 Dandae-ro, Cheonan, Chungnam 330-714, Republic of Korea
3
Department of Chemistry and Chemistry Institute for Functional Materials, Pusan National University,
Busan 609-735, Republic of Korea
E-mail: lottelya@naver.com
Troglitazone is a thiazolidinedione antidabetic agent which is the synthetic ligands for the peroxisome
proliferator-activated receptor γ (PPAR γ). However, it was withdrawn from the market in 2000 due to
liver injury in humans. In this study, we endeavor to discover surrogate biomarkers which are correlated
with hepatotoxicity induced by troglitazone using urinary proton nuclear magnetic resonance (1H NMR)
spectral data. A procedure of 1H NMR urinalysis using pattern recognition was proposed for early screening
of the hepatotoxicity of troglitazone with lipopolysachride in rats. The hepatotoxicity occurrence was tested
through clinical chemistry and histopathology. Troglitazone (2 g/kg) was single administered to rats
thereafter lipopolysaccharide (2ｘ106 EU/kg) was administered by intraperitoneal (i.p) injection 3 h after
drug dosing. Urine was collected 24 h pre-dosing (-1 Day) and at 2, 6, 10, 24 h post-dosing, respectively.
ALT (Alanine aminotransferase) and AST (aspartate aminotransferase) were the highest in 6 hours and
hepatic inflammation, in histopathology, was observed in 2 hours. 1H NMR spectroscopy showed evidently
different clustering between pre-dosing and post-dosing in global metabolic profiling through principal
component analysis (PCA) and orthogonal partial least-squares discriminant analysis (OPLS-DA). In targeted
profiling, endogenous metabolites of acetate, propionate, formate, taurine, succinate, malonate, alanine,
butyrate, and creatine were selected as putative biomarkers for hepatotoxicity by troglitazone. According to
these results, 1H NMR urinalysis can be used to predict or screen hepatotoxicity caused by troglitazone.
|
133
P1-29
Metabolic characterization of high-fat diet induced mouse model using
NMR spectroscopy
Hyun Ju Kim1, Myung Hee Nam1, Myung-Sook Choi2, and Eunjung Bang1
1
Korea Basic Science Institute, Seoul 136-701, Korea,
Food and Nutritional Genomics Research Center, Kyungpook National University, Daegu 702-701, Korea
E-mail: bej@kbsi.re.kr
2
Obesity is a multisystem disease that results from a failure of normal homeostatic mechanisms regulating
food intake, fat storage, and energy utilization. It is the predominant cause of diabetes, cardiovascular
disease, and liver disease. Increased in flux of fatty acid to the liver and potential alteration of their
metabolism in liver may result in hepatic disease.
In this study, we investigated the metabolic alterations of mouse liver induced by high fat diet (HFD)
using high resolution magic angle spinning (HR-MAS) NMR coupled with multivariate analysis such as
PCA, PLS-DA. We also feed some dietary supplements (antioxidant compound) to high fat diet induced
mouse to observe the obesity-preventive effect of the bioactive compound. These results can be used to
understand obesity and diet induced regulation of liver metabolism.
134
|
P1-30
Glucose metabolism of U87MG tumor cells using
spectrsocopy
13
C-NMR
Yun-Ju Lee, Gregory Hyung Jin Park, Chaejoon Cheong, and Hyeon-Man Baek
Division of Magnetic Resonance Research, Korea Basic Science Institute, Cheongwon, 363-883, Korea
E-mail: yjlee0118@kbsi.re.kr
Most of the attention in tumor metabolism has been focused on glycolysis and its role in providing
energy for the cell. However, there is a fundamental paradox between Warburg’s model of mitochondrial
impairment and the metabolic demands of tumor growth. We investigated the metabolic fate of glucose in
tumor cells by 13C- NMR spectroscopy. U87MG cell lines were cultured in supplemented DMEM at 37℃
in 5% CO2 and [U-13C]glucose DMEM at 37℃ in 5% CO2. Metabolites were extracted by lysing the cells
in ice-cold Methanol. Cell lysates were then spun down and the methanol/water phase lyophilized, dissolved
in D2O contain 0.25% TSP. 13C-13C coupling in lactate and alanine was observed on the 1H-decoupled
13
C-NMR spectrum of the cell extracts. The tumor cells also contained 13C-13C coupling in glutamate and
glutamine, demonstrating that glucose was metabolized to acetyl-CoA via pyruvate dehydrogenase and
oxidized further in the citric acid cycle. In this study, metabolic products of [U-13C]glucose were detected
easily by 13C-NMR experiment in the mammalian cell. The doublet of doublets or quartet in C4 of
glutamate and glutamine reflects 13C in both oxaloacetate and acetyl-CoA, the two substrates of citrate
synthase. This labeling pattern occurs with complete turnover of the citric acid cycle in functional
mitochondria.
Supported by grants from KBSI-#T33416.
|
135
P1-31
Global metabolomics and targeted steroid profiling reveal that rifampin,
a strong human PXR activator, alters endogenous urinary steroid markers
Bora Kim1, Ju-Yeon Moon2, Man Ho Choi2, Hyang Hee Yang1, SeungHwan Lee1,
1
1
1
1
1
Kyoung Soo Lim , Seo-Hyun Yoon , Kyung-Sang Yu , In-Jin Jang , and Joo-Youn Cho
1
Department of Clinical Pharmacology and Therapeutics, Seoul National University College of
Medicine and Hospital, Seoul, Korea,
2
Future Convergence Research Division, Korea Institute of Science and Technology, Seoul, Korea
E-mail: kbola@snu.ac.kr
Activation of the pregnane X receptor (PXR) is associated with increased the expression of metabolic
enzymes and transporters involved in the metabolism of xenobiotics and endobiotics and is commonly
targeted to the drug-drug interactions. The aim of this study is to identify endogenous biomarkers of PXR
activation in human, rifampicin, a strong PXR activator. Rifampicin was administered orally at the dose of
600 mg/day for 7 days in twelve healthy male volunteers and urinary metabolite profiling was performed
by ultra performance liquid chromatography time of flight mass spectrometry (UPLC-TOF-MS) for the
untargeted profiling and gas chromatography coupled with mass spectrometry (GC-MS) for the targeted
profiling in conjugation with multivariate statistical analysis. Multivariate analysis was performed between
control and rifampicin-induced urine using EZinfo (Umetrics Inc.) for the untargeted profiling and SIMCA
(Umetrics Inc.) for the targeted profiling. Candidates of untargeted metabolomic biomarkers were searched
from the human metabolome database and confirmed by tandem mass spectrometry of authentic compounds.
Principle components analysis (PCA) of untargeted metabolomic profiling yield a good separation of the
data sets from the control and rifampicin-treated groups. Untargeted urinary metabolites affected by
rifampicin treatment were as follows; hydroxytestosterone sulfate and glycochenodeoxycholate sulfate were
significantly increased and that androsterone sulfate, dehydroepiandrosterone (DHEA) sulfate, and p-cresol
sulfate levels were significantly decreased. In targeted metabolic profiling, rifampicin-induced urinary steroids
were as follows; 16a-OH-androstenedione (A-dione), 16a-OH-DHEA, 7α-DHEA , 7β-DHEA and
11β-OH-A-dione were increased, whereas DHEA, androsterone, etiocholanolone, estrone, β-cortolone, and
allo-tetrahydrocortisone were decreased. The analysis of the metabolic pathway and the metabolic ratio of
steroids enabled the estimation of the induction of CYP1A/3A/7B/11B/2C and the inhibition of
CYP17A/19A in response to PXR activation. These human urinary biomarkers may be useful for predicting
the extent of PXR activation, monitoring the activity of DMEs, and anticipating drug-drug interactions in
patients administered PXR-activating drugs. In addition, the combination of targeted and untargeted
metabolomic approach could provide promising biomarkers in clinical aspects.
136
|
P1-32
Global Metabolomics Profiling of Human Urine Reveals Change in
Endogenous Metabolites after Metformin and Pioglitazone Administration
Kumsun Cho1, Sung Kweon Cho2, Jae Yong Chung1, Seo-Hyun Yoon1, In-Jin Jang1, Joo-Youn Cho1
1
Department of Clinical Pharmacology and Therapeutics, Seoul National University College of Medicine and
Hospital, Seoul, Korea
2
Department of Pharmacology, Yonsei University College of Medicine, Seoul, Korea
E-mail: kscho615@snu.ac.kr
Both metformin and pioglitazone are used for the treatment of diabetes mellitus type 2 in monotherapy
and in combination with each other. The former activates AMP-activated protein kinase (AMPK), and the
latter stimulates PPAR-γ. Although their mechanisms of action were generally considered to be different,
they have not been fully understood. The aim of this study is to identify the changes of urinary
endogenous metabolites by metformin or pioglitazone treatment. Fourteen healthy male subjects were orally
administered metformin (1000 mg) on day 1 and day 17 and pioglitazone (30 mg/day) for 14 days from
day 3 to day 16. 12h-urine samples taken before and after administration of metformin were analyzed by
following a holistic LC-QTOF-MS-based metabolomics combined with multivariate data analyses (Mass
Profiler Professional, version B.02.01). Multivariate statistical data analyses satisfactorily classified samples
between control and metformin group, pioglitazone group, or both treatment groups. Urinary metabolites
affected by metformin treatment were as follows; in positive ion mode, while cortisol and hydroxycortisol
were decreased, 4-methoxyphenylacetic acid and retinyl β-glucuronide were increased, and metabolites found
in negative ion mode have not been fully identified. In pioglitazone treatment group compared to controls,
L-pipecolic acid and cholic acid glucuronide were increased in positive and negative ion mode, respectively.
In this study, the identified metabolites could show therapeutic effect of meformin or pioglitazone, however,
they didn't show apparent synergism of metformin and pioglitazone. Furthermore, the tendency of retinyl
β-glucuronide and cholic acid glucuronide expression in metformin or pioglitazone monotherapy group is
different. Therefore, they could be considered as biomarkers for explaining differences of mechanism of
action between metformin and pioglitazone.
|
137
P1-33
Metabolomic analysis of the enhanced effect of ursolic acid and
doxorubicin in human cancer cells using LC-QTOF/MS and
chemometric analysis
1
2
3
4
1
1
Dae-Young Lee , Mi-Kyoung Lee , Jae-Kwang Kim , Nam-In Baek , Hyung-Jun Noh , Seung-Eun Lee ,
1
1
1
1
1
Jehun Choi , Ji-Hyun Lee , Yoonpyo Hong , Seung-Yu Kim , and Geum-Soog Kim
1
Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, RDA, Eumseong
369-873, Korea
2
Department of Neurosurgery, Ajou University School of Medicine, Suwon 443-721, Korea
3
Department of Agricultural Bio-resources National Academy of Agricultural Science, RDA, Suwon 441-707, Korea
4
Graduate School of Biotechnology, Kyung Hee University, Yongin 446-701, Korea
E-mail: dylee0809@gmail.com
Ursolic acid (UA), the most abundant pentacyclic triterpenoid present in Cornus kousa Burg. (Cornaceae),
was known to have anticancer activities in prostate, breast and liver cancers. The dose-dependent
cardiotoxicities of doxorubicin (DOX) significantly limits its anticancer efficacies. One of the ways to
augment the efficacies of DOX at a relatively low cumulative dose is to use a chemical sensitizer.
Meanwhile, quenching ROS (superoxide anion, hydroxyl peroxide, and hydroxyl free radical) generated from
DOX in cardiomyocytes is thus one of the major pharmacological approaches to prevent
doxorubicin-induced cardiotoxicity. In this study, metabolomic technique was employed to evaluate the
mechanism of UA and DOX of anti-cancer activities using human breast cancer cells (MCF-7) by
LC-QTOF/MS analysis. LC-QTOF/MS spectra of cell extracts compared with those of UA and DOX treated
cell extracts gave bio marker candidates using chemometric analysis. As results, seven bio marker
candidates found and assumed to be involved metabolism of MCF-7 cells. And the developed a reliable
discrimination method between UA and DOX treated cells using LC-QTOF/MS based metabolomic
technology. UA and DOX treated cell extracts gave a marker which was appeared as different intensity of
TIC. An metabolite with masses m/z 611.1435 [M-H]- was identified as oxidized glutathione. Therefore, we
investigated the enhanced effect of UR and DOX in metabolic level and may bring benefit to clinical
chemotherapy.
Supported by Cooperative Research Program for Agricultural Science & Technology Development
(PJ008700), RDA, Korea.
138
|
P1-34
High-throughput screening for in vitro inhibitory effects of daidzein
and genistein, on human cytochrome P450 isoforms using
ultra-performance liquid chromatography/tandem mass spectrometry
1,2
1,2
1,2
1,2
1,2
Ji Yeon Hyun , Min Ho Bea , Jeong Ju Seo , Mi-Ri Gwon ,Yong-Chul Shin ,
1
1,2
Hae Won Lee , Young-Ran Yoon
1
Departmetnt of Biomedical Science, Kyungpook National University Graduate School, 680 Gukchaebosang-ro,
Jung-gu, Daegu 700-842, South Korea
2
Clinical Trial Center, Kyungpook National University Hospital, 130 Dongdeok-ro, Jung-gu, Daegu 700-721, South Korea
E-mail: htearsot@naver.com
A number of food-drug interactions have been identified following the co-administration of food with
prescribed Western medication. In this study, to evaluate potential food-drug interaction, the effects of
daidzein and genistein (soy isoflavones, one of the phytoestrogens) on the catalytic activities of nine major
CYP isoforms were investigated using an economically feasible and time-saving high-throughput screening
method. Two cocktail sets were incubated with human liver microsomes: Cocktail A consists of phenacetin
(CYP1A2), coumarin (CYP2A6), paclitaxel (CYP2C8), omeprazole (CYP2C19), dextromethorphan (CYP2D6),
and midazolam (CYP3A4). In contrast, the contents of Cocktail B involve bupropion (CYP2B6), tolbutamide
(CYP2C9), and chlorzoxazone (CYP2E1). After incubation, the supernatants of each reaction sample were
pooled and analyzed by using ultra-performance liquid chromatography-tandem mass spectrometry
(UPLC-MS/MS). The concentrations of the substrate metabolites were represented by substrate
metabolite/internal standard (chlorpropamide) ratio. According to data of this study, daidzein and genistein at
concentrations from 0.0001 to 1 uM showed notable inhibitory effects on CYP isoforms evaluated, except
CYP2C8 by daidzein and CYP2E1 by genistein. Accordingly, further in vivo human studies are needed to
confirm these results. The high-throughput screening method used in this study can be a useful tool in
evaluation and understanding of food-drug interactions.
This research was supported by grants from the Korea Healthy 21 R&D Project, Ministry of Health &
Welfare, and the Republic of Korea (A070001) & the National Project for Personalized Genomic Medicine
(A111218-PG02), the Ministry of Health & Welfare, Republic of Korea, and the Basic Science Research
Program through the National Research Foundation of Korea (NRF) funded by the Minisry of Education
Science and Technology (2010-0022996), Republic of Korea.
|
139
P1-35
Steady-state pharmacokinetic properties of tamsulosin
in healthy male volunteers
Sook Jin Seong, Joomi Lee, Mi-sun Lim, Sung Min Park, Jeonghyeon Park,
Jeong Ju Seo, Hae Won Lee, Young-Ran Yoon
Department of Biomedical Science and Clinical Trial Center, Kyungpook National University Graduate School and
Hospital, Daegu, Republic of Korea
BK21 program, Kyungpook National University School of Medicine, Daegu, Republic of Korea
E-mail: jy-nee@hanmail.net
Tamsulosin hydrochloride (CAS 128332-25-2), a third-generation α1-adrenoceptor antagonist, is prescribed
in a modified-release formulation for the treatment of lower urinary tract symptoms/benign prostatic
hyperplasia. Although the single-dose pharmacokinetics of tamsulosin have been widely evaluated in the
fasted and fed states, limited data are available investigating the multiple-dose pharmacokinetics of
tamsulosin, especially in the fasted state. The aim of the study was to investigate the pharmacokinetic (PK)
properties of a newly developed generic capsule formulation (test) and a branded capsule formulation
(reference) of tamsulosin after 0.2 mg/day oral doses administered to fasted healthy Korean male volunteers
for 5 days. In a randomized, open-label, multiple-dose, two-period, crossover study, all 44 subjects were
randomly assigned in a 1:1 ratio to receive a newly developed generic capsule formulation (test) or a
branded capsule formulation (reference) of tamsulosin 0.2 mg, followed by a 10-day washout period and
administration of the other formulation. Plasma concentrations of tamsulosin were assessed after
administration of five-day multiple doses, using HPLC/MS/MS. The main pharmacokinetic properties with
the test and reference formulations were as follows: Css,max , 9.0 (2.9) and 8.4 (2.6) ng/mL, respectively;
median (range) tmax, 4 (2-6) and 5 (2-7) hours; AUCτ, 93.7 (31.5) and 88.2 (29.3) ng ․ h/mL; and t½, 9.5
(2.6) and 10.0 (2.7) hours. The volume of distribution and clearance after oral administration of tamsulosin
were 0.5 L/kg, and 0.04 L/h/kg, respectively. The accumulation ratios for 0.2 mg once-daily dosing regimen
were 1.2. Clinical and laboratory adverse events were assessed. No serious AE was reported during the
study. Both formulations were well tolerated.
140
|
Poster 2.
Plant Metabolomics
|
141
P2-1
Mass spectrometry-based metabolite profiling of Korean black
raspberry (Rubus coreanus Miquel) at different ripening stages
Sarah Lee1, Tae Kyung Hyun2, Jae-Yean Kim2, Choong Hwan Lee1
1
Department of Bioscience and Biotechnology, Konkuk University, Seoul 143-701, Republic of Korea
Division of Applied Life Science (BK21-WCU program), Plant Molecular Biology and Biotechnology Research
Center, Gyeongsang National University, Jinju 660-701, Republic of Korea
E-mail: cybersarah@hanmail.net
2
Metabolite profiling of Korean black raspberry (Rubus coreanus Miquel) at different ripening stages was
performed using gas chromatography-ion trap-mass spectrometry (GC-IT-MS) and ultra performance liquid
chromatography-quadrupole-time of flight-mass spectrometry (UPLC-Q-TOF-MS). Multivariate analysis of the
data set via principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA)
clearly showed the metabolic changes according to ripening stages. In GC-MS analysis, most of the amino
acids, organic acids, fatty acids, and sucrose decreased, whereas fructose and glucose increased in relative
concentration according to ripening of fruits. In UPLC-Q-TOF-MS analysis, anthocyanins and phenolic
metabolites were the major metabolites distinguishing the different ripening of fruits. Anthocyanins such as
cyanidin 3-O-xylosylrutinoside and cyanidin 3-O-rutinoside increased, while proanthocyaninB and phenolic
metabolites such as catechin, epicatechin, quercetin 3-O-rutinoside, and quercetin-glucuronide decreased.
Trolox equivalent antioxidant capacity (TEAC) of fruits was also measured, and immature fruit was
possessed the highest antioxidant activity in different ripening stages. This study suggested that MS based
metabolite profiling of fruit ripening showed the changes of metabolites and their metabolic pathway, and
also phenolic metabolites such as catechin, epicatechin, and quercetin derivatives can explain their change of
antioxidant activity.
142
|
P2-2
Metabolite profiling of the short-term responses of rice leaves (Oryza
sativa cv. Ilmi) cultivated under different LED lights and its
correlations with antioxidant activities
1
1
2
Eun Sung Jung , Sarah Lee , Sun-Hyung Lim , Choog Hwan Lee
1
1
2
Department of Bioscience and Biotechnology, Konkuk University, Seoul 143-701, Republic of Korea
National Academy of Agricultural Science, Rural Development Administration, Suwon 441-707, Republic of Korea
E-mail: chlee123@konkuk.ac.kr
Metabolite profiling of rice leaves (Oryza sativa cv. Ilmi) was performed to investigate the short-term
responses to different light-emitting diode (LED) lights, blue (B), green (G), red (R), white (W), shade (S),
by using gas chromatography-ion trap-mass spectrometry (GC-IT-MS) and ultra-performance liquid
chromatography-quadrupole-time-of-flight-mass spectrometry (UPLC-Q-TOF-MS) with multivariate analysis.
Clear grouping patterns of each light-grown sample, except G and W, were shown in partial least
squares-discriminant analysis (PLS-DA). Thirty-two primary metabolites and eleven secondary metabolites
were selected and visualized using heatmap. Antioxidant activities of rice leaves followed the order B = W
= G > R > S and isoorientin-2′′-O-glucoside, isovitexin-2′′-O-glucoside, isoorientin-2′′-O-(6′′′-ρ-coumaroyl)-glucoside,
and isoscoparin-2′′-O-glucoside showed similar relative differences and had higher Pearson’s correlation
coefficients than other metabolites in correlation network. According to the orthogonal projection to latent
structures-discriminant analysis (OPLS-DA) between B and R, the levels of amino acids, organic acids, fatty
acids, and flavonoid glycosides were relatively high in B, whereas the glucose and fructose levels were
high in R.
|
143
P2-3
Metabolomics for the quality assessment of Lycium chinense fruits
1
1
1
1
1
Soo-Yun Park , Si Myung Lee , Hyo Jin Kim , Sun-Hyung Lim , Yunsoo Yeo ,
Hyun Suk Cho1, Sun-Hwa Ha1, Sang Un Park2, Jae Kwang Kim1
1
National Academy of Agricultural Science, Rural Development Administration, Suwon 441-707, Republic of Korea
2
Department of Crop Science, Chungnam National University, 220 Gung-dong, Yuseong-gu, Daejeon 305-764,
Republic of Korea
Lycium chinense has been used as a traditional medicine for centuries in Asia because of its positive
effects on health. However, its functional components have not been elucidated. This study determines the
levels of health-promoting lipophilic compounds, including carotenoids, tocopherols, and phytosterol, and
those of 42 hydrophilic metabolites, including sugars, organic acids, alcohols, amines, and amino acids, in
L. chinense fruit from 11 cultivars. The metabolite profiles were subjected to a principal component
analysis (PCA), Pearson correlation analysis, and hierarchical clustering analysis (HCA). PCA showed the
Cheongdang (LM-3) cultivar to be distinct from the others. The correlation results for a total of 55
compounds revealed strong correlations between the metabolites that participated on closely related
pathways. The Cheongdang cultivar appears to be most suited for functional food production because of its
high carotenoid, tocopherol, and phytosterol levels. These results indicate the usefulness of metabolite
profiling as a tool for assessing the quality of food.
144
|
P2-4
Fourier Transform Infrared (FT-IR) spectroscopy of genomic DNA
could discriminate F1 progenies of Chinese cabbage (Brassica rapa
subsp. pekinensis) depend on their paternal lineage
1,4
1
1
3
3
Seung Yeub Song , Eun Yee Jie , Myung Suk Ahn , In Ho Lee , Jun Ho Song ,
4
2
In Jung Kim , Suk Weon Kim
1
Greenbio Research Center, 2Biological Resource Center, Korea Research Institute of Bioscience and
Biotechnology(KRIBB),125 Gwahak-ro, Yuseong-gu, Daejeon, 305-806, Korea, 3Asia Seed Co. Research Institute of
Biotechnology Breeding, 518 beon-gil, Gyeongchungdae-ro, Janghowon-eup, Icheon-si, Gyeonggi-do, 467-906, Korea,
4
Department of Biotechnology, Jeju National University, Jeju 690-756, Korea
E-mail: kimsw@kribb.re.kr
FT-IR spectroscopy combined by multivariate analysis was used to determine whether two different F1
hybrid lines could be discriminated from their parental inbreed lines. Genomic DNA was isolated from
leaves of three parental lines and two F1 hybrid lines of Brassica campestris subsp. pekinensis. Purified
−1
genomic DNA was analyzed by FT-IR spectroscopy in the spectral region from 4000 to 400 cm . FT-IR
spectra showed that typical spectral differences were existed in the frequency regions of N-H stretching
−
(amide I), C=O stretching vibrations (amide II), and PO2 ionized asymmetric and symmetric stretching,
respectively. Principal component analysis (PCA) was able to discriminate F1 hybrid progenies depending
on their parental lineages, even though they have same maternal background. The partial least squares
discriminant analysis (PLS-DA) gave more clear discrimination between two parental and their progeny
hybrid lines. These FT-IR spectral differences might be directly related to subtle changes in base functional
group and backbone structures of genomic DNA. Considering these results, this technique could provide a
research foundation for the FT-IR spectral based rapid diagnosis, selection and discrimination of parental
line and their progenies. Furthermore this technique could be applied for testing the purity of seed in
hybrid seed industry.
|
145
P2-5
Metabolite profiling of Polygonatum odoratum var. pluriflorum from
1
different geographical origin using H NMR spectroscopy and
multivariate analysis
1,2
1
1
Miso Nam , Youngae Jung , Do Hyun Ryu , Geum-Sook Hwang
1,3
1
Integrated Metabolomics Research Group, Seoul Center, Korea Basic Science Institute, Seoul 136-713, Republic of Korea
Sungkyunkwan Univ. Natural Sciences Campus, Cheoncheon-dong, Jangan-gu, Suwon-si, Gyeonggi-do 440-746 Korea
3
Graduate School of Analytical Science and Technology, Chungnam University, Daejeon 305-764
E-mail: gshwang@kbsi.re.kr
2
Polygonatum odoratum var. pluriflorum is traditional Chinese medicine and used for multiple ways such
as medicine, vegetable and tea. It has been used to treat diabetes, intestinal problems, gout and rheumatism.
Polygonatum odoratum var. pluriflorum that is grown in three different regions (SanCheon, SaCheon,
SunCheon) of Korea were used in this study. Aqueous extracts were analyzed by 1H NMR spectroscopy
and metabolite profiling coupled with multivariate analysis was applied to characterize the differences origin.
PCA analysis showed significantly distinctions between origins. A one-way ANOVA was performed to
statistically certify the metabolite differences of polar extracts. The significant metabolites were sugars
(glucose, sucrose), amino acids (arginine, asparagine, glutamine, leucine, phenylalanine) and organic acids
(4-aminobutyrate, citrate, acetate, malate). Therefore, these data suggest that a metabolomic approach using
1
H NMR analysis is an effective analytical method to differentiate biochemical compositions among different
origins in plant.
146
|
P2-6
An integrated analysis for determining the geographical origin of
1
crops using ICP-AES/ICP-MS and H NMR analysis
Yong-Kook Kwona, Yeon-Sik Bongb, Kwang-Sik Leeb,c,*, Geum-Sook Hwanga,c,*
a
Integrated Metabolomics Research Group, Seoul Center, Korea Basic Science Institute, Seoul 136-713, Republic of Korea
b
Division of Earth&Environmental Sciences, Ochang Center, Korea Basic Science Institute, Ochang-eup,
Cheonwong-gun, Chungbuk 363-883, Republic of Korea
c
Graduate School of Analytical Science and Technology, Chungnam National University, Daejeon 305-764,
Republic of Korea
E-mail: gshwang@kbsi.re.kr
ICP-MS and 1H NMR are commonly used to determine the geographical origin of crops. In this study,
data from multielemental analysis performed by ICP-AES/ICP-MS and metabolomic data obtained from 1H
NMR were integrated to improve the reliability of determining the geographical origin of medicinal herbs.
Astragalus membranaceus and Paeonia albiflora with different origins in Korea and China were analyzed
by 1H NMR and ICP-AES/ICP-MS, and an integrated multivariate analysis was performed to characterize
the differences between their origins. Four classification methods were applied: linear discriminant analysis
(LDA), k-nearest neighbor classification (KNN), support vector machines (SVM), and partial least-squares
discriminant analysis (PLS-DA). Results were compared using leave-one-out cross-validation and external
validation. The integration of multielemental and metabolomic data was more suitable for determining
geographical origin than the use of each individual data set alone. The integration of the two analytical
techniques allowed diverse environmental factors such as climate and geology, to be considered. Our study
suggests that an appropriate integration of different types of analytical data is useful for determining the
geographical origin of food and crops with a high degree of reliability.
|
147
P2-7
Untargeted metabolite profiling to discriminate different species from
the genus Cirsium using UPLC-MS
In Jin Ha, Yong-Kook Kwon, Youngae Jung, Geom-Sook Hwang
Seoul Center, Korea Basic Science Institute, Seoul 136-713, Republic of Korea
E-mail: ijha@kbsi.re.kr
Cirsium species (family compositae) is one of several genera known commonly as thistle which is widely
distributed across Korea. Korean thistles (Cirsii Herba), Cirsium japonicum and other Cirsium species grown
in Korea, have been used as the important components of traditional herbal medicine to treat diverse kinds
of bleeding and inflammatory. However, those species are difficult to discriminate morphologically, but
discrimination of the dried and cut raw material is even more challenging. Therefore, this study was
designed to classify and identify closely related species in genus Cirsium. The comprehensive and
untargeted metabolite profiles of six Korean thistles were performed using ultraperformance liquid
chromatography (UPLC) in combination with a hybrid quadrupole time-of-flight mass spectrometer
(QTOF).The difference in metabolite profiles between species was observed by the principal component
analysis (PCA) and partial least squares discriminant analysis (PLS-DA), and the six species were
successfully classified. The species that have similar features were grouped into unique pattern in variable
clusters. Significant differentiation of metabolite biomarkers were detected in the Cirsium species. The
present study suggests that the LC-MS based untargeted metabolomic analysis is an efficient and powerful
tool for discriminating different species in medicinal herbs.
148
|
P2-8
Policosanol profiles of barley sprouts at different growth stages and
investigation of their adenosine monophosphate kinase activation
Woo Duck Seo, Sang-Ik Han, Seong-Hwan Oh, Ji-Eun Ra, Ji-Young Park, Kyung Hye Seo,
Mi-Jin Park, Byung-Joo Kim, Choon Woo Lee, Min Hee Nam
Departmen of Functional Crops, National Institute of Crop Science (NICS),
Rural Development Administration (RDA), Miryang, 627-803, Korea
E-mail: swd2002@korea.kr
AMPK is an intracellular sensor that modulates the energy balance within the cell. AMPK was activated
significantly by the hexane extract of barley sprouts. This AMPK activation emerges across the growth
stages of the sprout, becoming most significant (three times above the initial stages) 10 days after seeding.
After this time, the activation decreased between 13 days to 20 days post sprouting. Analysis of the hexane
extracts by GC-MS, showed the amounts of policosanols (C20–30) in the plant dramatically increased
between 5 days (109.7 mg/100g) to 10 days (343.7 mg/100g) post sprouting, and then levels fell back
down, reaching 76.4 mg/100g 20 days post sprouting. This trend is consistent with policosanols being the
active ingredient in the barley plants. We validate this by showing that hexacosanol is an activator of
AMPK. The richest cultivar for PCs was found to be Daejin cultivar. Cultivars had a significant effect on
total PC content (113.2 to 183.5 mg/100g) within the plant up to 5 days post sprouting. However this
dependence on cultivar was not so apparent at peak stages of PC production (10 days post sprouting). The
most abundant policosanol in barley sprout, hexacosanol contributed 62–80% of the total PC content at
every stage. These results are valuable to determine the optimal times of harvest to ensure that policosanols
from barley sprouts are within the required limits
Supported by grants from the "Cooperative Research Program for Agriculture Science & Technology
Development (Project No. PJ00850601)", Rural Development Administration, Republic of Korea
|
149
P2-9
Identification of component for blueberry, raspberry and blackberry
based on metabolomics
Wooseok Kim1,2 , Jeongjoo Pyo3, Bora Jung4, Myung-yoon Jun4, Hyun Jin Baek4, Jeongae Lee1,*
1
Molecular Recognition Research Center, Korea Institute of Science and Technology, Seoul 136-791, Korea
2
Department of Food Science and Biotechnology, Dongguk University, Seoul, 100-715, Korea
3
Dankook University High School, Seoul 135-852, Korea
4
Young In Scientific Co., Ltd. Seoul, 135-120, Korea
Email: frans@kfri.re.kr
Berries have antioxidant, antimicrobial, anticancer and anti-inflammatory activity as health functional food,
beverages, yogurts, and jam. And they were consumed by many peoples. In this study, we identified
components of blueberry, raspberry and blackberry using gas chromatography-mass spectrometer (GC-MS)
and mass profiler professional (MMP). Mass spectrometry based metabolite profiling in combination with
multivariate data analysis was introduced to quantity changes in metabolic patterns. Blueberry, raspberry and
blackberry were sequentially extracted with hexane, ethyl ether, ethyl acetate and methanol. After the
extracted berries were evaporated to dryness, resolved in solvent and then were injected on GC-MS. The
data of whole component of berries were processed by principle component analysis (PCA) as multivariate
analysis to determine whether separate classes of extraction solvents. PCA score plot were identified
through retention time and m/z data pair. At the results, PCA score plot obtained from hexane and ethyl
ether extracts was the good discrimination between berries. And then, loading plot analysis performed to
increase accuracy of data processing was collected 139 peaks. Among them, 28 peaks of main component
were categorized in the significant level using ANOVA test (p<0.001). Although we have not yet identified
the component of berries, tridecanal, methyl eugenol, 2-nonadecanol, 1-methylbutyl palmitate,
2-heptacosanone and 2-undecanone were searched using library. Methyl eugenol is known as antiallergic
reaction compound in blueberry, raspberry, and blackberry.
150
|
P2-10
Global metabolite profiling of the Medicago truncatula mutant under
salt condition using gas chromatography-mass spectrometry
Seok-Young Lee1, Sun-Hee Hyun1, So-Hyun Kim1, Hae Eun Park1, Shin Jung Park1,
1
1
1
1
1
1
Hyo Won Suh , Da Yeon Kim , Seul Gi Lee , Taek-Joo Oh , Xue Jiang , Jun-Sang Oh ,
1
2
1
Hye Min Ahn , Young-Woo Nam , Hyung-Kyoon Choi
1
2
College of Pharmacy, Chung-Ang University, Seoul 156-756, Republic of Korea
Department of Life Science, Sogang University, Seoul 121-742, Republic of Korea
E-mail: hykychoi@cau.ac.kr
Metabolite profiling of M. truncatula mutant was performed using gas chromatography-mass spectrometry
(GC-MS) coupled with multivariate statistical analysis. Comparative evaluation was carried out to investigate
the difference of metabolome according to various gene types, and cultivation conditions. Four groups of
M. truncatula are composed of wild type (WT)-control-shoot, WT-salt-shoot, mutant type
(MT)-control-shoot, and MT-salt-shoot. 1-Pyrroline-5-carboxylate synthase (P5CS) gene was genetically
manipulated in MT. P5CS is a key enzyme for the rate-limiting step of proline biosynthesis. Salinity is one
of the major environmental stress that changes the metabolites of plants by inducing formation of reactive
oxygen species (ROS). Metabolic profiles of each sample obtained from GC-MS analysis were further
analyzed using partial least squares-discriminant analysis (PLS-DA). This study shows the content of lipids,
amino acids, and other metabolites are significantly differed between wild type and P5CS3 gene mutant of
M. truncatula cultivated in control and salt conditions.
|
151
P2-11
Metabolic assessment of black raspberry (Rubus coreanus Miquel)
administration effect on 8-OHdG levels in plasma of smokers using
1
H-nuclear magnetic resonance spectrometry
1
1
1
1
1
Hyo Won Suh , Shin Jung Park , Sun-HeeHyun , So-Hyun Kim , Seung-Ok Yang ,
1
1
1
1
1
1
Hae EunPark , Seok-YoungLee , Da Yeon Kim , Seul Gi Lee , Taek-Joo Oh , Xue Jiang ,
Jun-Sang Oh1, Hye Min Ahn1, Joong-Hyuck Auh2, Soo-Muk Cho3, Jung-Hyun Kim4,
1
and Hyung-Kyoon Choi
1
College of Pharmacy, Chung-Ang University, Seoul 156-756, Republic of Korea
Department of Food Science and Technology, Chung-Ang University, Ansung 456-756, Republic of Korea
3
Department of Agrofood Resources, National Academy of Agricultural Science, Suwon 441-853
4
Republic of Korea Department of Home Education and Economics, Chung-Ang University, Seoul 156-756, Republic
of Korea
E-mail: hykychoi@cau.ac.kr
2
Black raspberry (BR) is well known to have health-beneficial effect such as antioxidant, antiproliferative,
and antiinflammatory activities. ¹H nuclear magnetic resonance (NMR) coupled with multivariate statistical
analysis were used to assess the effect of BR administration on 8-hydroxy-2'-deoxyguanosine (8-OHdG)
levels in plasma of smokers. In this study, to investigate the effect of BR on the level of 8-OHdG,
nineteen male smokers between 20 and 30 years old were treated with BR for 4 weeks. 8-OHdG is one of
the most popular markers for the evaluation of oxidative DNA damage. Urine and plasma samples were
collected from the volunteers before and after the BR administration. Twelve volunteers with the decreased
level of urinary 8-OHdG over 10% after BR administration were selected for plasma metabolic profiling
using ¹H NMR. By partial least squares-discriminant analysis (PLS-DA), the metabolic profiles between
preadministered and postadministered groups were discriminated. An independent t-test was performed to
compare the relative levels of the selected metabolites based on VIP values over 0.7. The relative levels of
isoleucine,
pyridoxine,
histamine,
proline,
lysine,
tyrosine,
phenylalanine,
acetate,
formate,
N,N-dimethylglycine, ethylmalonate, valine, and ornithine were lower in BR administered group, whereas
guanidoacetate, glycine, and trimethylamine N-oxide were lower in preadministered group. Decreased level
of histamine suggests the antiinflammatory modulation activity of BR administration in smokers.
152
|
P2-12
Enhancement of stigmasterol yield in Lemna minor whole plant culture
by co-treatment of methyl jasmonate and silver nitrate as elicitors
Hyo Won Suh1, Sun-Hee Hyun1, So-Hyun Kim1, Seok-Young Lee1, Da Yeon Kim1,
1
1
1
1
1
2
Seul Gi Lee , Taek-Joo Oh , Xue Jiang , Jun-Sang Oh , Hye Min Ahn , Byung-Kwan Cho ,
3
4
1
Hoo keun Lee , Choul-Gyun Lee ,and Hyung-Kyoon Choi
1
College of Pharmacy, Chung-Ang University, Seoul 156-756, Republic of Korea
Department of Biological Sciences, KAIST, Daejeon 305-701, Republic of Korea
3
College of Pharmacy, Gachon University, Incheon 406-840, Republic of Korea
4
Department of Biological Engineering, Inha University, Incheon 402-751, Republic of Korea
E-mail: hykychoi@cau.ac.kr
2
In this study, the effects of methyl jasmonate (MJ) and silver nitrate (SN) treatment on phytosterol yields
such as campesterol, stigmasterol, and β-sitosterol in Lemna minor whole plant cultures was investigated
using gas chromatography-mass spectrometry. MJ and SN were treated independently or together as
elicitors. MJ and SN treatment retarded the growth of L. minor whole plants; however, the phytosterol
yields were higher in each elicitation culture than in the control, and were higher at day 28 than at day 42
of treatment. Among the three phytosterols, the enhanced and highest yield of stigmasterol was achieved
when MJ and SN were treated together as elicitors, but there was no enhanced yield of campesterol and
β-sitosterol even under the co-treated condition.
|
153
P2-13
Neuraminidase Inhibitory Activity Polyphenolic Compounds from the
Barley sprouts (Hordeum vulgare L.)
Mi Jin Park, Sang-Ik Han, Seong-Hwan Oh, Ji-Eun Ra, Ji-Young Park, Kyung Hye Seo,
Mi-Jin Park, Byung-Joo Kim, Choon Woo Lee, Min Hee Nam, Woo Duck Seo
Departmen of Functional Crops, National Institute of Crop Science (NICS),
Rural Development Administration (RDA), Miryang, 627-803, Korea
E-mail: swd2002@korea.kr
Neuraminidase plays an important role in viral proliferation. The long-term efficacy of two commercially
available neuraminidase inhibitors, zanamivir and oseltamivir (Tamiflu), are often limited by adverse side
effects. Therefore, natural product-derived influenza agents have attacked considerable interest in treating
influenza. The 80% methanol extract of the sprout of barley sprouts were exhibited the greatest extraction
yield and neuraminidase inhibition rate (28.5% and 89.5%, respectively) and showed a potent neuraminidase
inhibitory activity (IC50 = 1.78 mg/mL). A series of polyphenol derivatives from barley Hordeum vulgare
L. were shown to display modest to high inhibition for neuraminidase with IC50 values of 23.2 – 64.7 µM.
All compounds were found to be non-competitive inhibitors. For quantitative analysis by UPLC, the main
polyphenolic component was saponarin (about 93%). It was assumed that sprouts extracts neuraminidase
inhibition activity of barley sprouts extract might be derived from saponarin. Interestingly, saponarin were
decreased with growth stage 10 days after sprouting. Therefore, the sprouted barley leaves in 10 days is
suitable to prevent the influenza epidemic infection.
Supported by grants from the "Cooperative Research Program for Agriculture Science & Technology
Development (Project No. PJ008506)", Rural Development Administration, Republic of Korea
154
|
P2-14
Plant metabolomics : Cheonhyehyang metabolite profiling and
identification by high resolution LC-MS/MS system
Jihyun Lee, Hee Jung Choi, Won Sil Choi and Ji Yoon Lee
National Instrumentation Center for Environmental Management, College of Agriculture and Life Sciences,
Seoul National University, Seoul 151-742, Korea
E-mail: jiyoon@snu.ac.kr
Cheonhehyang, also called as Setoka, was first introduced in 2002 from Japan. It is one kind of citrus
fruits and is well known for its taste and flavor that are better than tangerine’s. However, there is a
problem that its taste becomes bitter when exposed to cold temperature. Therefore, this study was done to
investigate why the taste of Cheonhehyang changes. We studied the amount of total phenolic compounds
and sugar content to analyse the two groups of Cheonhehyang that we classified for the experiment: one
with the normal taste and the other one that has the bitter taste. Also the differences between the two
groups were studied by using High resloution LC/Mass spectrometry and SEIVE program. PCA statistical
analysis was also used to compare the expression level of the compounds.
|
155
P2-15
Application of non-targeted metabolomic approach to identify origins
of Anemarrhena asphodeloides Bunge
Nahyun Kim1, Seungmok Ryu1, Donghyuk Lee2, Jae Won Lee2, Eun-Kyoung Seo3,
4
1
Je-Hyun Lee , and Dongho Lee
1
School of Life Sciences and Biotechnology, Korea University, Seoul 136-713, 2Department of Statistics, Korea
University, Seoul 136-701, 3Department of Herbal Crop Research, National Institute of Horticultural & Herbal Science,
Rural Development Administration, Eumseong 369-873, 4College of Pharmacy, Chung-Ang University, Seoul 156-756,
and 5College of Pharmacy, Chungbuk National University, Cheongju 361-763, Korea
E-mail: kimn@korea.ac.kr
An ultraperformance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF
MS) method was developed for metabolite profiling of Anemarrhena asphodeloides Bunge. All metabolites
were successfully analyzed by UPLC-QTOF MS, and a method validation process was applied to ensure the
validity of the analyzed data by showing the analytical stability and accuracy using a quality control
samples. Subsequently, the metabolic profile data were adopted to conduct multivariate statistical analyses
such as a principal component analysis and a hierarchical clustering analysis in order to determine
metabolite patterns that could be used to distinguish two different origins. Furthermore, metabolite selection
methods, t-test and significance analysis of microarrays, were applied to extract influential metabolites and
to identify key constituents for the efficient comparison of two groups. The UPLC-QTOF MS analysis
successfully classified 21 samples into two species groups, and the results suggest that this proposed
method is reliable, accurate, and effective for pattern analysis of A. asphodeloides showing the possibilities
that can be adapted for use to identify and provide quality control of diverse species of medicinal plants.
Supported by grants from the Korea Food and Drug Administration, Republic of Korea
156
|
P2-16
Determination of geographical origins of peony root (Paeonia lactiflora
Pall) and the estimation of mixing proportions of blended samples of
different origins
1
2
3
1
1
Jung A Um , Yun Sun Lee , Young Geun Choi , Dong Kyu Lee , Chang Ju Lim ,
4
5
5
1
6
Young A Youn , Hwa Dong Lee , Hi Jae Cho , Jeong Hill Park , Young Bae Seo ,
Hsun-chih Sean Kuo7, Johan Lim3, Tae Jin Yang2, Sung Won Kwon1, Jeongmi Lee4
1
College of pharmacy, Seoul National University, Seoul, Korea
Department of Plant Science, Plant Genomics and Breeding Institute, and Research Institute for Agriculture
and Life Sciences, College of Agriculture and Life Sciences, Seoul National University, Seoul, Korea.
3
Department of Statistics, Seoul National University, Seoul, Korea
4
School of Pharmacy, Sungkyunkwan University, Suwon, Korea
5
Korea Promotion Institute for Traditional Medicine Industry, Gyeongsan, Korea
6
Department of Herbalogy, College of Oriental Medicine, Daejeon University, Daejeon, Korea
7
Department of Statistics, National Chengchi University, Taipei Taiwan
E-mail: jlee0610@skku.edu
2
Because plant growing conditions such as cultivation soil and climate can affect plant metabolic
composition and consequently their medicinal efficacy, discriminating geographic origins is of great interest
in traditional Chinese medicine (TCM) research. In the present study, 60 authenticated samples of peony
root were collected from various regions in China and Korea, and their genetic diversity was investigated
using both chloroplast intergenic space (CIS) analysis and high resolution melting (HRM) analysis. All
samples were genetically indistinguishable, indicating that the DNA-based techniques were not appropriate
for origin discrimination. In contrast, 1H nuclear magnetic resonance (NMR) spectroscopy-based
metabolomics coupled with multivariate statistical analysis revealed a clear difference in the metabolic
profiles between the Chinese and the Korean peony samples. Orthogonal partial least squares-discrimination
analysis allowed the identification of potential marker metabolites responsible for classification, including
γ-aminobutyric acid, arginine, alanine, paeoniflorin, and albiflorin. The validity of the discrimination model
was tested by response permutation test and blind prediction test for internal and external validation,
respectively. Furthermore, a new statistical model to estimate the mixing proportions of blended samples
from China and Korea was established using constrained least square method for the first time. Prior to the
model construction, the uncontrolled variation between the testing and training set was eliminated by finding
a linear mapping between them, which shows the possibility of repetitive usage of time-honored data and
consequently deposition of the data . Consequently, the plot of the true mixing proportion versus the
estimated proportion exhibited the reliability and robustness of the constructed model. This study suggests
that 1H-NMR metabolomics could be applied in the TCM market for quality control purposes.
|
157
P2-17
Simultaneous determination of four phenolic compounds and
metabolite profiling of Gastrodia elata by LC/QTOFMS
JaeYoung Kwon1, Nahyun Kim1, DongHyuk Lee2, Ah-Reum Han3, Jae Won Lee2,
3
4
1
Eun-Kyoung Seo , Je-HyunLee , and Dongho Lee
1
School of Life Sciences and Biotechnology, Korea University, Seoul 136-713, Korea, 2Department of Statistics, Korea
University, Seoul 136-701, 3College of Pharmacy and Center for Cell Signaling & Drug Discovery Research, Ewha
Womans University, Seoul 120-750, 4College of Oriental Medicine, Dongguk University, Gyeongju 780-714
E-mail: kjy1207@korea.ac.kr
Qualitative and quantitative determination of four bioactive compounds including gastrodin, gastrodigenin,
p-hydroxybenzaldehyde, bis(4-hydroxybenzyl)ether isolated from Gastrodia elata has been developed using
high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (HPLC-Q-Tof MS). The
analysis of four standard compounds showed good linearity, intra- and inter-day precisions, and accuracy of
them. In addition, metabolite profiling of 23 different G.elata extracts was carried out to identify their
origins. Total analyzed metabolites were applied to various metabolite selection methods to determine an
optimum classification method with selected metabolites. This metabolite selection allowed discrimination of
the raw and steamed G.elata, and authentication of several biomarkers differently processed G.elata. Overall
the simultaneous determination of four components and subsequent pattern analysis can be used to identify
and provide quality control of G. elata.
Supported by grants from the Korea Food and Drug Administration, Republic of Korea.
158
|
P2-18
LC/MS-based metabolomic analysis of green tea cultivated with low
light and its anti-obesity effect
Lan-Sook Lee1*, Ji Hea Choi1,2*, Mi Jeong Sung1, Jin-Young Hur1, and Hyun-Jin Kim3
1
Korea Food Research Institute, 516 Baekhyun–dong, Bundang–gu, Seongnam, Gyeonggi 463-746, Korea
2
University of Science & Technology, 176 Gajung-dong, Yuseong-gu, Daejeon, Korea
3
Department of Food Science & Technology, Gyeongsang National University, Jinju, Gyeongsang, Korea
E-mail: funnywo@hanmail.net / sohee0809@hanmail.net
The effect of low light on their nutritional and sensory qualities of green tea (GT) was investigated by
LC/MS-based metabolomic analysis. The shade treatment increased quercetin-galactosylrutinoside,
kaempferol-glucosylrutinoside, epicatechin gallate, epigallocatechin gallate, tryptophan, phenylalanine, theanine,
glutamine, glutamate, and caffeine levels, but decreased quercetin-glucosylrutinoside, kaempferol-glucoside,
gallocatechin, and epigallocatechin levels. The change of these compounds positively affected the
anti-obesity effect of GT as well as the sensory and color quality. The obese characteristics involving body
and fat weight and obese-related metabolite profiles were positively affected by GT. Lean, obese and GT
groups were clearly discriminated on the PLS-DA scores plot and the major metabolites contributing to the
separation were identified as amino acids, acidic compounds, lipid metabolism intermediates. Based on these
metabolites, we proposed metabolomic pathway associated with high-fat diet induced obesity.
|
159
P2-19
Glycolipids platform establishment for the identification of MGDG,
DGDG, and SQDG in algae
Jung-Hoon Shin1), Jong Cheol Shon1), So-Hyun Kim2), Hyung-Kyoon Choi2)
1)
and Kwang-Hyeon Liu
1)
College of Pharmacy and Research Institute of Pharmaceutical Sciences, Kyungpook National University, Daegu, Korea
2)
College of Pharmacy, Chung-Ang University, Seoul, Korea
E-mail:dstlkh@gmail.com
Glycolipids are the most abundant thylakoid lipids of plant chloroplast. These Glycolipids are consisted of
monogalactosyldiacylglycerol (MGDG), digalactosyldiacylglycerol (DGDG), and sulfoquinovosyldiacylglycerol
(SQDG). They play an important role in photosynthetic light reactions. MS/MS analysis of glycolipids using
tandem mass spectrometry provided the characteristic fragmentation pattern, which are informative for the
structure identification of glycolipids. In this study, we developed glycolipids platform for the identification
of galactolipids such as MGDG, DGDG, and SQDG. The MS/MS fragmentation pattern of these glycolipids
were characterized and patternized through the interpretation of product ion scan mass spectra of them.
Based on the MS/MS fragmentation pattern of these glycolipids, an in silico glycolipids MS/MS library has
generated.
160
|
Poster 3.
Food and Microbial Metabolomics
|
161
P3-1
Alteration of media composition and light conditions change
morphology, metabolic profile, and beauvericin biosynthesis in
Cordyceps bassiana mycelium
1
1
1
1
1
Hyun, Sun-Hee , Seok-Young Lee , Shin Jung Park , Da Yeon Kim , So-Hyun Kim ,
1
1
1
1
1
1
Seul Gi Lee , Taek-Joo Oh , Xue Jiang , Jun-Sang Oh , Hye Min Ahn , Young-Jin Chun ,
Gi-Ho Sung2, Seong Hwan Kim3, and Hyung-Kyoon Choi1
1
College of Pharmacy, Chung-Ang University, Seoul 156-756, Korea
Mushroom Research Division, Department of Herbal Crop Research, National Institute of Horticultural
and Herbal Science, RDA, Suwon 441-707, Korea
3
Department of Microbiology, Dankook University, Cheonan 330-714, Korea
E-mail: hykychoi@cau.ac.kr
2
Metabolic alterations of Cordyceps bassiana mycelium were investigated under the following culture
medium and light conditions: dextrose agar supplemented with 0.5% yeast extract (SDAY) medium with
light (SL), SDAY medium without light (SD), nut medium without light (ND), and iron-supplemented
SDAY medium without light (FD). The levels of asparagine, aspartic acid, glutamic acid, glutamine,
histidine, lysine, ornithine, and proline were significantly higher under SD and SL conditions. The levels of
most of the alcohols, saturated fatty acids, unsaturated fatty acids, fatty acid esters, sterols, and terpenes
were higher under the ND condition than in the other conditions, but beauvericin was not detectable under
the ND condition. The FD condition was favorable for the enhanced production of aminomalonic acid,
malic acid, mannonic acid, and erythritol. Thus, the metabolic characteristics of C. bassiana can be
manipulated by varying the cultivation conditions, rendering this fungus potentially favorable as a
nutraceutical and medicinal resource.
162
|
P3-2
Culture condition-dependent metabolite profiling of Aspergillus
fumigatus with antifungal activity
Daejung Kang1, Gun Hee Son1, Hye Min Park1, Jiyoung Kim1, Jung Nam Choi1,
1
1
2
1*
Hyang Yeon Kim , Sarah Lee , Seung-Beom Hong, Choong Hwan Lee
1
Departmetnt of Biosciense and Biotechnology, Konkuk University, Seoul 143-701, Korea
2
Korean Agricultural Culture Collection, NAAS, RDA, Suwon 441-707, Korea
E-mail: ramgee@naver.com
Three sections of Aspergillus (five species, 21 strains) were classified according to culture
medium-dependent and time-dependent secondary metabolite profile-based chemotaxonomy. Secondary
metabolites were analysed by liquid chromatography–electrospray ionization tandem mass spectrometry
(LC-ESI-MS-MS) and multivariate statistical methods. From the Aspergillus sections that were cultured on
malt extract agar (MEA) and Czapek yeast extract agar (CYA) for 7, 12, and 16 d, Aspergillus sections
Fumigati (A. fumigatus), Nigri (A. niger), and Flavi (A. flavus, A. oryzae, and A. sojae) clustered separately
on the basis of the results of the secondary metabolite analyses at 16 d regardless of culture medium.
Based on orthogonal projection to latent structures discriminant analysis by partial least squares discriminant
analysis (PLS-DA), we identified the secondary metabolites that helped differentiate sections between A.
fumigatus and Aspergillus section Flavi to be gliotoxinG, fumigatinoxide, fumigatin, pseurotin A or D,
fumiquinazoline D, fumagillin, helvolic acid, 1,2-dihydrohelvolic acid, and 5,8-dihydroxy-9,12-octadecadienoic
acid (5,8-diHODE). Among these compounds, fumagillin, helvolic acid and 1,2-dihydrohelvolic acid of A.
fumigatus showed antifungal activities against Malasseziafurfur, which is lipophilic yeast that causes
epidermal skin disorders.
|
163
P3-3
Liquid chromatography-mass spectrometry-based chemotaxonomic
classification of Aspergillus spp. and evaluation of the biological
activity of its unique metabolite, neosartorin
Mee Youn Lee, Hye Min Park, Gun Hee Son, Choong Hwan Lee
Department of Bioscience and Biotechnology, Konkuk University, Seoul 143-701, Republic of Korea
E-mail: chlee123@konkuk.ac.kr
This work aimed to classify Aspergillus (8 species, 28 strains) by using a secondary metabolite
profile-based chemotaxonomical classification technique. Secondary metabolites were analyzed by liquid
chromatography ion trap mass spectrometry (LC-IT-MS) and multivariate statistical analysis. Most strains
were generally well separated from each section. A. lentulus was discriminated from other 7 species (A.
fumigatus, A. fennelliae, A. niger, A. kawachii, A. flavus, A. oryzae, and A. sojae) with partial least squares
discriminate analysis (PLS-DA) with five discriminate metabolites, including 4,6-dihydroxymellein, fumigatin,
5,8-dihydroxy-9-octadecenoic acid, cyclopiazonic acid, and neosartorin. Among them, neosartorin was
identified as A. lentulus-specific compound that showed anticancer activity, as well as antibacterial effects
on Staphylococcus epidermidis. This study showed that metabolite-based chemotaxonomic classification was
found to be one of the effective tools on a classification method of Aspergillus sp. with species-specific
activity.
164
|
P3-4
Metabolite and antioxidant activity changes of soybean by Aspergiilus
oryzae KACC40247 for the fermentation period
Sunmin Lee, Minho Seo, Deok Kun Oh, Choong Hwan Lee
Departmetnt of Biosciense and Biotechnology, Konkuk University, Seoul 143-701, Korea
Email: chlee123@konkuk.ac.kr
The metabolite profile of soybean fermented with Aspergillus oryzae KACC40247 was analyzed using
mass spectrometry-techniques, including LC-ESI-MS, and GC-TOF-MS. MS-based metabolite profiling of
soybean demonstrated the changes in metabolites according to the fermentation period. On the partial least
square-discriminate analysis, 14 secondary metabolites, 25 primary metabolites were detected based on the
fermentation period. During fermentation, the levels of sugars, several amino acids (aspartic acid,
pyroglutamic acid, gaba) and organic acids were gradually decreased, whereas several amino acids
(threonine, serine, glycine) were increased. Soybean extract containing genistein, daidzein, glycitein was used
as a substrate for hydroxyisoflavone production. The formation of hydroxyisoflavones were increased, other
isoflavones (acetyl glucosides, glucosides, aglycones) were decreased. Correlation analysis between
antioxidant activity and secondary metabolites was also revealed that antioxidant activity showed the
strongest positive correlation with hydroxyisoflavones. These data suggested that metabolomics approaches
on soybean fermentation can demonstrate metabolite changes associated with fermentation period.
|
165
P3-5
Bioethanol production by fermentation of whole pretreated slurry of
oil palm empty fruit bunches
Young Hoon Jung, Kyoung Heon Kim
School of Life Sciences and Biotechnology, Korea University, Seoul 136-713, Korea
E-mail: khekim@korea.ac.kr
Oil palm empty fruit bunches (EFB), one of the most abundant lignocelluloses, were investigated to show
low availability due to the high recalcitrance towards pretreatment and enzymatic saccharification. To
enhance the ethanol and saccharification yields, a whole slurry fermentation of pretreated EFB were
performed after general dilute sulfuric acid pretreatment. The pretreatment conditions applied here were the
incubation in a 1% (w/v) sulfuric acid solution at a high temperature for a short time like 190°C and 3
min ramping to the set temperature. An enzymatic digestibility of 88.5% of theoretical maximum yieldwas
obtained with acid-pretreated and washed EFB. When the whole slurryof acid-pretreated EFB was applied in
simultaneous saccharification and fermentation (SSF) using cellulase and Saccharomyces cerevisiae D5A after
pH adjustment to 4.8, only 52.5% of theoretical ethanol yield based on initial glucan in untreated EFB was
acquired after 72 h of fermentation. However, the ethanol yield from acid-pretreated EFB slurry which was
incubated with activated carbon before subjecting to SSF, was significantly increased to 87.5% after 48 h
of fermentation. When the yeast was cultured in ammonium-neutralized sulfuric acid containing media, a
noticeable inhibition of cell growth was found after 1% (w/v) sulfuric acid addition . These results implies
that the whole slurry SSF after dilute acid pretreatment with activated carbon adsorption warrants high
ethanol yields from lignocellulose which will eventually decrease total production cost.
Reference
1. Young Hoon Jung et al. Dilute acid pretreatment of lignocellulose for whole slurry ethanol
fermentation, Bioresour. Technol., 132:109-114 (2013)
166
|
P3-6
A rare sugar from red seaweeds, 3,6-anhydro-L-galactose, as a novel
health-benefiting functional substance
Eun Ju Yun, Kyoung Heon Kim
School of Life Sciences and Biotechnology, Korea University, Seoul 136-713, Korea
Email: khekim03@gmail.com
3,6-Anhydro-L-galactose (L-AHG), one of the main constituents of agarose with D-galactose, is known as
a rare sugar found in red macroalgae (Rhodophyta). Although agarase system and agarose metabolism were
reported by agar-degrading bacteria, no information is currently available on the mass production, metabolic
fate, or physiological activities of L-AHG. In this study, L-AHG was produced from agarose in the
following the steps: Acid pre-hydrolysis of agarose into agaro-oligosaccharides and enzymatic hydrolysis of
agaro-oligosaccharides into the monosaccharides, L-AHG and D-galactose, by an exo-agarase and a
neoagarobiose hydrolase. After the hydrolysis steps, L-AHG was purified by adsorption and gel permeation
chromatographies. The chemical structure of L-AHG was confirmed by nuclear magnetic resonance (NMR)
spectrometry. In a skin-whitening assay, α-melanocyte-stimulating hormone treated B16F10 melanoma cells
cultured with L-AHG showed significantly higher activity on melanin inhibition compared to the cells
treated with arbutin. L-AHG also showed anti-inflammatory activity, indicating the significant suppression of
lipopolysaccharide-induced nitrite production. Our results suggest that L-AHG can be considered as a
high-value metabolite found in an agarose metabolism with strong health benefits.
References
1. Eun Ju Yun et al. Enzymatic production of 3,6-anhydro-L-galactose from agarose and its purification
and in vitro skin-whitening and anti-inflammatory activities. Appl. Microbiol. Biotechnol.
97(7):2961-2970 (2013)
2. Hanseong Roh et al. Genome sequence of Vibrio sp. strain EJY3: an agarolytic marine bacterium
metabolizing 3,6-anhydro-L-galactose as a sole carbon source. J. Bacteriol. 194(10):2773 (2012)
|
167
P3-7
Biochemical characterization of cultivated Cordyceps bassiana mycelia
1
and fruiting bodies by H nuclear magnetic resonance spectroscopy
Sun-Hee Hyun1, So-Hyun Kim1, Hyo Won Suh1, Seok Young Lee1, Shin Jung Park1,
1
1
1
1
1
1
Da Yeon Kim , Seul Gi Lee , Il-Hwan Oh , Taek-Joo Oh , Xue Jiang , Jun-Sang Oh ,
1
2
3
1
Hye Min Ahn , Gi-Ho Sung , Seong Hwan Kim , Hyung-Kyoon Choi
1
College of Pharmacy, Chung-Ang University, Seoul 156-756, Republic of Korea 2Mushroom Research Division,
National Institute of Horticultural & Herbal Science, Rural Development Administration, Suwon 404-707, Republic of Korea
3
Department of Microbiology, Dankook University, Cheonan 330-714, Republic of Korea
E-mail: hykychoi@cau.ac.kr
In this study, nuclear magnetic resonance (NMR) techniques coupled with multivariate data analysis were
used for the metabolic profiling of mycelia and fruiting bodies of the entomopathogenic fungi, Cordyceps
bassiana according to developmental stages. A direct extraction method using two deutrated solvents of
D2O and CDCl3 was used to investigate the relative levels of identified metabolites in each extraction
condition in the mycelium and fruiting body formation stages. There was a clear separation among mycelia
and fruiting bodies with various developmental stages in partial least-squares discriminant analysis (PLS-DA)
derived score plots. During the transition from mycelia to fruiting bodies, the major metabolic change
observed was the conversion of glucose to mannitol, and beauvericin to phenylalanine and
1-hydroxyisovaleric acid. In the developmental stages of fruiting bodies studied, there was a clear separation
between stage 3 and the other stages in PLS-DA derived score plots. Nineteen compounds including 13
amino acids, 2 nucleosides, 3 organic acids, and glucose showed the highest levels in stage 3 fruiting
bodies. The flavonoid content in the fruiting bodies showed similar levels during stages 1, 2, and 3,
whereas the level at stage 4 was significantly decreased compared to the other stages. Results suggest that
the fruiting body of C. bassiana is richer in natural resources at stage 3 compared to the other fruiting
body stages due to its high abundance of compounds including total flavonoids. The metabolome
information acquired in this study can be useful criteria for the quality control of commercial use of C.
bassiana.
168
|
P3-8
Integrated Metabolomic and Transcriptomic Analyses to Understand
the Mucin Utilization of Vibrio vulnificus
Sun Haeng Jang, Jong Gyu Lim, Kyungku Jang, Sarah Lee, Choong Hwan Lee and Sang Ho Choi*
National Research Laboratory of Molecular Microbiology and Toxicology, Department of Agricultural Biotechnology,
Center for Food Safety and Toxicology, and Research institute for Agriculture and Life Sciences,
Seoul National University, Seoul 151-921, Republic of Korea
Department of Bioscience and Biotechnology, konkuk university, Seoul 143-701, Korea
Email: sardonyx28@hanmail.net
Mucin, a highly glycosylated large glycoprotein, serves simultaneously as a major component of intestinal
mucus layer that is the defensive frontline against enteropathogens and a source of nutrient to support the
survival of enteropathogens. To understand mucin utilization of V. vulnificus, the metabolome and
transcriptome profiles of V. vulnificus grown with mucin and glucose media were compared using a
GC-MS-based metabolomic and a RNA sequencing-based transcriptomic approaches. For metabolome
analysis, V. vulnificus was grown in M9 supplemented with 0.9% porcine stomach mucin or 0.4% glucose.
Bacterial cells grown to early-, mid-, late-log, and stationary phases were harvested and quenched with cold
MeOH. Dried cells were disrupted in a mixer mill and metabolites were extracted with a mixture of
solvents. Dried extracts were derivatized and then analyzed using GC-TOF-MS. For transcriptome analysis,
V. vulnificus was cultured under the same condition to matabolome analysis and RNA was isolated from
only mid-log phase. cDNA was generated from the enriched mRNA and sequenced using the Illumina
Genome Analyzer. Metabolome analysis revealed that the amounts of amino acids, fatty acids and pyruvate
were increased, while glycolysis intermediates such as glucose-6-p and fructose-6-p were decreased in the
cells grown with mucin. Transcriptome analysis also indicated that the expressions of genes encoding
oligopeptide transporters, fatty acid metabolic enzymes, and TCA cycle related enzymes were highly
induced by mucin. These suggest that V. vulnificus preferentially activates the catabolisms of amino acids
and fatty acids to produce TCA cycle intermediates in the presence of mucin. In addition, among the
metabolites from the log phase cells grown with mucin, N-acetylneuraminic acid (Neu5Ac), a terminal
carbohydrate of mucin was identified. Consistent with this, both of the level of glucosamine-6-p, a catabolic
intermediate of Neu5Ac, and the expression of genes encoding Neu5Ac metabolic enzymes were increased.
These indicate that V. vulnificus uptakes and metabolizes Neu5Ac as a nutrient. Interestingly, cells grown
with mucin had increased amounts of putrescine which is required for defense against oxidative stress,
invasion to epithelial cell, and virulence of several enteropathogens. Also, the expressions of genes encoding
putrescine biosynthesis enzymes were highly induced by mucin. The combined results suggest that V.
vulnificus utilizes mucin as a source of nutrient by activating the catabolic pathways of amino acids, fatty
acids, and Neu5Ac, and in the production of virulence related metabolites.
|
169
P3-9
Metabolism of Isoflavone glycosides by Human Intestinal Bacteria
Mihyang Kim, Nayoung Kim and Jaehong Han
Metalloenzyme Research Group and Department of Biotechnology, Chung-Ang University, Anseong, 456-756, Korea
E-mail: coterieus@wm.cau.ac.kr
Pueraria thunbergiana is used as an important medicine and cooking material in Asia. Many reports on
isoflavone glycosides content of P. thunbergiana and their biological activities have been published,
biotransformation of isoflavone glycosides in P. thunberginia by human intestinal microflora have never
been studied. The dried roots of P. thunbergiana were extracted in 80% methanol. Isoflavone glycosides of
the extract were separated by preparative HPLC and identified by LC/MS with standard compounds. The
major isoflavone glycosides in P. thunbergiana was puerarin and the identified isoflavone glycosides were
incubated with human feaces under the anaerobic conditions. The metabolites of the isoflavone glycosides
were isolated and indentified. Puerarin and daidzin were transformed to reduced metabolites with stronger
biological activity, such as daidzein , dihydrodaidzein and O-desmethylagolesin.
Supported by grants
2012R1A2A2A01013356)
170
|
from
National
Research
Foundation
of
Korea
(2012R1A1A-3011925
and
P3-10
Metabolite analysis of medicinal mushroom and their anti-aging
activities in vascular endothelial cell
Hyung-Jun Noh1, Geum-Sook Kim1, Seung-Eun Lee1, Jae-Han Jo1, Dae-Young Lee1,
1
1
2
Je-Hun Choi , Seung-Yu Kim , Jae-Ryong Kim
1
Department of Herbal Crop Research, NIHHS, RDA, Eumsung, Chungbuk, 369-873, Republic of Korea
2
College of medicine, Yeungnam University, Daegu 705-717, Republic of Korea
Email: jumpspace@korea.kr
Ganoderma lucidum has been used for food ingredients since early times. Recently it is being used as a
good pharmaceutical material or functional bio-material. In this meaning, this study was carried out to
investigate metabolites of fruiting body of Ganoderma lucidum and their anti-aging activities. Mycelia of all
strains were firstly inoculated into potato dextrose agar(PDA) and then transfered to a media of saw dust
which contained 20% rice bran. These mycelia of saw dust were then inoculated into oak tree in
polyethylene bags which has been sterilized for 8h at 120℃. Fruiting bodies were harvested after growing
1
three month in growth room. H NMR spectroscopy was used for metabolites of Ganoderma lucidum
extracts by 50% CD3OD/D2O direct extraction method and anti-aging activities were performed by cell
proliferation assay and SA-β-gal assay at human fibroblasts, human vein endothelia cells treated with
adriamycin for inducing aging.
Supported by grants from Rural Development Administration, Korea.
|
171
P3-11
Primary and secondary metabolites variation of soybean elicited
with Aspergillus sojae
K.M. Maria John,a Eun Sung Jung,a Sarah Lee,a Jong-Sang Kim,b Choong Hwan Leea,*
a
Department of Bioscience and Biotechnology, Konkuk University, Seoul 143-701, Republic of Korea
Department of Animal Science and Biotechnology, Kyungpook National University, Daegu 702-701, Republic of Korea
*Corresponding author: Choong Hwan Lee; Tel.: +82-2-2049-6177; Fax: +82-2-455-4291
E-mail: chlee123@konkuk.ac.kr
b
Fungal mediated secondary metabolite changes with particular reference to glyceollins in soybean were
reported earlier. Here, we report time dependent primary and secondary metabolite changes of soybean
elicited with Aspergillus sojae and their associations were discussed. Partial least squares discriminant
analysis showed that the patterns of fungus elicited soybean were clearly distinguished from untreated
samples based on its metabolite content. A. sojae depends on soybean for its carbon source resulting
gradual decrease in the glucose, fructose and myo-inositol levels. The stimulation in L-phenylalanine by A.
sojae increases the accumulation of naringenin from day 1 to 6 leading to the changes in genistein pool.
Even though the level of glucosides like daidzin, genistin and glycitin decreased during treatment other
isoflavones and coumestan levels enhanced. Due to the increase in glycinol, the resulting phytoalexins such
as glyceollin I and II were augmented by fungal treatment. The changes in secondary metabolites reflects
in total phenolic content and because of the increase in glyceollin I, II and glyceofuran reflect their radical
scavenging capacity; A. sojae elicited soybean registered a periodic increase in the radical scavenging
activity..
172
|
Poster 4.
Environmental Metabolomics
|
173
P4-1
Radiation metabolomics: an overview
Changhyun Roh, Jin-Kyu Kim
Division for Biotechnology, Advanced Radiation Technology Institute (ARTI),
Korea Atomic Energy Research Institute (KAERI), 1266 Shinjeong-dong, Jeongeup-si, Jeolabuk-do 580-185, Republic of Korea
E-mail: chroh@kaeri.re.kr
Radiation exposure triggers a complex network of molecular and cellular responses that impacts metabolic
processes and alters the levels of metabolites. Such metabolites have potential as biomarkers for radiation
dosimetry. This review provides an overview of radiation signaling and metabolism, of metabolomic
approaches used in the discovery phase, and of instrumentation with the potential to assess radiation injury
in the field. An understanding of the molecular and cellular effects of ionizing radiation (IR) have
depended profiling technologies such as genomics, transcriptomics, and proteomic platforms. Such efforts
have discovered IR-induced perturbations of DNA, RNA, and protein molecules and have been successful in
developing biomarkers that provide information regarding IR-induced phenomena such as the threshold dose.
Furthermore, integrating data from combinations of such platforms, in the spirit of emerging systems
biology, has given investigators the ability to reconstruct and analyze IR-responsive pathways. However,
pathways generated from such analyses remain incomplete without similar global measurements of
metabolites. Despite recent advances in metabolic profiling developments, changes in small-molecule
metabolites remain under-explored and under-exploited as yet. This is particularly unfortunate because, as
the end-products of transcriptional and proteomic signaling events, metabolites may represent the most
incisive and accurate indicators of the state of cellular physiology, toxicology, and disease progression, and
have led to appreciable advances by defining novel drug and carcinogen metabolites, as well as biomarkers
of disease. Here, we review the current status of radiation metabolomics that have contributed to a general
understanding of how metabolites and their biomarkers change under defined conditions. Our overview
provides a golden opportunity for the understanding of a radiation metabolomics in biodosimetry.
174
|
P4-2
1
H-NMR based profiling of organic components in leachate from
animal carcass disposal site over time.
Hyun-Whee Bae1,2, Geum-Sook Hwang1,2
1
Seoul center, Korea Basic Science Institute, Seoul, Republic of Korea
Graduate School of Analytical Science and Technology, Chungnam University, Daejeon, Republic of Korea
Email: gshwang@kbsi.re.kr
2
Leachate, is liquid generated by decomposition of animal carcass, has many environmental, sanitary and
food safety hazards. However, there is a lack of research on characteristic of leachate. In this study, we
performed 1H-NMR based profiling of cattle leachate from sandy soil and sandy loam soil followed by
multivariate data analysis. Principal component analysis (PCA) from NMR data is showed similar pattern
between samples prepared in two types of soils. As a result, major components including organic acids,
amino acids and phenols, were identified and quantified, demonstrating the microbial decomposition by soil
microbes and colonic bacteria from cattle carcass. This study suggests that NMR-based profiling can be
used to identify components in leachate and elucidate characteristic features of leachate to from different
types of soils over time.
|
175
P4-3
Urinary bisphenol-A concentrations in mother and baby pairs
1,2
2
1
1,*
Minji Bae , Dongho Lee , Bong Chul Chung , Jeongae Lee
1
Biomolecules Recognization Research Center, Korea Institute of Science and Technology, Seoul 136-791,
2
Department of Biotechnology, Korea University, Seoul 136-713
Email: 112018@kist.re.kr
Bisphenol-A(BPA), known as an endocrine disrupting substances in phenols, is used in production of
polycarbonate plastics, epoxy resins and in many plastic consumer products and is similar to effect of
estrogen. Due to the universal use of BPA in a wide range of products and the route of exposure of BPA
is food, water, dust, air and etc, low level human exposure to BPA arises. This study investigated
monitoring of urinary BPA, and it’s correlation with mother and baby pairs (n=551). The method involved
enzymatic hydrolysis, liquid-liquid extraction using methyl-t-butyl ether as an extraction solvent and
derivatization following by silylating agents for sensitive analysis. The selected ion-monitoring mode
response of BPA was linear with the correlation coefficient ranged from 0.9904 to 0.9999. BPA was
detected 91.1% in the mother and 98.4% in baby. The concentrations of BPA in baby were significantly
higher than in mother (p ≤ 2.14×
), but we did not observe correlations between the BPA
=0.0311). BPA concentration was not significantly associated with
concentration of mothers and babies (
aging, but the level of BPA in babies tended to increase up to 24 months and then decreased with aging.
In case of mother, BPA concentrations did not change with aging.
176
|
P4-4
1
H NMR-based Metabolomics of Fish Responses to Bisphenol A
Dahye Yoon, Jie Yu and Suhkmann Kim*
Department of Chemistry, Pusan National University, Busan, 609-735, Korea
Email: yoon_da_hye@pusan.ac.kr
Bisphenol A (BPA) is an important chemical to industry. BPA is used as a raw material for making
plastics and resins.[1] We are frequently exposed to BPA because there are many products made of
plastics. Moreover, wastes are created during the use of BPA to manufacture other products including
releases during handling, unloading, heating, as well as accidental spill.[2] BPA is widespread in our life
and environment, and BPA causes some effects on human and wildlife. BPA is considered as an endocrine
disruptor that modifies hormone functions by binding to the estrogen receptor.[1] However, there are not
enough studies about evident risk of BPA. In many studies, the vitellogenin(VTG) is used as a biomarker
for estrogenicity in oviparous vertebrates because BPA induces synthesis of VTG.[3] This study investigated
1
the effects of BPA on fish about the metabolic viewpoint using H NMR. Zebrafish (Brachydanio rerio)
and Chinese bleak (Aphyocypris chinensis) were used. Zebrafish has been extensively used to the toxicity
test. Chinese bleak is indigenous to Korea.
Recently, metabolomics is a rapidly growing study. Metabolites, as the end products of metabolism,
represent the functional responses of the body.[4] Although low concentration of BPA causes small effect,
metabolic changes can be detected by using NMR spectroscopy. NMR-based metabolomics has some
advantages. It is highly reproducible, it has short time to prepare the sample, and it takes short time to
measure and analyze the sample.[5]
Each fish was lyophilized after exposure to different concentrations of BPA and extracted using methanol,
1
chloroform and water solvent. The extracts of fish were measured using H NMR using 600MHz
equipment. Multivariate statistical analysis was conducted by SIMCA P+. Assignment and quantification of
metabolites were achieved using Chenomx.
References
[1] M. Ike et al. Environmental Toxicology 2002, 17, 457-61
[2] C. A. Staples et al. Chemosphere 1998, 36, 2149–2173
[3] Toxicological Profile for Bisphenol A
[4] C. Y. Lin et al. Metabolomics 2007, 3, 55-67
[5] A. Beltran et al. Analytical Chemistry 2012, 84, 5838-44
|
177
P4-5
Fish responses to carbamate pesticide methomyl exposure
1
by H NMR metabolomics
Siwon Kim, Heonho Lee, Seungkyu Cho and Suhkmann Kim*
Department of Chemistry, Pusan National University, Busan, 609-735, Korea
E-mail: ksw@pusan.ac.kr
Metabolomics, one of the techniques of system biology, is defined as "systematic study of the unique
chemical fingerprints (metabolic biomarkers) that cellular processes leave behind". Metabolites, as the end
products of metabolism, represent the functional responses of an organ. Their characterization can provide
insight into the underlying mechanisms of genomic or environmental actions on metabolism. 1H-NMR
spectroscopy is suitable for the detection of numerous endogenous metabolites in the biological sample
(biological fluids, tissue extracts, intact tissues) since all the metabolite molecules contain proton.[1] There
has been mounting concern regarding the adverse health effects of environmental contaminants in general
and carbamate in particular. Methomyl, a carbamate insecticide, has been widely used to control insect
pests in many countries around the world.[2] Also, methomyl is considered very toxic to mammals and it
is classified by the EPA as a restricted-use pesticide. Methomyl is highly soluble in water and can
therefore, easily cause ground water contamination in agricultural areas.[3] A large number of researches
used Zebrafish because genes of Zebrafish are very similar to human genes. AndChinese bleak,
Aphyocypris Chinensis, is a small cyprinid widely distributed the small streams and ponds in the Korea.[4]
In our studies,the responses of two kinds of fish exposure to methomyl at each of concentrations were
investigated with a 1H-NMR based metabolomics. Principal component analysis (PCA), Partial leasts quares
discriminant analysis (PLS-DA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were
achieved using SIMCA-P+.Assignment of 1H-NMR spectrum and quantification of metabolites were
accomplished by Chenomx.
References
[1] Linbao Zhang., et al, Clean-Soil,Air,Water, 2011, 39, 989-995
[2] Huixian Li., et al, Fish Physiol Biochem, 2008, 34, 2009-216
[3] M.Farre., et al, Anal Bional Chem, 2002, 373, 704-709
[4] Yeom., et al, Korean J.Lim nol., 2006, 39, 419-423
178
|
P4-6
1
H NMR-based Metabolomics approach to finding out toxicity
of the endocrine disruptor Perfluorinated compounds (PFCs)
to the Freshwater fish
Minji Lee, Sangmi Lee and Suhkmann Kim*
Department of Chemistry, Pusan National University, Busan, 609-735, Korea
E-mail :minjilee@pusan.ac.kr
Perfluorinated chemicals are used in many industrial and commercial processes, such as lubricants, fire
retardants, polymer additives, pesticides, and surfactants.[1] Concerns about perfluorinated chemicals,
particularly perfluorooctane sulfonic acid (PFOS), are growing because they can be bioaccumulated through
the food chain. In previous studies, juvenile carp exposed to PFOS, inflammation of liver cells and
disturbance of DNA metabolism homeostasis in liver tissue has been demonstrated. Also, exposure of
rainbow trout, fathead minnow to PFOS has been shown to increase hepatic fatty acyl-CoA oxidase activity
and oxidative damage as well as to alter hormone levels.[2] Nevertheless, it has hardly been addressed that
PFOS induction of biochemical effects in various organism and other interruptions on metabolic pathway.
Metabolomics relates to biological end points and provides a wealth of biological information on complex
systems.[3] Nuclear Magnetic Resonance (NMR) spectroscopy-based metabolomics detects the responses of
metabolites to toxicity has been successfully employed in various species relevant to ecotoxicology.[4]
Zebrafish, Danio rerio, is used an appropriate vertebrate model for investigating various environmental
pollutants.[5] Chinese bleak, Aphyocypris chinensis, well reflect the regional characteristics of Korea
environment because it resides surrounding of Kore. Here, the metabolic responses of two kinds of fish
were affected by PFOS were investigated with a 1H-NMR based metabolomics. Multivariate statistical
analysis results with NMR data, allow the identification of biomarkers and explain specific metabolic
pathways, were accomplished using SIMCA-P+12. Identification of 1H-NMR spectrum peaks and
quantification of metabolites were performed by Chenomx 7.1 software and the online HMDB
(www.hmdb.ca).
References
[1] K. Ji et al. Environmental Toxicology and Chemistry 2008, 27, 2159–2168
[2] Y. Du et al. Chemosphere 2009, 74, 723–729
[3] L. Zhang et al. Clean – Soil, Air, Water 2011, 39, 989–995
[4] M. Wang et al. Environmental Toxicology and Chemistry 2011, 9, 2073–2080
[5] K. Ohara1 et al. Ichthyological Research 2003, 50, 86–89
|
179
P4-7
Identification of the derivated species from Dendropanax morbifera
and lacquer films by pyrolysis-gas chromatography/mass spectrometry
Yun Gyong Ahn and Geum-Sook Hwang
Seoul Center, Korea Basic Science Institute, Seoul 136-701, Korea
Email: ygahn@kbsi.re.kr
Identification of lacquer film species from ancient coating materials is needed to maintaine their surfaces
without loss of their original beauty for a long time and understand the historical background of
manufacturing techniques. A pyrolysis-gas chromatography/mass spectrometry(py-GC/MS) was applied to
identify the origine of films in ancient coating materials. The pyrolysis products, which reflect the source
from which they originate were detected distinctively at 500˚C. This is a rapid technique that does not
require large amounts of sample or any sample preparation. Sesquiterpenes are a class of terpenes that
consist of three isoprene units were identified as cadienes, selinenes, cubebenes from the raw material of
dendropanax Dendropanax morbifera. On the other hand, Alkanes(tetra~heptadecanes), alkenes
(tri~heptadecenes), allkyphenols, catechols and fatty acids were detected from the raw material of the
lacquer film. Based on these results, the origine of historic coatings artifacts was identified using
py-GC/MS by comparison with their pyrolysis products.
Supported by the Creative Allied Project grant from KRCF
180
|
P4-8
Health impact assessment of urinary di(2-ethylhexyl) phthalate
metabolites in mother-child pairs.
Ji-won On
1
1,2
, Na Rae Song1,3, Jeongae Lee1, Sang-won Lee2, Heesoo Pyo1
Molecular Recognition Research Center, Korea Institute of Science and Technology, Seoul, 136-791, Korea
2
Department of Chemistry, Korea University
3
Department of Chemistry and Biochemistry, University of California, Los Angeles
Email: onjwon@naver.com
Di(2-ethylhexyl) phthalate (DEHP) is one of the common phthalate plasticizers used primarily in soft
polyvinyl chloride including personal-care products, industrial plastics, and medical devices. The DEHP is
rapidly metabolized to its monoester, mono(2-ethylhexyl)phthalate(MEHP), which is further metabolized by
various
hydroxylation
and
oxidation
to
the
secondary
metabolites
such
as
mono(2-ethyl-5-hydroxyhexyl)phthalate (5OH-MEHP) and mono(2-ethyl-5-oxo-hexyl)phthalate (5oxo-MEHP).
Recently, it is known that DEHP causes adverse effects on endocrine functions and reproductive organs of
humans. The objectives of this study were to examine the excretion of urinary DEHP metabolites and to
assess human health risk from DEHP exposure. The total of 954 urine samples including 258 mother and
child pairs were analyzed using isotope dilution gas chromatography-mass spectrometry. The median levels
of DEHP metabolite concentrations increased in the order of male adults (51.8 µg/L, 42.2 µg/g creatinine),
female adults (55.7 µg/L, 61.6 µg/g creatinine), mothers (67.4 µg/L, 67.3 µg/g creatinine) and children
(185.2 µg/L, 258.5 µg/g creatinine), respectively. The result shows no significant correlation in exposure to
DEHP between the mother and child pairs except for MEHP (p-value ≤ 0.01). Relative metabolic rate
(RMR) were calculated to examine the difference in DEHP exposure levels to the primary and secondary
st
metabolites. MEHP hydroxylation to 5-OH-MEHP (1 step, RMR1) followed by 5-OH-MEHP oxidation to
nd
5-oxo-MEHP (2 step, RMR2). The RMR1 and RMR2 of children, mothers and adults were very significant
to each other (both p-values ≤ 0.0005) and children had the highest value of RMR1. Each group had
similar value of RMR2 and the values showed a significant difference in male and female adults (p-value
≤ 0.05). Children group had particularly faster relative metabolic rate in the MEHP hydroxylation to
5-OH-MEHP than mother. Combined with sample volunteer’s health information, the above study would be
a useful source that can be referred to make an assessment of human metabolic system.
|
181
182
|
| 발행일
2013년 4월
| 발행처
(사)한국대사체학회
143-701 서울특별시 광진구 능동로 120
건국대학교 생명과학관부속동(학군단) 210
Tel：02-444-4290,
Fax：02-455-4291
E-mail：metabol@komets.or.kr www.komets.or.kr
| 디자인·인쇄 동양기획
Tel：02-2272-6826,
E-mail：dy98@unitel.co.kr
Fax：02-2273-2790