5^th International Conference and Exhibition on Metabolomics May 16-18, 2016 Osaka, Japan

Yoriko Heianza has completed her PhD from University of Tsukuba and has been conducting Post-doctoral studies in Tulane University School of Public Health and Tropical Medicine. She has published more than 30 papers in peer-reviewed medical journals.

Taurine metabolism disturbance is closely linked to obesity, insulin resistance and diabetes. Previous evidence suggested the preventative effects of taurine on diabetes might be through regulating the expression levels of diabetes related genes. In current study, we aimed to estimate whether blood taurine levels modify the overall genetic susceptibility to diabetes on improvement of insulin sensitivity in a randomized dietary trial. We analyzed data from 711 overweight or obese participants (80% Caucasians) who had genetic variants as well as blood taurine levels measured, from the Preventing Overweight Using Novel Dietary Strategies (POUNDS Lost) trial, a 2-year weight-loss dietary intervention trial. We calculated a genetic risk score (GRS) based on 31 diabetes-associated variants, and estimated the long-term (2 years) improvements of glycemic control traits. We found that baseline plasma taurine levels significantly modified the effects of diabetes GRS on changes in fasting glucose, insulin, and homeostatic model assessment of insulin resistance (HOMA-IR) over 2-year diet interventions (P for interaction = 0.04, 0.01, 0.002, respectively), regardless of weight loss. High baseline taurine levels were associated with a less reduction in both fasting glucose and HOMA-IR among the participants with the lowest tertile of diabetes GRS (both P=0.02), and with a greater reduction in both insulin and HOMA-IR among those with the highest tertile of diabetes GRS (both P=0.04). Our data suggest that blood taurine levels might differentially modulate the genetic effects of diabetes on improvement of insulin sensitivity among overweight/obese patients on weight-loss diets.

Chihiro Mitsui was graduated from Hokkaido University in 2008. After completing initial Clinical Residency Program, she has working as a Research Fellow at Sagamihara National Hospital Clinical Research Center. She has published more than 10 papers in reputed journals.

Background: Aspirin exacerbated respiratory disease (AERD) is characterized by respiratory reactions upon ingestion of cyclooxygenase (COX)-1 inhibitors and cysteinyl leukotriene (cysLT) overproduction. This hypersensitivity reaction is induced by aspirin at low doses that irreversibly inhibit COX-1 in platelets but not COX-2 in endothelial cells and leukocytes. Method: First, platelet activation markers [the expression levels of P-selectin (CD62P), CD63, CD69, and GPIIb/IIIa (PAC-1) in peripheral platelets, the percentage of circulating platelet adherent leukocytes and the levels of plasma soluble P-selectin (sP-selectin) and soluble CD40 ligand (sCD40L)] were examined in stable patients with AERD (n=30), aspirin tolerant asthma (ATA; n=21) and idiopathic chronic eosinophilic pneumonia (CEP; n=10). Furthermore, the levels of plasma sP-selectin and sCD40L in AERD (n=24) and ATA patients (n=7) and surface markers on platelets in AERD patients (n=8) were also assessed during the aspirin challenge test. Results: In stable condition, the expression levels of all surface markers on platelets (P-selectin, P=0.022; CD63, P=0.001; CD69, P=0.029; and PAC-1, P=0.014), the percentage of platelet adherent eosinophils (P=0.025) and the levels of plasma sP-selectin (P=0.017) and sCD40L (P=0.013) were significantly higher in AERD patients than in ATA patients. The expression levels of CD63 and CD69 on platelets and the plasma sCD40L level in AERD patients were higher than those in CEP patients (P<0.001, P=0.008, and P=0.028, respectively). In contrast, there were no significant differences in the expression levels of these markers among ATA patients, CEP patients and controls. In the aspirin challenge test, the levels of platelet activation markers did not change both in AERD and ATA patients. P-selectin and CD63 expressions on platelets and plasma sP-selectin and sCD40L levels positively correlated with the percentage of platelet adherent eosinophils. Among these markers, p-selectin expression and plasma sP-selectin levels positively correlated with urinary leukotriene E4 concentration. Additionally, plasma sP-selectin and sCD40L levels negatively correlated with lung function. Conclusions: Peripheral platelets were activated to greater extent in stable AERD patients than in stable ATA patients, CEP patients and controls. Platelet activation was involved in cysLT overproductions and persistent airflow limitations in AERD in stable disease condition.

Wan-Rou Shih is a Master’s degree student at the School of Institute of Pharmaceutical Sciences and Technology, Central Taiwan University of Science and Technology, Taichung, Taiwan.

Fenofibrate is a lipid-lowering agent that has been shown to suppress tumor progression in many cancers. In the present study, we demonstrated that treatment of human U87 glioma cells with fenofibrate decreased the protein levels of HIF-1under normoxia and hypoxia conditions. The suppression of HIF-1 was reversed by pretreatment with proteosome inhibitor. The reduction of HIF-1level was associated with decreased expression levels of glucose transporter Glut-1, and lactate dehydrogenase A. Treatment with fenofibrate also decreased the expression of pyruvate dehydrogenase kinase-1 and pyruvate dehydrogenase phosphorylation which subsequently reactivated mitochondrial oxidative phosphorylation and increased mitochondrial ATP production in U87 cells. Treatment of cells with fenofibrate decreased cell viability, which can be prevented by pretreatment with l-N-aceytylcystein, an anti-oxidant. These data suggest that reactivation of mitochondrial energy metabolism by ferrous glycinate may increase ROS accumulation, and the excessive ROS cause mitochondrial damage and subsequently results in cell death.

Minjoo Kim has completed her PhD from Yonsei University. She is currently a Post-doctoral student in Research Center for Silver Science, Yonsei University. Her researches are focussed on medical nutrition therapy in metabolic diseases, aging, nutrition-related metabolomics, etc. She has published more than 24 papers in reputed journals and has registered 2 patents (also, 9 patent applications)

The aim of this study was to identify age-related metabolite changes associated with elevated serum alanine aminotransferase (ALT) and to explore possible underlying mechanisms. The study included 602 healthy, nondiabetic subjects (aged 30−65 years); 393 individuals had normal ALT levels at baseline. Fifty individuals developed elevated ALT levels after 3 years. The remaining 340 subjects with normal ALT were matched to the elevated-ALT group (n=50) for age, gender, BMI, fasting glucose, and ALT to form the control group (n=50). At the 3-year follow-up, the elevated-ALT group exhibited greater increases in waist circumference, free fatty acid, ALT, aspartate aminotransferase (AST), γ-glutamyltransferase (GGT), bilirubin, oxidized LDL, Lp-PLA2 activity, urinary 8-epiprostaglandin F2α (8-epi-PGF2α), and brachial-ankle pulse-wave velocity (ba-PWV) compared to the control group after adjusting for baseline. The elevated-ALT group exhibited greater increases in plasma L-valine (q=0.014), L-leucine (q=0.007), L-phenylalanine (q=0.006), and decanoylcarnitine (q<0.001). Mean ALT levels increased in a linear fashion with increasing changes in these four metabolites, which correlated with changes in AST, GGT, Lp-PLA2 activity, urinary 8-epi-PGF2α, and ba-PWV. Mean ALT changes did not significantly correlate with HOMA-insulin resistance. These findings suggest that increased plasma levels of L-valine, L-leucine, Lphenylalanine, and decanoylcarnitine precede insulin resistance during periods of elevated ALT and occur in conjunction with enhanced risk factors for cardiovascular disease, such as Lp-PLA2 activity, oxidative stress, and arterial stiffness.

Miso Kang is a PhD student working at Yonsei University Clinical Nutrigenetics/Nutrigenomics Laboratory with Jong Ho Lee. She consulted on a variety of projects, involving qualitative and quantitative analysis to achieve acquisitions, restructurings, and starategic realignments with her team. She has published 1 paper in reputed journal.

We investigated that increase in circulating levels of low-density lipoprotein cholesterol (LDL-cholesterol) by aging may cause alterations in plasma metabolites. Among 596 healthy, nondiabetic subjects (aged 30–65 years), 390 individuals had normal levels of LDL-cholesterol at baseline, 53 subjects were in high levels of fasting LDL-cholesterol after 3 years. The 337 individuals, who retained normal LDL-cholesterol levels after 3-year follow-up, were matched for age, gender, BMI, and fasting LDL-cholesterol, and the matched group was included as the control group (n = 75). At the 3-year follow-up, total-cholesterol, LDL-cholesterol, oxidized LDL (ox-LDL), lipoprotein-associated phospholipase A2 (Lp-PLA2) activity, and urinary 8-epi-prostaglandin F2a were increased in the high-LDL group than those of the control group. The high-LDL group also showed significant decreases of sphingomyelin (SM) (d18:0/16:1) and phosphatidylcholine (PC) (18:0/20:4), associated with an increases of LDL-cholesterol, and significant increases of palmitic amide and lactosylceramide. Mean changes of SM (d18:0/16:1), C17 sphinganine, PC (18:0/20:4), and lysoPCs (C16:1, C16:0, C17:0, C18:1, C18:0, C20:4, C20:3, and C22:6) were statistically different between the control and the high-LDL groups. Overall, the changes of ox-LDL positively correlated with changes of LDL-cholesterol, Lp-PLA2 activity, palmitic amide, oleamide, lysoPCs, and C17 sphinganine, and negatively correlated with changes of SM (d18:0/16:1). Increase in LDL-cholesterol induced by aging is associated with increased oxidative stress and disturbed sphingolipid metabolism.

Yan-xin Zhang is pursuing PhD at the School of Chemical Engineering & Technology, Harbin Institute of Technology, China. She has been engaged in the study of metabolomics in Chinese Academy of Agricultural Sciences for two years. The research is to evaluate the hazards of pesticides and pesticide additives on the body by using metabolomics, supported by the special program for basic work of the Ministry of Science and Technology of China. In order to fully complete the study, she has participated in the training of high resolution mass spectrometry by AB SCIEX and multivariate analysis (SIMCA software) by UMETRICS.

Metabolomics was conducted in urine using HPLC-quadrupole time-of-flight mass spectrometry (QTOF-MS). Pesticide-related metabolic biomarkers were investigated by comparing 76 farmers in the agricultural area with 30 office workers as long-term exposure to low level pesticides research. Comparison of urine metabolites of 76 farmers before and after spraying with pesticides as acute exposure research was carried out. A series of metabolites were analyzed using multivariate analysis. To further find discrepant metabolites reflecting metabolic characteristics between controls and exposure groups both in long-term and acute exposure study, principle component analysis (PCA) and partial least-squares discrimination analysis (PLS-DA) were implemented. High resolution MS/MS analyses, METLIN and MASSBANK database system were performed for the identification of the metabolites of interest. Finally, the most statistically significant changes in long-term exposure were observed for kynurenine and glycine significantly increased while dopamine, 5-hydroxytryptamine, tryptophan, hippuric acid, taurine, and citrate decreased among the farmers. 5 metabolites were identified as potential urinary biomarkers of acute pesticide-induced toxicity including dopamine, serotonin, tyrosine, tryptophan, taurine and hippuric acid (downward trend), and canine urea and creatine (upward trend). To discuss the mechanism of the different ways of pesticide exposure, oxidative stress (as determined by urinary 8-oxo-deoxyguanosine levels) and inflammatory markers were elevated in individuals with long-term pesticide exposure, supporting the hypothesis that xenoestrogen accumulation was causing mitochondrial dysfunction. These changes of metabolites in acute exposure may relate with the nervous system and liver function damage. This study demonstrated that metabolomics could be used to investigate pesticide-related biomarkers causing metabolic changes.

Yu-Syuan Lin is a graduate student at the School of Department of Medical Laboratory Sciences and Biotechnology, College of Medical Science and Technology, Taipei Medical University, Taipei, Taiwan.

Hypoxia-inducible factor (HIF)-1has been shown to play important roles in regulating cancer metastasis, invasion and drug resistant. In present study, we showed that treatment of human U87 glioma cells with ferrous glycinate reduced HIF-1 accumulation, which is dependent upon prolyl hydroxylase activity. Suppression of HIF-1by ferrous glycinate reduced MMP-2 and MMP-9 secretion in the media. Ferrous glycinate decreased cell invasiveness, which was associated with decreased expression of transcription factors Twist and Nestin, and the mesenchymal marker, Vimentin. Consistently, ferrous glycinate increased expression of differentiation marker, the glial acidic fibrillary protein. Taken together, these suggest that ferrous glycinate may reverse the epithelial-mesenchymal transition (EMT). In addition, treatment with ferrous glycinate reversed drug resistance to 1,3-bis(2-chloroethyl)-1-nitrosourea, a second line chemotherapeutic agent for malignant gliomas. Given ferrous glycinate may reverse the EMT and drug resistance in malignant gliomas cells, these data suggest that ferrous glycinate may be used in the treatment of malignant gliomas.

Minkyung Kim is a Doctoral student working at Nutrigenetics/Nutrigenomics lab in the Department of Food and Nutrition at Yonsei University with Professor Jong Ho Lee. She has published 4 papers in reputed journals and presented 5 papers at international conferences.

Prehypertension can develop hypertension and atherosclerosis which represents early cardiovascular disease. Therefore, it is essential to understand the role of prehypertension in metabolic alterations. We compared plasma metabolites in subjects who maintained prehypertension (n=51) and age- and gender-matched controls who maintained normal blood pressure (n=51) during 3-year follow-up period. The prehypertension group indicated significantly higher intensity in lysophosphatidylcholines (lysoPCs; C14:0, C16:1, C16:0, C18:2, C18:1, C18:0, C20:5, C20:4, C20:3, and C22:6), and higher levels in Lp-PLA2 activity, oxidized LDL, interleukin-6, 8-epi-PGF2α, and brachial-ankle pulse wave velocity (ba-PWV) than control group. Statistical significance were remained after adjusting BMI, waist hip ratio, smoking and drinking, lipid profiles, fasting blood glucose, and insulin. The most important plasma metabolites to discriminate two groups was lysoPC (16:0), which showed positive and independent correlations with blood pressure. In contrast to control group, lysoPC (16:0) showed positive correlations with oxidized LDL, Lp-PLA2 activity, interleukin-6, 8-epi-PGF2α, and ba-PWV after adjusting for confounding factors in the prehypertension group. These results suggest that maintaining prehypertension status could induce alteration in plasma metabolites and these are associated with oxidative stress, inflammation, and Lp-PLA2 activity.

Byung Hwa Jung has completed her PhD from Seoul National University in 2000 and Post-doctoral studies from University of North Carolina at Chapel Hill, School of Pharmacy in 2004. She is a Principal Researcher and Center Head of Molecular Recognition Research Center at Korea Institute of Science and Technology (KIST). She is also an Adjunct Professor of University of Science and Technology. She has published more than 60 papers in reputed journals and serving as an Editorial Board Member of repute.

A quantitative analytical method was developed for the simultaneous detection of tryptophan and its metabolites (indole-3-lactic acid, anthranilic acid, serotonin, nicotinic acid, kynurenic acid, kynurenine and 3-indoxyl sulfate) in both human serum and gastric juice using liquid chromatography-tandem mass spectrometry. Serum and gastric juice samples were prepared with a simple protein precipitation procedure after optimizing solvents and pH conditions used. The stability conditions were also optimized and finally achieved a >70% recovery for all of the analytes. To simultaneously detect tryptophan and its metabolites using mass spectrometry, polarity switching was performed to alternate positive and negative ionization, which enabled both tryptophan and its metabolites to be detected in a single run. Acceptable levels of accuracy (within 20% in LOQ and 15% in other QCs) and precision (within 20% in LOQ and 15% in other QCs) were obtained in the intra- and inter-day validations of serum and gastric juice. The method successfully applied to identify the changes in tryptophan metabolism in patients with gastritis (control) and those with gastric cancer. By comparing the level differences in the serum and gastric juice, it was found that the kynurenine pathway of tryptophan metabolism was activated in gastric cancer and the metabolic ratio of kynurenine/tryptophan, which reflects the enzyme activity of indoleamine-2,3-dioxygenase, was associated with the observed metabolic changes. Finally, the analysis of tryptophan metabolites levels, especially kynurenic acid, in serum and gastric juice might serve as biomarkers for the diagnosis of gastric cancer.

Pan Zou is a PhD student at the School of Chemical Engineering & Technology, Harbin Institute of Technology, China. She joined the group of Pr. Wang Jing at Chinese Academy of Agricultural Sciences, and then worked at Academy of Military Medical Sciences as visit student. She has participated in several projects supported by National Nature Science Fund, and published 3 papers in the last two years. Her research focuses on the hazards of pesticides and pesticide additives.

The metabonomic strategy has been used based on liquid chromatography/time-of-flight mass spectrometry (LC/TOF-MS). As mobile phase, Acetonitrile or methanol usually has been used alone. Here, three types of organic phases including acetonitrile, methanol and the acetonitrile/methanol cocktail under a gradient elution system have been particularly compared for metabonomic analysis. The results showed that the number of peaks in the cocktail was significantly greater than acetonitrile and methanol in positive mode, and no significant difference was found in negative mode. The detected ion were much more under the cocktail as organic phases in both positive and negative modes. The total number of ion eluted were 1585 in positive ion mode and 955 in negative ion mode, of which 91% and 99% by the cocktail. Mixture of organic phases increased response and resolution of ions than the others. Moreover, cocktail further was observed for detection of complementary marker ions. Some typical compounds that are common in urine such as Thornasterol B, Hydroeortisone, Glycocholic acid were unique existence in cocktail. Cocktail organic phases has advantage in detecting more abundant metabolite. Finally rat urine from control group and dosage group of Nonylphenol (NP) was reanalyzed using both the acetonitrile, methanol and cocktail as organic mobile phase in HPLC-QTOF-MS systems in positive ion mode.

Jhong-Huei Jheng is a graduate student at the School of Medical Laboratory Sciences and Biotechnology, College of Medical Science and Technology, Taipei Medical University, Taipei, Taiwan

HIF-1is a transcription factor and play a crucial role in cancer metabolism, which are characterized with aerobic glycolysis and suppressing of mitochondrial energy metabolism (Warburg effect). In the present study, we demonstrated that incubation of A549 cells with ferrous glycinate decreased the protein levels of HIF-1under either nomoxia or hypoxia conditions, which was further reversed by pretreatment with proteosome inhibitor, or prolyl hydroxylase inhibitor. The reduction of HIF-1level was associated with decreased expression levels of glucose transporter, Glut-1, and glycolytic enzymes including hexokinase-2, and lactate dehydrogenase A. On the other hand, treatment with ferrous glycinate reactivated oxidative phosphorylation by decreasing the expression of pyruvate dehydrogenase kinase-1 and pyruvate dehydrogenase phosphorylation that subsequently increased mitochondrial membrane potential and ATP production, suggesting ferrous glycinate regulate energy metabolism in lung adenocarcinoma cells. These results indicate that ferrous glycinate may serve as a therapeutic adjuvant for related human lung cancers.

Song Vogue Ahn has completed his MD and PhD from Yonsei University. He is working as an Associate Professor at the Department of Preventive Medicine, Yonsei University Wonju College of Medicine and Director of the Institute of Genomic Cohort, Yonsei University. He has published more than 35 papers in reputed journals and has been serving as an Editorial Board Member of the repute.

It has been reported that stable isotopic ratios can be used as biomarkers for animal protein intake. A high protein or meat intake could be a risk factor for metabolic disorders. We investigated whether the carbon or nitrogen stable isotopic ratio of human hair is associated with cardiovascular risk factors. We conducted a cross-sectional study within the Korean Genomic Rural Cohort Study in 2011. Hair samples from 399 subjects (233 men and 166 women) aged 40 and over were measured for carbon [δ13C={(13C/12C)sample/(13C/12C)Pee dee Belemnite}-1] and nitrogen [δ15N={(15N/14N)sample/(15N/14N)Air}-1] stable isotopic ratios. Cardiovascular risk factors, including anthropometrics, blood pressure, serum adiponectin, leptin, gamma-glutamyltransferase were measured. Men showed a negative correlation between δ15N and serum adiponectin, a positive correlation between δ15N and serum leptin. Women showed a positive correlation between δ15N and serum leptin. On multivariable models, δ15N was negatively associated with serum adiponectin in men and positiveely associated with serum leptin in women. The δ15N value of human hair might be a surrogate marker associated with cardiovascular risk factors.

Seung Han Baek has completed his PhD in 2014 at Cancer Biology Lab, Dept. of Biology, College of Life Science and Biotechnology, Yonsei University which is in connection with Yonsei University Research Institute of Science for Aging. At present he is pursuing his Post-doctoral research at Dr. Min Songs lab Department of Library & Information Science, Yonsei University and is working on bio-text and bio-medical text mining.

The objective of this study is to check metabolic profiles of metabolically healthy and unhealthy overweight individuals in order to verify metabolic biomarkers or pathways regulating the different metabolic characteristics of overweight. Metabolic characteristics were obtained from 109 overweight (25kg/m2 ≤ BMI < 30kg/m2) subjects aged 30-65 year. Among them, 34 individuals, whose veceral fat adiposity at L4 is over 100 cm2, were included as metabolically unhealthy overweight group. Metabolically healthy overweight group (n=34), matched for age, gender, and BMI, set as a control group. Despite similar body fat % and total fat mass, measured by using DEXA, metabolically unhealthy overweight group showed higher visceral fat area at L1 and L4 than metabolically healthy overweight group. Metabolically unhealthy overweight subjects had higher blood pressure, lipid profiles, hs-CRP, malondialdehyde, and oxidized LDL, HOMI-IR, and had lower HDL-cholesterol and adiponectin. In metabolic profile analysis, methoxybenzepropanoic acid, α-linolenic acid, docosahexaenoic acid, dodecenoylcarnintine (C12:1), dodecanoylcarnintine (C12), tetradecenoylcarnitine (C14:1), hexadecenoylcarnitine (C16:1), palmitoylcarnitine (C16) and urobillinogen were higher in metabolically unhealthy overweight group than those of metabolically healthy overweight group. In addition, the two groups were distinguished by glycerophospholipid pathway, which showed higher levels of lysophosphatidylethanolamine (lysoPE) (22:6), lysophosphatidylcholine (lysoPC) (22:6) and lysoPC (22:5) in metabolically unhealthy overweight group. In conclusion, inidviduals who have same age, gender, BMI, body fat %, and total fat mass are able to be classified according to metabolic status including long-chain fatty acids, medium chain (C12:1, C12) acylcarnitine (AC), and long chain (C14;1, C16:1, C16) AC levels and glycerophospholipid pathway.

Tamara Al Abdi has completed her postgraduate studies in 2006 from Leeds Metropolitan University in the U.K and is a qualified state registered dietitian with the Academy of nutrition and dietetics and the British Dietetic Assoication. She has been the lecturer and clinical coordinator of the supervised practice program at the human nutrition department in college of health sciences at Qatar university since 2010. Her research interest is in clinical dietetics and practice in the Middle East as well as promoting the role of dietitians in Qatar.

Little is known regarding attitudes of healthcare professional students towards team-based care in the Middle East. As modernization of health systems is rapidly occurring across the Gulf Cooperation Council countries, it is important for students to engage in interprofessional education (IPE) activities. The objective of this study was to assess pre-clinical student’s attitudes towards interprofessional healthcare teams after completion of their first IPE activity. A previously validated questionnaire was distributed to 25 pharmacy and 17 nutrition students at Qatar University after participation in an IPE event. Questions related to quality of team based care and physician centricity. Results showed high agreement regarding high quality care provided by teams yet students were unsure of the value of team-based care when considering required time for implementation. Results provide baseline data for future studies to assess student attitudes throughout the professional programs and give valuable insight for future IPE program design in the Middle East.

Wu Ching-Shuang has achieved his PhD degree in 2004 from Kaohsiung Medical University. He has been promoted as Professor in 2013. He is now the Chairperson of Department of Medical Laboratory Science and Biotechnology in Kaohsiung Medical University

Our objective was to investigate and compare the effects of heat-killed (HK) and live Lr263 on insulin resistance and its related complications in high-fat diet (HFD)-induced rats. Male Sprague-Dawley rats were fed with a HFD with either HK or live Lr263 for 12 weeks. The increase in weight gain, serum glucose, insulin, and lipid profile in serum and liver observed in HFD group were significantly reduced after HK or live Lr263 administration. Feeding of HK or live Lr263 reversed the decreased number of probiotic bacteria and increased number of pathogenic bacteria induced by high-fat treatment. The decreased intestinal barrier in HFD group was markedly reversed by HK or live Lr263 treatments. The elevations of pro-inflammatory associated genes expressions in both adipose and hepatic tissues by high-fat administration were markedly decreased by HK or live Lr263 treatments. The increased macrophage infiltration noticed in adipose tissue after high-fat treatment was effectively suppressed by HK or live Lr263 consumption. The insulin resistance associated gene expressions in both adipose and hepatic tissues which were downregulated in HFD group were markedly enhanced after HK or live Lr263 feeding. HK or live Lr263 consumption significantly decreased hepatic lipogenic gene expressions stimulated by high-fat treatment. Administration of HK or live Lr263 significantly reduced hepatic oil red O staining and ameliorated hepatic steatosis observed in high-fat treated rats. Our data suggested that similar to live Lr263, HK Lr263 exerted significant effectiveness on attenuating obesity-induced metabolic abnormalities by reducing insulin resistance and hepatic steatosis formation.

Gwang-Woo Jeong received his Ph. D. in analytical chemistry from Syracuse University. After that he worked at the Korea Basic Science Institute as a senior research staff. He is currently Professor of Radiology at Chonnam National University Hospital and Chonnam National University Medical School. His research interests include magnetic resonance(MR) spectroscopy, morphometric and functional MR imaging techniques in connection with neurological diseases and mental disorders, as well as development of new techniques in MR-related fields and their applications to in vivo and in vitro MR studies of biological systems

In patient with generalized anxiety disorder (GAD), the dorsolateral prefrontal cortex (DLPFC) is related to cognitive function and emotional regulation in GAD. However, it is unclear how brain metabolite in the DLPFC is changed in conjunction with the major symptoms of GAD. This study assessed the metabolic change in the DLPFC and clarify the correlation between brain metabolite changes in the DLPFC and anxiety levels in GAD using 1H-MR spectroscopy.Thirty subjects comprised 15 patients with GAD (mean age, 37.1±11.0 years)and 15 age-matched healthy controls (mean age, 36.5±7.7 years).The patients with GAD had the average duration of illness for 5.2±6.9 years with anxiety severity (Generalized Anxiety Disorder Scale 7was 11.7±4.2). The 1H-MR spectra were acquired by using PRESS from a localized voxel in the right DLPFC. The differential metabolic concentration between two groups was analyzed by the Mann-Whitney U test.The concentration ratio of Cho/Cr was significantly lower in patients with GAD than in the healthy controls: 0.73±0.08 vs. 0.80±0.09(p=0.034). Also, the ratio of Cho/NAA was significantly lower in patients than in controls: 0.44±0.07 vs. 0.47±0.07 (p=0.024). It is important to note that the Cho concentrations were negatively correlated with anxiety level in patients with GAD.However, there was no correlation in healthy controls.It is concluded that 1H-MRS provides valuable information on the metabolites change in the DLPFC and its correlation with anxiety level in patients with GAD.

Hye Jin Yoo is a graduate school student in PhD course at Yonsei University. She is woking in Nutrigenetics/Nutrigenomics laboratory, in Department of Food and Nutriton, lead by Prof. Jong Ho Lee. Her research focus is clinical nutrition. She has published 1 paper in reputed journal.

We investigated whether metabolic profiles of PBMC and plasma were affected by decrease in PBMC and plasma lipoprotein-associated phospholipase A2 (Lp-PLA2) activities induced by dietary intervention. 70 nonobese subjects aged 40–70 years (18.5 ≤ BMI < 30 kg/m2) with prediabetes or newly-diagnosed type-2 diabetes were randomly divided according to diet that they consumed; control group (n=35) have a usual refined-rice diet, whole-grain group (n=35) have a diet replaced refined rice with whole grains and legumes. All subject consumed the diet for 3 meals per day during 12 weeks. After 12 weeks invervention, changes in fasting glucose, HbA1c, HOMA-IR index, malondialdehyde, oxidized LDL (ox-LDL), LDL particle size, and plasma and PBMC Lp-PLA2 activity in the whole-grain group were significantly different from those of the control group. The PBMC levels of L-leucine, oleamide, lysoPC (16:0), and lysoPC (18:0) in the whole-grain group showed greater decrease compared to those of the control group. Changes in plasma metabolites were not significantly different between the two groups. Changes in PBMC Lp-PLA2 activity positively correlated with changes in glucose, ox-LDL, and metabolites including L-leucine, oleamide, lysoPC (16:0), lysoPC (18:0). Moreover, Changes in PBMC Lp-PLA2 activity negatively correlated with changes in LDL particle size. This study indicated that dietary intervention affects on PBMC Lp-PLA2 activity and metabolites as compared to those of plasma metabolites in subjects with prediabetic or type-2 diabetes.

I-Chuan Sheih has completed her PhD in 2009 and Post-doctoral studies for two and a half years in National Chung Hsing University. She is a Project Director of Ministry of Science and Technology, MOST, Project No MOST 104-2221-E-233-005 and the Chair of Department of Food and Beverage Management. She has published many SCI papers in reputed journals. She has been a researcher for six years in Food Industry Research Development Institute and a RD for four years in the Resource Microbiology Institute.

Many peptides, in addition to nutritional supplements, were found to have various bioactivities, such as anti-oxidation, anti-cancer activity, etc. In our previous study, it was attained a multifunctional hendeca-peptide with a molecular mass of 1309 Da from algae protein hydrolysate. In order to enhance the physiological effectiveness of the peptide used as medicine and health food in the future, we modified the original skeleton of algae peptides for various bioactivities detection, and utilized the technology of amino acids synthesis and bioinformatics knowledge to explore the relationship of amino acids diversity. The results showed the synthetic tripeptide (PRF) was much more protective capacity than that of full length peptide in quenching hydroxyl radicals. Besides, we further detected the antioxidative study on oxidation-induced DNA damage to elucidate the prospective antioxidative role of peptide in humans. The result showed that the super coil (SC) form in DNA was completely converted to the open circular (OC) form due to the hydroxyl radical damage, and the tripeptide had the protective capacity against hydroxyl radicals. On the other hands, the superoxide dismutase (SOD) is one of the defense mechanisms in the living cell for cytoprotection against this reactive oxygen. In this study, the synthetic hexapeptide(YGPNRP) were much more effective than that of full length peptide in quenching superoxide radicals, and they were close to the value of natural antioxidant Trolox. The hexapeptide also have quite angiotensin-converting enzyme inhibitor activity. These results suggested the peptides might have the potential to serve as food additives with health claims in the future.

Fatin Najwa Ramli has completed her study in Bachelor of Science in Nutrition and Community Health from Universiti Putra Malaysia in 2014. Currently, she is postgraduate candidate in MSc (Nutritional Science) program also from Universiti Putra Malaysia under the supervision of Prof Dr. Azrina Azlan. She has published one paper on comparison of vitamin C content by diffrent methods in International Food Research Journal during her undergraduate studies and submitted a short review article on various analytical method used in metabolomics analysis.

Overweight and obesity are rapidly growing health problem and indeed it has become astrong risk factor in many types of chronic diseases. The aim of this study was to evaluate the anti-obesity effect of saffron extract and its pure bioactive compound, crocin on obesity in in-vivo high fat diet (HFD) induced model in comparison to orlistat as the positive control. Obese model was successfully established in male Sprague Dawley rats fed by a HFD (40%) for 12 weeks. The obese rats were administered with saffron extract (40 and 80 mg/kg), crocin (40 and 80 mg/kg) and orlistat (20mg/kg) by mixing with HFD for consecutive 8 weeks. Changes in metabolic profiles of serum were determined by using 1H-NMR based metabolomics approach combined with multivariate statistical analysis. The Orthogonal projections to latent structures discriminant analysis (OPLS-DA) on the serum 1H-NMR profiles showed a clear discrimination between obese rats from a normal control rats. This discrimination showed that obese rats serum had higher content of glucose, lactate, β-hydroxybutyrate, alanine and creatinine with lower content of taurine, betaine and trimetylamine N-oxide than those in normal control group. The metabolic profile of serum in obese rat after the administration of saffron extract (80mg/kg) and crocin (80mg/kg) were deviated from negative control. This results indicated that administration of saffron extract (80mg/kg) and crocin (80mg/kg) could effectively improve disturbed metabolism in obese rats induced by HFD.These study findings could demonstrated the potential anti obesity effect of saffron extract and crocin at pre-clinical study.

Mei-Chen Lo is a PhD student at the School of Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei, Taiwan. She has published 3 papers in reputed journals.

We proposed that advanced glycated end-products (AGEs) reduce insulin secretion from beta cellsby damaging mitochondrial function and inducing mitophagy. Mitochondrial morphology and autophagy were examined in pancreatic islets from diabeticdb/db mice and the insulinoma cell line RIN-m5F, both of which were treated with Nε-(carboxymethyl) lysine (CML)-conjugated bovine serum albumin, which is a major component of AGEs. In addition, the effects of the antioxidant alpha-lipoic acid (ALA) on mitochondria in AGEs-damaged cells were evaluated. Diabetic db/db mice exhibited increased numbers of mitochondria and autophagosomes; however, administration of ALA for 12 weeks induced greater numbers of mitochondria with well-organised cristae and fewer autophagosomes. CML treatment of RIN-m5F cells increased autophagosome formation, promoted mitochondrial dysfunction, and decreased insulin secretion. We determined that CML lowered the mitochondrial membrane potential, reduced ATP production, and increased the production of reactive oxygen species (ROS) and lipid peroxides, which was accompanied by mitochondrial DNA deletions. Elevated fission protein dynamin-related protein 1 (Drp1) levels and mitochondrial fragmentation indicated the disruption of the balance between mitochondrial fusion and fission in CML-treated cells. Additionally, elevated Parkin and PTEN-induced putative kinase 1 (PINK1) expression suggested that fragmented mitochondria were associated with increased mitophagic activity in CML-treated cells. Lastly, we observed that ALA treatment attenuated CML-induced mitophagy. Our results suggest that AGEs may damage beta cells and affect insulin secretion in the diabetic disease.

Sik-Won Choi has completed his PhD from Chunbuk National University, Repubic of Korea and Post-doctoral studies from Korea Research Institute of Chemical Technology (KRICT). He is young scientist, a premier bone metabolic disorder such as osteoporosis, and rhumatiod arthritis. He has published more than 26 papers in reputed journals. At present, his mission is identify bioactive small molecules in natural product-based crop resource and elucidate their mode of action.

A decrease of bone mass is a major risk factor for fracture. Several natural products have traditionally been used as herbal medicines to prevent and/or treat bone disorders including osteoporosis. Praeruptorin A is isolated from the dry root extract of Peucedanum praeruptorum Dunn and has several biological activities, but its anti-osteoporotic activity has not been studied yet. In this study, Praeruptorin A inhibited the RANKL-stimulated osteoclast differentiation accompanied by inhibition of p38 and Akt signaling, which could be the reason for praeruptorin A-downregulated expression levels of c-Fos and NFATc1, transcription factors that regulates osteoclast-specific genes, as well as osteoclast fusion-related molecules. The anti-osteoclastogenic effect of praeruptorin A was rescued by overexpression of NFATc1. Praeruptorin A strongly prevented the RANKL-induced Ca2+ oscillation without any changes in the phosphorylation of PLCγ. In conclusion, Praeruptorin A could exhibit its anti-osteoclastogenic activity by inhibiting p38/Akt-c-Fos-NFATc1 signaling and PLCγ-independent Ca2+ oscillation.

Mr. Yan-Ju Lin completed his master’s degree in National Cheng-Kung University, Tainan, Taiwan. He joined Industrial Technology Research Institute as a Researcher in 2004. His research is focused on transcriptome analysis and RNA technology development. He owned a patent on antisense oligonucleotide-related applications issued by the United States Patent and Trademark Office.

Metabolic diseases, such as obesity and diabetes cause multitudinous health problems. Drugs used to treat obesity show limited efficacy (around 5-7% weight loss) and cause some significant side effects. Therefore, new drugs with novel biological mechanisms may improve the efficacy and safety of the treatments. Galectin-12 is preferentially expressed by adipocytes and negatively regulates lipolysis in mice. Galectin-12 knock-out mice had a significant reduction in adiposity, with ∼40% reduction in whole-body lipid content, reduced adipocyte size, and ameliorated insulin resistance. Due to the anti-obesity potential of galectin-12, we designed and modified galectin-12 siRNA as a new therapeutic tool to metabolic diseases. One of the designed galectin-12 siRNA, ITRI-3, inhibits the expression of endogenous galectin-12 in adipocytes efficiently in vitro, and knock-down galectin-12 mRNA in adipose tissue (13 mg/kg;~ 60% knock down) in vivo. We further tested the efficacy of ITRI-3 in a diet-induced obesity (DIO) mouse model. It shows that ITRI-3 inhibits 50% of endogenous galectin-12 expression, reduces adipocyte size and also increases lipolysis of adipose tissue with 9 mg/kg dosage after 42 day treatment. Although in a long term experiment (88 days), no significant body weight loss was observed, these findings revealed the potential of ITRI-3 for treatment of human metabolic disorders. Application of ITRI-3 to reduce spot fat in mouse model is under investigation.

Aigar Ottas has completed his Master’s degree in Molecular and Cell Biology. Currently, he is a PhD student of Medicine at University of Tartu, Estonia.

Psoriasis is classified as a chronic inflammatory skin disease that affects 2-3% of the population. The disease is associated with significant mortality and morbidity. Psoriasis manifests itself as scaly inflamed plaques on the skin. The mechanisms underlying psoriasis remain unclear. It is known that an interplay between environmental and genetic factors initiate events leading to the activation of dendritic cells in the skin. This stimulate the migration and differentiation of effector T cells (Th17 and Th1) to the skin. The release of inflammatory cytokines that follows promoting further stimulation of keratinocyte proliferation, continued immune cell recruitment and the sustainment of a state of chronic inflammation. Past work on psoriasis has focused mainly on the genetic background and the proteins that have an effect on the disease. Using methods of analysis that are prevalent in metabolomics can yield new information on the etiology of psoriasis. In this study 20 serum samples from patients with psoriasis were matched with healthy controls and measured in a random order on a QTRAP 3200 coupled to a Shimadzu liquid chromatograph through a ZIP-pHILIC column for the untargeted analysis. In the targeted analysis, the AbsoluteIDQ p180 kit from Biocrates was used to quantitate a number of metabolites in the serum ranging from carbohydrates, biogenic amines, amino acids, acylcarnitines to phospholipids. The main results from this work indicate a strong dysregulation of the urea cycle in patients with psoriasis. This can be noted from a higher concentration of urea, ornithine, arginine and glutamine in patients with psoriasis.

Sara Amer completed her Bachelor’s degree in Pharmaceutical Sciences in 2013 from Misr International University, Egypt and is a teaching assistant at the University. She currently works as a research intern at the RIKEN Quantitative Biology Center (QBiC) in Dr. Masujima’s Single Cell Lab.

Floating cells have not been the target of single-cell mass spectrometry due to low adhesion properties. In this study, we have successfully trapped and analyzed a single white blood cell from a single drop of blood. A single lymphocyte (identified by morphology and structure) was monitored live and picked up using a nanospray tip attached to a micromanipulator. The cell was then injected with minimal pretreatment into the Orbitrap mass analyzer and the metabolomics profile was established. On the other hand, organelle analysis has become an essential target for single-cell analysis. Exosomes are granules excreted by most, if not all, living cells, although their specific role and function is still poorly understood. Difficulties in isolating exosomes arise from their small size (20–200 nm in diameter) and low concentration. We have efficiently isolated and analyzed exosomes of the blood through our nanospray tip methodology, and in doing so, have taken the first step in compiling the first metabolomics profile of exosomes.

Yasmine Abouleila completed her Bachelor’s degree in Pharmaceutical Sciences in 2013 from Misr International University, Cairo, Egypt. She currently works as a research intern at the RIKEN Quantitative Biology Center (QBiC) as well as a teaching assistant at Misr International University.

Tumor cells are often found in the peripheral blood of cancer patients, even those with localized cancer or who have had a primary tumor removed. Such cells are called circulating tumor cells (CTC). CTCs are cells that have shed into the vasculature from a primary tumor and circulate through the bloodstream, and they are thought to cause metastatic disease. CTCs, requiring only a blood sample instead of biopsies, might contribute greatly to early detection of cancer. However, difficulty usually arises from the isolation of these cells since they have very low concentration (few cells per 10 mL). Live single-cell mass spectrometry was used on CTC cells isolated by flow cytometry from a 10-year-old neuroblastoma patient. The single-cell metabolomics profile was established using mass spectrometry. Interestingly, a few catechol amine metabolites were detected which are specific to this type of tumor cell. The catechol amine metabolites were confirmed by MS/MS and their fragmentation patterns were matched with the Met frag data base.

Ahmed Ali completed his Bachelor’s degree in Pharmaceutical Sciences in 2013 from Misr International University, Egypt. He is currently working on his Master’s degree. He is a research intern in the RIKEN QBiC, single-cell analysis laboratory as well as a full-time teaching assistant in Misr International University in Egypt.

A single HepG2 cell was visualized by three-dimensional (3D) holographic and tomographic microscopy. While the cell was alive, it was digitally stained according to the refractive index value of each component. A micromanipulator was connected to the body of the microscope by a common base plate, and attached to the micromanipulator, was a 1 μm internal diameter nanospray tip that was used to suck out the content of the single HepG2 cell monitored in 3D.Volume analysis was carried out by 3D holographic image analysis, and the volume of the cytoplasm, nucleus, endoplasmic reticulum and a single granule was calculated. Moreover, a 3D image was taken of the cell before and after suction of the cytoplasm to determine the exact volume of the cytoplasm taken by the Nanospray tip for metabolomic analysis. Finally, the cytoplasm contents were analyzed using mass spectrometry and several unique metabolites were identified. Metabolites specific to the cytosol of liver cells were also detected, and MS/MS confirmed their respective fragmentation patterns.