Researcher Profile and Settings

Alias Name

HUI SP

Affiliation

• Faculty of Health Sciences　Health Sciences　Medical Laboratory Science

Job Title

• Professor

URL

http://kaken.nii.ac.jp/d/r/90337030.ja.html

Research funding number

• 90337030

Research Interests

• Medium chain fatty acids   Lipid hydroperoxide   酸化リポ蛋白   分析化学   臨床化学   過酸化脂質   

Research Areas

• Other / Other / Laboratory medicine
• Other / Other

Educational Organization

•  Bachelor's degree program　Departments of Health Sciences, School of Medicine
•  Master's degree program　Graduate School of Health Sciences
•  Doctoral (PhD) degree program　Graduate School of Health Sciences

Academic & Professional Experience

• 2014/08 - Today Hokkaido University Faculty of Health Sciences
• 2012/04 - 2014/07 Hokkaido University
• 2010/11 - 2012/03 北海道大学 大学院保健科学研究院 健康科学分野
• 2006/03 - 2010/09 北海道医療大学 薬学部 生命物理科学講座
• 2001/02 - 2006/03 北海道医療大学 薬学部 生命物理科学講座 助教
• 1999/01 - 2001/01 日本学術振興会 外国人特別研究員 （北海道医療大学 薬学部 生命物理科学講座）

Education

• 1994/04 - 1997/06  Hokkaido University  School of Medicine
• 1981/09 - 1986/07  中国西安医科大学

Association Memberships

• THE MASS SPECTOMETRY SOCIETY OF JAPAN   JAPANESE SOCIETY FOR BIOMEDICAL MASS SPECTROMETRY   Society for Free Radical Biology and Medicine, USA   JAPANESE SOCIETY OF LABORATORY MEDICINE   SOCIETY FOR FREE RADICAL RESEARCH JAPAN   THE JAPAN SOCIETY FOR ANALYTICAL CHEMISTRY   日本臨床化学会   THE PHARMACEUTICAL SOCIETY OF JAPAN   日本未病システム学会   

Research Activities

Published Papers

• Dissecting new lipids and their composition in herbal tea using untargeted LC/MS.

  Lipsa Rani Nath, Siddabasave Gowda B Gowda, Divyavani Gowda, Fengjue Hou, Hitoshi Chiba, Shu Ping Hui

  Food chemistry 447 138941 - 138941 2024/07/30 

  Herbal teas and beverages have gained global attention because they are rich in natural bioactive compounds, which are known to have diverse biological effects, including antioxidant and anticarcinogenic properties. However, the lipidomic profiles of herbal teas remain unclear. In this study, we applied an untargeted lipidomics approach using high-performance liquid chromatography coupled with linear ion trap-Orbitrap mass spectrometry to comprehensively profile, compare, and identify unknown lipids in four herbal teas: dokudami, kumazasa, sugina, and yomogi. A total of 341 molecular species from five major classes of lipids were identified. Multivariate principal component analysis revealed distinct lipid compositions for each of the herbs. The fatty acid α-linolenic acid (FA 18:3) was found to be abundant in kumazasa, whereas arachidonic acid (FA 20:4) was the most abundant in sugina. Interestingly, novel lipids were discovered for the first time in plants; specifically, short-chain fatty acid esters of hydroxy fatty acids (SFAHFAs) with 4-hydroxy phenyl nonanoic acid as the structural core. This study provides insight into the lipidomic diversity and potential bioactive lipid components of herbal teas, offering a foundation for further research into their health-promoting properties and biological significance.
• A facile method for monitoring sphingomyelin synthase activity in HeLa cells using liquid chromatography/mass spectrometry.

  Punith M Sundaraswamy, Yusuke Minami, Jayashankar Jayaprakash, Siddabasave Gowda B Gowda, Hiroyuki Takatsu, Divyavani Gowda, Hye-Won Shin, Shu-Ping Hui

  The Analyst 2024/05/07 

  Sphingomyelin synthase (SMS) is a sphingolipid-metabolizing enzyme involved in the de novo synthesis of sphingomyelin (SM) from ceramide (Cer). Recent studies have indicated that SMS is a key therapeutic target for metabolic diseases such as fatty liver, type 2 diabetes, atherosclerosis, and colorectal cancer. However, very few SMS inhibitors have been identified because of the limited sensitivity and selectivity of the current fluorescence-based screening assay. In this study, we developed a simple cell-based assay coupled with liquid chromatography/tandem mass spectrometry (LC-MS/MS) to screen for SMS inhibitors. HeLa cells stably expressing SMS1 or SMS2 were used for the screening. A non-fluorescent unnatural C6-Cer was used as a substrate for SMS to produce C6-SM. C6-Cer and C6-SM levels in the cells were monitored and quantified using LC-MS/MS. The activity of ginkgolic acid C15:1 (GA), a known SMS inhibitor, was measured. GA had half-maximal inhibitory concentrations of 5.5 μM and 3.6 μM for SMS1 and SMS2, respectively. To validate these findings, hSMS1 and hSMS2 proteins were optimized for molecular docking studies. In silico analyses were conducted to assess the interaction of GA with SMS1 and SMS2, and its binding affinity. This study offers an analytical approach for screening novel SMS inhibitors and provides in silico support for the experimental findings.
• Regio-specific lipid fingerprinting of edible sea cucumbers using LC/MS

  Yusuke Minami, Siddabasave Gowda B. Gowda, Divyavani Gowda, Hitoshi Chiba, Shu-Ping Hui

  Food Research International 184 114253 - 114253 2024/05 

  Sea cucumbers are a rich source of bioactive compounds and are gaining popularity as nutrient-rich seafood. They are consumed as a whole organism in Pacific regions. However, limited data are available on the comparison of their lipid composition and nutritional value. In this study, untargeted liquid chromatography/mass spectrometry was applied to comprehensively profile lipids in the skin, meat, and intestinal contents of three color-distinct edible sea cucumbers. Multivariate principal component analysis revealed that the lipid composition of the intestinal contents of red, black, and blue sea cucumbers differs from that of skin, and meats. Polyunsaturated fatty acids (PUFAs) are abundant in the intestinal contents, followed by meats of sea cucumber. Lipid nutritional quality assessments based on fatty acid composition revealed a high P:S ratio, low index of atherogenicity, and high health promotion indices for the intestinal contents of red sea cucumber, suggesting its potential health benefits. In addition, hierarchical cluster analysis revealed that the intestinal contents of sea cucumbers were relatively high in PUFA-enriched phospholipids and lysophospholipids. Ceramides are abundant in black skin, blue meat, and red intestinal content samples. Overall, this study provides the first insights into a comprehensive regio-specific profile of the lipid content of sea cucumbers and their potential use as a source of lipid nutrients in food and nutraceuticals.
• Inhibition of Lipid Accumulation and Oxidation in Hepatocytes by Bioactive Bean Extracts

  Dya Fita Dibwe, Emi Kitayama, Saki Oba, Nire Takeishi, Hitoshi Chiba, Shu-Ping Hui

  Antioxidants 2024/04/25
• Sex-Specific Effect of Ethanol on Colon Content Lipidome in a Mice Model Using Nontargeted LC/MS.

  Jayashankar Jayaprakash, Siddabasave Gowda B Gowda, Pradeep K Shukla, Divyavani Gowda, Lipsa Rani Nath, Hitoshi Chiba, Radhakrishna Rao, Shu-Ping Hui

  ACS omega 9 (14) 16044 - 16054 2024/04/09 

  Consumption of alcohol has widespread effects on the human body. The organs that are most significantly impacted are the liver and digestive system. When alcohol is consumed, it is absorbed in the intestines and processed by the liver. However, excessive alcohol use may affect gut epithelial integrity, microbiome composition, and lipid metabolism. Despite past studies investigating the effect of ethanol on hepatic lipid metabolism, the focus on colonic lipid metabolism has not been well explored. In this study, we investigated the sex-specific effect of ethanol on the colonic content lipidome in a mouse model using nontargeted liquid chromatography-mass spectrometry. Comprehensive lipidome analysis of colonic flush samples was performed using ethanol-fed (EF) and pair-fed (PF) mice of each sex. Partial least-squares discriminant analysis revealed that ethanol altered colonic lipid composition largely in male mice compared with female mice. A significant increase in free fatty acids, ceramides, and hexosylceramides and decreased phosphatidylglycerols (PG) was observed in the EF group compared to the PF group in male mice. Phosphatidylethanolamine (PE) levels were increased significantly in the EF group of both sexes compared to the PF group. The volcanic plot shows that PG (O-15:1/15:0) and PE (O-18:2/15:0) are common markers that are increased in both sexes of the EF group. In addition, decreased fatty acid esters of hydroxy fatty acids (FAHFA) were observed specifically in the EF group of female mice. Overall, a significant variation in the mice colonic content lipidome between the EF and PF groups was observed. Target pathways, such as sphingolipid metabolism in males, FAHFA in females, and PE metabolism in both sexes, were suggested. This study provides new insight into the sex-dependent lipid change associated with alcohol-induced gut-microbiota dysfunction and its potential health impacts.
• Glucosylceramide flippases contribute to cellular glucosylceramide homeostasis.

  Natsuki Kita, Asuka Hamamoto, Siddabasave Gowda B Gowda, Hiroyuki Takatsu, Kazuhisa Nakayama, Makoto Arita, Shu-Ping Hui, Hye-Won Shin

  Journal of lipid research 65 (3) 100508 - 100508 2024/03 

  Lipid transport is an essential cellular process with importance to human health, disease development, and therapeutic strategies. Type IV P-type ATPases (P4-ATPases) have been identified as membrane lipid flippases by utilizing nitrobenzoxadiazole (NBD)-labeled lipids as substrates. Among the 14 human type IV P-type ATPases, ATP10D was shown to flip NBD-glucosylceramide (GlcCer) across the plasma membrane. Here, we found that conversion of incorporated GlcCer (d18:1/12:0) to other sphingolipids is accelerated in cells exogenously expressing ATP10D but not its ATPase-deficient mutant. These findings suggest that 1) ATP10D flips unmodified GlcCer as well as NBD-GlcCer at the plasma membrane and 2) ATP10D can translocate extracellular GlcCer, which is subsequently converted to other metabolites. Notably, exogenous expression of ATP10D led to the reduction in cellular hexosylceramide levels. Moreover, the expression of GlcCer flippases, including ATP10D, also reduced cellular hexosylceramide levels in fibroblasts derived from patients with Gaucher disease, which is a lysosomal storage disorder with excess GlcCer accumulation. Our study highlights the contribution of ATP10D to the regulation of cellular GlcCer levels and maintaining lipid homeostasis.
• Kaempferol Improves Cardiolipin and ATP in Hepatic Cells: A Cellular Model Perspective in the Context of Metabolic Dysfunction-Associated Steatotic Liver Disease

  Akiko Sakurai, Toshihiro Sakurai, Hsin-Jung Ho, Hitoshi Chiba, Shu-Ping Hui

  Nutrients 2024/02/11
• Synthesis and quantification of short-chain fatty acid esters of hydroxy fatty acids in rat intestinal contents and fecal samples by LC-MS/MS

  Siddabasave Gowda B. Gowda, Fengjue Hou, Divyavani Gowda, Hitoshi Chiba, Kentaro Kawakami, Satoru Fukiya, Atsushi Yokota, Shu-Ping Hui

  Analytica Chimica Acta 1288 342145 - 342145 0003-2670 2024/02
• Lysosomal membrane integrity in fibroblasts derived from patients with Gaucher disease.

  Asuka Hamamoto, Natsuki Kita, Siddabasave Gowda B Gowda, Hiroyuki Takatsu, Kazuhisa Nakayama, Makoto Arita, Shu-Ping Hui, Hye-Won Shin

  Cell structure and function 49 (1) 1 - 10 0386-7196 2024/01/23 

  Gaucher disease (GD) is a recessively inherited lysosomal storage disorder characterized by a deficiency of lysosomal glucocerebrosidase (GBA1). This deficiency results in the accumulation of its substrate, glucosylceramide (GlcCer), within lysosomes. Here, we investigated lysosomal abnormalities in fibroblasts derived from patients with GD. It is noteworthy that the cellular distribution of lysosomes and lysosomal proteolytic activity remained largely unaffected in GD fibroblasts. However, we found that lysosomal membranes of GD fibroblasts were susceptible to damage when exposed to a lysosomotropic agent. Moreover, the susceptibility of lysosomal membranes to a lysosomotropic agent could be partly restored by exogenous expression of wild-type GBA1. Here, we report that the lysosomal membrane integrity is altered in GD fibroblasts, but lysosomal distribution and proteolytic activity is not significantly altered.Key words: glucosylceramide, lysosome, Gaucher disease, lysosomotropic agent.
• Identification of a β-Carboline Alkaloid from Chemoselectively Derived Vanilla Bean Extract and Its Prevention of Lipid Droplet Accumulation in Human Hepatocytes (HepG2)

  Dya Fita Dibwe, Nire Takeishi, Saki Oba, Akiko Sakurai, Toshihiro Sakurai, Takayuki Tsukui, Hitoshi Chiba, Shu-Ping Hui

  Molecules 2023/12/09
• Regiospecific analysis of lipidome in the brain from mammals of land and aquatic habitats-by liquid chromatography-mass spectrometry

  Siddabasave Gowda B. Gowda, Divyavani Gowda, Fengjue Hou, Chandra Shekhar, Hitoshi Chiba, Nina Patzke, Shu-Ping Hui

  Heliyon 9 (12) e22959 - e22959 2405-8440 2023/12
• Flazin improves mitochondrial dynamics in renal tubular epithelial cells under oxidative stress

  Xun-Zhi Wu, Hsin-Jung Ho, Miki Eguchi, Zhen Chen, Hitoshi Chiba, Shu-Ping Hui

  Food Bioscience 2023/12
• Crosstalk between Lipids and Non-Alcoholic Fatty Liver Disease

  Divyavani Gowda, Chandra Shekhar, Siddabasave Gowda B. Gowda, Yifan Chen, Shu-Ping Hui

  Livers 2023/11/23
• The elucidation of plasma lipidome profiles during severe influenza in a mouse model.

  Marumi Ohno, Siddabasave Gowda B Gowda, Toshiki Sekiya, Naoki Nomura, Masashi Shingai, Shu-Ping Hui, Hiroshi Kida

  Scientific reports 13 (1) 14210 - 14210 2023/08/30 

  Although influenza virus infection has been shown to affect lipid metabolism, details remain unknown. Therefore, we elucidated the kinetic lipid profiles of mice infected with different doses of influenza virus A/Puerto Rico/8/34 (H1N1) (PR8) by measuring multiple lipid molecular species using untargeted lipidomic analysis. C57BL/6 male mice were intranasally infected with PR8 virus at 50 or 500 plaque-forming units to cause sublethal or lethal influenza, respectively. Plasma and tissue samples were collected at 1, 3, and 6 days post-infection (dpi), and comprehensive lipidomic analysis was performed using high-performance liquid chromatography-linear trap quadrupole-Orbitrap mass spectrometry, as well as gene expression analyses. The most prominent feature of the lipid profile in lethally infected mice was the elevated plasma concentrations of phosphatidylethanolamines (PEs) containing polyunsaturated fatty acid (PUFA) at 3 dpi. Furthermore, the facilitation of PUFA-containing phospholipid production in the lungs, but not in the liver, was suggested by gene expression and lipidomic analysis of tissue samples. Given the increased plasma or serum levels of PUFA-containing PEs in patients with other viral infections, especially in severe cases, the elevation of these phospholipids in circulation could be a biomarker of infection and the severity of infectious diseases.
• Plasmalogen Profiling in Porcine Brain Tissues by LC-MS/MS.

  Yue Wu, Yifan Chen, Min Zhang, Hitoshi Chiba, Shu-Ping Hui

  Foods (Basel, Switzerland) 12 (16) 2023/08/08 

  Plasmalogen, a functional glycerophospholipid, is known for its beneficial nutritional effects, such as anti-oxidation and anti-inflammation. As the porcine brain is a plasmalogen-rich resource, this study aimed to explore its potential for plasmalogen-based health food product development, with special attention on whether and how the industrial production processes influence the plasmalogen content and composition. In the present work, plasmalogens from different porcine brain products were investigated using liquid chromatography-tandem mass spectrometry. The results indicated that all the porcine brain products showed abundant total plasmalogens, of which more than 95% were ethanolamine plasmalogen species. Acetone precipitation, ethanol extraction, and drying did not significantly affect the plasmalogen content, whereas repeated freeze-thaw cycles in the production process led to noticeable loss. The chemometric investigation suggested that raw products and glycerophospholipid products exhibited different profiles; furthermore, the concentration step seemed to impact the plasmalogen composition. The nutritional assessment revealed that porcine brain products showed favorable values of multiple indexes, including PUFA/SFA ratio, n-6/n-3 ratio, thrombogenicity index, and unsaturation index, suggesting a health-beneficial value. The current study not only shows the feasibility of producing porcine brain-derived plasmalogens, but also provides possible strategies for developing and quality-controlling dietary plasmalogen supplements and healthcare products.
• Lipid Composition Analysis and Characterization of Acyl Sterol Glycosides in Adzuki and Soybean Cultivars by Non-Targeted LC-MS.

  Rachana M Gangadhara, Siddabasave Gowda B Gowda, Divyavani Gowda, Ken Inui, Shu-Ping Hui

  Foods (Basel, Switzerland) 12 (14) 2023/07/21 

  Beans, a globally significant economic and nutritional food crop, are rich in polyphenolic chemicals with potential health advantages, providing high protein, fiber, minerals, and vitamins. However, studies on the global profiling of lipids in beans are limited. We applied a non-targeted lipidomic approach based on high-performance liquid chromatography coupled with linear ion trap-Orbitrap mass spectrometry (HPLC/LTQ-Orbitrap-MS) to comprehensively profile and compare the lipids in six distinct bean cultivars, namely, adzuki red beans-adzuki cultivar (ARB-AC), adzuki red beans-Benidainagon cultivar (ARB-BC), adzuki red beans-Erimoshouzu cultivar (ARB-EC), soybean-Fukuyutaka cultivar 2021 (SB-FC21), soybean-Fukuyutaka cultivar 2022 (SB-FC22), and soybean-Oosuzu cultivar (SB-OC). MS/MS analysis defined 144 molecular species from four main lipid groups. Multivariate principal component analysis indicated unique lipid compositions in the cultivars except for ARB-BC and ARB-EC. Evaluation of the concentrations of polyunsaturated fatty acid to saturated fatty acid ratio among all the cultivars showed that SB-FC21 and SB-FC22 had the highest value, suggesting they are the most beneficial for health. Furthermore, lipids such as acyl sterol glycosides were detected and characterized for the first time in these bean cultivars. Hierarchical cluster correlations revealed the predominance of ceramides in ARB-EC, lysophospholipids in SB-FC21, and glycerophospholipids in SB-OC. This study comprehensively investigated lipids and their compositions in beans, indicating their potential utility in the nutritional evaluation of beans as functional foods.
• A Pacific Oyster-Derived Antioxidant, DHMBA, Protects Renal Tubular HK-2 Cells against Oxidative Stress via Reduction of Mitochondrial ROS Production and Fragmentation.

  Hsin-Jung Ho, Natsumi Aoki, Yi-Jou Wu, Ming-Chen Gao, Karin Sekine, Toshihiro Sakurai, Hitoshi Chiba, Hideaki Watanabe, Mitsugu Watanabe, Shu-Ping Hui

  International journal of molecular sciences 24 (12) 2023/06/13 

  The kidney contains numerous mitochondria in proximal tubular cells that provide energy for tubular secretion and reabsorption. Mitochondrial injury and consequent excessive reactive oxygen species (ROS) production can cause tubular damage and play a major role in the pathogenesis of kidney diseases, including diabetic nephropathy. Accordingly, bioactive compounds that protect the renal tubular mitochondria from ROS are desirable. Here, we aimed to report 3,5-dihydroxy-4-methoxybenzyl alcohol (DHMBA), isolated from the Pacific oyster (Crassostrea gigas) as a potentially useful compound. In human renal tubular HK-2 cells, DHMBA significantly mitigated the cytotoxicity induced by the ROS inducer L-buthionine-(S, R)-sulfoximine (BSO). DHMBA reduced the mitochondrial ROS production and subsequently regulated mitochondrial homeostasis, including mitochondrial biogenesis, fusion/fission balance, and mitophagy; DHMBA also enhanced mitochondrial respiration in BSO-treated cells. These findings highlight the potential of DHMBA to protect renal tubular mitochondrial function against oxidative stress.
• Sleep deprivation in obesogenic setting alters lipidome and microbiome toward suboptimal inflammation in acute heart failure.

  Ganesh V Halade, Yusuf Mat, Siddabasave Gowda B Gowda, Shalini Jain, Shu-Ping Hui, Hariom Yadav, Vasundhara Kain

  FASEB journal : official publication of the Federation of American Societies for Experimental Biology 37 (5) e22899  2023/05 

  Sleep is a fundamental medicine for cardiac homeostasis, and sleep-deprived individuals are prone to higher incidences of heart attack. The lipid-dense diet (obesogenic diet-OBD) is a cumulative risk factor for chronic inflammation in cardiovascular disease; thus, understanding how sleep fragmentation (SF) in an obesity setting impacts immune and cardiac health is an unmet medical need. We hypothesized whether the co-existence of SF with OBD dysregulates gut homeostasis and leukocyte-derived reparative/resolution mediators, thereby impairing cardiac repair. Two-month-old male C57BL/6J mice were randomized first into two groups, then four groups; Control, control + SF, OBD, and OBD + SF mice subjected to myocardial infarction (MI). OBD mice had higher levels of plasma linolenic acid with a decrease in eicosapentaenoic and docosahexaenoic acid. The OBD mice had lower Lactobacillus johnsonii indicating a loss of probiotic microbiota. SF in OBD mice increased Firmicutes/Bacteroidetes ratio indicative of a detrimental change in SF-directed microbiome. OBD + SF group increased in the neutrophil: lymphocyte ratio suggestive of suboptimal inflammation. As a result of SF, resolution mediators (RvD2, RvD3, RvD5, LXA4 , PD1, and MaR1) decreased and inflammatory mediators (PGD2 , PGE2 , PGF2a , 6k-PGF1a ) were increased in OBD mice post-MI. At the site of infarction, the proinflammatory cytokines Ccl2, IL1β, and IL-6 were amplified in OBD + SF indicating a robust proinflammatory milieu post-MI. Also, brain circadian genes (Bmal1, Clock) were downregulated in SF-subjected control mice, but remained elevated in OBD mice post-MI. SF superimposed on obesity dysregulated physiological inflammation and disrupted resolving response thereby impaired cardiac repair and signs of pathological inflammation.
• Mass spectrometric approaches in discovering lipid biomarkers for COVID-19 by lipidomics: Future challenges and perspectives.

  Siddabasave Gowda B Gowda, Chandra Shekhar, Divyavani Gowda, Yifan Chen, Hitoshi Chiba, Shu-Ping Hui

  Mass spectrometry reviews 2023/04/27 

  Coronavirus disease 2019 (COVID-19) has emerged as a global health threat and has rapidly spread worldwide. Significant changes in the lipid profile before and after COVID-19 confirmed the significance of lipid metabolism in regulating the response to viral infection. Therefore, understanding the role of lipid metabolism may facilitate the development of new therapeutics for COVID-19. Owing to their high sensitivity and accuracy, mass spectrometry (MS)-based methods are widely used for rapidly identifying and quantifying of thousands of lipid species present in a small amount of sample. To enhance the capabilities of MS for the qualitative and quantitative analysis of lipids, different platforms have been combined to cover a wide range of lipidomes with high sensitivity, specificity, and accuracy. Currently, MS-based technologies are being established as efficient methods for discovering potential diagnostic biomarkers for COVID-19 and related diseases. As the lipidome of the host cell is drastically affected by the viral replication process, investigating lipid profile alterations in patients with COVID-19 and targeting lipid metabolism pathways are considered to be crucial steps in host-directed drug targeting to develop better therapeutic strategies. This review summarizes various MS-based strategies that have been developed for lipidomic analyzes and biomarker discoveries to combat COVID-19 by integrating various other potential approaches using different human samples. Furthermore, this review discusses the challenges in using MS technologies and future perspectives in terms of drug discovery and diagnosis of COVID-19.
• Oxidized Low-Density Lipoproteins Trigger Hepatocellular Oxidative Stress with the Formation of Cholesteryl Ester Hydroperoxide-Enriched Lipid Droplets.

  Iku Sazaki, Toshihiro Sakurai, Arisa Yamahata, Sumire Mogi, Nao Inoue, Koutaro Ishida, Ami Kikkai, Hana Takeshita, Akiko Sakurai, Yuji Takahashi, Hitoshi Chiba, Shu-Ping Hui

  International journal of molecular sciences 24 (5) 4281 - 4281 2023/02/21 

  Oxidized low-density lipoproteins (oxLDLs) induce oxidative stress in the liver tissue, leading to hepatic steatosis, inflammation, and fibrosis. Precise information on the role of oxLDL in this process is needed to establish strategies for the prevention and management of non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH). Here, we report the effects of native LDL (nLDL) and oxLDL on lipid metabolism, lipid droplet formation, and gene expression in a human liver-derived C3A cell line. The results showed that nLDL induced lipid droplets enriched with cholesteryl ester (CE) and promoted triglyceride hydrolysis and inhibited oxidative degeneration of CE in association with the altered expression of LIPE, FASN, SCD1, ATGL, and CAT genes. In contrast, oxLDL showed a striking increase in lipid droplets enriched with CE hydroperoxides (CE-OOH) in association with the altered expression of SREBP1, FASN, and DGAT1. Phosphatidylcholine (PC)-OOH/PC was increased in oxLDL-supplemented cells as compared with other groups, suggesting that oxidative stress increased hepatocellular damage. Thus, intracellular lipid droplets enriched with CE-OOH appear to play a crucial role in NAFLD and NASH, triggered by oxLDL. We propose oxLDL as a novel therapeutic target and candidate biomarker for NAFLD and NASH.
• Extraction and Quantitation of Phytosterols from Edible Brown Seaweeds: Optimization, Validation, and Application.

  Zhen Chen, Nianqiu Shen, Xunzhi Wu, Jiaping Jia, Yue Wu, Hitoshi Chiba, Shuping Hui

  Foods (Basel, Switzerland) 12 (2) 2023/01/05 

  Brown seaweeds are known as important marine food sources, from which phytosterols have been recognized as functional food components with multiple health-beneficial effects. However, studies on phytosterol extraction and quantitation from edible brown seaweeds are limited. In the present work, extraction methods for seaweed phytosterols were compared and optimized by one-factor-at-one-time method and response surface methodology. Moreover, the quantitation method of total sterols and major sterol components, including fucosterol, saringosterol, and ostreasterol, was established and validated using 1H NMR. Furthermore, the developed extraction and determination methods were applied to investigate three common edible seaweeds from Japan (Hijiki, Wakame, and Kombu). As a result, the finally optimized conditions were ultrasound-assisted extraction with CHCl3-MeOH 2:3 for 15 min followed by saponification with 1.65 mL of 1.85 M KOH for 14.5 h. Based on the developed methods, phytosterols in three seaweeds were compared, and Hijiki showed an abundant total sterol amount (2.601 ± 0.171 mg/g DW), significantly higher than Wakame (1.845 ± 0.137 mg/g DW) and Kombu (1.171 ± 0.243 mg/g DW). Notably, the composition of the sterol components varied in different seaweeds. These findings might help the nutritional investigation and functional food development concerning phytosterols from seaweeds.
• Distinctive serum lipidomic profile of IVIG-resistant Kawasaki disease children before and after treatment.

  Zhen Chen, Shuji Sai, Kiyoshi Nagumo, Yue Wu, Hitoshi Chiba, Shu-Ping Hui

  PloS one 18 (3) e0283710  2023 

  Kawasaki Disease (KD) is an acute inflammatory disorder associated with systemic vasculitis. Intravenous immunoglobulin (IVIG) is an effective therapy for KD, yet, about 20% of cases show IVIG resistance with persistent inflammation. The lipid profile in IVIG-resistant KD patients and the relationship between lipid characteristics and IVIG resistance remain unknown. In this study, serum samples from twenty KD patients with different IVIG responses (sensitive, intermediate, or resistant) were collected both before and after treatment, and lipidomic analysis was performed using high-performance liquid chromatography-mass spectrometry. As a result, before treatment, six lipid species were found as the most variant features, in which all the top decreased lipids in the IVIG-resistant group were lysophosphatidylcholine (LPC) and lysophosphatidylethanolamine (LPE), suggesting the potential to be IVIG-resistant markers in pretreatment diagnosis. During treatment, lipidomic changes showed a weaker response in the IVIG-resistant group. After treatment, LPC and LPE species exhibited lower in the IVIG-resistant group and negative correlation with the inflammatory markers, indicating that the unique metabolism may occur among IVIG-responsiveness. These results might contribute to diagnosing IVIG-resistant patients more accurately for alternative therapy and to a better understanding of how lipid metabolism is associated with IVIG sensitiveness/resistance in KD.
• Temporal lipid profiling in the progression from acute to chronic heart failure in mice and ischemic human hearts.

  Siddabasave Gowda B Gowda, Divyavani Gowda, Fengjue Hou, Hitoshi Chiba, Vibhu Parcha, Pankaj Arora, Ganesh V Halade, Shu-Ping Hui

  Atherosclerosis 363 30 - 41 0021-9150 2022/12 

  BACKGROUND AND AIMS: Myocardial infarction (MI) is a leading cause of heart failure (HF). After MI, lipids undergo several phasic changes implicated in cardiac repair if inflammation resolves on time. However, if inflammation continues, that leads to end stage HF progression and development. Numerous studies have analyzed the traditional risk factors; however, temporal lipidomics data for human and animal models are limited. Thus, we aimed to obtain sequential lipid profiling from acute to chronic HF. METHODS: Here, we report the comprehensive lipidome of the hearts from diseased and healthy subjects. To induce heart failure in mice, we used a non-reperfused model of coronary ligation, and MI was confirmed by echocardiography and histology, then temporal kinetics of lipids in different tissues (heart, spleen, kidney), and plasma was quantitated from heart failure mice and compared with naïve controls. For lipid analysis in mouse and human samples, untargeted liquid chromatography-linear trap quadrupole orbitrap mass spectrometry (LC-LTQ-Orbitrap MS) was performed. RESULTS: In humans, multivariate analysis revealed distinct cardiac lipid profiles between healthy and ischemic subjects, with 16 lipid species significantly downregulated by 5-fold, mainly phosphatidylethanolamines (PE), in the ischemic heart. In contrast, PE levels were markedly increased in mouse tissues and plasma in chronic MI, indicating possible cardiac remodeling. Further, fold change analysis revealed site-specific lipid biomarkers for acute and chronic HF. A significant decrease in sulfatides (SHexCer (34:1; 2O)) and sphingomyelins (SM (d18:1/16:0)) was observed in mouse tissues and plasma in chronic HF. CONCLUSIONS: Overall, a significant decreased lipidome in human ischemic LV and differential lipid metabolites in the transition of acute to chronic HF with inter-organ communication could provide novel insights into targeting integrative pathways for the early diagnosis or development of novel therapeutics to delay/prevent HF.
• Simultaneous free fatty acid elevations and accelerated desaturation in plasma and oocytes in early postpartum dairy cows under intensive feeding management.

  Eri Furukawa, Zhen Chen, Tomoaki Kubo, Yue Wu, Koichiro Ueda, Madalitso Chelenga, Hitoshi Chiba, Yojiro Yanagawa, Seiji Katagiri, Masashi Nagano, Shu-Ping Hui

  Theriogenology 193 20 - 29 0093-691X 2022/11 

  A severe negative energy balance and high circulating free fatty acids (FFA) in postpartum cows impair fertility. The lipotoxicity of FFA has been shown to decrease the quality of bovine oocytes in vitro. Therefore, excess FFA in cells is converted to triacylglycerol (TAG), a non-toxic form, to avoid lipotoxicity. We recently reported that the TAG content in oocytes was higher in postpartum lactating cows subjected to grazing management than in heifers (Theriogenology 176: 174-182, 2021). The present study investigated the compositions of the energy metabolism-related lipids, FFA and TAG, in the plasma and oocytes of cows at different lactation stages under indoor intensive feeding management in order to obtain insights into lipotoxicity in oocytes, particularly those in early postpartum cows. Blood and oocytes were collected from 20 lactating cows categorized into the following lactation groups: 20-30 days in milk (DIM) (n = 5), 40-50 DIM (n = 5), 60-80 DIM (n = 5), and 130-160 DIM (n = 5). Daily energy balance data were obtained for 3 weeks prior to oocyte collection using the ovum pick up (OPU) method. The contents and compositions of FFA and TAG in plasma and oocytes were analyzed using liquid chromatography-mass spectrometry. As expected, plasma FFA was high at 20-30 DIM, decreased by 50 DIM, and was maintained at a low level for the remainder of the experimental period. Similar changes were observed in oocyte FFA and TAG with DIM as plasma FFA. Oocyte FFA positively correlated with plasma FFA (P < 0.05), but negatively correlated with the mean energy balance 1 and 21 days before OPU (P < 0.05). Relationships were noted between the composition and content of FFA in plasma and oocytes, with the FFA 16:1/16:0 and 18:1/18:0 ratios positively correlating with the total amount of FFA (P < 0.05). Elevated oocyte FFA in cows in the early postpartum period under intensive feeding management suggested that oocytes were at a high risk of FFA lipotoxicity. Furthermore, the present results implied that the severe negative energy balance in the previous few weeks was closely related to increases in oocyte FFA, which supports the importance of long-term cow feeding management for preserving the quality of oocytes in the early postpartum period. The present results provide insights into the effects of high circulating FFA on the fertility of postpartum cows.
• Detection and characterization of lipids in eleven species of fish by non-targeted liquid chromatography/mass spectrometry

  Siddabasave Gowda B. Gowda, Yusuke Minami, Divyavani Gowda, Hitoshi Chiba, Shu-Ping Hui

  Food Chemistry 393 133402 - 133402 0308-8146 2022/11
• Analysis of Antioxidant Lipids in Five Species of Dietary Seaweeds by Liquid Chromatography/Mass Spectrometry

  Siddabasave Gowda B. Gowda, Chen Yifan, Divyavani Gowda, Yui Tsuboi, Hitoshi Chiba, Shu-Ping Hui

  Antioxidants 11 (8) 1538 - 1538 2022/08/08 

  Seaweeds are a good source of bioactive lipids and are known for their nutritional benefits, making them a valuable food source. Despite their dietary significance and nutritional importance, there are limited reports on comprehensive lipidome analysis of lipids with antioxidant properties. Therefore, this study aimed to compare the lipid profiles of five commonly consumed Japanese dietary seaweeds using non-targeted liquid chromatography/mass spectrometry (LC/MS). A total, of 304 molecular species from four major lipid classes were detected and characterized by MS/MS analysis. Multivariate statistical analysis revealed distinct lipid molecular compositions in kombu and sea mustard compared to hijiki, mozuku, and laver seaweeds. Kombu has been shown to contain large amounts of antioxidants, such as polyunsaturated fatty acids (PUFAs), and a high health promotion index compared to other seaweeds. Hierarchical cluster correlations indicated the predominance of glycerophospholipids (GPs) and glycerolipids (GLs) in sea mustard and kombu. As a result, dietary seaweeds have great potential as antioxidants and health-promoting foods for human consumption due to their high levels of PUFA-rich GPs and GLs. Unsaturated triacylglycerols are predominant in hijiki, whereas other health-beneficial lipids, such as monogalactosyldiacylglycerol and sulfoquinovosyl diacylglycerols, are predominant in sea mustard. This study provides a detailed characterization of lipids and their comparative fingerprints in seaweeds, demonstrating the potential use of dietary seaweeds in biotechnological and industrial applications involving the development of functional food products.
• Oxidative Stress and Lipid Dysregulation in Lipid Droplets: A Connection to Chronic Kidney Disease Revealed in Human Kidney Cells.

  Zhen Chen, Rojeet Shrestha, Xiaoyue Yang, Xunzhi Wu, Jiaping Jia, Hitoshi Chiba, Shu-Ping Hui

  Antioxidants (Basel, Switzerland) 11 (7) 2022/07/18 

  Chronic kidney disease (CKD), which is defined as a condition causing the gradual loss of kidney function, shows renal lipid droplet (LD) accumulation that is associated with oxidative damage. There is a possibility that an LD abnormality in quality plays a role in CKD development. This study aimed to explore the chemical composition of LDs that are induced in human kidney cells during exposure to free fatty acids as an LD source and oxidized lipoproteins as oxidative stress. The LDs were aspirated directly from cells using nanotips, followed by in-tip microextraction, and the LD lipidomic profiling was conducted using nanoelectrospray mass spectrometry. As a result, the free fatty acids increased the LD lipid content and, at the same time, changed their composition significantly. The oxidized lipoproteins caused distorted proportions of intact lipids, such as triacylglycerols (TG), phosphatidylcholines (PC), phosphatidylethanolamines (PE), and cholesteryl esters (CE). Notably, the oxidized lipids, including the hydroperoxides of TG, PC, and PE, exhibited significant elevations in dose-dependent manners. Furthermore, the dysregulation of intact lipids was paralleled with the accumulation of lipid hydroperoxides. The present study has revealed that the oxidation of lipids and the dysregulation of the lipid metabolism coexisted in LDs in the kidney cells, which has provided a potential new target for diagnosis and new insights into CKD.
• Determination of short-chain fatty acids by N,N-dimethylethylenediamine derivatization combined with liquid chromatography/mass spectrometry and their implication in influenza virus infection.

  Divyavani Gowda, Yonghan Li, Siddabasave Gowda B Gowda, Marumi Ohno, Hitoshi Chiba, Shu-Ping Hui

  Analytical and bioanalytical chemistry 414 (22) 6419 - 6430 2022/07/16 

  Short-chain fatty acids (SCFAs) are the end products of the fermentation of complex carbohydrates by the gut microbiota. Although SCFAs are recognized as important markers to elucidate the link between gut health and disease, it has been difficult to analyze SCFAs with mass spectrometry technologies due to their poor ionization efficiency and high volatility. Here, we present a novel and sensitive method for the quantification of SCFAs, including C2-C6 SCFAs and their hydroxy derivatives, by liquid chromatography/tandem mass spectrometry (LC-MS/MS) upon N,N-dimethylethylenediamine (DMED) derivatization with a run time of 10 min. Moreover, the quantification method of DMED-derivatized SCFAs in intestinal contents using isotope-labeled internal standards was also established. The method validation was performed by analyzing spiked intestinal samples; the limits of detection and quantification of SCFAs with this method were found to be 0.5 and 5 fmol, respectively; the recovery was greater than 80% and good linearity (0.9932 to 0.9979) of calibration curves was obtained over the range from 0.005 to 5000 pmol/μL; the intraday and interday precisions were achieved in the range of 1-5%. Furthermore, the validated method was applied to analyze SCFAs in the cecum and colon contents of mice infected with the influenza virus. The results showed that the concentration of most of the SCFAs tested here decreased significantly in a time-dependent manner after the infection, suggesting a possibility that SCFAs in intestinal samples could be used as severe disease markers. Overall, we here successfully developed a simple, fast, and sensitive method for SCFA analysis by LC-MS/MS combined with DMED derivatization. The method for the quantification of SCFAs will be a useful tool for both basic research and clinical studies.
• Profiling of lysophosphatidylethanolamine molecular species in human serum and in silico prediction of the binding site on albumin.

  Nao Inoue, Toshihiro Sakurai, Yusuke Yamamoto, Hitoshi Chiba, Shu-Ping Hui

  BioFactors (Oxford, England) 48 (5) 1076 - 1088 2022/06/10 

  Lysophosphatidylethanolamine (LPE) is a major lysophospholipid produced by phospholipids and binds to human serum albumin (HSA). LPEs may play various roles in vivo depending on the differences in their acyl chains. However, only few reports have been published on the biological functions of LPEs. Hence, we determined the exact relative abundance of the major LPEs in the serum of healthy participants (n = 8) using liquid chromatography-tandem mass spectrometry. Consequently, LPE 18:2 (24.1 ± 5.2%) was found to be the most abundant in serum. To understand the distribution of LPEs, the serum separated via gel-filtration high-performance liquid chromatography was subjected to quantitative measurement. LPEs were more observed in the albumin fraction than the lipoprotein fraction. We also performed a fluorescence displacement assay and an in silico molecular docking experiment using AutoDock to confirm the affinity and binding sites of the LPEs on HSA. The binding affinities of the LPEs for drug sites 1 and 2 on HSA were relatively low, with Ki values of approximately 11 and 3.8 μM, respectively. AutoDock analysis revealed the conformation of the LPEs bound to drug sites and the possibility of LPEs binding to other HSA sites. These findings could help to elucidate the biological and pathological functions of LPEs.
• Quantitative Evaluation on the Degradation Process of the Pulmonary Surfactant Monolayer When Exposed to Low-Level Ozone of Ambient Environment.

  Lin Qiao, Zhen Chen, Chunji Takada, Hitoshi Chiba, Ken-Ichi Inoue, Shu-Ping Hui, Shen Ye

  Analytical chemistry 94 (24) 8651 - 8658 2022/05/23 

  Ozone is a potent environmental oxidant with high chemical reactivity and is present in the ambient environment at a low level of a few tens of ppb. However, only limited information is known about low-level ozone's influence on the respiratory system. In the present study, we systematically investigated the degradation of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), which is one of the major components of the pulmonary surfactant (PS), enabling breath function of the lung exposed to low ambient-level ozone (40 ± 10 ppb). Using the liquid chromatography-mass spectrometry technique combined with the Langmuir-Blodgett approach, we first tracked the degradation process of POPC molecules by determining the degradation products during exposure to the ambient environment. As a result, we found that the POPC molecules can be readily degraded from the C═C moiety in 45 min, yielding an aldehyde-type product of 1-palmitoyl-2-(9'-oxo-nonanoyl)-sn-glycero-3-phosphocholine (POnPC) and a trace amount of an acid-type one of 1-palmitoyl-2-azelaoyl-sn-glycero-3-phosphocholine (PAzPC), as well as a pair of secondary ozonide (SOZ) isomers. Furthermore, with prolonged exposure, the SOZ stayed constant but the yield of PAzPC significantly increased with the decrease in POnPC. The low-level ozone-induced oxidation mechanisms for unsaturated lipids are discussed based on the quantitative analyses of these experimental observations. The present results demonstrate that the ground-level ozone is strong enough to induce dramatic oxidation damage to the unsaturated lipids of the PS. These oxidized species may trigger the lung's inflammatory response and be used as biomarkers for oxidative stress and inflammation.
• Food-Derived β-Carboline Alkaloids Ameliorate Lipid Droplet Accumulation in Human Hepatocytes.

  Dya Fita Dibwe, Saki Oba, Nire Takeishi, Toshihiro Sakurai, Takayuki Tsukui, Hitoshi Chiba, Shu-Ping Hui

  Pharmaceuticals (Basel, Switzerland) 15 (5) 2022/05/05 

  Lipid droplet accumulation (LDA) in hepatocytes is the initial stage of nonalcoholic fatty liver disease (NAFLD). In the search for natural compounds for the prevention of NAFLD, a series of β-carboline alkaloid derivatives, inspired by flazin and its derivative, newly identified in Crassostrea gigas Thunberg. extracts, were examined for LDA inhibition (LDAI) activity in oleic acid-loaded hepatocytes (HepG2). Eight compounds with a piperidine or pyridine C-ring were chemically synthesized (1-8). Among them, compounds 2 and 4 (flazin) with a carboxy group at C-3 and furfuryl alcohol moiety at C-1 showed low cytotoxicity and they exhibited significant LDAI activity. Compound 2 with piperidine C-ring was identified for the first time in C. gigas extract, and ameliorated the lipid accumulation with the LDAI value of 25.4%. Active compounds 2 and 4 significantly inhibited triacylglycerol species accumulation in cells. These compounds upregulated ATGL and downregulated SREBP1, FASN, and SCD1 genes, suggesting that they activated lipolysis and suppressed lipogenesis, respectively. These results suggest that β-carboline alkaloids, especially compounds 2 and 4, might be potentially useful for preventing NAFLD.
• Flazin as a Lipid Droplet Regulator against Lipid Disorders.

  Xunzhi Wu, Zhen Chen, Yue Wu, Yifan Chen, Jiaping Jia, Nianqiu Shen, Hitoshi Chiba, Shu-Ping Hui

  Nutrients 14 (7) 2022/04/03 

  Lipid disorders are closely related to numerous metabolic diseases, and lipid droplets (LDs) have been considered as a new target for regulating lipid metabolism. Dietary intervention and nutraceuticals provide safe and long-term beneficial effects for treating metabolic diseases. Flazin is a diet-derived bioactive constituent mainly existing in fermented foods, of which the lipid metabolism improvement function has not been studied. In this study, the effect of flazin on lipid regulation at both cell level and organelle level was investigated. Lipidomic profiling showed that flazin significantly decreased cellular triglyceride (TG) by 12.0-22.4% compared with modeling groups and improved the TG and free fatty acid profile. LD staining revealed that flazin efficiently reduced both cellular neutral lipid content by 17.4-53.9% and LD size by 10.0-35.3%. Furthermore, nanoelectrospray ionization mass spectrometry analysis proved that flazin exhibited a preferential suppression of LD TG and regulated LD morphology, including a size decrease and surface property improvement. An evaluation of related gene expression suggested the mechanism to be lipolysis promotion and lipogenesis inhibition. These findings indicated that flazin might be an LD regulator for reversing lipid metabolism disturbance. Moreover, the strategy proposed in this study may contribute to developing other nutraceuticals for treating lipid disorder-related metabolic diseases.
• Effect of charcoal grilling on polycyclic aromatic hydrocarbons (PAHs): content, composition, and health risk in edible fish in Japan.

  Zijun Gao, Zhen Chen, Shu-Ping Hui

  Analytical sciences : the international journal of the Japan Society for Analytical Chemistry 38 (3) 515 - 523 2022/03 

  The content and composition of polycyclic aromatic hydrocarbons (PAHs) induced by charcoal grilling of fish were determined in this study. Using HPLC-DAD, the simultaneously quantitative method for 16 priority PAHs was developed and applied to three common Japanese fish: mackerel, pacific saury, and sardine. Charcoal grilling largely increased the content of both total and representative PAHs. Moreover, all of the three fish showed a similar PAH composition under the effect of charcoal grilling: in raw samples, naphthalene was observed as the dominant PAH, while in charcoal-grilled samples, phenanthrene, fluoranthene, and pyrene served as the major PAHs. Furthermore, a health risk assessment showed that charcoal grilling resulted in high levels of the toxic equivalence quotients, the daily dietary intake exposure to PAHs, and the incremental lifetime cancer risk in three fish, which outclassed the raw samples and exceeded the recommended limitations. The results suggested that charcoal grilling-induced accumulation of PAHs and their potential health risk on human health should be of great concern, which might contribute to the dietary guidance and risk management of food-contained PAHs.
• LC/MS analysis of storage-induced plasmalogen loss in ready-to-eat fish.

  Zhen Chen, Jiaping Jia, Yue Wu, Hitoshi Chiba, Shu-Ping Hui

  Food chemistry 383 132320 - 132320 2022/02/02 

  Plasmalogens are functional and oxidation-sensitive phospholipids abundant in fish. Chilling and freezing are common storage methods for maintaining the quality of fish, but their effect on plasmalogen preservation has not been studied. Therefore, plasmalogen loss in ready-to-eat tuna meat during storage under different conditions was investigated. LC/MS was used to analyze the time- and temperature-dependent changes of plasmalogens, which was the most evident for the species with an ethanolamine headgroup and polyunsaturated fatty acyl chains. Moreover, a series of oxidized plasmalogen molecules were identified, and their storage-induced accumulation was observed. Plasmalogen loss was strongly correlated with total lipid oxidation and phospholipid degradation. Repeated freeze-thaw cycles were found to accelerate the loss of plasmalogens, whereas the different thawing methods did not. The present study provides a deeper understanding of changes in lipid nutrients from fish meat during storage and demonstrates the importance of using advanced strategies to maintain food quality.
• Lysophosphatidylethanolamine Affects Lipid Accumulation and Metabolism in a Human Liver-Derived Cell Line.

  Yusuke Yamamoto, Toshihiro Sakurai, Zhen Chen, Nao Inoue, Hitoshi Chiba, Shu-Ping Hui

  Nutrients 14 (3) 2022/01/28 

  The physiological functions of lysophosphatidylethanolamine (lysoPE) have not been fully elucidated. In this study, the effects of lysoPE on lipogenesis and lipolysis were investigated in a cultured human liver-derived cell line. The intracellular lipid profile was investigated in detail using liquid chromatography-tandem mass spectrometry (LC-MS/MS) to better understand the underlying mechanism. The expression of genes related to lipid metabolism and catabolism was analyzed using real-time PCR. LysoPE supplementation induced cellular lipid droplet formation and altered triacylglycerol (TAG) profiles. Furthermore, lysoPE downregulated expression of the TAG hydrolyzation regulation factor ATGL, and reduced the expression of fatty acid biosynthesis-related genes SREBP1 and SCD1. LC-MS/MS-based lipidomic profiling revealed that the addition of lysoPE 18:2 increased the PE species containing linoleic acyl, as well as the CE 18:2 species, likely due to the incorporation of linoleic acyl from lysoPE 18:2. Collectively, these findings suggest that lysoPE 18:2 is involved in lipid droplet formation by suppressing lipolysis and fatty acid biosynthesis. Thus, lysoPE might play a pathological role in the induction of fatty liver disease.
• Simple and Sensitive Method for the Quantitative Determination of Lipid Hydroperoxides by Liquid Chromatography/Mass Spectrometry.

  Chongsheng Liang, Siddabasave Gowda B Gowda, Divyavani Gowda, Toshihiro Sakurai, Iku Sazaki, Hitoshi Chiba, Shu-Ping Hui

  Antioxidants (Basel, Switzerland) 11 (2) 2022/01/25 

  Lipid hydroperoxides (LOOH) are the initial products of the peroxidation of unsaturated lipids and play a crucial role in lipid oxidation due to their ability to decompose into free radicals and cause adverse effects on human health. Thus, LOOHs are commonly considered biomarkers of oxidative stress-associated pathological conditions. Despite their importance, the sensitive and selective analytical method for determination is limited, due to their low abundance, poor stability, and low ionizing efficiency. To overcome these limitations, in this study, we chemically synthesized eight fatty acid hydroperoxides (FAOOH), including FA 18:1-OOH, FA 18:2-OOH, FA 18:3-OOH, FA 20:4-OOH, FA 20:5-OOH, FA 22:1-OOH, FA 22:6-OOH as analytes, and FA 19:1-OOH as internal standard. Then, they were chemically labeled with 2-methoxypropene (2-MxP) to obtain FAOOMxP by one-step derivatization (for 10 min). A selected reaction monitoring assisted targeted analytical method was developed using liquid chromatography/tandem mass spectrometry (LC-MS/MS). The MxP-labelling improved the stability and enhanced the ionization efficiency in positive mode. Application of reverse-phase chromatography allowed coelution of analytes and internal standards with a short analysis time of 6 min. The limit of detection and quantification for FAOOH ranged from 0.1-1 pmol/µL and 1-2.5 pmol/µL, respectively. The method was applied to profile total FAOOHs in chemically oxidized human serum samples (n = 5) and their fractions of low and high-density lipoproteins (n = 4). The linoleic acid hydroperoxide (FA 18:2-OOH) and oleic acid hydroperoxide (FA 18:1-OOH) were the most abundant FAOOHs in human serum and lipoproteins. Overall, our validated LC-MS/MS methodology features enhanced detection and rapid separation that enables facile quantitation of multiple FAOOHs, therefore providing a valuable tool for determining the level of lipid peroxidation with potential diagnostic applications.
• Adverse Effects of Chrysene on Human Hepatocytes via Inducement of Oxidative Stress and Dysregulation of Xenobiotic Metabolism

  Zijian Zhu, Zhen Chen, Toshihiro Sakurai, Hitoshi Chiba, Shu-Ping Hui

  POLYCYCLIC AROMATIC COMPOUNDS 1040-6638 2021/12 

  Chrysene is a four-ring polycyclic aromatic hydrocarbon, which has been demonstrated to induce adverse effects through the inducement of oxidative stress. However, the possible mechanisms and potential pathways of chrysene-induced oxidative damages are scarcely investigated. In the present study, oxidative stress and xenobiotic metabolism disorder were investigated in human hepatocytes. Chrysene could cause cell viability reduction and cellular reactive oxygen species accumulation in a dose-dependent manner from 10 to 400 ng/mL. Moreover, the mRNA expression detoxification phases, including phase I and phase II as xenobiotic metabolizing enzymes, along with phase III, were investigated by real-time PCR. The results showed that chrysene upregulated the phase I enzymes cytochrome P-450 1A1, 1A2, and epoxide hydrolase 1, while at the same time downregulated the phase II enzymes glutathione S-transferase alpha 1, NAD(P)H: quinone oxidoreductase 1, and the phase III transporters multiple drug resistant 1 and multidrug resistance-associated protein 2. Furthermore, the inflammatory markers tumor necrosis factor alpha, interleukin-6, interleukin-8, and cyclooxygenase-2 were increased, whereas the antioxidant transcription factor nuclear factor erythroid 2-related factor 2 and its regulated enzymes glutathione peroxidase1, heme oxygenase-1, and superoxide dismutase 2 were decreased. These data suggested that chrysene expressed oxidative stress to hepatocytes, resulting in cytotoxicity and inflammation, which might be due to the damage of the antioxidant system and xenobiotic metabolism.
• Oxidative Stress Linked Organ Lipid Hydroperoxidation and Dysregulation in Mouse Model of Nonalcoholic Steatohepatitis: Revealed by Lipidomic Profiling of Liver and Kidney.

  Yue Wu, Zhen Chen, Hirotoshi Fuda, Takayuki Tsukui, Xunzhi Wu, Nianqiu Shen, Natsuki Saito, Hitoshi Chiba, Shu-Ping Hui

  Antioxidants (Basel, Switzerland) 10 (10) 2021/10/12 

  Nonalcoholic steatohepatitis (NASH) is a prevalent disease related to lipid metabolism disorder and oxidative stress. Lipid hydroperoxidation is known to be a critical driving force of various disorders and diseases. However, the combination of both intact and hydroperoxidized lipids in NASH has not yet been studied. In this work, the liver and kidney samples from NASH-model mice were comprehensively investigated by using the LC/MS-based lipidomic analysis. As a result, triglycerides showed the amount accumulation and the profile alteration for the intact lipids in the NASH group, while phosphatidylethanolamines, lysophosphatidylethanolamines, plasmalogens, and cardiolipins largely depleted, suggesting biomembrane damage and mitochondria dysfunction. Notably, the lipid hydroperoxide species of triglyceride and phosphatidylcholine exhibited a significant elevation in both the liver and the kidney of the NASH group and showed considerable diagnostic ability. Furthermore, the relationship was revealed between the lipid metabolism disturbance and the lipid hydroperoxide accumulation, which played a key role in the vicious circle of NASH. The present study suggested that the omics approach to the lipid hydroperoxide profile might be the potential diagnostic marker of NASH and other oxidative stress-related diseases, as well as the evaluative treatment index of antioxidants.
• Defining the kinetic effects of infection with influenza virus A/PR8/34 (H1N1) on sphingosine-1-phosphate signaling in mice by targeted LC/MS.

  Divyavani Gowda, Marumi Ohno, Siddabasave Gowda B Gowda, Hitoshi Chiba, Masashi Shingai, Hiroshi Kida, Shu-Ping Hui

  Scientific reports 11 (1) 20161 - 20161 2021/10/11 

  Influenza remains a world-wide health concern, causing 290,000-600,000 deaths and up to 5 million cases of severe illnesses annually. Noticing the host factors that control biological responses, such as inflammatory cytokine secretion, to influenza virus infection is important for the development of novel drugs. Sphingosine-1-phosphate (S1P) is a bioactive sphingolipid metabolite and has essential biological functions in inflammation. However, the kinetic effects of influenza virus infection on physiological S1P levels and their signaling in multiple tissues remain unknown. In this study, we utilized a mouse model intranasally infected with 50 or 500 plaque forming units (PFU) of A/Puerto Rico/8/34 (H1N1; PR8) virus to investigate how S1P levels and expression of its regulating factors are affected by influenza virus infection by the liquid-chromatography/mass spectrometry and real-time PCR, respectively. The S1P level was significantly high in the plasma of mice infected with 500 PFU of the virus than that in control mice at 6 day-post-infection (dpi). Elevated gene expression of sphingosine kinase-1 (Sphk1), an S1P synthase, was observed in the liver, lung, white adipose tissue, heart, and aorta of infected mice. This could be responsible for the increased plasma S1P levels as well as the decrease in the hepatic S1P lyase (Sgpl1) gene in the infected mice. These results indicate modulation of S1P-signaling by influenza virus infection. Since S1P regulates inflammation and leukocyte migration, it must be worth trying to target this signaling to control influenza-associated symptoms.
• Postpartum cows showed high oocyte triacylglycerols concurrently with high plasma free fatty acids.

  Eri Furukawa, Zhen Chen, Hiroki Ueshiba, Yue Wu, Hitoshi Chiba, Yojiro Yanagawa, Seiji Katagiri, Masashi Nagano, Shu-Ping Hui

  Theriogenology 176 174 - 182 0093-691X 2021/10/02 

  Impaired oocyte quality is one of the main causes of low fertility in modern high-yielding dairy cows. One of the potential factors of the impaired oocyte quality is the effects of free fatty acids (FFA). In fact, high FFA supplementation to culture media exacerbated oocyte developmental competence in vitro. Meanwhile, artificially induced high blood FFA levels in heifers did not affect the lipid composition of oocytes in vivo; however, the oocyte lipid profile of postpartum cows has not yet been investigated. Therefore, the profile of lipids involved in energy metabolism, including FFA and triacylglycerols (TAG), and their relationship between plasma and oocytes were compared among cows at different lactation stages. Heifers were used as a control group that was not affected by lactation. Plasma and oocytes were collected from heifers (n = 4) and 14 Holstein cows categorized to the early lactation stage: 25-47 days in milk (DIM) (n = 6), peak lactation stage: 61-65 DIM (n = 4), and middle lactation stage: 160-202 DIM (n = 4). The FFA and TAG profiles of plasma and oocytes were examined by liquid chromatography mass spectrometry. Plasma FFA positively correlated with oocyte TAG (P < 0.05). Plasma FFA and oocyte TAG were significantly higher in cows in the early lactation stage than in heifers (P < 0.05), while the peak and middle lactation stage groups had intermediate levels. The proportion of oleic acid in plasma increased concurrently with elevations in total FFA, while the compositions of oocyte FFA and TAG fatty acyls were constant regardless of plasma FFA concentration or oocyte TAG content. The present results suggest that high postpartum plasma FFA concentrations affect the quantity of oocyte TAG. Taken together with the adverse effects of high FFA concentrations on oocyte developmental competence in vitro, oocyte quality in postpartum cows may be impaired due to high circulating FFA concentrations. These results provide a more detailed understanding of the effects of postpartum high circulating FFA concentrations on the low fertility of cows.
• Serum 25-hydroxyvitamin D3 Levels and Diabetes in a Japanese Population: The DOSANCO Health Study.

  Koshi Nakamura, Shu-Ping Hui, Shigekazu Ukawa, Emiko Okada, Takafumi Nakagawa, Akihiro Imae, Hiroaki Okabe, Zhen Chen, Yusuke Miura, Hitoshi Chiba, Akiko Tamakoshi

  Journal of epidemiology 33 (1) 31 - 37 2021/09/28 

  BACKGROUND: Both decreased insulin sensitivity and impaired insulin secretion are common in Asian populations with diabetes, in contrast to Western populations. There is limited evidence regarding the association between insulin response in diabetes in Asian populations and serum 25-hydroxyvitamin D3 (25[OH]D3) insufficiency. METHODS: The present cross-sectional study compared the prevalence of diabetes, defined as a fasting plasma glucose level ≥126 mg/dL and/or a HbA1c level ≥6.5%, among 480 participants aged 35-79 years not taking anti-diabetes medications, based on serum 25(OH)D3 levels. A logistic regression model was used to calculate the odds ratios for diabetes in each serum 25(OH)D3 group. Furthermore, this study examined the association between serum 25(OH)D3 levels and the index of homeostasis model assessment of insulin resistance (HOMA-IR) using a linear regression model. RESULTS: The prevalence of diabetes was 7.29% in the study population, and was higher in lower serum 25(OH)D3 quartile groups. The odds ratios for diabetes in the first, second, and third serum 25(OH)D3 quartile groups (25[OH]D3: ≤18.10, 18.11-22.90, and 22.91-28.17 ng/mL) were 4.02 (95% confidence interval [CI], 1.25-12.92), 2.50 (95% CI, 0.77-8.10), and 1.91 (95% CI, 0.60-6.09), respectively, with the fourth quartile group (≽28.18 ng/mL) serving as the reference group, after adjusting for sociodemographic, lifestyle, physical and environmental factors. Serum 25(OH)D3 levels showed an inverse association with log-transformed HOMA-IR after adjusting for similar factors (standardized β = -0.08; 95% CI, -0.14 to -0.02). CONCLUSIONS: Serum 25(OH)D3 levels were inversely associated with diabetes prevalence in a general Japanese population, with a slight inverse association between serum 25(OH)D3 levels and HOMA-IR.
• Comparison of dimension reduction methods on fatty acids food source study.

  Yifan Chen, Yusuke Miura, Toshihiro Sakurai, Zhen Chen, Rojeet Shrestha, Sota Kato, Emiko Okada, Shigekazu Ukawa, Takafumi Nakagawa, Koshi Nakamura, Akiko Tamakoshi, Hitoshi Chiba, Hideyuki Imai, Hiroyuki Minami, Masahiro Mizuta, Shu-Ping Hui

  Scientific reports 11 (1) 18748 - 18748 2021/09/21 

  Serum fatty acids (FAs) exist in the four lipid fractions of triglycerides (TGs), phospholipids (PLs), cholesteryl esters (CEs) and free fatty acids (FFAs). Total fatty acids (TFAs) indicate the sum of FAs in them. In this study, four statistical analysis methods, which are independent component analysis (ICA), factor analysis, common principal component analysis (CPCA) and principal component analysis (PCA), were conducted to uncover food sources of FAs among the four lipid fractions (CE, FFA, and TG + PL). Among the methods, ICA provided the most suggestive results. To distinguish the animal fat intake from endogenous fatty acids, FFA variables in ICA and factor analysis were studied. ICA provided more distinct suggestions of FA food sources (endogenous, plant oil intake, animal fat intake, and fish oil intake) than factor analysis. Moreover, ICA was discovered as a new approach to distinguish animal FAs from endogenous FAs, which will have an impact on epidemiological studies. In addition, the correlation coefficients between a published dataset of food FA compositions and the loading values obtained in the present ICA study suggested specific foods as serum FA sources. In conclusion, we found that ICA is a useful tool to uncover food sources of serum FAs.
• HPLC with spectrophotometric or mass spectrometric detection for quantifying very-long chain fatty acids in human plasma and its association with cardiac risk factors.

  Rojeet Shrestha, Zhen Chen, Zijun Gao, Yifan Chen, Emiko Okada, Shigekazu Ukawa, Takafumi Nakagawa, Koshi Nakamura, Akiko Tamakoshi, Hitoshi Chiba, Shu-Ping Hui

  Annals of clinical biochemistry 58 (5) 400 - 410 0004-5632 2021/09 

  BACKGROUND: We developed and compared two liquid chromatography methods, one with UV/Visible spectrophotometric detection (HPLC) and the other with mass spectrometric detection (LC-MS), for quantifying very-long chain fatty acids (VLCFA) in human plasma. Association of VLCFA with various cardiovascular risk factors were evaluated. METHOD: Fasting blood samples were collected from 541 human volunteers (242 men and 299 women; mean age ±SD, 58.9 ± 12.4 years), including 429 and 112 individuals with and without hypertriglyceridemia, respectively. Esterified VLCFA were saponified and derivatized with 2-nitrophenylhydrazine. Separation of VLCFA species was achieved with C4 Mightysil column (HPLC) and Ascentis Express Phenyl-Hexyl column (LC-MS) followed by spectrophotometric and selected-reaction monitoring mode of mass spectrometric detection, respectively. RESULTS: The HPLC assay of VLCFA was precise with intra-assay imprecision of 2.5% to 6.9% and inter-assay imprecision of 3.2% to 9.5%. Moreover, there was an excellent correlation (r > 0.96) between HPLC and LC-MS methods. The 95 percentile reference intervals (RI; upper limit) of VLCFA were determined to be 41.3 µmol/L in healthy volunteers. Plasma VLCFA were significantly correlated with triglycerides (Spearman's ρ = 0.306, P < 0.001) and total cholesterol (Spearman's ρ = 0.251, P < 0.001). All species of VLCFA were significantly elevated in hypertriglyceridaemic individuals compared with control. CONCLUSION: We established LC-based assays of VLCFA with either spectrophotometry or mass spectrometry as a detection system. Hypertriglyceridaemia is significantly associated with elevated concentration of each species of VLCFA.
• Sphingosine-1-phosphate interactions in the spleen and heart reflect extent of cardiac repair in mice and failing human hearts.

  SiddabasaveGowda B Gowda, Divyavani Gowda, Vasundhara Kain, Hitoshi Chiba, Shu-Ping Hui, Charles E Chalfant, Vibhu Parcha, Pankaj Arora, Ganesh V Halade

  American journal of physiology. Heart and circulatory physiology 321 (3) H599-H611  2021/09/01 

  Sphingosine-1-phosphate (S1P) is a bioactive mediator in inflammation. Dysregulated S1P is demonstrated as a cause of heart failure (HF). However, the time-dependent and integrative role of S1P interaction with receptors in HF is unclear after myocardial infarction (MI). In this study, the sphingolipid mediators were quantified in ischemic human hearts. We also measured the time kinetics of these mediators post-MI in murine spleen and heart as an integrative approach to understand the interaction of S1P and respective S1P receptors in the transition of acute (AHF) to chronic HF (CHF). Risk-free 8-12 wk male C57BL/6 mice were subjected to MI surgery, and MI was confirmed by echocardiography and histology. Mass spectrometry was used to quantify sphingolipids in plasma, infarcted heart, spleen of mice, and ischemic and healthy human heart. The physiological cardiac repair was observed in mice with a notable increase of S1P quantity (pmol/g) in the heart and spleen significantly reduced in patients with ischemic HF. The circulating murine S1P levels were increased during AHF and CHF despite lowered substrate in CHF. The S1PR1 receptor expression was observed to coincide with the respective S1P quantity in mice and human hearts. Furthermore, selective S1P1 agonist limited inflammatory markers CCL2 and TNF-α and accelerated reparative markers ARG-1 and YM-1 in macrophages in the presence of Kdo2-Lipid A (KLA; potent inflammatory stimulant). This report demonstrated the importance of S1P/S1PR1 signaling in physiological inflammation during cardiac repair in mice. Alteration in these axes may serve as the signs of pathological remodeling in patients with ischemia.NEW & NOTEWORTHY Previous studies indicate that sphingosine-1-phosphate (S1P) has some role in cardiovascular disease. This study adds quantitative and integrative systems-based approaches that are necessary for discovery and bedside translation. Here, we quantitated sphinganine, sphingosine, sphingosine-1-phosphate (S1P) in mice and human cardiac pathobiology. Interorgan S1P quantity and respective systems-based receptor activation suggest cardiac repair after myocardial infarction. Thus, S1P serves as a therapeutic target for cardiac protection in clinical translation.
• Effect of charcoal grilling on polycyclic aromatic hydrocarbons (PAHs): content, composition, and health risk in edible fish in Japan.

  Zijun Gao, Zhen Chen, Shu-Ping Hui

  Analytical sciences : the international journal of the Japan Society for Analytical Chemistry 2021/08/20
• Detection and Structural Characterization of SFAHFA Homologous Series in Mouse Colon Contents by LTQ-Orbitrap-MS and Their Implication in Influenza Virus Infection.

  Siddabasave Gowda B Gowda, Divyavani Gowda, Marumi Ohno, Chongsheng Liang, Hitoshi Chiba, Shu-Ping Hui

  Journal of the American Society for Mass Spectrometry 32 (8) 2196 - 2205 2021/08/04 

  Fatty acid esters of hydroxy fatty acids (FAHFAs) are a new class of endogenous lipids with promising physiological functions in mammals. We previously introduced a new type of lipids to this family called short-chain fatty acid esters of hydroxy fatty acids (SFAHFAs), branching specific to the C2 carbon of a long-chain fatty acid (≥C20). In this study, we discovered a homologous series of SFAHFAs comprising C16-C26 hydroxy fatty acids esterified with short-chain fatty acids (C2-C5) in mouse colon contents. The detected SFAHFAs were characterized by high-resolution mass spectrometry with MSn analysis. The double-bond position of monounsaturated SFAHFAs was determined by the epoxidation reaction of samples with m-chloroperoxybenzoic acid and their MSn analysis. Further, the measurement of SFAHFA concentration in the colon contents of mice infected with influenza A/Puerto Rico/8/34 (H1N1; PR8) virus revealed a significant increase in their levels compared to native control. A strong correlation was observed between hydroxy fatty acid and SFAHFAs. Detection, characterization, and profiling of these new SFAHFA levels in relation with pandemic H1N1; PR8 influenza virus will contribute to the in-depth study of their function and metabolism.
• Docosahexaenoic Acid Esters of Hydroxy Fatty Acid Is a Novel Activator of NRF2.

  Siddabasave Gowda B Gowda, Takayuki Tsukui, Hirotoshi Fuda, Yusuke Minami, Divyavani Gowda, Hitoshi Chiba, Shu-Ping Hui

  International journal of molecular sciences 22 (14) 2021/07/15 

  Fatty acid esters of hydroxy fatty acids (FAHFAs) are a new class of endogenous lipids with interesting physiological functions in mammals. Despite their structural diversity and links with nuclear factor erythroid 2-related factor 2 (NRF2) biosynthesis, FAHFAs are less explored as NRF2 activators. Herein, we examined for the first time the synthetic docosahexaenoic acid esters of 12-hydroxy stearic acid (12-DHAHSA) or oleic acid (12-DHAHOA) against NRF2 activation in cultured human hepatoma-derived cells (C3A). The effect of DHA-derived FAHFAs on lipid metabolism was explored by the nontargeted lipidomic analysis using liquid chromatography-mass spectrometry. Furthermore, their action on lipid droplet (LD) oxidation was investigated by the fluorescence imaging technique. The DHA-derived FAHFAs showed less cytotoxicity compared to their native fatty acids and activated the NRF2 in a dose-dependent pattern. Treatment of 12-DHAHOA with C3A cells upregulated the cellular triacylglycerol levels by 17-fold compared to the untreated group. Fluorescence imaging analysis also revealed the suppression of the degree of LDs oxidation upon treatment with 12-DHAHSA. Overall, these results suggest that DHA-derived FAHFAs as novel and potent activators of NRF2 with plausible antioxidant function.
• Lipidomic analysis of non-esterified furan fatty acids and fatty acid compositions in dietary shellfish and salmon by UHPLC/LTQ-Orbitrap-MS.

  Siddabasave Gowda B Gowda, Yusuke Minami, Divyavani Gowda, Daisuke Furuko, Hitoshi Chiba, Shu-Ping Hui

  Food research international (Ottawa, Ont.) 144 110325 - 110325 2021/06 

  Lipids such as furan fatty acids (F-acids) are the valuable minor bioactive components of food such as fatty fish and plants. They are reported to have positive health benefits, including antioxidant and anti-inflammatory activities. Despite their importance, limited studies are focusing on F-acid determination in dietary seafood. This study aimed to identify and profile non-esterified F-acids and free fatty acids in total lipid extract of seafood such as shellfish and salmon. The lipidomic analysis using liquid chromatography-linear trap quadrupole-orbitrap mass spectrometry led to identifying seven types of free F-acids in shellfish (n = 5) and salmon (n = 4). The identified F-acids were confirmed by their high-resolution masses and acquired mass spectra. The relative concentrations of F-acids in shellfish range from 0.01 to 10.93 mg/100 g of the fillet, and in salmon, 0.01 to 14.21 mg/100 g of the fillet. The results revealed the highest abundance of F-acids in Sakhalin surf clam, Japanese scallop, and a fatty salmon trout. Besides, relative levels of saturated, monounsaturated, and polyunsaturated fatty acids (PUFAs) in these seafoods were compared with each other, suggesting basket clams and salmon trout to have significantly higher levels of PUFAs. The dietary seafoods enriched with F-acids and PUFAs may have possible health benefits. Hence, the applied technique could be a promising tool for rapid detection and analysis of non-esterified fatty acids in food.
• The Phenolic Antioxidant 3,5-dihydroxy-4-methoxybenzyl Alcohol (DHMBA) Prevents Enterocyte Cell Death under Oxygen-Dissolving Cold Conditions through Polyphyletic Antioxidant Actions.

  Moto Fukai, Takuya Nakayabu, Shintaro Ohtani, Kengo Shibata, Shingo Shimada, Soudai Sakamoto, Hirotoshi Fuda, Takayuki Furukawa, Mitsugu Watanabe, Shu-Ping Hui, Hitoshi Chiba, Tsuyoshi Shimamura, Akinobu Taketomi

  Journal of clinical medicine 10 (9) 2021/05/04 

  Cold preservation in University of Wisconsin (UW) solution is not enough to maintain the viability of the small intestine, due to the oxidative stress. The novel phenolic antioxidant 3,5-dihydroxy-4-methoxybenzyl alcohol (DHMBA) has dual properties to reduce oxidative stress, radical scavenging, and antioxidant protein induction, in other cells. This study was designed to determine whether DHMBA reduces cold preservation injury of enterocytes, and to identify the effector site. Enterocytes were subjected to 48-h cold preservation under atmosphere in UW solution (±DHMBA), and then returned to normal culture to replicate reperfusion of the small intestine after cold preservation. At the end of cold preservation (ECP) and at 1, 3, 6, and 72 h after rewarming (R1h, R3h, R6h, and R72h), we evaluated cell function and the injury mechanism. The results showed that DHMBA protected mitochondrial function mainly during cold preservation, and suppressed cell death after rewarming, as shown by the MTT, ATP, mitochondrial membrane potential, LDH, and lipid peroxidation assays, together with enhanced survival signals (PI3K, Akt, p70S6K) and induction of antioxidant proteins (HO-1, NQO-1, TRX-1). We found that DHMBA mitigates the cold-induced injury of enterocytes by protecting the mitochondria through direct and indirect antioxidative activities.
• Branched-chain amino acids and l-carnitine attenuate lipotoxic hepatocellular damage in rat cirrhotic liver.

  Yasuyuki Tamai, Zhen Chen, Yue Wu, Jun Okabe, Yoshinao Kobayashi, Hitoshi Chiba, Shu-Ping Hui, Akiko Eguchi, Motoh Iwasa, Masaaki Ito, Yoshiyuki Takei

  Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie 135 111181 - 111181 2021/03 

  Branched-chain amino acids (BCAA) reverse malnutrition and l-carnitine leads to the reduction of hyperammonemia and muscle cramps in cirrhotic patients. BCAA and l-carnitine are involved in glucose and fatty acid metabolism, however their mechanistic activity in cirrhotic liver is not fully understood. We aim to define the molecular mechanism(s) and combined effects of BCAA and l-carnitine using a cirrhotic rat model. Rats were administered carbon tetrachloride for 10 weeks to induce cirrhosis. During the last 6 weeks of administration, cirrhotic rats received BCAA, l-carnitine or a combination of BCAA and l-carnitine daily via gavage. We found that BCAA and l-carnitine treatments significantly improved hepatocellular function associated with reduced triglyceride level, lipid deposition and adipophilin expression, in cirrhotic liver. Lipidomic analysis revealed dynamic changes in hepatic lipid composition by BCAA and l-carnitine administrations. BCAA and l-carnitine globally increased molecular species of phosphatidylcholine. Liver triacylglycerol and phosphatidylcholine hydroperoxides were significantly decreased by BCAA and l-carnitine. Furthermore, serum and liver ATP levels were significantly increased in all treatments, which were attributed to the elevation of mature cardiolipins and mitochondrial component gene expressions. Finally, BCAA and l-carnitine dramatically reduced hepatocellular death. In conclusion, BCAA and l-carnitine treatments attenuate hepatocellular damage through the reduction of lipid peroxides and the overall maintenance of mitochondrial integrity within the cirrhotic liver. These effectiveness of BCAA and l-carnitine support the therapeutic strategies in human chronic liver diseases.
• A mouse model of short-term, diet-induced fatty liver with abnormal cardiolipin remodeling via downregulated Tafazzin gene expression.

  Toshihiro Sakurai, Zhen Chen, Arisa Yamahata, Takahiro Hayasaka, Hiroshi Satoh, Hirotaka Sekiguchi, Hitoshi Chiba, Shu-Ping Hui

  Journal of the science of food and agriculture 101 (12) 4995 - 5001 2021/02/05 

  BACKGROUND: Cardiolipin (CL) helps maintain mitochondrial structure and function. Here we investigated whether a high carbohydrate diet (HCD) fed to mice for a short period (5 days) could modulate the CL level, including that of monolysoCL (MLCL) in the liver. RESULTS: Total CL in the HCD group was 22% lower than that in the normal chow diet (NCD) group (P < 0.05). The CL72:8 level strikingly decreased by 93% (P < 0.0001), whereas total nascent CLs (CLs other than CL72:8) increased (P < 0.01) in the HCD group. The total MLCL in the HCD group increased by 2.4-fold compared with that in the NCD group (P < 0.05). Tafazzin expression in the HCD group was significantly downregulated compared with that in the NCD group (P < 0.05). A strong positive correlation between nascent CL and total MLCL (r = 0.955, P < 0.0001), and a negative correlation between MLCL and Tafazzin expression (r = -0.593, P = 0.0883) were observed. CONCLUSION: A HCD modulated the fatty acid composition of CL and MLCL via Tafazzin in the liver, which could lead to mitochondrial dysfunction. This model may be useful for elucidating the relationship between fatty liver and mitochondrial dysfunction. © 2021 Society of Chemical Industry.
• Quantitative and Comparative Investigation of Plasmalogen Species in Daily Foodstuffs.

  Yue Wu, Zhen Chen, Jiaping Jia, Hitoshi Chiba, Shu-Ping Hui

  Foods (Basel, Switzerland) 10 (1) 2021/01/08 

  Plasmalogens are an animal-derived functional phospholipid increasingly known as a safe and effective nutritional ingredient, however, the quantitation and comparison of plasmalogen species in foods is limited. In the present work, determination methods for dietary plasmalogens using liquid chromatography-tandem mass spectroscopy under positive and negative ionization modes were compared. The negative-mode method, which showed better selectivity, sensitivity, and accuracy, was then applied in 14 kinds of livestock, poultry, and seafood samples. Livestock and poultry showed abundant total plasmalogen (530.83-944.94 nmol/g), higher than fish (46.08-399.75 nmol/g) and mollusk (10.00-384.76 nmol/g). While fish and mollusk samples expressed healthier fatty acyl composition, with higher eicosapentaenoyl and more beneficial n-6/n-3 ratio than the land animal meats, especially for squid and octopus, with eicosapentaenoyl of 98.4% and 94.5%, respectively. The correlations among plasmalogen species varied in different foodstuffs with distinguishing patterns, suggesting the customizable strategies for achieving targeted plasmalogen species. These findings not only provided fundamental comparison of plasmalogen among daily foodstuffs, but also contributed to extend the dietary plasmalogen sources for health food development.
• Analysis of serum lysophosphatidylethanolamine levels in patients with non-alcoholic fatty liver disease by liquid chromatography-tandem mass spectrometry.

  Yusuke Yamamoto, Toshihiro Sakurai, Zhen Chen, Takayuki Furukawa, Siddabasave Gowda B Gowda, Yue Wu, Kazuhiro Nouso, Yuki Fujii, Yuki Yoshikawa, Hitoshi Chiba, Shu-Ping Hui

  Analytical and bioanalytical chemistry 413 (1) 245 - 254 2021/01 

  Lysophosphatidylethanolamines (LysoPEs) are the partial hydrolysis products of phosphatidylethanolamine. Despite the unique in vitro bioactivities of LysoPEs, there are limited reports on the pathophysiological role of LysoPEs in the serum, due to the lack of sensitive analytical methods for determination of each molecular species in clinical samples. Herein, we developed a highly sensitive quantitative method to profile the serum LysoPE species by liquid chromatography-tandem mass spectrometry (LC-MS/MS) with selected reaction monitoring (SRM). The internal standard (IS), chemically synthesized in-house, and the lineup of seven major LysoPE species were used in this study. The limits of detection and quantification for each LysoPE species ranged within 0.5-3.3 pmol/mL and 1.0-5.0 pmol/mL, respectively. The combined concentrations of LysoPEs in the serum from healthy subjects (n = 8) and the patients with non-alcoholic fatty liver diseases (NAFLD) including simple steatosis (SS, n = 9) and non-alcoholic steatohepatitis (NASH, n = 27) were 18.030 ± 3.832, 4.867 ± 1.852, and 5.497 ± 2.495 nmol/mL, respectively. The combined and individual concentrations of LysoPEs, except for LysoPE 18:0, significantly decreased in the patients with NAFLD compared with those for the healthy subjects. However, no significant difference was observed between the SS and NASH groups. Our proposed LC-MS/MS method is valid and has advantages of small sample volume, high sensitivity, and simultaneous absolute quantitation for multiple molecular species. This method may enable diagnostic evaluation and elucidation of the as-yet uncovered pathophysiological role of LysoPEs.
• Comparison of anthocyanin distribution in berries of Haskap (Lonicera caerulea subsp. edulis (Turcz. ex. Herder) Hultén), Miyama-uguisukagura (Lonicera gracilipes Miq.), and their interspecific hybrid using imaging mass spectrometry.

  Ryohei Fujita, Takahiro Hayasaka, Shigeki Jin, Shu-Ping Hui, Yoichiro Hoshino

  Plant science : an international journal of experimental plant biology 300 110633 - 110633 2020/11 

  Haskap (Lonicera caerulea subsp. edulis), a shrub with violet-blue fruits, is distributed mainly in Hokkaido, Japan. Miyama-uguisukagura (Lonicera gracilipes), a species related to Haskap, produces red fruits. Interspecific hybridization of Miyama-uguisukagura and Haskap was performed to introduce novel characteristics in the resulting hybrids. The shape and color of the interspecific hybrid fruits differed from those of the parent fruits. A comparison of anthocyanin distribution among these three fruit types by imaging mass spectrometry (IMS) revealed the presence of five different anthocyanins. The average cyanidin 3,5-diglucoside and peonidin 3,5-diglucoside intensities in the interspecific hybrid fruit were higher than those of the parent fruits, whereas the average pelargonidin 3-glucoside, cyanidin 3-glucoside, and peonidin 3-glucoside intensities were the highest in Haskap. All anthocyanins were mainly accumulated in the inner and outer skins of Haskap and interspecific hybrid fruits, and in the skin of Miyama-uguisukagura fruits. The order of signal intensities of all anthocyanins among the three fruits was unchanged in different regions. Additionally, a comparison of IMS and LC/MS data from our previous study confirmed the possibility of comparing multiple fruits in the same plate by IMS. Thus, we elucidated anthocyanin distribution patterns of the interspecific hybrid and parent fruits by IMS.
• Low-Density Lipoprotein (LDL)-Triglyceride and Its Ratio to LDL-Cholesterol as Diagnostic Biomarkers for Nonalcoholic Steatohepatitis.

  Yuki Fujii, Kazuhiro Nouso, Hiroshi Matsushita, Kazuya Kariyama, Toshihiro Sakurai, Yuji Takahashi, Hitoshi Chiba, Shu-Ping Hui, Yasuki Ito, Motoko Ohta, Hiroyuki Okada

  The journal of applied laboratory medicine 5 (6) 1206 - 1215 2020/11/01 [Refereed][Not invited]
   

  BACKGROUND: Nonalcoholic fatty liver disease (NAFLD) is the most common type of liver disease, but it is difficult to distinguish its pathogenic phenotype, nonalcoholic steatohepatitis (NASH), from nonalcoholic fatty liver (NAFL) without a liver biopsy. We analyzed serum lipids, including low-density lipoprotein triglyceride (LDL-TG), to elucidate their usefulness for diagnosing NASH. PATIENTS AND METHODS: Serum samples obtained from 35 NASH and 9 NAFL biopsy-confirmed patients and 6 healthy volunteers (HLT) were studied for 13 lipid-related markers and compared between HLT, NAFL, and NASH groups. The relationship between histological findings and the lipid markers was also analyzed. RESULTS: There were significant differences in triglyceride, LDL-TG, the ratio of LDL-TG to the LDL-cholesterol (LDL-TG/LDL-C), small dense LDL-C, and apolipoprotein E between the three groups. Among the 5 lipid components, serum LDL-TG level and the ratio of LDL-TG to the LDL-cholesterol (LDL-TG/LDL-C) were significantly elevated in NASH. The median concentrations of LDL-TG in HLT, NAFL, and NASH were 9, 15, and 20 mg/dL (P < 0.001), and those of LDL-TG/LDL-C were 0.097, 0.102, and 0.173 (P < 0.001), respectively. Although the degree of steatosis was not correlated with the LDL-TG/LDL-C, the ratio was significantly higher in patients with lobular inflammation (P = 0.071), ballooning (P = 0.031), and fibrosis (P < 0.001). The area under the receiver operating characteristic curve of the ratio for distinguishing NASH from NAFL was 0.857. The rest of studied markers showed no significant utility. CONCLUSION: Serum LDL-TG levels and the LDL-TG/LDL-C ratio might serve as simple and noninvasive diagnostic biomarkers for NASH.
• Comprehensive lipidomic profiling in serum and multiple tissues from a mouse model of diabetes.

  Zhen Chen, Qiangrong Liang, Yue Wu, Zijun Gao, Satoru Kobayashi, Joy Patel, Cairong Li, Fei Cai, Youhua Zhang, Chongsheng Liang, Hitoshi Chiba, Shu-Ping Hui

  Metabolomics : Official journal of the Metabolomic Society 16 (11) 115 - 115 2020/10/16 

  INTRODUCTION: Diabetes mellitus is a serious metabolic disorder causing multiple organ damage in human. However, the lipidomic profiles in different organs and their associations are rarely studied in either diabetic patients or animals. OBJECTIVES: To evaluate and compare the characteristics of lipid species in serum and multiple tissues in a diabetic mouse model. METHODS: Semi-quantitative profiling analyses of intact and oxidized lipids were performed in serum and multiple tissues from a diabetic mouse model fed a high fat diet and treated with streptozotocin by using LC/HRMS and MS/MS. The total content of each lipid class, and the tissue-specific lipid species in all tissue samples were determined and compared by multivariate analyses. RESULTS: The diabetic mouse model displayed characteristic differences in serum and multiple organs: the brain and heart showed the largest reduction in cardiolipin, while the kidney had more alterations in triacylglycerol. Interestingly, the lipidomic differences also existed between different regions of the same organ: cardiolipin species with highly polyunsaturated fatty acyls decreased only in atrium but not in ventricle, while renal cortex showed longer fatty acyl chains for both increased and decreased triacylglycerol species than renal medulla. Importantly, diabetes caused an accumulation of lipid hydroperoxides, suggesting that oxidative stress was induced in all organs except for the brain during the development of diabetes. CONCLUSIONS: These findings provided novel insight into the organ-specific relationship between diabetes and lipid metabolism, which might be useful for evaluating not only diabetic tissue injury but also the effectiveness of diabetic treatments.
• Chemical Labeling Assisted Detection and Identification of Short Chain Fatty Acid Esters of Hydroxy Fatty Acid in Rat Colon and Cecum Contents.

  Siddabasave Gowda B Gowda, Divyavani Gowda, Chongsheng Liang, Yonghan Li, Kentaro Kawakami, Satoru Fukiya, Atsushi Yokota, Hitoshi Chiba, Shu-Ping Hui

  Metabolites 10 (10) 2020/10/08 

  Branched fatty acid esters of hydroxy fatty acids (FAHFAs) are novel endogenous lipids with important physiological functions in mammals. We previously identified a new type of FAHFAs, named short-chain fatty acid esterified hydroxy fatty acids (SFAHFAs), with acetyl or propyl esters of hydroxy fatty acids of carbon chains, C ≥ 20. However, sensitive determination of SFAHFAs is still a challenge, due to their high structural similarity and low abundance in biological samples. This study employs one-step chemical derivatization following total lipid extraction using 2-dimethylaminoethylamine (DMED) for enhanced detection of SFAHFAs. The labeled extracts were subjected to ultrahigh performance liquid chromatography coupled to linear ion trap quadrupole-Orbitrap mass spectrometry (UHPLC/LTQ-Orbitrap MS). Our results demonstrated that the detection sensitivities of SFAHFAs increased after DMED labeling, and is highly helpful in discovering six additional novel SFAHFAs in the cecum and colon contents of WKAH/HKmSlc rats fed with normal and high-fat diet (HFD). The identified DMED labeled SFAHFAs were characterized by their detailed MS/MS analysis, and their plausible fragmentation patterns were proposed. The concentrations of SFAHFAs were significantly reduced in the cecum of HFD group compared to the control. Hence, the proposed method could be a promising tool to apply for the enhanced detection of SFAHFAs in various biological matrices, which in turn facilitate the understanding of their sources, and physiological functions of these novel lipids.
• IVIG感受性川崎病患児と抵抗性患児との間にみられる脂質の特徴の違い(Differential lipid characteristics in Kawasaki disease between IVIG sensitive and resistant children)

  陳 震, 齋 秀二, Wu Yue, 千葉 仁志, 惠 淑萍

  臨床化学 49 (Suppl.1) 215 - 215 0370-5633 2020/10
• Identification of short-chain fatty acid esters of hydroxy fatty acids (SFAHFAs) in a murine model by nontargeted analysis using ultra-high-performance liquid chromatography/linear ion trap quadrupole-Orbitrap mass spectrometry.

  Siddabasave Gowda B Gowda, Chongsheng Liang, Divyavani Gowda, Fengjue Hou, Kentaro Kawakami, Satoru Fukiya, Atsushi Yokota, Hitoshi Chiba, Shu-Ping Hui

  Rapid communications in mass spectrometry : RCM 34 (17) e8831  2020/09/15 [Refereed][Not invited]
   

  RATIONALE: Fatty acid esters of hydroxy fatty acids (FAHFAs) are recently discovered endogenous lipids with outstanding health benefits. FAHFAs are known to exhibit antioxidant, antidiabetic and anti-inflammatory properties. The number of known long-chain FAHFAs in mammalian tissues and dietary resources increased recently because of the latest developments in high-resolution tandem mass spectrometry techniques. However, there are no reports on the identification of short-chain fatty acid esterified hydroxy fatty acids (SFAHFAs). METHODS: Intestinal contents, tissues, and plasma of rats fed with high-fat diet (HFD) and normal diet (ND) were analyzed for fatty acids, hydroxy fatty acids, and FAHFAs using ultra-high-performance liquid chromatography (UHPLC) and linear trap quadrupole-Orbitrap mass spectrometry (LTQ Orbitrap MS) with negative heated electrospray ionization. RESULTS: Untargeted analysis of total lipid extracts from murine samples (male 13-week-old WKAH/HKmSlc rats) led to the identification of several new SFAHFAs of acetic acid or propanoic acid esterified long-chain (>C20)-hydroxy fatty acids. Furthermore, MS3 analysis revealed the position of the hydroxyl group in the long-chain fatty acid as C-2. The relative amounts of SFAHFAs were quantified in intestinal contents and their tissues (Cecum, small intestine, and large intestine), liver, and plasma of rats fed with HFD and ND. The large intestine showed the highest abundance of SFAHFAs with a concentration range from 0.84 to 57 pmol/mg followed by the cecum with a range of 0.66 to 28.6 pmol/mg. The SFAHFAs were significantly altered between the HFD and ND groups, with a strong decreasing tendency under HFD conditions. CONCLUSIONS: Identification of these novel SFAHFAs can contribute to a better understanding of the chemical and biological properties of individual SFAHFAs and their possible sources in the gut, which in turn helps us tackle the role of these lipids in various metabolic diseases.
• Plasmalogen fingerprint alteration and content reduction in beef during boiling, roasting, and frying.

  Yue Wu, Zhen Chen, Hitoshi Chiba, Shu-Ping Hui

  Food chemistry 322 126764 - 126764 2020/08/30 [Refereed][Not invited]
   

  Plasmalogens are dietary phospholipids with beneficial health effects. In this work, plasmalogen characteristics and changes in beef during boiling, frying, and roasting were comprehensively investigated by liquid-chromatography-mass spectrometry. The alteration of plasmalogen fingerprint during cooking processes was found by untargeted omics approach, in which time of boiling, temperature of roasting, and meat core/surface of frying were responsible for the observed variations. Moreover, the targeted determination of representative plasmalogen species showed significant loss with a temperature- and time-dependent manner in roasting and frying. And frying even showed an extra loss in meat surface compared with core. Furthermore, an artificial neural network-based predictive model elucidated the dynamics of plasmalogen species during cooking. Finally, batter-coating pretreatment was performed to show its protection against plasmalogens loss during frying. These results might provide a potential strategy to better control and improve the quality of functional foodstuffs during cooking processes.
• Correction to: Deoxynivalenol-induced alterations in the redox status of HepG2 cells: identification of lipid hydroperoxides, the role of Nrf2-Keap1 signaling, and protective effects of zinc

  Wageh Sobhy Darwish, Zhen Chen, Yonghan Li, Hui Tan, Hitoshi Chiba, Shu-Ping Hui

  Mycotoxin Research 2020/08
• RETRACTED ARTICLE: Deoxynivalenol-induced alterations in the redox status of HepG2 cells: identification of lipid hydroperoxides, the role of Nrf2-Keap1 signaling, and protective effects of zinc

  Wageh Sobhy Darwish, Zhen Chen, Yonghan Li, Hui Tan, Hitoshi Chiba, Shu-Ping Hui

  Mycotoxin Research 2020/08
• Untargeted Lipidomic Analysis of Plasma from High-fat Diet-induced Obese Rats Using UHPLC-Linear Trap Quadrupole-Orbitrap MS.

  Siddabasave Gowda B Gowda, Zi-Jun Gao, Zhen Chen, Takayuki Abe, Shota Hori, Satoru Fukiya, Satoshi Ishizuka, Atsushi Yokota, Hitoshi Chiba, Shu-Ping Hui

  Analytical sciences : the international journal of the Japan Society for Analytical Chemistry 36 (7) 821 - 828 2020/07/10 [Refereed][Not invited]
   

  High-fat diet (HFD)-induced obesity is a primary risk factor for serious health problems. Although much research has been performed at the genomic level, lipidomic studies were limited. In this study, we aim to obtain a comprehensive profile of circulating plasma lipids, which are altered in rodent rat obesity by untargeted liquid chromatography-mass spectrometry. Rats fed with HFD for 8 weeks had increased body weight, liver and adipose tissue weight. The analysis results revealed that polyunsaturated fatty acids (PUFAs) and their corresponding phosphatidylcholine, phosphatidylinositol, and phosphatidylserine were significantly decreased in rats fed with HFD. In contrast, less unsaturated and ether type phosphatidylglycerols were increased. The triacylglycerides (TAGs) having saturated FA were increased in the HFD condition, whereas TAGs having PUFA were decreased. The levels of many plasma lipids were altered, and interestingly PUFA derived lipids were negatively associated with obesity. This signifies the importance of a PUFAs enriched diet to overwhelm obesity associated diseases.
• A Simple and Efficient Method for Synthesis of sn-Glycero-Phosphoethanolamine.

  Siddabasave Gowda B Gowda, Hirotoshi Fuda, Yusuke Yamamoto, Hitoshi Chiba, Shu-Ping Hui

  Lipids 55 (4) 395 - 401 2020/07 [Refereed][Not invited]
   

  An efficient three-step strategy for the convenient synthesis of Sn-glycero-3-phosphoethanolamine (GroPEtn) from a commercially available 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine (DPPE) is reported. Direct hydrolysis of DPPE produces a complex inseparable mixture, hence a protection and deprotection strategy is employed to prepare GroPEtn. The primary amine of DPPE is protected with a highly stable acid-labile trityl group, followed by strong base hydrolysis of N-trityl-DPPE gives N-trityl-GroPEtn. Further a mild, rapid, and efficient deprotection method is established using trifluoroacetic acid to remove N-trityl moiety, affords GroPEtn as a single product. This is the first semisynthetic approach and efficient method to produce GroPEtn with a total yield of 66% in three steps. GroPEtn did not show any cytotoxicity against human kidney (HK-2) cells and reporter gene assay for activation of Keap1-Nrf2-mediated antioxidant defense mechanism showed no significant effects.
• Oxidized lipid species in lipoproteins: Significance and analytical considerations

  Rojeet Shrestha, Hitoshi Chiba, Shu-Ping Hui

  Medical Mass Spectrometry 4 (1) 1 - 22 2432-745X 2020/06 [Refereed][Not invited]
  
• Discovery of Eicosapentaenoic Acid Esters of Hydroxy Fatty Acids as Potent Nrf2 Activators.

  Siddabasave Gowda B Gowda, Hirotoshi Fuda, Takayuki Tsukui, Hitoshi Chiba, Shu-Ping Hui

  Antioxidants (Basel, Switzerland) 9 (5) 2020/05/08 [Refereed][Not invited]
   

  Branched fatty acid esters of hydroxy fatty acids (FAHFAs) are a recently discovered class of biologically active lipids with anti-inflammatory and anti-diabetic properties. Despite the possible link between endogenous FAHFA levels and nuclear factor erythroid 2-related factor 2 (Nrf2), their possible function as antioxidants and the mechanisms involved in this are unknown. Here, we investigate FAHFAs' plausible antioxidant potential with reference to their effect on the Nrf2 levels, oxidative stress, and lipid droplet oxidation in human hepatocytes (C3A). Six authentic FAHFAs were chemically synthesized and performed activity-based screening by reporter gene assay. Among them, eicosapentaenoic acid (EPA) esterified 12-hydroxy stearic acid (12-HSA) and 12-hydroxy oleic acid (12-HOA) FAHFAs showed less cytotoxicity compared to their free fatty acids and potent activators of Nrf2. To define their mode of action, relative levels of nuclear Nrf2 were determined, which found a higher amount of Nrf2 in nucleus of cells treated with 12-EPAHSA compared to the control. Furthermore, 12-EPAHSA increased the expression of Nrf2-dependent antioxidant enzyme genes (NQO1, GCLM, GCLC, SOD-1, and HO-1). Fluorescence imaging analysis of linoleic-acid-induced lipid droplets (LDs) in C3A cells treated with 12-EPAHSA revealed the strong inhibition of small-size LD oxidation. These results suggest that EPA-derived FAHFAs as a new class of lipids with less cytotoxicity, and strong Nrf2 activators with plausible antioxidant effects via the induction of cytoprotective proteins against oxidative stress, induced cellular damage.
• Identification of novel biomarkers of hepatocellular carcinoma by high-definition mass spectrometry: Ultrahigh-performance liquid chromatography quadrupole time-of-flight mass spectrometry and desorption electrospray ionization mass spectrometry imaging.

  Koshi Nagai, Baasanjav Uranbileg, Zhen Chen, Amane Fujioka, Takahiro Yamazaki, Yotaro Matsumoto, Hiroki Tsukamoto, Hitoshi Ikeda, Yutaka Yatomi, Hitoshi Chiba, Shu-Ping Hui, Toru Nakazawa, Ritsumi Saito, Seizo Koshiba, Junken Aoki, Daisuke Saigusa, Yoshihisa Tomioka

  Rapid communications in mass spectrometry : RCM 34 Suppl 1 (Suppl 1) e8551  2020/04 [Refereed][Not invited]
   

  RATIONALE: Hepatocellular carcinoma (HCC) is a highly malignant disease for which the development of prospective or prognostic biomarkers is urgently required. Although metabolomics is widely used for biomarker discovery, there are some bottlenecks regarding the comprehensiveness of detected features, reproducibility of methods, and identification of metabolites. In addition, information on localization of metabolites in tumor tissue is needed for functional analysis. Here, we developed a wide-polarity global metabolomics (G-Met) method, identified HCC biomarkers in human liver samples by high-definition mass spectrometry (HDMS), and demonstrated localization in cryosections using desorption electrospray ionization MS imaging (DESI-MSI) analysis. METHODS: Metabolic profiling of tumor (n = 38) and nontumor (n = 72) regions in human livers of HCC was performed by an ultrahigh-performance liquid chromatography quadrupole time-of-flight MS (UHPLC/QTOFMS) instrument equipped with a mixed-mode column. The HCC biomarker candidates were extracted by multivariate analyses and identified by matching values of the collision cross section and their fragment ions on the mass spectra obtained by HDMS. Cryosections of HCC livers, which included both tumor and nontumor regions, were analyzed by DESI-MSI. RESULTS: From the multivariate analysis, m/z 904.83 and m/z 874.79 were significantly high and low, respectively, in tumor samples and were identified as triglyceride (TG) 16:0/18:1(9Z)/20:1(11Z) and TG 16:0/18:1(9Z)/18:2(9Z,12Z) using the synthetic compounds. The TGs were clearly localized in the tumor or nontumor areas of the cryosection. CONCLUSIONS: Novel biomarkers for HCC were identified by a comprehensive and reproducible G-Met method with HDMS using a mixed-mode column. The combination analysis of UHPLC/QTOFMS and DESI-MSI revealed that the different molecular species of TGs were associated with tumor distribution and were useful for characterizing the progression of tumor cells and discovering prospective biomarkers.
• Multivariate Analysis for Molecular Species of Cholesteryl Ester in the Human Serum.

  Yifan Chen, Shu-Ping Hui, Yusuke Miura, Sota Kato, Toshihiro Sakurai, Zhen Chen, Emiko Okada, Shigekazu Ukawa, Takafumi Nakagawa, Koshi Nakamura, Akiko Tamakoshi, Hitoshi Chiba, Hiroyuki Minami, Masahiro Mizuta

  Analytical sciences : the international journal of the Japan Society for Analytical Chemistry 36 (3) 373 - 378 0910-6340 2020/03/10 [Refereed][Not invited]
   

  Cholesteryl ester (CE) is an ester of cholesterol and fatty acid (FA). Plasma CE reflects complicated metabolisms of cholesterol, phospholipids, lipoproteins, and dietary FAs. An informatics approach could be useful for analysis of the CE species. In this study, two basic dimension reduction methods, principal component analysis (PCA) and factor analysis, were applied to serum CE species determined by LC-MS/MS in a Japanese population (n = 545). PCA and factor analysis both reflected the size (concentration), food source, fat solubility, and biological aspect of the CE species. In a comparison between PCA (PC4) and factor analysis (factor 4), the latter was found to be more suggestive from a biological aspect of n-6 FAs. Cholesteryl docosahexaenoate (DHA) was found to be unique by a factor analysis, possibly relevant to the unique accumulation of DHA in the brain. An informatics approach, especially factor analysis, might be useful for the analysis of complicated metabolism of CE species in the serum.
• Lipidomic profiling of dairy cattle oocytes by high performance liquid chromatography-high resolution tandem mass spectrometry for developmental competence markers.

  Zhen Chen, Yue Wu, Masashi Nagano, Kouki Ueshiba, Eri Furukawa, Yusuke Yamamoto, Hitoshi Chiba, Shu-Ping Hui

  Theriogenology 144 56 - 66 2020/03/01 [Refereed][Not invited]
   

  A comparative lipidomic profiling analysis of dairy cattle oocytes with different developmental competences was performed using a combination of high performance liquid chromatography-high resolution tandem mass spectrometry and multivariate statistical analysis. Significant lipidomic changes were identified in degenerating oocytes. Total triacylglycerol in the degenerating oocytes was 1.8-fold higher than that in the normal oocytes; however, total cardiolipin was 53.5% lesser than that in the normal oocytes, which indicated attenuation of energy metabolism. Compared to those in the normal oocytes, triacylglycerols in the degenerating oocytes were composed of longer and more unsaturated acyl chains. In contrast, the acyl chains in free fatty acids present in the degenerating oocytes were shorter and with lesser degree of unsaturation compared to those in the normal oocytes. Moreover, a significant decrease in degenerating oocytes were found in total phosphatidylinositol (14.8 ± 7.6 pmol vs. 24.8 ± 5.5 pmol), total phosphatidylcholine (20.8 ± 8.7 pmol vs. 33.5 ± 7.2 pmol), and total plasmalogen ethanolamine (9.0 ± 4.7 pmol vs. 16.8 ± 5.2 pmol), which indicated dysfunction of lipid-metabolizing enzymes in oocytes during degeneration. Thus, increase of triacylglycerols together with the decrease of certain phospholipid species could be potential markers of oocyte developmental competence. In addition to providing a new approach to investigate the lipidomic changes in oocyte development, the lipidomic profiling in the present study has revealed insights that hold potential to unravel the role of lipid metabolism in oocyte developmental competence in cattle.
• Identification of molecular species of phosphatidylcholine hydroperoxides in native and copper-oxidized triglyceride-rich lipoproteins in humans.

  Rojeet Shrestha, Zhen Chen, Yusuke Miura, Yusuke Yamamoto, Toshihiro Sakurai, Hitoshi Chiba, Shu-Ping Hui

  Annals of clinical biochemistry 57 (1) 95 - 98 0004-5632 2020/01 [Refereed][Not invited]
  
• Identification of cadmium-produced lipid hydroperoxides, transcriptomic changes in antioxidant enzymes, xenobiotic transporters, and pro-inflammatory markers in human breast cancer cells (MCF7) and protection with fat-soluble vitamins.

  Wageh Sobhy Darwish, Zhen Chen, Yonghan Li, Yue Wu, Hitoshi Chiba, Shu-Ping Hui

  Environmental science and pollution research international 27 (2) 1978 - 1990 2020/01 [Refereed][Not invited]
   

  Cadmium (Cd) is a toxic metal that is regarded as a metallohormone with estrogen-like properties. The present study aimed at identification of lipid hydroperoxides produced in human breast cancer (MCF7) exposed to cadmium (Cd) at environmentally relevant levels. Cd induced cytotoxicity and oxidative stress and produced a series of 26 lipid hydroperoxide species including 14 phosphatidylcholine hydroperoxides (PC-OOH), 9 triacylglycerol hydroperoxides (TG-OOH), and 3 cholesteryl ester hydroperoxides (CE-OOH). Among these hydroperoxides, PC34:2-OOH, PC34:3-OOH, TG60:14-OOH, TG48:5-OOH, TG60:15-OOH, and CE20:4-OOH were produced in a dose-dependent manner, suggesting these as possible biomarkers for Cd exposure in MCF7 cells. In addition, Cd led to significant decreases in the gene expressions of antioxidants, detoxification enzymes, and xenobiotic transporters. In a protection trial, co-exposure of MCF7 cells to fat-soluble vitamins including vitamin A, D, and E reduced Cd-induced cytotoxicity, lipid peroxidation, oxidative stress, and inflammatory responses. Fat-soluble vitamins upregulated antioxidant and detoxification enzymes, and xenobiotic transporters. Therefore, dietary supplementation of such micronutrients is recommended for people at risk for exposure to Cd.
• Flazin as a Promising Nrf2 Pathway Activator.

  Hirotoshi Fuda, Satoshi Miyanaga, Takayuki Furukawa, Satomi Umetsu, Sae Joko, Yuning Roan, Hirotaka Suzuki, Shu-Ping Hui, Mitsugu Watanabe, Hitoshi Chiba

  Journal of agricultural and food chemistry 67 (46) 12844 - 12853 0021-8561 2019/11/20 [Refereed][Not invited]
   

  Flazin is a β-carboline-derived alkaloid found in Japanese fermented foods. Here, the potential of flazin as an antioxidant food was studied with particular reference to its effect on the Kelch-like ECH-associated protein 1 (Keap1)-nuclear factor erythroid 2-related factor 2 (Nrf2) system in human hepatocytes (C3A). Flazin and flazin analogues including the decarboxylated derivative perlolyrine were chemically synthesized and compared with each other and with chlorogenic acid and curcumin. Among these compounds, flazin showed the lowest cytotoxicity (IC50 < 500 μM) and the highest capacity to activate the Keap1-Nrf2 system. It provided the largest (>3-fold of the control) cytoprotection ability against a pro-oxidant, although its radical absorbance capacity was relatively low. Flazin increased the expressions of Nrf2-dependent phase II enzyme genes and their products (NQO1, GSTP, and GSH proteins). The strong cytoprotection ability of flazin associated with low log P (0-3) is shared by sulforaphane and 3,5-dihydroxy-4-methoxybenzyl alcohol, suggesting the potential value of flazin and flazin-rich foods for the prevention of oxidation-related health disorders.
• A phase II, open-label clinical trial on the combination therapy with medium-chain triglycerides and ghrelin in patients with chronic obstructive pulmonary disease.

  Keisuke Miki, Seigo Kitada, Mari Miki, Shu-Ping Hui, Rojeet Shrestha, Kenji Yoshimura, Kazuyuki Tsujino, Hiroyuki Kagawa, Yohei Oshitani, Hiroshi Kida, Ryoji Maekura, Kenji Kangawa

  The journal of physiological sciences : JPS 69 (6) 969 - 979 1880-6546 2019/11 [Refereed][Not invited]
   

  The aim of this study was to investigate the effect of activated ghrelin with dietary octanoic acids or medium-chain triglyceride (MCT) administration to underweight patient with chronic obstructive pulmonary disease (COPD). Eleven severe and very severe COPD patients received a 5-day treatment with edible MCT. Sequentially, 10 patients received a 3-week combination treatment with MCT and intravenous acyl ghrelin. Five-day MCT treatment increased endogenous acyl ghrelin (p = 0.0049), but the total ghrelin level was unchanged. MCT-ghrelin combination treatment improved the peak oxygen uptake (p = 0.0120) during whole treatment course. This effect was attributed to the resultant improvements in cardiac function by O2 pulse, and to the difference between inspired and expired oxygen concentration rather than minute ventilation. Addition of dietary MCT to ghrelin treatment improved the aerobic capacity of underweight COPD patients, likely by mechanisms of increased O2 delivery through improvements in primary cardiocirculatory and muscular crosstalk.
• Circulating Apolipoprotein L1 is associated with insulin resistance-induced abnormal lipid metabolism.

  Kenji Nishimura, Taichi Murakami, Toshihiro Sakurai, Masashi Miyoshi, Kiyoe Kurahashi, Seiji Kishi, Masanori Tamaki, Tatsuya Tominaga, Sumiko Yoshida, Kojiro Nagai, Hideharu Abe, Shu-Ping Hui, Kazuhiko Kotani, Toshio Doi

  Scientific reports 9 (1) 14869 - 14869 2019/10/16 [Refereed][Not invited]
   

  Circulating ApolipoproteinL1 (ApoL1) is a component of pre-β-high-density lipoprotein (HDL), however little is known about the relationship of ApoL1 with cardiometabolic factors. Considering previous studies reporting the correlation of ApoL1 to triglyceride, we have hypothesized that ApoL1 associates with insulin-related metabolism. The current study examined their associations in 126 non-diabetic subjects and 36 patients with type 2 diabetes (T2DM). Non-diabetic subjects demonstrated triglyceride (standardized coefficients [s.c.] = 0.204, p < 0.05), body mass index (s.c. =0.232, p < 0.05) and HDL cholesterol (s.c. = -0.203, p < 0.05) as independent determinant of ApoL1 levels, and the significant elevation of ApoL1 in metabolic syndrome. Lipoprotein fractionation analysis revealed the predominant distribution of ApoL1 in large HDL fraction, and the significant increase of ApoL1 in large LDL fraction in high ApoL1 samples with insulin resistance. In T2DM, ApoL1 was higher in T2DM with metabolic syndrome, however ApoL1 was lower with β cell dysfunction. Insulin significantly promotes ApoL1 synthesis and secretion in HepG2 cells. In conclusion, circulating ApoL1 may be associated with abnormal HDL metabolism in insulin resistant status. This may suggest a regulation of insulin signal on the ApoL1 level, leading to offer a novel insight to the ApoL1 biology.
• Development of a simultaneous quantitation for short-, medium-, long-, and very long-chain fatty acids in human plasma by 2-nitrophenylhydrazine-derivatization and liquid chromatography-tandem mass spectrometry.

  Zhen Chen, Zijun Gao, Yue Wu, Rojeet Shrestha, Hiromitsu Imai, Naoto Uemura, Ken-Ichi Hirano, Hitoshi Chiba, Shu-Ping Hui

  Journal of chromatography. B, Analytical technologies in the biomedical and life sciences 1126-1127 121771 - 121771 1570-0232 2019/09/15 [Refereed][Not invited]
   

  Fatty acids (FA) have been important in clinical diagnosis for long, which makes the increasing need for a fast, reliable, and economic approach to determine FA of short-, medium-, long-, and very long-chain by widely available equipment and with high-throughput capacity. In the present work, 2‑nitrophenylhydrazine derivatization coupling with LC-MS/MS detection was utilized to simultaneously quantitate 18 FAs ranging from C4 to C26 in human plasma. The sample preparation protocol was optimized and extracting with diethyl ether‑potassium phosphate buffer twice was found as the highest efficiency along with economic feasibility. Under the optimized conditions, all the FA showed excellent linearity (R2 > 0.999 for each), sufficient sensitivity (LOD 0.2-330 fmol and LOQ 2.3-660 fmol for all), favorable accuracy (recovery ranged from 98.1 ± 3.6% to 104.9 ± 5.5% with coefficient of variation no >8.6% for all), and negligible matrix effect. In the clinical application on 30 healthy subjects, compared with the previous HPLC-UV method, the developed method showed high reliability, as well as reduced time and reagent costs. The established method showed the potential to apply to not only diagnostic practice, but also nutritional and epidemiological studies.
• Novel Fluorescence-Based Method To Characterize the Antioxidative Effects of Food Metabolites on Lipid Droplets in Cultured Hepatocytes.

  Tsukui T, Chen Z, Fuda H, Furukawa T, Oura K, Sakurai T, Hui SP, Chiba H

  Journal of agricultural and food chemistry 67 (35) 9934 - 9941 0021-8561 2019/09 [Refereed][Not invited]
   

  A fluorescence microscopic method for characterizing size, quantity, and oxidation of lipid droplets (LDs) in HepG2 cells was developed. LDs were induced by palmitic (PA), oleic (OA), or linoleic acids (LA) and stained with two fluorescent probes for neutral lipids and lipid peroxides. Each fatty acid increased the number of LDs and oxidized LDs (oxLDs) and the degree of LD oxidation time dependently, as well as increased intracellular triglyceride hydroperoxides. LDs induced by LA without 2,2'-azobis(2-amidinopropane)dihydrochloride (AAPH) showed the most significant oxidation degree over PA and OA, especially in large LDs (area ≥ 3 μm2, oxLD/LD = 52.3 ± 21.7%). Under this condition, two food-derived antioxidants were evaluated, and both of them significantly improved the LD characteristics. Moreover, chlorogenic acid reduced the quantity of large LDs by 74.0-87.6% in a dose-dependent manner. The proposed method provides a new approach to evaluate the effect of dietary antioxidants on LD characteristics.
• Identification of novel biomarkers of hepatocellular carcinoma by a high definition mass spectrometry; UHPLC-QTOF/MS and DESI-MSI.

  Nagai K, Uranbileg B, Chen Z, Fujioka A, Yamazaki T, Matsmoto Y, Tsukamoto H, Ikeda H, Yatomi Y, Chiba H, Hui SP, Nakazawa T, Saito R, Koshiba S, Aoki J, Saigusa D, Tomioka Y

  Rapid communications in mass spectrometry : RCM 34 0951-4198 2019/08 [Refereed][Not invited]
   

  Rationale Hepatocellular carcinoma (HCC) is a highly malignant disease for which the development of prospective or prognostic biomarkers is urgently required. Although metabolomics is widely used for biomarker discovery, there are some bottlenecks regarding the comprehensiveness of detected features, reproducibility of methods, and identification of metabolites. In addition, information on localization of metabolites in tumor tissue is needed for functional analysis. Here, we developed a wide-polarity global metabolomics (G-Met) method, identified HCC biomarkers in human liver samples by high-definition mass spectrometry (HDMS), and demonstrated localization in cryosections using desorption electrospray ionization MS imaging (DESI-MSI) analysis. Methods Metabolic profiling of tumor (n = 38) and nontumor (n = 72) regions in human livers of HCC was performed by an ultrahigh-performance liquid chromatography quadrupole time-of-flight MS (UHPLC/QTOFMS) instrument equipped with a mixed-mode column. The HCC biomarker candidates were extracted by multivariate analyses and identified by matching values of the collision cross section and their fragment ions on the mass spectra obtained by HDMS. Cryosections of HCC livers, which included both tumor and nontumor regions, were analyzed by DESI-MSI. Results From the multivariate analysis, m/z 904.83 and m/z 874.79 were significantly high and low, respectively, in tumor samples and were identified as triglyceride (TG) 16:0/18:1(9Z)/20:1(11Z) and TG 16:0/18:1(9Z)/18:2(9Z,12Z) using the synthetic compounds. The TGs were clearly localized in the tumor or nontumor areas of the cryosection. Conclusions Novel biomarkers for HCC were identified by a comprehensive and reproducible G-Met method with HDMS using a mixed-mode column. The combination analysis of UHPLC/QTOFMS and DESI-MSI revealed that the different molecular species of TGs were associated with tumor distribution and were useful for characterizing the progression of tumor cells and discovering prospective biomarkers.
• Determination of polycyclic aromatic hydrocarbon content in heat-treated meat retailed in Egypt: Health risk assessment, benzo[a]pyrene induced mutagenicity and oxidative stress in human colon (CaCo-2) cells and protection using rosmarinic and ascorbic ac

  Darwish WS, Chiba H, El-Ghareeb WR, Elhelaly AE, Hui SP

  Food chemistry 290 114 - 124 0308-8146 2019/08 [Refereed][Not invited]
   

  This study was undertaken to estimate the concentrations of the formed polycyclic aromatic hydrocarbons (PAHs) in heat-treated (boiled, pan-fried and grilled) meats collected from Egypt. Dietary intakes and cancer risks of PAHs among Egyptian adults were calculated. Benzo[a]pyrene (B[a]P)-induced mutagenicity and oxidative stress in human colon (CaCo-2) cell line and mechanisms behind such effects were also investigated. Finally, protection trials using rosmarinic (RMA) and ascorbic acids (ASA) were carried out. The results indicated formation of PAHs at high levels in the heat-treated meats. Calculated incremental life time cancer risk among Egyptian adults were 7.05179E-07, 7.00604 E-06 and 1.86069 E-05 due to ingestion of boiled, pan-fried and grilled meats, respectively. B[a]P-exposed CaCo-2 cells had high abilities for mutagenicity (490.05 ± 21.37 His + revertants) and production of reactive oxygen species. RMA and ASA protected CaCo-2 cells via reduction of B[a]P-induced mutagenicity and oxidative stress and upregulation of phase II detoxification enzymes and xenobiotic transporters.
• Choline and Ethanolamine Plasmalogens Prevent Lead-Induced Cytotoxicity and Lipid Oxidation in HepG2 Cells.

  Wu Y, Chen Z, Darwish WS, Terada K, Chiba H, Hui SP

  Journal of agricultural and food chemistry 67 (27) 7716 - 7725 0021-8561 2019/07 [Refereed][Not invited]
  
• Triglyceride deposit cardiomyovasculopathy: a rare cardiovascular disorder.

  Ming Li, Ken-Ichi Hirano, Yoshihiko Ikeda, Masahiro Higashi, Chikako Hashimoto, Bo Zhang, Junji Kozawa, Koichiro Sugimura, Hideyuki Miyauchi, Akira Suzuki, Yasuhiro Hara, Atsuko Takagi, Yasuyuki Ikeda, Kazuhiro Kobayashi, Yoshiaki Futsukaichi, Nobuhiro Zaima, Satoshi Yamaguchi, Rojeet Shrestha, Hiroshi Nakamura, Katsuhiro Kawaguchi, Eiryu Sai, Shu-Ping Hui, Yusuke Nakano, Akinori Sawamura, Tohru Inaba, Yasuhiko Sakata, Yoko Yasui, Yasuyuki Nagasawa, Shintaro Kinugawa, Kazunori Shimada, Sohsuke Yamada, Hiroyuki Hao, Daisaku Nakatani, Tomomi Ide, Tetsuya Amano, Hiroaki Naito, Hironori Nagasaka, Kunihisa Kobayashi

  Orphanet journal of rare diseases 14 (1) 134 - 134 2019/06/11 [Refereed][Not invited]
   

  Triglyceride deposit cardiomyovasculopathy (TGCV) is a phenotype primarily reported in patients carrying genetic mutations in PNPLA2 encoding adipose triglyceride lipase (ATGL) which releases long chain fatty acid (LCFA) as a major energy source by the intracellular TG hydrolysis. These patients suffered from intractable heart failure requiring cardiac transplantation. Moreover, we identified TGCV patients without PNPLA2 mutations based on pathological and clinical studies. We provided the diagnostic criteria, in which TGCV with and without PNPLA2 mutations were designated as primary TGCV (P-TGCV) and idiopathic TGCV (I-TGCV), respectively. We hereby report clinical profiles of TGCV patients. Between 2014 and 2018, 7 P-TGCV and 18 I-TGCV Japanese patients have been registered in the International Registry. Patients with I-TGCV, of which etiologies and causes are not known yet, suffered from adult-onset severe heart disease, including heart failure and coronary artery disease, associated with a marked reduction in ATGL activity and myocardial washout rate of LCFA tracer, as similar to those with P-TGCV. The present first registry-based study showed that TGCV is an intractable, at least at the moment, and heterogeneous cardiovascular disorder.
• Analysis of molecular species of phosphatidylcholine hydroperoxide in human native and copper-oxidized triglyceride-rich lipoproteins

  Shrestha Rojeet, Chen Z, Miura Y, Yamamoto Y, Sakurai T, Chiba H, Hui SP

  Clinica Chimica Acta 439 S82 - S83 0009-8981 2019/06 [Refereed][Not invited]
  
• Composition of plasmalogens in serum lipoproteins from patients with non-alcoholic steatohepatitis and their susceptibility to oxidation

  Akiko Ikuta, Toshihiro Sakurai, Megumi Nishimukai, Yuji Takahashi, Atsushi Nagasaka, Shu-Ping Hui, Hiroshi Hara, Hitoshi Chiba

  Clinica Chimica Acta 493 1 - 7 0009-8981 2019/06 [Refereed][Not invited]
  
• Interaction between LDL-mimetic liposomes and acid-treated carbon nanotube electrode during Cu2+-mediated oxidation

  Seiji Takeda, Koichi Mukasa, Shu-Ping Hui, Hitoshi Chiba

  Biochemical and Biophysical Research Communications 513 (1) 275 - 279 0006-291X 2019/05 [Refereed][Not invited]
  
• Serum 25-hydroxyvitamin D3 levels and poor sleep quality in a Japanese population: the DOSANCO Health Study.

  Koshi Nakamura, Shu-Ping Hui, Shigekazu Ukawa, Emiko Okada, Takafumi Nakagawa, Hiroaki Okabe, Zhen Chen, Yusuke Miura, Hitoshi Chiba, Akiko Tamakoshi

  Sleep medicine 57 135 - 140 1389-9457 2019/05 [Refereed][Not invited]
   

  OBJECTIVE: The present cross-sectional study investigated the relationship between serum 25-hydroxyvitamin D3 (25[OH]D3) levels and the presence of poor sleep quality in a community-based Japanese adult population. METHODS: Poor sleep quality, defined as poor subjective sleep quality and/or use of sleep medications, was assessed using a self-administered questionnaire. The prevalence of poor sleep quality was compared among 512 Japanese participants aged 35-79 years, based on serum 25(OH)D3 levels, which were determined using tandem mass spectrometry. A logistic regression model was used to calculate the odds ratios (ORs) for the presence of poor sleep quality in each group with the highest quartile of 25(OH)D3 serving as the reference group. RESULTS: Poor sleep quality was reported by 33.2% of the total study population. The prevalence of poor sleep quality was higher in the first quartile group (25[OH]D3: 2.08-18.13 ng/mL) than in the second, third and fourth quartile groups (18.14-23.07 ng/mL, 23.08-28.32 ng/mL, and 28.33-78.83 ng/mL, respectively). The ORs for poor sleep quality were 1.86 (95% confidence interval, 1.08-3.20) for the first quartile group, 0.73 (0.41-1.29) for the second quartile group, and 0.73 (0.42-1.27) for the third quartile group after adjusting for age, sex, and sociodemographic, lifestyle, physical and environmental factors, while the ORs were 1.68 (0.96-2.95), 0.69 (0.39-1.24), and 0.65 (0.37-1.15) after further adjustment for overall health status and depression status. CONCLUSIONS: The first quartile group of serum 25(OH)D3 was associated with the presence of poor sleep quality.
• Separating and Profiling Phosphatidylcholines and Triglycerides from Single Cellular Lipid Droplet by In-Tip Solvent Microextraction Mass Spectrometry.

  Zhao Y, Chen Z, Wu Y, Tsukui T, Ma X, Zhang X, Chiba H, Hui SP

  Analytical chemistry 91 (7) 4466 - 4471 0003-2700 2019/04 [Refereed][Not invited]
  
• Estimation of cadmium content in Egyptian foodstuffs: health risk assessment, biological responses of human HepG2 cells to food-relevant concentrations of cadmium, and protection trials using rosmarinic and ascorbic acids.

  Darwish WS, Chiba H, Elhelaly AE, Hui SP

  Environmental science and pollution research international 26 (15) 15443 - 15457 0944-1344 2019/04 [Refereed][Not invited]
   

  Cadmium (Cd) is an environmental pollutant that can get entry into human body via ingestion of contaminated foods causing multiple organ damage. This study aimed at monitoring Cd residues in 20 foodstuffs of animal origin that are commonly consumed in Egypt. Health risk assessment was conducted via calculation of Cd dietary intakes and non-carcinogenic target hazard quotient. An in vitro approach was performed to investigate the constitutive effects of Cd on human hepatoma (HepG2) cells under food-relevant concentrations. Trials to reduce Cd-induced adverse effects on HepG2 cells were done using rosmarinic (RMA) and ascorbic acids (ASA). The achieved results indicated contamination of the tested foodstuffs with Cd at high levels with potential human health hazards. Cd at food-relevant concentrations caused significant cytotoxicity to HepG2 cells. This may be attributed to induction of oxidative stress and inflammation, as indicated by the overexpression of stress and inflammatory markers. At the same time, Cd downregulated xenobiotic transporters and upregulated the proliferation factors. Co-exposure of HepG2 cells to Cd and micronutrients such as RMA and ASA led to recovery of cells from the oxidative damage, and subsequently cell viability was strongly improved. RMA and ASA ameliorated the biological responses of HepG2 cells to Cd exposure.
• Determination of total, free and esterified short-chain fatty acid in human serum by liquid chromatography-mass spectrometry.

  Chen Z, Wu Y, Shrestha R, Gao Z, Zhao Y, Miura Y, Tamakoshi A, Chiba H, Hui SP

  Annals of clinical biochemistry 56 (2) 190 - 197 0004-5632 2019/03 [Refereed][Not invited]
   

  BACKGROUND: Short-chain fatty acids are primarily absorbed through the portal vein during lipid digestion, which is utilized as the energy source, as well as prevent type 2 diabetes and some cancers. However, reports on the determination of these short-chain fatty acids in human serum are limited. METHODS: Blood samples from human subjects ( n = 547, male/female = 246/301, age 58.85 ± 12.57) were collected. Saponification was applied to obtain total fatty acid. After derivatization by 2-nitrophenylhydrazine, fatty acid 4:0 and fatty acid 6:0 were measured by liquid chromatography-mass spectrometry. RESULTS: The developed method exhibited good linearity (R2 = 0.9996 for both). All the coefficients of variation of reproducibility and accuracy for fatty acid 4:0 and fatty acid 6:0 ranged 3.0%-6.1%, with the average recoveries of 87.8%-102.4% and 92.2%-98.2%, respectively. In all the samples, the concentration of fatty acid 4:0 (162.4 ± 76.4 μmol/L) was significantly higher than fatty acid 6:0 (2.0 ± 2.5 μmol/L, P < 0.001). Furthermore, the esterified form was predominant in both fatty acid 4:0 and fatty acid 6:0 (98.2% and 82.4% of total fatty acids, respectively). Besides, short-chain fatty acids showed no significant differences with regard to sex or age differences. CONCLUSION: This developed liquid chromatography-mass spectrometry method is convenient and reliable, which might be useful for monitoring the variations of short-chain fatty acids in blood.
• Dietary salmon milt extracts attenuate hepatosteatosis and liver dysfunction in diet-induced fatty liver model.

  Sakurai T, Hayasaka T, Sekiguchi H, Satoh H, Chen Z, Chiba H, Hui SP

  Journal of the science of food and agriculture 99 (4) 1675 - 1681 0022-5142 2019/03 [Refereed][Not invited]
  
• Microbiome Alteration in Type 2 Diabetes Mellitus Model of Zebrafish.

  Fumiyoshi Okazaki, Liqing Zang, Hiroko Nakayama, Zhen Chen, Zi-Jun Gao, Hitoshi Chiba, Shu-Ping Hui, Takahiko Aoki, Norihiro Nishimura, Yasuhito Shimada

  Scientific reports 9 (1) 867 - 867 2019/01/29 [Refereed][Not invited]
   

  Understanding the gut microbiota in metabolic disorders, including type 2 diabetes mellitus (T2DM), is now gaining importance due to its potential role in disease risk and progression. We previously established a zebrafish model of T2DM, which shows glucose intolerance with insulin resistance and responds to anti-diabetic drugs. In this study, we analysed the gut microbiota of T2DM zebrafish by deep sequencing the 16S rRNA V3-V4 hypervariable regions, and imputed a functional profile using predictive metagenomic tools. While control and T2DM zebrafish were fed with the same kind of feed, the gut microbiota in T2DM group was less diverse than that of the control. Predictive metagenomics profiling using PICRUSt revealed functional alternation of the KEGG pathways in T2DM zebrafish. Several amino acid metabolism pathways (arginine, proline, and phenylalanine) were downregulated in the T2DM group, similar to what has been previously reported in humans. In summary, we profiled the gut microbiome in T2DM zebrafish, which revealed functional similarities in gut bacterial environments between these zebrafish and T2DM affected humans. T2DM zebrafish can become an alternative model organism to study host-bacterial interactions in human obesity and related diseases.
• Examining the effect of regioisomerism on the physico-chemical properties of lysophosphatidylethanolamine-containing liposomes using fluoro probes.

  Yamamoto Y, Furukawa T, Takeda S, Kashida H, Chiba H, Hui SP

  Chemistry and physics of lipids 216 9 - 16 0009-3084 2018/11 [Refereed][Not invited]
  
• Lipidomic Profiling on Oxidized Phospholipids in Type 2 Diabetes Mellitus Model Zebrafish.

  Zhen Chen, Liqing Zang, Yue Wu, Hiroko Nakayama, Yasuhito Shimada, Rojeet Shrestha, Yaoyao Zhao, Yusuke Miura, Hitoshi Chiba, Shu-Ping Hui, Norihiro Nishimura

  Analytical sciences : the international journal of the Japan Society for Analytical Chemistry 34 (10) 1201 - 1208 0910-6340 2018/10/10 [Refereed][Not invited]
   

  Type 2 diabetes mellitus is a serious metabolic disorder in the world. Oxidative stress, as a key role on the pathogenesis of diabetes, also results in the oxidation of phospholipids. However, studies on phospholipid oxidation in diabetes, especially directly focusing on oxidized and degraded phospholipid species, are quite limited. In this study, phospholipid profiles of diabetic zebrafish plasma were characterized by LC-HRMS and MS/MS, and the total amounts of each lipid class were compared. Furthermore, the key molecular species as biomarkers in distinguishing control and diabetic samples were investigated by orthogonal partial least squares discriminant analysis. Among the identified 114 phospholipid species in total, there were 11 hydroperoxides, 7 aldehydes, and 19 lysophospholipids found significantly elevated along with the increasing blood glucose, which were known as oxidation or degradation products. Furthermore, lysophosphatidylcholine 20:5 and lysophosphatidylcholine 22:6 were assessed as potential biomarkers in diabetic zebrafish. The current work would not only help to gain further insights into diabetes, but also contribute to find new clinical parameters for the screening of the promising antioxidant agents for its therapies.
• Determination of Total, Free, and Esterified Short-Chain Fatty Acid in Human Serum by LC-MS/MS.

  Chen Z, Wu Y, Shrestha R, Gao Z, Zhao Y, Miura Y, Tamakoshi A, Chiba H, Hui SP

  Annals of clinical biochemistry {SAGE} Publications 4563218801393  0004-5632 2018/09 [Refereed][Not invited]
  
• Determination of Serum 25-Hydroxyvitamin D₃ by LC/MS/MS and Its Monthly Variation in Sapporo Indoor Workers.

  Okabe H, Shimizu C, Yamamoto M, Kikuchi R, Minami A, Chen YF, Imai H, Mizuta M, Chen Z, Chiba H, Hui SP

  Analytical sciences : the international journal of the Japan Society for Analytical Chemistry 34 (9) 1043 - 1047 0910-6340 2018/09 [Refereed][Not invited]
   

  25-Hydroxyvitamin D3 (25(OH)D3) as the metabolite of vitamin D, is connected with various of diseases, and important to people with limited sunshine. Thus, the investigation of serum 25-hydroxyvitamin D and its variation in these people is necessary. In this study, a simple, precise, and accurate method for serum 25(OH)D3 determination by LC/MS/MS was developed. Serum samples were obtained monthly for one year from 11 male and 11 female indoor workers in Sapporo, Japan, and the overall 25(OH)D3 concentration was 12.9 ± 4.7 ng/mL. The 25(OH)D3 in females was significantly lower than that in males (14.0 ± 5.0 vs. 11.9 ± 4.3 ng/mL). The serum 25(OH)D3 concentration in males and females were both strongly correlated to UV-B radiation (r2 = 0.8477 and 0.7384, respectively), with a two-month's lag. Also the monthly change in 25(OH)D3 in males was more significant than that in females.
• A two-step homogeneous assay for apolipoprotein E-containing high-density lipoprotein-cholesterol.

  Takahashi Y, Ito Y, Sakurai T, Wada N, Nagasaka A, Fujikawa M, Chiba H, Hui SP

  Annals of clinical biochemistry 56 (1) 4563218795212 - 132 0004-5632 2018/08 [Refereed][Not invited]
  
• Effects of enzymes on elastic modulus of low-density lipoproteins were investigated using atomic force microscopy

  Seiji Takeda, Toshihiro Sakurai, Shu-Ping Hui, Hirotoshi Fuda, Hitoshi Chiba

  Biochemical and Biophysical Research Communications 501 (3) 607 - 611 1090-2104 2018/06/27 [Refereed][Not invited]
   

  Oxidation of low-density lipoproteins (LDLs) induces development of cardiovascular disease. Recently, reports of studies using atomic force microscopy (AFM) have described that the elastic modulus of metal-induced oxidized LDLs is lower than the modulus before oxidation. However, the mechanisms of change of the elastic modulus have not been well investigated. We postulated that disorder of the LDL structure might decrease the elastic modulus. This study measured the elastic modulus of LDLs before and after enzyme treatment with V8 protease, α-chymotrypsin, and phospholipase A2. After LDLs were obtained from serum by ultracentrifugation, LDLs or enzyme-treated LDLs were physically absorbed. They were crowded on a mica surface. Although V8 protease and α-chymotrypsin did not induce the elastic modulus change, treatment with PLA2 decreased the elastic modulus. The LDL particle size did not change during the enzyme treatment. Results suggest that disordering of the lipid structure of the LDL might contribute to the elastic modulus change. Results show that AFM might be a useful tool to evaluate disorders of complex nanoscale particle structures from lipids and proteins such as lipoproteins.
• Microwave-assisted Derivatization of Fatty Acids for Its Measurement in Milk Using High-Performance Liquid Chromatography.

  Rojeet Shrestha, Yusuke Miura, Ken-Ichi Hirano, Zhen Chen, Hiroaki Okabe, Hitoshi Chiba, Shu-Ping Hui

  Analytical sciences : the international journal of the Japan Society for Analytical Chemistry 34 (5) 575 - 582 1348-2246 2018 [Refereed][Not invited]
   

  Fatty acid (FA) profiling of milk has important applications in human health and nutrition. Conventional methods for the saponification and derivatization of FA are time-consuming and laborious. We aimed to develop a simple, rapid, and economical method for the determination of FA in milk. We applied a beneficial approach of microwave-assisted saponification (MAS) of milk fats and microwave-assisted derivatization (MAD) of FA to its hydrazides, integrated with HPLC-based analysis. The optimal conditions for MAS and MAD were determined. Microwave irradiation significantly reduced the sample preparation time from 80 min in the conventional method to less than 3 min. We used three internal standards for the measurement of short-, medium- and long-chain FA. The proposed method showed satisfactory analytical sensitivity, recovery and reproducibility. There was a significant correlation in the milk FA concentrations between the proposed and conventional methods. Being quick, economic, and convenient, the proposed method for the milk FA measurement can be substitute for the convention method.
• Identification and Quantitation of C=C Location Isomers of Unsaturated Fatty Acids by Epoxidation Reaction and Tandem Mass Spectrometry

  Yaoyao Zhao, Hansen Zhao, Xu Zhao, Jia Jia, Qiang Ma, Sichun Zhang, Xinrong Zhang, Hitoshi Chiba, Shu-Ping Hui, Xiaoxiao Ma

  ANALYTICAL CHEMISTRY 89 (19) 10270 - 10278 0003-2700 2017/10 [Refereed][Not invited]
   

  Unsaturated fatty acids (FAs) serve as nutrients, energy sources, and signaling molecules for organisms, which are the major components for a large variety of lipids. However, structural characterization and quantitation of unsaturated FAs by mass spectrometry remain an analytical challenge. Here, we report the coupling of epoxidation reaction of the C-C in unsaturated FAs and tandem mass spectrometry (MS) for rapid and accurate identification and quantitation of C-C isomers of FAs in a shotgun lipidomics approach. Epoxidation of the C-C leads to the production of an epoxide which, upon collision induced dissociation (CID), produces abundant diagnostic ions indicative of the C-C location. The total intensity of the same set of diagnostic ions for one specific FA C-C isomer was also used for its relative and absolute quantitation. The simple experimental setup, rapid reaction kinetics (<2 min), high reaction yield (>90% for monounsaturated FAs), and easy-to-interpret tandem MS spectra enable a promising methodology particularly for the analysis of unsaturated FAs in complex biological samples such as human plasma and animal tissues.
• An Efficient Glycoblotting-Based Analysis of Oxidized Lipids in Liposomes and a Lipoprotein

  Takayuki Furukawa, Hiroshi Hinou, Seiji Takeda, Hitoshi Chiba, Shin-Ichiro Nishimura, Shu-Ping Hui

  CHEMBIOCHEM 18 (19) 1903 - 1909 1439-4227 2017/10 [Refereed][Not invited]
   

  Although widely occurring lipid oxidation, which is triggered by reactive oxygen species (ROS), produces a variety of oxidized lipids, practical methods to efficiently analyze oxidized lipids remain elusive. Herein, it is shown that the glycoblotting platform can be used to analyze oxidized lipids. Analysis is based on the principle that lipid aldehydes, one of the oxidized lipid species, can be captured selectively, enriched, and detected. Moreover, 3-methyl-1-p-tolyltriazene (MTT) methylates phosphoric and carboxylic acids, and this MTT-mediated methylation is, in combination with conventional tandem mass spectrometry (MS/MS) analysis, an effective method for the structural analysis of oxidized lipids. By using three classes of standards, liposomes, and a lipoprotein, it is demonstrated that glycoblotting represents a powerful approach for focused lipidomics, even in complex macromolecules.
• Profiling of cardiolipins and their hydroperoxides in HepG2 cells by LC/MS

  Zhen Chen, Yue Wu, Yi-Shing Ma, Yuu Kobayashi, Yao-Yao Zhao, Yusuke Miura, Hitoshi Chiba, Shu-Ping Hui

  ANALYTICAL AND BIOANALYTICAL CHEMISTRY 409 (24) 5735 - 5745 1618-2642 2017/09 [Refereed][Not invited]
   

  Cardiolipin (CL) exists as crucial functional phospholipid in mitochondria. The oxidation of CL is concerned with mitochondrial dysfunction and various diseases. As main oxidation products, CL hydroperoxide (CL-OOH) plays a key role in intermediating oxidative reaction. Thus, direct analysis of CL-OOH is of great interest. In the present study, CL and CL-OOH profiles were analyzed in oxidized HepG2 cell lipid via HPLC-Orbitrap MS/MS. Furthermore, the contents of individual molecular species were compared between intact and AAPH-oxidized HepG2 cells. In total, 46 CL and 18 CL-OOH were identified from oxidized cell lipids, while 21 CL and 9 CL-OOH were detected in AAPH-treated cells. Most CL depleted significantly after AAPH inducement, with percentages varying from 8.3% (CL70:7) to 73.7% (CL72:4), depending on fatty acyl composition. While almost all the CL-OOH remarkably increased, among them 68:6-, 72:6-, and 72:7-OOHs were only detected in AAPH-treated cells. CL68:5- and CL68:4-OOH were the most abundant species, while CL70:5-OOH among all the species expressed the highest oxidation percentage of the corresponding CL. Our results showed practical separation, identification, and semi-quantitation of CL-OOH species, which could contribute to approaches to lipidomic analysis of CL and CL-OOH, as well as tracing biomarkers in mitochondrial oxidative stress diagnosis.
• Comparison of chemical structures and cytoprotection abilities between direct and indirect antioxidants

  Sae Joko, Mitsugu Watanabe, Hirotoshi Fuda, Seiji Takeda, Takayuki Furukawa, Shu-Ping Hui, Rojeet Shrestha, Hitoshi Chiba

  JOURNAL OF FUNCTIONAL FOODS 35 245 - 255 1756-4646 2017/08 [Refereed][Not invited]
   

  To study structural and biological relationships in antioxidants, the cytoprotection, cytotoxicity, and Keap1-Nrf2 pathway activation potentials of nine natural antioxidants were compared. Sulforaphane, quercetin, isoliquiritigenin, curcumin, lycopene, and 3,5-dihydroxy-4-methoxybenzyl alcohol (DHMBA) activated the Keap1-Nrf2 pathway, whereas chlorogenic acid, gallic acid, rosmarinic acid, and cyanidin-3-O-glucoside did not. Hence, the former group was considered indirect antioxidants, and the latter direct antioxidants. Except for DHMBA and sulforaphane, the indirect antioxidants exhibited higher logP values and cytoprotectiveness, albeit with higher cytotoxicities, compared to the direct antioxidants. Exceptionally, DHMBA and sulforaphane showed lower logP values and higher cytoprotectiveness without cytotoxicity, indicating that indirect antioxidants might be more beneficial than direct antioxidants for cytoprotection, although caution towards cytotoxicity should be exercised regarding those with high logP values. In this regard, indirect antioxidants with low logP values like DHMBA and sulforaphane might serve as promising ingredients in functional foods. (C) 2017 Elsevier Ltd. All rights reserved.
• Absolute quantification of cholesteryl esters using liquid chromatography-tandem mass spectrometry uncovers novel diagnostic potential of urinary sediment

  Yusuke Miura, Takayuki Furukaw, Miho Kobayashi, Rojeet Shrestha, Ryoji Takahashi, Chikara Shimizu, Hitoshi Chiba, Shu-Ping Hui

  STEROIDS 123 43 - 49 0039-128X 2017/07 [Refereed][Not invited]
   

  Background: Urine has been utilized as a source of biomarkers in renal disease. However, urinary lipids have not attracted much attention so far. Here we studied urinary cholesteryl ester (CE) and its relevance in renal disease. Methods: Quantitative analysis of CE molecular species in serum, urinary supernatant, and urinary sediment from patients with renal disease (N = 64) and non-renal disease (N = 23) was carried out using liquid chromatography-tandem mass spectrometry (LC-MS/MS) and deuterated CEs as internal standards. Results: Validation study showed good precision and accuracy of LC MS/MS. Many CE species were detected in the urinary sediment and supernatant in the renal disease group, whereas only a few CE species were detected in the other group. In the renal disease group, the sum of the concentrations of all CE species showed a significant correlation between the sediment and the supernatant from urinary samples (r = 0.876, p < 0.001); however, the composition of CEs was significantly different between them. Further, the composition of CEs of the supernatant was similar to that of the serum. Conclusions: Our LC-MS/MS analysis uncovered a distinct CE profile in urinary sediment from patients with renal disease, suggesting a possible contribution of CEs in urothelial cells to the development of renal disease.
• Selective improvement of peptides imaging on tissue by supercritical fluid wash of lipids for matrix-assisted laser desorption/ionization mass spectrometry

  Shoko Matsushita, Noritaka Masaki, Kohei Sato, Takahiro Hayasaka, Eiji Sugiyama, Shu-Ping Hui, Hitoshi Chiba, Nobuyuki Mase, Mitsutoshi Setou

  ANALYTICAL AND BIOANALYTICAL CHEMISTRY 409 (6) 1475 - 1480 1618-2642 2017/02 [Refereed][Not invited]
   

  There is a high analytical demand for improving the detection sensitivity for various peptides in matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) because exhaustive distribution analyses of various peptides could help to reveal the function of peptides in vivo. To improve the sensitivity of peptide detection, we used supercritical fluid of CO2 (scCO(2)) as washing solvent for a pretreatment to remove lipids. We evaluated whether our wash method using scCO(2) with an entrainer improved the detection of peptides and suppressed lipid detection in MALDI-IMS. Our analysis revealed that the signal intensities of peptides such as m/z 3339.8, 3530.9, 4233.3, 4936.7, and 4963.7 were increased in scCO(2)-washed samples. The greatest improvement in the signal-to-noise ratio (S/N) was found at m/z 4963.7, which was identified as thymosin beta 4, with the S/N reaching almost 190-fold higher than the control. Additionally, all of the improved signals were associated with the morphologic structure. Our method allows us to analyze the distribution of molecules, especially in the region of m/z 3000-5200. For these improvements, the polarity difference between scCO(2) and the matrix solution used was considered as a key. A wider variety of molecules can be analyzed in the future due to this improvement of the detection sensitivity by optimizing the polarity of scCO(2) with various entrainers.
• Change in Plasma Total, Esterified and Non-esterified Capric Acid Concentrations during a Short-term Oral Administration of Synthetic Tricaprin in Dogs.

  Rojeet Shrestha, Ken-Ichi Hirano, Akira Suzuki, Satoshi Yamaguchi, Yusuke Miura, Yi-Fan Chen, Masahiro Mizuta, Hitoshi Chiba, Shu-Ping Hui

  Analytical sciences : the international journal of the Japan Society for Analytical Chemistry 33 (11) 1297 - 1303 0910-6340 2017 [Refereed][Not invited]
   

  We studied change in the plasma total, esterified and non-esterified capric acid (FA10:0) and its effect on longer fatty acid concentrations during the short-term oral administration of synthetic tricaprin in dogs. We administered 150 and 1500 mg tricaprin/kg body weight per day orally to dogs for 7 consecutive days. Blood samples were collected at 0, 0.5, 1, 2, 4, 8, and 24 h on the 1st and 7th days for measuring the total-, esterified- and non-esterified-FA10:0. The total-FA10:0 concentration increased in a dose-dependent manner, reaching a peak at 1 h on the 1st day and at 2 to 4 h on the 7th day; it then mostly disappeared within 24 h. The mean esterified FA10:0 concentration was found be 75.5 and 60.3% of total-FA10:0 in dogs fed 150 and 1500 mg of tricaprin/kg body weight, respectively. The plasma level of FA10:0 depends on the duration and dose of tricaprin administration, but are rapidly cleared from circulation within several hours.
• Rapid tin-mediated access to a lysophosphatidylethanolamine (LPE) library: Application to positional LC/MS analysis for hepatic LPEs in non-alcoholic steatohepatitis model mice

  Takayuki Furukawa, Hirotoshi Fuda, Satoshi Miyanaga, Chinatsu Watanabe, Hitoshi Chiba, Shu-Ping Hui

  CHEMISTRY AND PHYSICS OF LIPIDS 200 133 - 138 0009-3084 2016/10 [Refereed][Not invited]
   

  Even though lysophospholipids have attracted much interest in recent years on account of their unique bioactivity, research related to lysophospholipids is usually hampered by problems associated with standard sample preparation and discrimination of regioisomers. Herein, we demonstrate a quick tin chemistry-based synthetic route to lysophosphatidylethanolamines (LPEs) and its application in the positional analysis of hepatic LPEs in non-alcoholic steatohepatitis (NASH) model mice. We found that the preference of hepatic LPE regioisomer largely depends on the unsaturation of acyl chain in both control and NASH model mice. In addition, hepatic C18:2-LPE and C20:5-LPE levels were significantly lower in the NASH model mice than those in the control. The LC/MS technique based on the library of LPE regioisomers allows an accurate observation of hepatic LPE metabolism and might provide useful information to elucidate yet ambiguous pathogenesis of NASH. (C) 2016 Elsevier Ireland Ltd. All rights reserved.
• A fatty acid profiling method using liquid chromatography-high resolution mass spectrometry for improvement of assisted reproductive technology

  Akiko Yagi, Satoshi Miyanaga, Rojeet Shrestha, Seiji Takeda, Seiichi Kobayashi, Hitoshi Chiba, Hirobumi Kamiya, Shu-Ping Hui

  CLINICA CHIMICA ACTA 456 100 - 106 0009-8981 2016/05 [Refereed][Not invited]
   

  Background: Liquid chromatography-high resolution mass spectrometry (LC-HRMS) can be useful to improve in vitro fertilization (IVF). This study aims to find out an association between embryonic growth and embryonic uptake of free fatty acid (FFA) from culture media by using LC-HRMS. Methods: Embryos (n = 55) from 15 couples undergoing IVF were studied. An embryo was cultivated for up to 6 days in a 20 mu l-medium drop under mineral oil, and classified by a morphological grading system into the good-growth group (n = 32; good quality blastocysts) and the poor-growth group (n = 23; non-blastocysts). The control study was set up without embryo. Extracted ion chromatogram of FFAs was collected in negative ion mode for each medium sample obtained after use. Results: The percent change from control to sample in mass area for docosahexaenoic acid showed a decrease in the good-growth group than that in the poor-growth group (p < 0.05). Decrease in %change of docosahexaenoic acid might indicate proper embryonic growth. Similar but insignificant change was observed for other essential FFA5, but not for non-essential FFA5. Conclusion: The proposed metabolomic approach using LC-HRMS might be a powerful tool for non-invasive evaluation of embryonic growth. (C) 2016 Elsevier B.V. All rights reserved.
• Imaging Mass Spectrometry Reveals a Decrease of Cardiolipin in the Kidney of NASH Model Mice

  Takahiro Hayasaka, Hirotoshi Fuda, Shu-Ping Hui, Hitoshi Chiba

  ANALYTICAL SCIENCES 32 (4) 473 - 476 0910-6340 2016/04 [Refereed][Not invited]
   

  Non-alcoholic steatohepatitis (NASH) can be complicated with chronic kidney disease (CKD). In this study, changes in the distribution of biomolecules in the kidney were studied in NASH model mice with the use of imaging mass spectrometry (IMS). The mass spectra and ion images of IMS showed that the signals of cardiolipin (CL) species were decreased in the kidney cortex of the NASH mice. The decrease of CL might therefore suggest the kidney involvement of NASH.
• Synthesis of (2 beta,3 alpha,6-H-2(3))cholesteryl linoleate and cholesteryl oleate as internal standards for mass spectrometry

  Yusuke Miura, Shu-Ping Hui, Rojeet Shrestha, Takahisa Hiruma, Seiji Takeda, Hirotoshi Fuda, Shigeo Ikegawa, Ken-ichi Hirano, Hitoshi Chiba

  STEROIDS 107 1 - 9 0039-128X 2016/03 [Refereed][Not invited]
   

  The accurate analysis of trace component in complex biological matrices requires the use of reliable standards. For liquid chromatography/mass spectrometry analysis, the stable isotope-labeled derivatives of the analyte molecules are the most appropriate internal standards. We report here the synthesis of (2 beta,3 alpha,6-H-2(3))cholesteryl linoleate and oleate containing three non-exchangeable deuterium in the steroid ring. The principal reactions used were: (1) trans diaxial opening of 2 alpha,3 alpha-epoxy-6-oxo-5 alpha-cholestane with LiAlD4 and subsequent oxidation of the resulting (2 beta,6 alpha-H-2(2))-3 alpha,6 beta-diol with Jones' reagent, followed by reduction of the resulting (2 beta-H-2)-3,6-dione with NaBD4 leading to the (2 beta,3 alpha,6 alpha-H-2(3))-3 beta,6 beta-dihydroxy-5 alpha-cholestane, (2) selective protection of the 3 beta-hydroxy group as the tert-butyldimethylsilyl ether, (3) dehydration of the 6 beta-hydroxy group with POCI3 and removal of tert-butyldimethylsilyloxy groups with 5 M HCl in acetone, and (4) esterification of the resultant (2 beta,3 alpha,6-H-2(3))cholesterol with linoleic and oleic acids using 1-(3-dimethylaminopropyI)-3-ethylcarbodiim ide. The isotopic purity was found to be satisfactory by mass spectrometry, and nuclear magnetic resonance properties of the new compounds were tabulated. The labeled compounds can be used as internal standards in liquid chromatography/mass spectrometry assays for clinical and biochemical studies. (C) 2015 Elsevier Inc. All rights reserved.
• Development of homogeneous assay for simultaneous measurement of apoE-deficient, apoE-containing, and total HDL-cholesterol

  Yuji Takahashi, Yasuki Ito, Norio Wada, Atsushi Nagasaka, Masato Fujikawa, Toshihiro Sakurai, Rojeet Shrestha, Shu-Ping Hui, Hitoshi Chiba

  CLINICA CHIMICA ACTA 454 135 - 142 0009-8981 2016/02 [Refereed][Not invited]
   

  Background: Pathophysiological role for high-density lipoprotein (HDL) subclasses remains to be elucidated. Homogeneous assay for simultaneous measurements of apoE-deficient HDL-cholesterol (HDL-C), apoE-containing HDL-.C, and total HDL-C is desired, because apoE plays important roles in lipid metabolism. Methods: The proposed assay consists of a primary reaction to remove non-HDL-C, a secondary reaction to measure apoE-deficient HDL-C, and a tertiary reaction to measure apoE-containing HDL-C. The assay is completed within 10 min. For control study, 13% polyethylene glycol precipitation assay and phosphotungstate-dextran sulfate -magnesium precipitation assay were carried out. Results: Good correlations between the control assays and the proposed assay was obtained in serum samples from patients without liver disease (n = 33): r = 0.987, 0.957, and 0.991 for apoE-deficient, apoE-containing, and total HDL-C, respectively. ApoE-containing HDL-C by the proposed method in healthy individuals (n = 12) and patients with hyper-HDL-cholesterolemia (n = 5) were 0.11 +/- 0.03 and 0.26 +/- 0.05 mmol/l (4.1 +/- 1.3 and 10.1 +/- 2.0 mg/dl), respectively. ApoE-containing HDL-C increased rapidly at >2.59 mmol/l (100 mg/dl) of total HDL-C, suggesting a unique regulating mechanism of apoE-containing HDL-C. Conclusions: The established homogeneous assay might be useful for clinical and epidemiological studies on apoE-deficient and apoE-containing HDL subclasses. (C) 2016 Elsevier B.V. All rights reserved.
• Improvement of Mitochondrial Function and Lipid Utilization by 3,5-dihydroxy-4-methoxybenzyl Alcohol, an Oyster-derived polyphenol, in Oleate-loaded C2C12 Myotubes.

  Ma YS, Yoshida S, Kobayashi Y, Kawanishi N, Furukawa T, Fuda H, Hui SP, Chiba H

  Journal of Food and Nutrition Research Science & Education Publishing 4 (8) 498 - 507 2333-1119 2016 [Refereed][Not invited]
   

  Anti-oxidative effects of the Pacific oyster-derived phenolic antioxidant, 3,5-dihydroxy-4-methoxybenzyl alcohol (DHMBA), has been documented in hepatocytes. Additionally, DHMBA-rich oyster extracts significantly attenuated obesity in a non-alcoholic steatohepatitis mouse model. Whether the administration of DHMBA might improve muscular mitochondrial function was investigated. The mouse C2C12-derived myotubes were loaded with oleic acid (400μM) and cultured for 24 hours in the presence of DHMBA (500μM) with or without electrical stimulation (ES), where ES was given as exercise mimic. The fatty acid uptake, lipid accumulation, and mitochondrial function were subsequently accessed. DHMBA and ES increased fatty acid uptake, TG contents, mitochondrial membrane potential, intracellular level of H2O2, and mitochondrial O2 consumption rate. Intracellular ATP content was significantly increased when both DHMBA and ES were loaded at the same time, suggesting their synergic action. Phosphorylated AMPKα, AMPKβ1, and acetyl-CoA carboxylase were increased by DHMBA, indicating a possible role for DHMBA for activation of metabolic adaptation system and consequent increase of fatty acid oxidation. In conclusion, DHMBA solely or in collaboration with exercise might possibly serve as a fitness food for obese persons by stimulating muscular fatty acid utilization and mitochondrial energy production. This assumption must be verified by animal experiment.
• Oyster extracts attenuate pathological changes in non-alcoholic steatohepatitis (NASH) mouse model

  Mitsugu Watanabe, Hirotoshi Fuda, Hiroaki Okabe, Sae Joko, Yusuke Miura, Shu-Ping Hui, Yimin, Naohiro Hamaoka, Emiko Miki, Hitoshi Chiba

  JOURNAL OF FUNCTIONAL FOODS 20 516 - 531 1756-4646 2016/01 [Refereed][Not invited]
   

  The phenolic compound 3,5-dihydroxy-4-methoxybenzyl alcohol (DHMBA) is a natural antioxidant recently isolated from the Pacific oyster. DHMBA, up to a concentration of 500 mu M, has demonstrated a strong in vitro hepatocyte-protective effect from oxidative stress without any cytotoxicity. This study investigated the in vivo potential of DHMBA-rich oyster extracts (DOE) for prevention or attenuation of non-alcoholic steatohepatitis (NASH). NASH-model mice, developed by supplementation of a high-fat diet for 23 weeks and intravenous injections of oxidised low-density lipoproteins, exhibited obesity, insulin resistance, hepatic steatosis, inflammation, fibrosis, and apoptosis. These changes were significantly moderated by supplementation of DOE. The search for an underlying mechanism determined that DOE significantly improved the redox status of DNA, proteins, and lipids. Moreover, DOE suppressed the increase of hepatic expression of PPARy and CD36 (fatty acid transporter) in the NASH-model mice. DOE might serve as a functional food for people at elevated risk for NASH. (C) 2015 The Authors. Published by Elsevier Ltd.
• Mass Spectrometric Quantification of Amphipathic, Polyphenolic Antioxidant of the Pacific Oyster (Crassostrea Gigas)

  Hiroaki Okabe, Shu-Ping Hui, Hirotoshi Fuda, Takayuki Furukawa, Seiji Takeda, Rojeet Shrestha, Yusuke Miura, Mitsugu Watanabe, Hitoshi Chiba

  ANALYTICAL SCIENCES 31 (12) 1341 - 1344 0910-6340 2015/12 [Refereed][Not invited]
   

  A novel amphipathic phenolic compound, 3,5-dihydroxy-4-methoxybenzyl alcohol (DHMBA), that can be isolated from the Pacific oyster (Crassostrea gigas) has been found to protect human hepatocytes against oxidative stress. This study aims to establish a method for the measurement of DHMBA for industrial application. Liquid chromatography-tandem mass spectrometry using deuterated DHMBA as an internal standard and a polar end-capped ODS (Hypersil GOLD aQ) as the solid phase was validated. The limit of detection was 0.04 pmol (S/N = 5), and the limit of quantitation was 0.1 pmol (S/N = 10). The calibration curve was linear throughout the range of 0.1 - 16 pmol (r(2) = 0.9995). This method successfully quantified DHMBA in oysters from 11 sea areas in Japan. The results showed that the yield of DHMBA was variable from 9.8 to 58.8 mu g g(-1) whole oyster meat wet weight but not affected by the seawater temperature. The proposed LC-MS/MS method is useful in quantitative studies for DHMBA and potentially for other amphipathic substances.
• Elastic modulus of low-density lipoprotein as potential indicator of its oxidation

  Seiji Takeda, Agus Subagyo, Shu-Ping Hui, Hirotoshi Fuda, Rojeet Shrestha, Kazuhisa Sueoka, Hitoshi Chiba

  ANNALS OF CLINICAL BIOCHEMISTRY 52 (6) 647 - 653 0004-5632 2015/11 [Refereed][Not invited]
   

  Background Evaluation of low-density lipoprotein oxidation is important in the risk assessment of cardiovascular disease. Atomic force microscope is widely used to evaluate the physical properties including stiffness on a single-particle scale. In this study, the effect of low-density lipoprotein oxidation on the low-density lipoprotein stiffness was investigated using an atomic force microscope. Methods Isolated low-density lipoprotein particles with or without oxidation were densely bound to an Au substrate on mica, and then pressed and deformed by the atomic force microscope tip. The stiffness of each low-density lipoprotein particle was estimated as the elastic modulus obtained by the force curve analysis. Biochemical change of low-density lipoprotein due to oxidation was studied by electrophoresis. Results and conclusion The elastic modulus of low-density lipoprotein particles ranged between 0.1 and 2MPa. The oxidation of low-density lipoprotein increased the number of low-density lipoprotein particles with smaller elastic moduli, indicating the decrease in low-density lipoprotein stiffness. The elastic modulus of low-density lipoprotein might be potentially useful to evaluate low-density lipoprotein oxidation.
• Identification of molecular species of oxidized triglyceride in plasma and its distribution in lipoproteins

  Rojeet Shrestha, Shu-Ping Hui, Yusuke Miura, Akiko Yagi, Yuji Takahashi, Seiji Takeda, Hirotoshi Fuda, Hitoshi Chiba

  CLINICAL CHEMISTRY AND LABORATORY MEDICINE 53 (11) 1859 - 1869 1434-6621 2015/10 [Refereed][Not invited]
   

  Background: The role of triglycerides carried in the triglyceride-rich lipoproteins (TRL) in the progression of atherosclerosis is uncertain. Identification of oxidized triglycerides and its possible association with atherosclerosis were largely ignored. Here we applied mass spectrometric approach to detect and identify triglyceride hydroperoxides (TGOOH) in human plasma and lipoproteins. Methods: EDTA plasma was collected from healthy human volunteers (n = 9) after 14-16 h of fasting. Very low-density lipoprotein (VLDL) (d<1.006) and intermediatedensity lipoprotein (IDL) (d=1.006-1.019) were isolated from the plasma (n=6) by sequential ultracentrifugation in KBr, followed by the isolation of LDL and high-density lipoprotein (HDL) using size-exclusion high-performance liquid chromatography (HPLC). Total lipids from the plasma and isolated lipoproteins were extracted, and analyzed for the detection and identification of TGOOH using liquid chromatography/LTQ ion trap Orbitrap mass spectrometry. All the processes, from specimen collection to the mass spectrometric analysis, were carried out at 4 degrees C in the presence of antioxidant to prevent oxidation of lipoproteins. Results: We identified 11 molecular species of TGOOH in either plasma or VLDL and IDL, of which TGOOH-18:1/18:2/16:0, TGOOH-18:1/18:1/16:0, TGOOH-16:0/18:2/16:0, TGOOH-18:1/18:1/18:1, and TGOOH-16:0/20:4/16:0 were most dominant. These TGOOH molecules are carried by TRL but not by LDL and HDL. Mean concentration of TGOOH in plasma, VLDL and IDL were, respectively, 56.1 +/- 25.6, 349.8 +/- 253.6 and 512.5 +/- 173.2 mu mol/mol of triglycerides. Conclusions: This is the first report to identify several molecular species of oxidized triglycerides in TRL. Presence of oxidized triglyceride may contribute to the atherogenicity of TRL. Further work is needed to elucidate the association of the oxidized triglyceride in atherosclerosis.
• Plasma capric acid concentrations in healthy subjects determined by high-performance liquid chromatography

  Rojeet Shrestha, Shu-Ping Hui, Hiromitsu Imai, Satoru Hashimoto, Naoto Uemura, Seiji Takeda, Hirotoshi Fuda, Akira Suzuki, Satoshi Yamaguchi, Ken-ichi Hirano, Hitoshi Chiba

  ANNALS OF CLINICAL BIOCHEMISTRY 52 (5) 588 - 596 0004-5632 2015/09 [Refereed][Not invited]
   

  Background Capric acid (FA10:0, decanoic acid) is a medium-chain fatty acid abundant in tropical oils such as coconut oil, whereas small amounts are present in milk of goat, cow, and human. Orally ingested FA10:0 is transported to the liver and quickly burnt within it. Only few reports are available for FA10:0 concentrations in human plasma. Methods Fasting (n=5, male/female=3/2, age 319.3 years old) and non-fasting (n=106, male/female=44/62, age 21.93.2 years old) blood samples were collected from apparently healthy Japanese volunteers. The total FA10:0 in the plasma were measured by high-performance liquid chromatography after derivatization with 2-nitrophenylhydrazine followed by UV detection. Results Inter and intra-assay coefficient of variation of FA10:0 assay at three different concentrations ranged in 1.7-3.9 and 1.3-5.4%, respectively, with an analytical recovery of 95.2-104.0%. FA10:0 concentration was below detection limit (0.1 mu mol/L) in each fasting human plasma. FA10:0 was not detected in 50 (47.2%) of 106 non-fasting blood samples, while 29 (27.4%) plasma samples contained FA10:0 less than or equal to 0.5 mu mol/L (0.4 +/- 0.1), and 27 (25.5%) contained it at more than 0.5 mu mol/L (0.9 +/- 0.3). Conclusion A half of the non-fasting plasma samples contained detectable FA10:0. This simple, precise, and accurate high-performance liquid chromatography method might be useful for monitoring plasma FA10:0 during medium-chain triglycerides therapy.
• Synergistic Costimulatory Effect of Chlamydia pneumoniae with Carbon Nanoparticles on NLRP3 Inflammasome-Mediated Interleukin-1 beta Secretion in Macrophages

  Junji Matsuo, Shinji Nakamura, Seiji Takeda, Kasumi Ishida, Tomohiro Yamazaki, Mitsutaka Yoshida, Hitoshi Chiba, Shu-Ping Hui, Hiroyuki Yamaguchi

  INFECTION AND IMMUNITY 83 (7) 2917 - 2925 0019-9567 2015/07 [Refereed][Not invited]
   

  The obligate intracellular bacterium Chlamydia pneumoniae is not only a causative agent of community-acquired pneumonia but is also associated with a more serious chronic disease, asthma, which might be exacerbated by air pollution containing carbon nanoparticles. Although a detailed mechanism of exacerbation remains unknown, the proinflammatory cytokine interleukin- 1 beta (IL-1 beta) is a critical player in the pathogenesis of asthma. C. pneumoniae induces IL-1 beta in macrophages via NACHT, LRR, and PYD domain-containing protein 3 (NLRP3) inflammasome activation and Toll-like receptor 2/4 (TLR2/4) stimulation. Carbon nanoparticles, such as carbon nanotubes (CNTs), can also evoke the NLRP3 inflammasome to trigger IL-1 beta secretion from lipopolysaccharide-primed macrophages. This study assessed whether costimulation of C. pneumoniae with CNTs synergistically enhanced IL-1 beta secretion from macrophages, and determined the molecular mechanism involved. Enhanced IL-1 beta secretion from C. pneumoniae-infected macrophages by CNTs was dose and time dependent. Transmission electron microscopy revealed that C. pneumoniae and CNTs were engulfed concurrently by macrophages. Inhibitors of actin polymerization or caspase-1, a component of the inflammasome, significantly blocked IL-1 beta secretion. Gene silencing using small interfering RNA (siRNA) targeting the NLRP3 gene also abolished IL-1 beta secretion. Other inhibitors (K+ efflux inhibitor, cathepsin B inhibitor, and reactive oxygen species-generating inhibitor) also blocked IL-1 beta secretion. Taken together, these findings demonstrated that CNTs synergistically enhanced IL-1 beta secretion from C. pneumoniae-infected macrophages via the NLRP3 inflammasome and caspase-1 activation, providing novel insight into our understanding of how C. pneumoniae infection can exacerbate asthma.
• Anti-apoptotic effects of novel phenolic antioxidant isolated from the Pacific oyster (Crassostrea gigas) on cultured human hepatocytes under oxidative stress

  Hirotoshi Fuda, Mitsugu Watanabe, Shu-Ping Hui, Sae Joko, Hiroaki Okabe, Shigeki Jin, Seiji Takeda, Emiko Miki, Takayuki Watanabe, Hitoshi Chiba

  FOOD CHEMISTRY 176 226 - 233 0308-8146 2015/06 [Refereed][Not invited]
   

  The antioxidant, and hepatoprotective properties of 3,5-dihydroxy-4-methoxybenzyl alcohol (DHMBA), a natural phenolic antioxidant isolated from the Pacific oyster, were defined using cultured human hepatocyte-derived cells (C3A). DHMBA showed no cytotoxicity at 62.5-500 mu M, as well as chlorogenic acid (CGA), vitamin C, and vitamin E. However, butylated hydroxytoluene, eicosapentaenoic acid, docosahexaenoic acid and catechin reduced cell viability. In the presence of the prooxidant 2,2'-azobis(2-amidino-propane) dihydrochloride (AAPH), DHMBA at 125-500 mu M improved cell viability, whereas CGA had no effect. DNA ladder formation and flow-cytometric studies indicated that DHMBA inhibited MPH-induced apoptosis and necrosis. CGA was ineffective. Thus, DHMBA is a novel, potent antioxidant, effectively protecting cultured hepatocytes from apoptosis and necrosis caused by oxidative stress. Additionally, the concentration of DHMBA was determined by mass spectrometry to be 24.4 mu mol/kg wet oyster meat, and three polyphenols (gentisic acid, daidzein, and matairesinol) were newly identified in the oyster extracts. (C) 2014 Elsevier Ltd. All rights reserved.
• Serum choline plasmalogens-those with oleic acid in sn-2-are biomarkers for coronary artery disease

  Megumi Nishimukai, Ryouta Maeba, Akiko Ikuta, Naoya Asakawa, Kiwamu Kamiya, Shiro Yamada, Takashi Yokota, Mamoru Sakakibara, Hiroyuki Tsutsui, Toshihiro Sakurai, Yuji Takahashi, Shu-Ping Hui, Hitoshi Chiba, Tomoki Okazaki, Hiroshi Hara

  CLINICA CHIMICA ACTA 437 147 - 154 0009-8981 2014/11 [Refereed][Not invited]
   

  Background: Identifying risk factors is crucial for preventing cardiovascular events, but there are no widely accepted predictive biomarkers. In our previous study of Japanese asymptomatic cohorts, we performed global analysis of serum ether glycerophospholipids (Egp) molecular profiles, and found that choline plasmalogens (PlsCho; 1-O-alk-1'-enyl-2-acyl-sn-glycero-3-phosphocholine), particularly those containing oleic acid (18:1) in the sn-2 position, were strongly associated with a wide range of risk factors for metabolic syndrome/atherosclerosis. Methods: We determined serum concentrations of Egp molecular species of coronary artery disease patients (n = 50; 31 males and 19 females) by LC/MS/MS, and plasmalogen (Pls; 1-O-alk-1'-enyl-2-acyl-sn-glycerophospholipids) contents in lipoprotein fractions by HPLC using radioactive iodine. Results: We found that the serum concentrations of ether choline glycerophospholipids (EgpCho), particularly PlsCho, were not only significantly lower in males with significant coronary stenosis but also associated with atherosclerosis-related parameters, and their association was stronger than either high-density lipoprotein cholesterol or adiponectin. In addition, serum PlsCho containing 18:1 or linoleic acid (18:2) in sn-2 showed the highest correlations with a wide range of atherogenic parameters among PlsCho molecular species. Conclusion: These results verify our previous findings that serum PlsCho, particularly those containing 18:1 in sn-2, may serve as reliable biomarkers for atherosclerosis. (C) 2014 Elsevier B.V. All rights reserved.
• Identification of molecular species of cholesteryl ester hydroperoxides in very low-density and intermediate-density lipoproteins

  Rojeet Shrestha, Shu-Ping Hui, Toshihiro Sakurai, Akiko Yagi, Yuji Takahashi, Seiji Takeda, Shigeki Jin, Hirotoshi Fuda, Hitoshi Chiba

  ANNALS OF CLINICAL BIOCHEMISTRY 51 (6) 662 - 671 0004-5632 2014/11 [Refereed][Not invited]
   

  Background Oxidation of lipoproteins is thought to play a crucial role in atherogenesis. Role for triglyceride-rich lipoproteins in atherogenesis is unclear. Thus, we aimed to investigate whether cholesteryl ester hydroperoxides (CEOOH) are present in very low-density lipoproteins (VLDL) and intermediate-density lipoproteins (IDL) by using highly sensitive liquid chromatography/mass spectrometry. Methods Total lipids were extracted from the plasma of healthy donors (n=6) and their fractions of VLDL and IDL. Additional three plasma samples were analysed freshly for CEOOH. Detection and identification of CEOOH was conducted by liquid chromatography/LTQ ion trap mass spectrometry/Orbitrap high mass accuracy mass spectrometry. Authentic standards of CEOOH were used for unequivocal identification on the basis of their mass spectra. Results We identified six molecular CEOOH species overall, namely, Ch18:1-OOH, Ch18:2-OOH, Ch18:3-OOH, Ch20:4-OOH, Ch20:5-OOH and Ch22:6-OOH. Of them, Ch18:2-OOH, Ch20:5-OOH, Ch20:4-OOH and Ch22:6-OOH were detected in all IDL samples, while only Ch20:4-OOH was detected in all VLDL samples. All of CEOOH species except for Ch18:3-OOH were detected in plasma, with constant detection of Ch20:5-OOH, and Ch22:6-OOH in all plasma samples. Conclusion The presence of CEOOH species in VLDL and IDL was confirmed with the analytical sensitivity of 0.1pmol, showing the constant appearance of more CEOOH species in IDL than VLDL. This finding might add biochemical evidences of atherogenicity of these lipoproteins. Clinical utility of measuring CEOOH level in these lipoproteins need to be investigated for the risk assessment of the cardiovascular disease.
• Changes of lipoproteins in phenylalanine hydroxylase-deficient children during the first year of life

  Hironori Nagasaka, Hirokazu Tsukahara, Yoshiyuki Okano, Ken-ichi Hirano, Toshihiro Sakurai, Shu-Ping Hui, Toshihiro Ohura, Hiromi Usui, Tohru Yorifuji, Satoshi Hirayama, Akira Ohtake, Takashi Miida

  CLINICA CHIMICA ACTA 433 (C) 1 - 4 0009-8981 2014/06 [Refereed][Not invited]
   

  Background: Influence of hyperphenylalaninemia on lipoproteins in early life remains unclear. Methods: We enrolled 24 phenylalanine hydroxylase (PAH)-deficient children who were classified into a phenylketonuria (PKU) group (n = 12) lacking PAH activity and a benign hyperphenylalaninemia (HPA) group (n = 12) having partial PAH activity, and their 11 non-affected siblings. We measured serum total-cholesterol, low-density lipoprotein (LDL)-cholesterol, and high-density lipoprotein (HDL)-cholesterol levels together with apolipoproteins for the first year of life, and compared them with those of 30 age-matched healthy controls. Results: The affected groups invariably had lower cholesterol levels than non-affected groups. At birth, HDL-cholesterol decrease was greatest and predominated over the LDL-cholesterol decrease: total cholesterol, 28/36% decrease to the control level in HPA/PKU; HDL-cholesterol, 33/51%; LDL-cholesterol, 20/28%. At 3 months, the opposite changes were observed: total cholesterol, 16/28%; HDL-cholesterol, 13/23%; LDL-cholesterol, 16/33%. At 12 months, LDL were still significantly lower in both groups (8/18%, p < .05 and .001), although HDL was significantly decreased only in the PKU group (15%, p < .05). Apolipoprotein A-I/A-II and B changed respectively in accordance with HDL-cholesterol and LDL-cholesterol changes. Despite similar phenylalanine levels, the PKU group invariably had lower cholesterol concentrations than the HPA group had. Conclusion: Irrespective of phenylalanine concentrations, lipoprotein synthesis in PAH-deficient children, particularly in PKU children, was suppressed in early life. (C) 2014 Elsevier B.V. All rights reserved.
• Serum choline plasmalogens, particularly those with oleic acid in sn-2, are associated with proatherogenic state

  Megumi Nishimukai, Ryouta Maeba, Yuya Yamazaki, Toru Nezu, Toshihiro Sakurai, Yuji Takahashi, Shu-Ping Hui, Hitoshi Chiba, Tomoki Okazaki, Hiroshi Hara

  JOURNAL OF LIPID RESEARCH 55 (5) 956 - 965 0022-2275 2014/05 [Refereed][Not invited]
   

  Serum plasmalogens (Pls) (1-O-alk-1'-enyl-2-acyl glycerophospholipids) are of particular interest for studies on metabolic disorders associated with oxidative stress and chronic inflammation. Serum levels of Pls are known to correlate positively with HDL-cholesterol (HDL-C); however, few studies have examined serum Pls molecular species in association with pathophysiological conditions and their clinical significance. To clarify these, we determined serum levels of individual ether glycerophospholipids in Japanese asymptomatic cohorts (n = 428; 362 male and 66 female subjects) by LC/MS/MS, and examined their correlations with clinical parameters. We found that the proportion of choline Pls (PlsCho) among total serum phospholipids was significantly lower in the male group over 40 years old and was associated with multiple risk parameters more strongly than HDL-C. The abundance of serum PlsCho with oleic acid (18:1) in sn-2 exhibited the strongest positive correlation with serum concentrations of adiponectin and HDL-C, while being inversely associated with waist circumference and the serum levels of TG and small dense LDL-cholesterol. The characterization of serum ether glycerophospholipids verified the specificity of PlsCho, particularly the ones with 18:1 in sn-2, as a sensitive biomarker for the atherogenic state.
• Evaluation of Antioxidant Activity of Natural and Synthetic Compounds Against LDL Oxidation Using CNT Electrode

  Futaba Ohkawa, Seiji Takeda, Shu-Ping Hui, Toshihiro Sakurai, Shigeki Jin, Hirotoshi Fuda, Kazuhisa Sueoka, Hitoshi Chiba

  IEEE SENSORS JOURNAL 14 (2) 532 - 537 1530-437X 2014/02 [Refereed][Not invited]
   

  In this paper, a novel method for evaluating antioxidant activity against low density lipoprotein (LDL) oxidation (anti-LDLox) using a carbon nanotube (CNT) based electrode was investigated. Although ORAC and DPPH are known methods to evaluate antioxidant activity, they do not directly reflect the capacities of anti-LDLox. A method has been reported for measuring oxidized LDL using a CNT electrode. We evaluated anti-LDLox by measuring the amount of oxidized LDL using the electrode after incubation of LDL with antioxidants during copper-mediated oxidation. Thiobarbituric acid reactive substances (TBARS) were also determined for comparison with the CNT electrode. There was a correlation of the CNT electrode with the TBARS and the ORAC, whereas no correlation was found between the CNT electrode and the DPPH. We demonstrate here that a CNT electrode method has a potential for the determination of the antioxidant activity of various natural and synthetic compounds by detecting oxidized LDL.
• Peripheral leukocyte anomaly detected with routine automated hematology analyzer sensitive to adipose triglyceride lipase deficiency manifesting neutral lipid storage disease with myopathy/triglyceride deposit cardiomyovasculopathy

  Akira Suzuki, Hironori Nagasaka, Yasuhiro Ochi, Kazuhiro Kobayashi, Hiroshi Nakamura, Daisaku Nakatani, Satoshi Yamaguchi, Shinobu Yamaki, Atsushi Wada, Yoshihisa Shirata, Shu-Ping Hui, Tatsushi Toda, Hiroshi Kuroda, Hitoshi Chiba, Ken-Ichi Hirano

  Molecular Genetics and Metabolism Reports 1 (1) 249 - 253 2214-4269 2014 [Refereed][Not invited]
   

  Adipose triglyceride lipase (ATGL) deficiency manifesting neutral lipid storage disease with myopathy/triglyceride deposit cardiomyovasculopathy presents distinct fat-containing vacuoles known as Jordans' anomaly in peripheral leucocytes. To develop an automatic notification system for Jordans' anomaly in ATGL-deficient patients, we analyzed circulatory leukocyte scattergrams on automated hematology analyzer XE-5000. The BASO-WX and BASO-WY values were found to be significantly higher in patients than those in non-affected subjects. The two parameters measured by automated hematology analyzer may be expected to provide an important diagnostic clue for homozygous ATGL deficiency. © 2014 The Authors.
• Genetic mutations in adipose triglyceride lipase and myocardial up-regulation of peroxisome proliferated activated receptor-gamma in patients with triglyceride deposit cardiomyovasculopathy

  Ken-ichi Hirano, Tatsuya Tanaka, Yoshihiko Ikeda, Satoshi Yamaguchi, Nobuhiro Zaima, Kazuhiro Kobayashi, Akira Suzuki, Yasuhiko Sakata, Yasushi Sakata, Kunihisa Kobayashi, Tatsushi Toda, Norihide Fukushima, Hatsue Ishibashi-Ueda, Daniela Tavian, Hironori Nagasaka, Shu-Ping Hui, Hitoshi Chiba, Yoshiki Sawa, Masatsugu Hori

  BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 443 (2) 574 - 579 0006-291X 2014/01 [Refereed][Not invited]
   

  Adipose triglyceride lipase (ATGL, also known as PNPLA2) is an essential molecule for hydrolysis of intracellular triglyceride (TG). Genetic ATGL deficiency is a rare multi-systemic neutral lipid storage disease. Information regarding its clinical profile and pathophysiology, particularly for cardiac involvement, is still very limited. A previous middle-aged ATGL-deficient patient in our institute (Case 1) with severe heart failure required cardiac transplantation (CTx) and exhibited a novel phenotype, "Triglyceride deposit cardiomyovasculopathy (TGCV)". Here, we tried to elucidate molecular mechanism underlying TGCV. The subjects were two cases with TGCV, including our second case who was a 33-year-old male patient (Case 2) with congestive heart failure requiring CTx. Case 2 was homozygous for a point mutation in the 5' splice donor site of intron 5 in the ATGL, which results in at least two types of mRNAs due to splicing defects. The myocardium of both patients (Cases 1 and 2) showed up-regulation of peroxisome proliferated activated receptors (PPARs), key transcription factors for metabolism of long chain fatty acids (LCFAs), which was in contrast to these molecules' lower expression in ATGL-targeted mice. We investigated the intracellular metabolism of LCFAs under human ATGL-deficient conditions using patients' passaged skin fibroblasts as a model. ATGL-deficient cells showed higher uptake and abnormal intracellular transport of LCFA, resulting in massive TG accumulation. We used these findings from cardiac specimens and cell-biological experiments to construct a hypothetical model to clarify the pathophysiology of the human disorder. In patients with TGCV, even when hydrolysis of intracellular TG is defective, the marked up-regulation of PPAR gamma and related.genes may lead to increased uptake of LCFAs, the substrates for TG synthesis. This potentially vicious cycle of LCFAs could explain the massive accumulation of TG and severe clinical course for this rare disease. (C) 2013 Elsevier Inc. All rights reserved.
• Measurement of single low-density lipoprotein particles by atomic force microscopy

  Toshihiro Sakurai, Seiji Takeda, Jun-ya Takahashi, Yuji Takahashi, Norio Wada, Suchin Trirongjitmoah, Takeshi Namita, Shigeki Jin, Akiko Ikuta, Hiroaki Furumaki, Shu-Ping Hui, Hirotoshi Fuda, Masato Fujikawa, Koichi Shimizu, Hitoshi Chiba

  ANNALS OF CLINICAL BIOCHEMISTRY 50 (6) 564 - 570 0004-5632 2013/11 [Refereed][Not invited]
   

  Background The size of lipoprotein particles is relevant to the risk of coronary artery disease (CAD). Methods We investigated the feasibility of atomic force microscopy (AFM) for evaluating the size of large low-density lipoprotein (LDL) and small dense LDL (sd-LDL) separated by ultracentrifugation. The measurements by AFM in tapping mode were compared to those by electron microscopy (EM). Results There was a significant difference in particle sizes determined by AFM between large LDL (20.61.9nm, mean +/- SD) and sd-LDL (16.2 +/- 1.4nm) obtained from six healthy volunteers (P<0.05). The particle sizes determined by EM for the same samples were 23.2 +/- 1.4nm for large LDL and 20.4 +/- 1.4nm for sd-LDL. The difference between large LDL and sd-LDL detected by EM was also statistically significant (P<0.05). In addition, the particle sizes of each lipoprotein fraction were significantly different between AFM and EM: P<0.05 for large LDL and P<0.05 for sd-LDL. Conclusions AFM can differentiate between sd-LDL and large LDL particles by their size, and might be useful for evaluating risk for CAD.
• Evaluation of Oxidized-Low-Density Lipoproteins Using Kelvin Force Microscopy

  Seiji Takeda, Futaba Ohkawa, Shu-Ping Hui, Toshihiro Sakurai, Shigeki Jin, Hirotoshi Fuda, Kazuhisa Sueoka, Hitoshi Chiba

  IEEE SENSORS JOURNAL 13 (9) 3449 - 3453 1530-437X 2013/09 [Refereed][Not invited]
   

  Oxidized-low-density lipoprotein (ox-LDL), which is generated by LDL oxidation, is a risk factor of cardiovascular disease development. LDL has positive charges on the surface attributable to amino groups of lysine moieties on Apo B-100. Its positive charge is decreased by oxidation. For this paper, the topography and surface potential of LDL and ox-LDL are measured using atomic force microscopy and Kelvin probe force microscopy (KPFM). Although it is difficult to ascertain a difference between topographic images of LDL and ox-LDL, the KPFM images of ox-LDL differ from those obtained using LDL. The averaged potential of the surface decreases concomitantly with increasing oxidation time of the LDL. We demonstrate a method to evaluate the oxidation level of LDL using KPFM.
• Quantification of urinary 18-hydroxycortisol using LC-MS/MS

  Shigeki Jin, Norio Wada, Yuji Takahashi, Shu-Ping Hui, Toshihiro Sakurai, Hirotoshi Fuda, Seiji Takeda, Masato Fujikawa, Katsuyuki Yanagisawa, Shigeo Ikegawa, Takao Kurosawa, Hitoshi Chiba

  ANNALS OF CLINICAL BIOCHEMISTRY 50 (5) 450 - 456 0004-5632 2013/09 [Refereed][Not invited]
   

  Background: Urinary 18-hydroxycortisol has been investigated as a marker of aldosterone-producing adenoma (APA). The aim of this study was to develop and validate a method for the measurement of 18-hydroxycortisol using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Methods: Urine was collected over a 24-hour period in patients with APA (n=11), idiopathic hyperaldosteronism (IHA, n=9), and essential hypertension (EH, n=6). 18-Hydroxycortisol was extracted in solid-phase, and measured by LC-MS/MS based on selected reaction monitoring. Results: The method allowed quantification of 18-hydroxycortisol with a lower quantification limit of 0.26nmol/L, intra- and inter-assay coefficients of variation of <3.4% and a range of analytical recovery of 98.0-103.7%. Urinary 18-hydroxycortisol excretion for APA, IHA and EH were determined as 725 (SD 451), 102 (SD 68) and 88 (SD 76) nmol/day, respectively. Conclusions: The proposed method met the basic analytical requirements and was considered to be useful in the screening and differential diagnosis of APA.
• Immunological detection of large oxidized lipoproteins in hypertriglyceridemic serum

  Toshihiro Sakurai, Norio Wada, Yuji Takahashi, Ayako Ichikawa, Akiko Ikuta, Hiroaki Furumaki, Shu-Ping Hui, Shigeki Jin, Seiji Takeda, Hirotoshi Fuda, Masato Fujikawa, Chikara Shimizu, Hironori Nagasaka, Hiroyuki Furukawa, Seiichi Kobayashi, Hitoshi Chiba

  ANNALS OF CLINICAL BIOCHEMISTRY 50 (5) 465 - 472 0004-5632 2013/09 [Refereed][Not invited]
   

  Background: Triglyceride-rich, low-density lipoproteins (TG-rich LDL) have been reported as an oxidized lipoprotein species in patients with severe liver disease. Using TG-rich LDL as an immunogen, we obtained a monoclonal antibody (G11-6) that reacted with TG-rich LDL from patients with liver disease and with metal-oxidized LDL only in the early process of the oxidation reaction. This study determined the G11-6-reactive lipoproteins in hypertriglyceridemic serum. Methods: Serum samples from healthy volunteers (n=12) and hypertriglyceridemic patients (n=9) were fractionated by gel filtration and subjected to a sandwich enzyme-linked immunosorbent assay (ELISA) using G11-6 and polyclonal anti-apolipoprotein B antibodies. Results: Small LDL and larger lipoproteins reacted with G11-6. G11-6-reactive small LDL was identified in both the healthy subjects and hypertriglyceridemic patients, whereas G11-6-reactive larger lipoproteins were found only in the hypertriglyceridemic patients. Conclusions: G11-6 is a useful tool for detecting increased large oxidized lipoproteins in hypertriglyceridemic patients.
• Evaluation of various electrode materials for detection of oxidized low-density lipoproteins

  Seiji Takeda, Shu-Ping Hui, Hirotoshi Fuda, Shigeki Jin, Toshihiro Sakurai, Atsushi Ishii, Koichi Mukasa, Kazuhisa Sueoka, Hitoshi Chiba

  Journal of Biomedical Nanotechnology 9 (2) 303 - 306 1550-7033 2013/02 [Refereed][Not invited]
   

  Measurement of oxidized low-density lipoprotein (LDL) generated by oxidative stress of various kinds might be useful for evaluating the risk of cardiovascular disease. We evaluated some electrode materials to detect oxidized LDL electrochemically. Some carbon nanotube dispersions were studied as electrode materials. Native LDL was isolated from normal human serum using ultracentrifugation. Oxidized LDL was prepared by treating the native LDL with CuSO4. Electrodes were fabricated by depositing the nanotube dispersion on a gold electrode, with subsequent drying. The potential change of the electrode against a reference electrode was monitored before and after adding native LDL or oxidized LDL. Only acid-treated carbon nanotubes were able to discriminate both LDL preparations, perhaps because of the carboxylic acid groups introduced on the nanotube by acid treatment. Copyright © 2013 American Scientific Publishers All rights reserved.
• Effects of acid oxidation on carbon nanotube based electrodes for detection of oxidized LDL

  Seiji Takeda, Toshihiro Sakurai, Futaba Ohkawa, Shigeki Jin, Shu-Ping Hui, Hirotoshi Fuda, Koichi Mukasa, Hitoshi Chiba, Kazuhisa Sueoka

  2013 IEEE SENSORS 1038 - 1041 1930-0395 2013 [Refereed][Not invited]
   

  Carbon nanotubes are sometimes treated with an acid or mixture of acids to increase their solubility. From results of our study of single-walled carbon nanotubes, we reported that a CNT-based electrode can detect oxidation of low density-lipoproteins. As described herein, we investigated the effects of acid treatment time on electrode sensitivity for detection of oxidized low-density lipoproteins. Response capabilities of CNT-based electrodes for detecting ox-LDL were improved by acid treatment. However, further increased treatment time degraded their sensitivity. Although detailed mechanisms related to the response are under investigation, results show that an optimum condition exists for acid treatment condition to detect ox-LDL using an electrode.
• Analysis of triacylglycerol hydroperoxides in human lipoproteins by Orbitrap mass spectrometer

  Shu-Ping Hui, Toshihiro Sakurai, Seiji Takeda, Shigeki Jin, Hirotoshi Fuda, Takao Kurosawa, Hitoshi Chiba

  Analytical and Bioanalytical Chemistry 405 (14) 4981 - 4987 1618-2642 2013 [Refereed][Not invited]
   

  Herein, we represent a simple method for the detection and characterization of molecular species of triacylglycerol monohydroperoxides (TGOOH) in biological samples by use of reversed-phase liquid chromatography with a LTQ Orbitrap XL mass spectrometer (LC/LTQ Orbitrap) via an electrospray ionization source. Data were acquired using high-resolution, high-mass accuracy in Fourier-transform mode. Platform performance, related to the identification of TGOOH in human lipoproteins and plasma, was estimated using extracted ion chromatograms with mass tolerance windows of 5 ppm. Native low-density lipoproteins (nLDL) and native high-density lipoproteins (nHDL) from a healthy donor were oxidized by CuSO4 to generate oxidized LDL (oxLDL) and oxidized HDL (oxHDL). No TGOOH molecular species were detected in the nLDL and nHDL, whereas 11 species of TGOOH molecules were detected in the oxLDL and oxHDL. In positiveion mode, TGOOH was found as [M + NH4]+. In negativeion mode, TGOOH was observed as [M + CH3COO]-. TGOOH was more easily ionized in positive-ion mode than in negative-ion mode. The LC/LTQ Orbitrap method was applied to human plasma and three molecular species of TGOOH were detected. The limit of detection is 0.1 pmol (S/N=10:1) for each synthesized TGOOH. © Springer-Verlag Berlin Heidelberg 2013.
• 過酸化脂質の分析―質量分析法を中心に―

  HUI Shu‐Ping, 千葉仁志

  JSBMS Lett 37 (3) 38 - 43 1881-5464 2012/11/30 [Not refereed][Not invited]
  
• A phenolic antioxidant from the Pacific oyster (Crassostrea gigas) inhibits oxidation of cultured human hepatocytes mediated by diphenyl-1-pyrenylphosphine

  Mitsugu Watanabe, Hirotoshi Fuda, Shigeki Jin, Toshihiro Sakurai, Shu-Ping Hui, Seiji Takeda, Takayuki Watanabe, Takao Koike, Hitoshi Chiba

  FOOD CHEMISTRY 134 (4) 2086 - 2089 0308-8146 2012/10 [Refereed][Not invited]
   

  3,5-Dihydroxy-4-methoxybenzyl alcohol (DHMBA), an antioxidant isolated from the Pacific oyster (Crassostrea gigas), was studied in a cell-based fluorometric antioxidant assay using human hepatocyte-derived cells (C3A) and diphenyl-1-pyrenylphosphine (DPPP) as a fluorescent probe. In comparison with two hydrophilic antioxidants. DHMBA showed the stronger inhibition of DPPP-mediated fluorescence than chlorogenic acid and L-ascorbic acid: at a concentration of 320 mu M of DPPP, the inhibition was 26.4 +/- 2.6%, 11.1 +/- 1.2%, and 0 +/- 2.0% for DHMBA, chlorogenic acid, and L-ascorbic acid, respectively (mean +/- SD, n = 4). Their relative oxygen radical absorbance capacities (ORAC) were dissociated with their cell-based antioxidant activities: 1.47 +/- 0.40, 4.57 +/- 0.30, and 0.53 +/- 0.13 mu mol TE/mu mol for DHMBA, chlorogenic acid, and L-ascorbic acid, respectively (mean +/- SD, n = 4). The amphiphilicity of DHMBA was better than chlorogenic acid and L-ascorbic acid might underlie this dissociation. Since the C3A cells are human hepatoma-derived cells, DHMBA might be useful in the prevention and treatment of liver diseases by involving an oxidation process. (C) 2012 Elsevier Ltd. All rights reserved.
• LC‐MS/MSで測定した血清,尿18‐hydroxycortisolによる原発性アルドステロン症の診断

  和田典男, 神繁樹, 藤谷昴文, 惠淑萍, 柳澤克之, 黒澤隆夫, 千葉仁志

  日本内分泌学会雑誌 88 (2) 695  0029-0661 2012/09/20 [Not refereed][Not invited]
  
• Novel monoclonal antibody recognizing triglyceride-rich oxidized LDLs associated with severe liver disease and small oxidized LDLs in normal subjects

  Toshihiro Sakurai, Ayako Ichikawa, Hiroyuki Furukawa, Norio Wada, Atsushi Nagasaka, Yuji Takahashi, Masato Fujikawa, Akiko Ikuta, Hiroaki Furumaki, Maiko Shiga, Chikara Shimizu, Shu-Ping Hui, Shigeki Jin, Seiji Takeda, Hirotoshi Fuda, Hironori Nagasaka, Seiichi Kobayashi, Hitoshi Chiba

  ANNALS OF CLINICAL BIOCHEMISTRY 49 (Pt5) 456 - 462 0004-5632 2012/09 [Refereed][Not invited]
   

  Background: Triglyceride-rich low-density lipoproteins (TG-rich LDLs) in the plasma of patients with severe liver disease are reported to change macrophages into foam cells in vitro. Methods: Male BALB/c mice were immunized with TG-rich LDLs isolated from the plasma of a patient with severe liver disease. The resulting monoclonal antibody (G11-6) was used in a sandwich enzyme-linked immunosorbent assay (ELISA) in combination with polyclonal anti-apolipoprotein B antibodies. The time course of copper-mediated LDL oxidation was monitored using this ELISA. The results were compared with those of the two commercial ELISAs for oxidized LDLs using DLH or ML25, thiobarbituric acid reactive substances and the optical absorbance for the conjugated dienes generated in lipid peroxides. Furthermore, the lipoprotein fractions separated by gel filtration were tested with this ELISA in healthy volunteers (n = 11) and patients (n = 3) with liver disease. Results: G11-6 reacted with oxidized LDLs during only the early phase of copper oxidation, being distinct from the other monoclonal antibodies and methods. G11-6 was confirmed to react with TG-rich LDLs in patients, while it reacted with small LDL particles in normal controls. Conclusions: The monoclonal antibody G11-6 is useful for detecting oxidized small LDLs in normal controls and oxidized TG-rich LDLs in patients with severe liver disease.
• Methods for determination of lipid peroxides

  惠 淑萍, 千葉 仁志

  細胞 ニューサイエンス社 44 (8) 370 - 373 1346-7557 2012/07 [Not refereed][Not invited]
  
• Detection and characterization of cholesteryl ester hydroperoxides in oxidized LDL and oxidized HDL by use of an Orbitrap mass spectrometer

  Shu-Ping Hui, Toshihiro Sakurai, Futaba Ohkawa, Hiroaki Furumaki, Shigeki Jin, Hirotoshi Fuda, Seiji Takeda, Takao Kurosawa, Hitoshi Chiba

  ANALYTICAL AND BIOANALYTICAL CHEMISTRY 404 (1) 101 - 112 1618-2642 2012/07 [Refereed][Not invited]
   

  Oxidation of cholesteryl esters in lipoproteins by reactive oxygen species yields cholesteryl ester hydroperoxides (CEOOH). In this study, we developed a novel method for identification and characterization of CEOOH molecules in human lipoproteins by use of reversed-phase liquid chromatography with an hybrid linear ion trap-Orbitrap mass spectrometer (LC-LTQ Orbitrap). Electrospray ionization tandem mass spectrometric analysis was performed in both positive-ion and negative-ion modes. Identification of CEOOH molecules was completed by use of high-mass-accuracy (MA) mass spectrometric data obtained by using the spectrometer in Fourier-transform (FT) mode. Native low-density lipoproteins (nLDL) and native high-density lipoproteins (nHDL) from a healthy donor were oxidized by CuSO4, furnishing oxidized LDL (oxLDL) and oxidized HDL (oxHDL). No CEOOH molecules were detected in the nLDL and the nHDL, whereas six CEOOH molecules were detected in the oxLDL and the oxHDL. In positive-ion mode, CEOOH was detected as [M + NH4](+) and [M + Na](+) ions. In negative-ion mode, CEOOH was detected as [M + CH3COO](-) ions. CEOOH were more easily ionized in positive-ion mode than in negative-ion mode. The LC-LTQ Orbitrap method was applied to human plasma and six species of CEOOH were detected. The limit of detection was 0.1 pmol (S/N = 5:1) for synthesized CEOOH.
• Quantitative determination of phosphatidylcholine hydroperoxides during copper oxidation of LDL and HDL by liquid chromatography/mass spectrometry

  Shu-Ping Hui, Yudai Taguchi, Seiji Takeda, Futaba Ohkawa, Toshihiro Sakurai, Shinobu Yamaki, Shigeki Jin, Hirotoshi Fuda, Takao Kurosawa, Hitoshi Chiba

  ANALYTICAL AND BIOANALYTICAL CHEMISTRY 403 (7) 1831 - 1840 1618-2642 2012/06 [Refereed][Not invited]
   

  1-Palmitoyl-2-linoleoylphosphatidylcholine monohydroperoxide (PC 16:0/18:2-OOH) and 1-stearoyl-2-linoleoylphosphatidylcholine monohydroperoxide (PC 18:0/18:2-OOH) were measured by liquid chromatography/mass spectrometry (LC/MS) using nonendogenous 1-palmitoyl-2-heptadecenoylphosphatidylcholine monohydroperoxide as an internal standard. The calibration curves for synthetic PC 16:0/18:2-OOH and PC 18:0/18:2-OOH, which were obtained by direct injection of the internal standard into the LC/MS system, were linear throughout the calibration range (0.8-12.8 pmol). Within-day and between-day coefficients of variation were less than 10%, and the recoveries were between 86% and 105%. The limit of detection (LOD) and the limit of quantification (LOQ) were determined using synthetic standards. The LOD (signal-to-noise ratio 3:1) was 0.01 pmol, and the LOQ (signal-to-noise ratio 6:1) was 0.08 pmol for both PC 16:0/18:2-OOH and PC 18:0/18:2-OOH. With use of this method, the concentrations of PC 16:0/18:2-OOH and PC 18:0/18:2-OOH in the lipoprotein fractions during copper-mediated oxidation were determined. We prepared oxLDL and oxHDL by incubating native LDL and native HDL from human plasma (n = 10) with CuSO4 for up to 4 h. The time course of the PC 16:0/18:2-OOH and PC 18:0/18:2-OOH levels during oxidation consisted of three phases. For oxidized LDL, both compounds exhibited a slow lag phase and a subsequent rapidly increasing propagation phase, followed by a gradually decreasing degradation phase. In contrast, for oxidized HDL, both compounds initially exhibited a prompt propagation phase with a subsequent plateau phase, followed by a rapid degradation phase. The analytical LC/MS method for phosphatidylcholine hydroperoxides might be useful for the analysis of biological samples.
• Detection of oxidized LDL using a carbon nanotube electrode

  Seiji Takeda, Shu-Ping Hui, Keisuke Fukuda, Hirotoshi Fuda, Shigeki Jin, Toshihiro Sakurai, Atsushi Ishii, Koichi Mukasa, Kazuhisa Sueoka, Hitoshi Chiba

  SENSORS AND ACTUATORS B-CHEMICAL 166 (5) 833 - 836 0925-4005 2012/05 [Not refereed][Not invited]
   

  Oxidized low-density lipoproteins (LDLs) play a key role in cardiovascular disease development, but no convenient measurement method is available for them. Using a carbon nanotube (CNT) electrode, we measured oxidized LDL using amperometric detection. Treating SWCNT with a mixture of acids produced a CNT dispersion that yielded nanotube-based electrodes after deposition on a gold electrode and drying. Current was monitored in the nanotube electrode before and after adding LDL or oxidization of LDL Oxidized LDL changed the current more than 10 nA, although LDL addition induced no significant change. Our CNT electrode enables simple detection of oxidized LDL. (C) 2012 Elsevier B.V. All rights reserved.
• 血中,尿中18‐hydroxycortisol測定による原発性アルドステロン症の鑑別診断

  和田典男, 神繁樹, 惠淑萍, 柳澤克之, 黒澤隆夫, 千葉仁志

  日本内分泌学会雑誌 88 (1) 302  0029-0661 2012/04/01 [Not refereed][Not invited]
  
• 過酸化脂質およびプラズマローゲンに関する臨床化学的研究

  HUI Shu‐Ping

  臨床化学 41 (1) 54 - 61 0370-5633 2012/01/31 [Not refereed][Not invited]
  
• Isolation and Characterization of a Phenolic Antioxidant from the Pacific Oyster (Crassostrea gigas)

  Mitsugu Watanabe, Hirotoshi Fuda, Shigeki Jin, Toshihiro Sakurai, Futaba Ohkawa, Shu-Ping Hui, Seiji Takeda, Takayuki Watanabe, Takao Koike, Hitoshi Chiba

  JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY 60 (3) 830 - 835 0021-8561 2012/01 [Refereed][Not invited]
   

  Using an oxygen radical absorbance capacity (ORAC) assay, antioxidant activity was detected in the ethanol extract of the Pacific oyster, which was purified by sequential extraction with organic solvents. The ethyl acetate fraction showed the strongest antioxidant activity and was further purified, yielding a single compound [as assessed by thin-layer chromatography (TLC) reverse-phase high-performance liquid chromatography (HPLC)]. This compound was identified as 3,5-dihydroxy4-metboxybenzyl alcohol on the basis of H-1 and C-13 nuclear magnetic resonance (NMR), heteronuclear multiple-bond correlz.tion (HMBC), and electrospray ionization-mass spectrometry (ESI-MS) spectral analyses, a conclusion that was confirmed bycheinical synthesis. The concentration of the compound was 6.7 mg/100 g of whole oyster meat wet weight. This azaphi'philic gritiosidant retarded the copper mediated oxidation of low density lipoproteins (LDLs) and the generation of,tr.ickarbitonc acid.teactive substances. Furthermore, the compound showed substantial antioxidant activity using the ORAC and 2,2-diphenyl-1-picrythy'drazyl (DPPH) assays compared to natural antioxidants. Although the same compound was previously found in brown algae, its presence in other organisms and antioxidant activity are reported here for the first time.
• Application of Kelvin force microscopy for evaluation of oxidized low-density lipoprotein

  Seiji Takeda, Futaba Ohkawa, Toshihiro Sakurai, Shigeki Jin, Hirotoshi Fuda, Shu-Ping Hui, Hitoshi Chiba, Kazuhisa Sueoka

  2012 IEEE SENSORS PROCEEDINGS 1751 - 1754 1930-0395 2012 [Refereed][Not invited]
   

  Investigation of the properties of oxidized low-density lipoprotein is important because it is a risk factor of cardiovascular disease development. Low-density lipoprotein is known to have a positive charge on the surface attributable to amino groups of lysine moieties. Its positive charge can be decreased by oxidation. For this study, the topography and surface potential of LDL and ox-LDL were measured using atomic force microscopy and Kelvin probe force microscopy. The Kelvin probe force microscopy images of ox-LDL differ from those obtained using n-LDL. We demonstrated a method to evaluate the oxidation level of LDL using AFM and KPFM.
• 過酸化脂質及び抗酸化脂質に関する研究

  惠淑萍

  臨床化学 40 79 - 80 0370-5633 2011/07/31 [Not refereed][Not invited]
  
• CD36 deficiency predisposing young children to fasting hypoglycemia

  Hironori Nagasaka, Tohru Yorifuji, Tomozumi Takatani, Yoshiyuki Okano, Hirokazu Tsukahara, Hidekatsu Yanai, Ken-ichi Hirano, Shu-Ping Hui, Satoshi Hirayama, Tetsuya Ito, Hitoshi Chiba, Takashi Miida

  METABOLISM-CLINICAL AND EXPERIMENTAL 60 (6) 881 - 887 0026-0495 2011/06 [Refereed][Not invited]
   

  Fatty acid (FA) beta-oxidation defects cause hypoglycemia. Our aim was to determine if CD36-a membrane transporter for long-chain FAs-deficiency predisposes children to hypoglycemia. After overnight fasting, we measured parameters for carbohydrate and FA metabolisms at 12-, 14-, and 16-hour fasting points in 51 preschool children with histories of episodic hypoglycemia and 49 age-matched healthy controls. Simultaneously, the expressions of CD36 on platelets and monocytes were examined to determine the phenotypes. Six of the 51 hypoglycemic children and none of the 49 control children were diagnosed as having type I CD36 deficiency. Four and 3 children were diagnosed as having type II CD36 deficiency, respectively. Hypoglycemia was often recurrent in the type I CD36 group. At any fasting point, the type I CD36 group showed significantly lower blood glucose and insulin concentrations than the other groups: glucose, P < .001 vs control group and P < .01 or P < .001 vs type II/wild-type CD36 hypoglycemic groups; insulin, P < .001 vs control group and P < .01 vs type II/wild-type CD36 hypoglycemic groups. Free FA concentration in the type I group was always 1.5- to 2.0-fold higher than that in the other groups, whereas the total ketone body concentration was consistently about two thirds of that in the other groups. Among the type II, wild-type, and control groups, there were no significant differences in the parameters except that the wild-type group showed significantly lower FFA concentration (P < .05). These results suggested that type I CD36 deficiency but not type II CD36 deficiency predisposes preschool children to hypoglycemia. (C) 2011 Elsevier Inc. All rights reserved.
• Liquid chromatography-mass spectrometric determination of plasmalogens in human plasma

  Shu-Ping Hui, Hitoshi Chiba, Takao Kurosawa

  ANALYTICAL AND BIOANALYTICAL CHEMISTRY 400 (7) 1923 - 1931 1618-2642 2011/06 [Refereed][Not invited]
   

  A new liquid chromatography-mass spectrometry (LC/MS) method has been developed for the quantitative analysis of plasmalogens in human plasma using a nonendogenous plasmalogen (1-O-1'-(Z)-tricosenyl-2-oleoyl-rac-glycero-3-phosphocholine, PLS 23:0/18:1) as an internal standard. 1-O-1'-(Z)-Tricosenyl glyceryl ether was prepared by reacting lithioalkoxyallyl with 1-iodoeicosane as the key intermediate in the formation of PLS 23:0/18:1. In LC/MS analyses, PLS 23:0/18:1 generated significant fragment ions in positive and negative modes. In positive ion mode, the [M+H](+) of PLS 23:0/18:1 yielded unique fragments with cleavages at the sn-1 and sn-2 positions of the glycerol backbone. In negative ion mode, the [M+CH(3)COO](-) of PLS 23:0/18:1 resulted in characteristic fragmentation at the sn-2 and sn-3 positions. 1-O-1'-(Z)-Hexadecenyl-2-linoleoyl-rac-glycero-3-phosphocholine (PLS 16:0/18:2) and 2-arachidonoyl-O-1'-(Z)-hexadecenyl-rac-glycero-3-phosphocholine (PLS 16:0/20:4) were chemically synthesized as PLS 23:0/18:1. The calibration curves obtained for PLS 16:0/18:2 and PLS 16:0/20:4 were linear throughout the calibration range (0.04-1.60 pmol). The LOD (S/N = 5:1) was 0.008 pmol and the LOQ (S/N = 6:1) was 0.01 pmol for both PLS 16:0/18:2 and PLS 16:0/20:4. Plasma concentrations of PLS 16:0/18:2 and PLS 16:0/20:4 were 4.0 +/- 1.3 mu M and 3.5 +/- 1.2 mu M (mean +/- SD), respectively, in five healthy volunteers.
• Chiral Separation, Determination of Absolute Configuration, and High-performance Liquid Chromatography Detection of Enantiomeric 3-hydroxyhexadecanoyl-CoA

  Shirou Tsuchida, Koutarou Kawamoto, Kana Nunome, Naoya Hamaue, Shu-Ping Hui, Tsuyoshi Murai, Takashi Aoki, Takao Kurosawa

  JOURNAL OF OLEO SCIENCE 60 (2) 87 - 92 1345-8957 2011/02 [Refereed][Not invited]
   

  The enoyl-coenzyme A (CoA) hydratase catalyzes the hydration of 2-enoyl-CoA to yield 3-hydroxyacyl-CoA in mitochondrial and peroxisomal beta-oxidation. However, the stereospecificities of these hydratases differ from each other. To provide clear evidence of the stereospecificities of hydratases, the absolute configuration of 3-hydroxyhexadecanoyl-CoAs was determined, and they were subjected to a high-performance liquid chromatography (HPLC) using a chiral separation column. The retention time of 3(R)-hydroxyhexadecanoyl-CoA was shorter than that of 3(S)-hydroxyhexadecanoyl-CoA. The HPLC analysis carried out using a chiral separation column is considered to be useful for the study of enoyl-CoA hydratase.
• Cellular response after crush injury in adult zebrafish spinal cord.

  Hui SP, Dutta A, Ghosh S

  Developmental dynamics : an official publication of the American Association of Anatomists 11 239 (11) 2962 - 2979 1058-8388 2010/11 [Refereed][Not invited]
  
• Measurement of lipoprotein particle sizes using dynamic light scattering

  Toshihiro Sakurai, Suchin Trirongjitmoah, Yuka Nishibata, Takeshi Namita, Masahiro Tsuji, Shu-Ping Hui, Shigeki Jin, Koichi Shimizu, Hitoshi Chiba

  ANNALS OF CLINICAL BIOCHEMISTRY 47 (5) 476 - 481 0004-5632 2010/09 [Refereed][Not invited]
   

  Background: A simple method for the measurement of LDL particle sizes is needed in clinical laboratories because a predominance of small, dense LDL (sd LDL) has been associated with coronary heart disease. We applied dynamic light scattering (DLS) to measure lipoprotein particle sizes, with special reference to sd LDL. Methods: Human serum lipoproteins isolated by a combination of ultracentrifugation and gel chromatography, or by sequential ultracentrifugation, were measured for particle size using DLS. Results: The sizes of polystyrene beads, with diameters of 21 and 28 nm according to the manufacturer, were determined by DLS as 19.3 +/- 1.0 nm (mean +/- SD, n = 11) and 25.5 +/- 1.0 nm, respectively. The coefficients of variation for the 21 and 28 nm beads were 5.1% and 3.8% (within-run, n = 11), and 2.9% and 6.2% (between-run, n = 3), respectively. The lipoprotein sizes determined by DLS for lipoprotein fractions isolated by chromatography were consistent with the elution profile. Whole serum, four isolated lipoprotein fractions (CM + VLDL + IDL, large LDL, sd LDL and HDL) and a non-lipoprotein fraction isolated by sequential ultracentrifugation were determined by DLS to be 13.1 +/- 7.5, 37.0 +/- 5.2, 21.5 +/- 0.8, 20.3 +/- 1.1, 8.6 +/- 1.5 and 8.8 +/- 2.0 nm, respectively. Conclusions: The proposed DLS method can differentiate the sizes of isolated lipoprotein particles, including large LDL and sd LDL, and might be used in clinical laboratories in combination with convenient lipoprotein separation.
• Analyses for phosphatidylcholine hydroperoxides by LC/MS

  Shu-Ping Hui, Hitoshi Chiba, Shigeki Jin, Hironori Nagasaka, Takao Kurosawa

  JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES 878 (20) 1677 - 1682 1570-0232 2010/06 [Refereed][Not invited]
   

  A new liquid chromatography mass spectrometry (LC/MS) method has been developed for the qualitative and quantitative analyses of phosphatidylcholine hydroperoxides (PC-OOH) in human plasma using a synthetic hydroperoxide (1-stearoyl-2-erucoyl-PC monohydroperoxide, PC 18 0/22 1-OOH) as an internal standard 1-Stearoyl-2-linoleoyl-PC monohydroperoxide (PC 18 0/18 2-OOH) was identified in plasma by LC/MS by comparison with an authentic standard The calibration curves obtained for 1-palmitoyl-2-linoleoyl-PC monohydroperoxide. PC 16 0/18 2-OOH and PC 18.0/18 2-OOH were linear throughout the calibration range (0 1-1 0 pmol) The limit of detection (LOD) (S/N = 3 1) was 0 01 pmol, and the limit of quantification (LOQ) (S/N = 6 1) was 0 1 pmol for both PC 16 0/18 2-OOH and PC 18.0/18.2-OOH Plasma concentrations of PC 16 0/18 2-OOH and PC 18 0/18 2-OOH were 89 and 32 nM, respectively, in a healthy volunteer (C) 2010 Elsevier B V All rights reserved
• Measurement of the Transport Activities of Bile Salt Export Pump Using Chemiluminescence Detection Method

  Kana Yamaguchi, Tsuyoshi Murai, Hikaru Yabuuchi, Shu-Ping Hui, Takao Kurosawa

  YAKUGAKU ZASSHI-JOURNAL OF THE PHARMACEUTICAL SOCIETY OF JAPAN 130 (5) 755 - 761 0031-6903 2010/05 [Refereed][Not invited]
   

  Monovalent bile acids, such as taurine- and glycine-conjugated bile acids, are excreted into bile by bile salt export pumps (BSEP, ABCB11). Human BSEP (hBSEP) is physiologically important because it was identified as the gene responsible for the genetic disease: progressive familial intrahepatic cholestasis type 2 (PFIC-2). The evaluation of the inhibitory effect of hBSEP transport activity provides significant information for predicting toxic potential in the early phase of drug development. The role and function of hBSEP have been investigated by the examination of the ATP-dependent transport of radioactive isotopically (RI) -labeled bile acid such as a tritium labeled taurocholic acid, in membrane vesicles obtained from hBSEP-expressing cells. The chemiluminescence detection method using 3 alpha-hydroxysteroid dehydrogenase (3 alpha-HSD) had been developed for a simple analysis of bile acids in human biological fluids. This method is extremely sensitive and it may be applicable for the measurements of bile acid transport activities by hBSEP vesicles without using RI-labeled bile acid. The present paper deals with an application of the chemiluminescence detection method using 3 alpha-HSD with enzyme cycling method to the measurement of ATP-dependent transport activities of taurocholic acid (T-CA) in membrane vesicles obtained from hBSEP-expressing Sf9 cells. Calibration curves for T-CA was linear over the range from 10 to 400 pmol/ml. The values of the kinetic parameters for hBSEP vesicles obtained by the chemiluminescence detection method were comparable with the values of that obtained by liquid chromatography-mass spectrometry method. This assay method was highly useful for the measurements of bile acid transport activities.
• 生化学的検査[2]A.脂質関係 過酸化脂質

  HUI ShuPing

  日本臨床 68 120 - 124 0047-1852 2010/01/28 [Not refereed][Not invited]
  
• Lipid peroxide

  惠 淑萍

  Japanese journal of clinical medicine 日本臨床社 68 (0) 120 - 124 0047-1852 2010/01 [Not refereed][Not invited]
  
• Measurement of Bile Salt Export Pump Transport Activities using a Fluorescent Bile Acid Derivative

  Kana Yamaguchi, Tsuyoshi Murai, Hikaru Yabuuchi, Shu-Ping Hui, Takao Kurosawa

  DRUG METABOLISM AND PHARMACOKINETICS 25 (2) 214 - 219 1347-4367 2010 [Refereed][Not invited]
   

  A novel fluorescent bile acid derivative, 4-N,N-dimethylaminosulfony1-2,1.3-benzoxadiazole-conjugated bile acid was synthesized as a probe to develop a rapid screening method for function analysis of bile salt export pump (BSEP. ABCB 11) The transport properties of the synthetic fluorescent bile acid derivative in membrane vesicles obtained from hBSEP-expressing Sf9 cells were examined using the liquid chromatography-electrospray ionization-mass spectrometry method The Michaelis-Menten constant and maximum uptake rate for the synthetic fluorescent bile acid derivative by hBSEP were 23 1 +/- 1 6 mu M and 623 2 +/- 22 4 pmol/min/mg protein, respectively. These kinetic parameters of the synthetic fluorescent bile acid derivative were comparable with those of an unlabeled bile acid, taurocholic acid. Moreover, we examined inhibitory effects of various drugs on hBSEP-mediated uptake of the fluorescent bile acid derivative using a fluorescence detection method The relative uptake activities (percent of control) for the fluorescent bile acid derivative in the presence of an inhibitor were in accordance with previous findings using (3)H-labeled taurocholic acid Our results suggest that the synthetic fluorescent bile acid derivative may be useful for evaluation of the inhibitory effects of various drugs on hBSEP-mediated uptake.
• Improved LC/MS Analytical Method for PC Hydroperoxides in Human Plasma

  Shuping Hui, Hitoshi Chiba, Takao Kurosawa

  FREE RADICAL BIOLOGY AND MEDICINE 49 S211 - S211 0891-5849 2010 [Refereed][Not invited]
  
• Increased Plasma TG Hydroperoxide in Advanced Liver Disease

  Shuping Hui, Hitoshi Chiba, Takao Kurosawa

  FREE RADICAL BIOLOGY AND MEDICINE 47 S188 - S188 0891-5849 2009 [Refereed][Not invited]
  
• Fluctuation of lipoprotein metabolism linked with bile acid-activated liver nuclear receptors in Alagille syndrome

  Hironori Nagasaka, Takashi Miida, Ken-ichi Hirano, Akemi Ota, Kei Murayama, Tohru Yorifuji, Kunihiko Kobayashi, Tornommi Takatani, Hirokazu Tsukahara, Shu-Ping Hui, Masaki Takayanagi, Hitoshi Chiba

  ATHEROSCLEROSIS 198 (2) 434 - 440 0021-9150 2008/06 [Refereed][Not invited]
   

  Alagille syndrome (AGS) is a rare hereditary disorder exhibiting fluctuating cholestasis and dyslipidemia. Farnesoid X receptor (FXR) and liver X receptor (LXR) are hepatic nuclear receptors that regulate bile acid and lipoprotein metabolism. To investigate whether cholestasis is related to dyslipidemia and hepatic nuclear receptor expression in AGS patients, we determined the blood levels of total bile acid (TBA) and lipoprotein parameters, and examined hepatic nuclear receptor expression in three AGS children and their three incomplete AGS parents repeatedly over several years. In the AGS children, TBA level showed significant positive correlations with low-density lipoprotein-cholesterol, apolipoprotein E (apoE)-rich high-density lipoprotein-cholesterol (HDL-C), apoA-I, apoE, and cholesteryl ester transfer protein (CETP) concentrations, but negative correlation with apoE-poor HDL-C concentration. Western blot analysis of liver biopsy specimens revealed that FXR and LXR expression increased in parallel with TBA level. CETP- and ATP-binding cassette transporter A1 expression also increased with TBA level, while scavenger receptor class B type-I expression showed the opposite response. However, apoA-I expression was similar to the control level at any TBA level. In the incomplete AGS parents, TBA and lipoprotein parameters showed little fluctuation. In summary, cholestasis is closely related to dyslipidemia and hepatic nuclear receptor expression in AGS patients. (C) 2008 Elsevier Ireland Ltd. All rights reserved.
• Reduced apolipoprotein E-rich high-density lipoprotein level at birth is restored to the normal range in patients with familial Hypercholesterolemia in the first year of life

  Hironori Nagasaka, Takashi Miida, Kenichi Hirano, Akemi Ota, Tohru Yorifuji, Tomozumi Takatani, Hirokazu Tsukahara, Masaki Takayanagi, Shu-Ping Hui, Kunihiko Kobayashi, Hitoshi Chiba

  JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM 93 (3) 779 - 783 0021-972X 2008/03 [Refereed][Not invited]
   

  Background: High-density lipoprotein (HDL) consists of apolipoprotein E (apoE)-rich and apoE-poor HDL particles. ApoE-rich HDL level is high at birth but decreases after birth with reciprocal elevation in low-density lipoprotein (LDL)-cholesterol. Objectives: The objective of the study was to clarify whether apoE-rich HDL decreases after birth in children with familial hypercholesterolemia (FH), a disorder caused by impaired LDL clearance. Methods: We measured apoE-rich HDL-cholesterol and LDL-cholesterol during the first year of life in 10 FH children (one homozygote and nine heterozygotes), 12 non-FH siblings, and 75 healthy controls. Results: At birth, apoE-rich HDL-cholesterol was undetectable in a homozygous FH child and lower in heterozygous FH children than non-FH siblings and controls ( 4 +/- 2 vs. 12 +/- 4 and 11 +/- 4 mg/dl, P < 0.001). At 3-4 months, apoE-rich HDL-cholesterol increased in homozygous and heterozygous FH children and decreased in non-FH siblings and controls. At 12 months, apoE-rich HDL-cholesterol levels were similar among these four groups (6-7 mg/dl). In contrast, LDL-cholesterol concentration was always twice as high in heterozygous FH children as non-FH siblings and controls ( at birth, 50 +/- 15 vs. 25 +/- 7 and 25 +/- 5 mg/dl, P < 0.001; at 3 - 4 months of age, 159 +/- 29 vs. 71 similar to 16 and 73 +/- 15 mg/dl, P < 0.001; at 12 months of age, 156 +/- 29 vs. 75 +/- 18 and 76 +/- 17 mg/dl, P < 0.001). Conclusion: ApoE- rich HDL level is low at birth in FH children and increases to the normal level in the first year of life, opposite to the change in normal children.
• An improved HPLC assay for phosphatidylcholine hydroperoxides (PCOOH) in human plasma with synthetic PCOOH as internal standard

  Shu-Ping Hui, Hitoshi Chiba, Toshihiro Sakurai, Chitose Asakawa, Hironori Nagasaka, Tsuyoshi Murai, Hajime Ide, Takao Kurosawa

  JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES 857 (1) 158 - 163 1570-0232 2007/09 [Refereed][Not invited]
   

  Quantitative and qualitative analyses of 1-palmitoyl-2-linoleoyl-phosphatidylcholne monohydroperoxide [PC 16:0/18:2-OOH] and 1-stearoyl-2-linoleoyl-phosphatidylcholine monohydroperoxide [PC 18:0/18:2-OOH] in human plasma were improved by chemiluminescence HPLC using synthetic 1-stearoyl-2-erucoyl-phosphatidylcholine monohydroperoxide (PC 18:0/22:1-OOH) as internal standard. The calibration curves of synthetic PC 16:0/18:2-OOH and PC 18:0/18:2-OOH, obtained by their direct injections with the IS into the HPLC system, were linear throughout the calibration range (10-1000 pmol). Within-day and between-day coefficients of variation were below 8%, and the recoveries were between 84% and 101%. Plasma concentrations of PC 16:0/18:2-OOH and PC 18:0/18:2-OOH were 102 +/- 59 nM (mean +/- SD) and 36 +/- 120 nM. respectively, in the 33 healthy volunteers. The present method might help understanding incompletely understood pathway of plasma phosphatidylcholine hydroperoxides. (c) 2007 Elsevier B.V. All rights reserved.
• Attenuated aerobic exercise capacity in CD36 deficiency

  Hidekatsu Yanai, Ichiro Watanabe, Kojiro Ishii, Mie Morimoto, Hironobu Fujiwara, Shigeru Yoshida, Shu-Ping Hui, Kazuhiko Matsuno, Hitoshi Chiba

  JOURNAL OF MEDICAL GENETICS 44 (7) 445 - 447 0022-2593 2007/07 [Refereed][Not invited]
   

  Background: An important role of CD36 in muscle fatty acid (FA) uptake has been shown in CD36-knockout or CD36-overexpressed mice. FA is a predominant substrate in energy production during light exercise below the anaerobic threshold (AT). We studied whether aerobic exercise capacity in humans could be affected by CD36 deficiency. Methods: We investigated the ventilatory threshold (VT) and serum FA changes in normal participants (n = 22) and participants with CD36 deficiency (n = 12) during pedalling on a cycle ergometer. Results: In participants with CD36 deficiency, FA levels were not reduced at peak work rate, whereas FA levels decreased by about 50% in normal participants. Participants with CD36 deficiency showed significantly lower VT than normal participants. A significant correlation was observed between VT and percentage changes in FA at peak work rate. Conclusion: This study found reduced FA utilisation and an attenuated aerobic exercise capacity in CD36 deficiency, indicating that CD36-mediated FA oxidation is an important determinant for aerobic exercise capacity in humans.
• Depletion of high-density lipoprotein and appearance of triglyceride-rich low-density lipoprotein in a Japanese patient with FIC1 deficiency manifesting benign recurrent intrahepatic cholestasis

  Hironori Nagasaka, Hitoshi Chiba, Shu-Ping Hui, Hajime Takikawa, Takashi Miida, Masaki Takayanagi, Tohru Yorifuji, Makoto Hasegawa, Akemi Ota, Ken-ichi Hirano, Hideaki Kikuchi, Hirokazu Tsukahara, Kunihiko Kobayashi

  JOURNAL OF PEDIATRIC GASTROENTEROLOGY AND NUTRITION 45 (1) 96 - 105 0277-2116 2007/07 [Refereed][Not invited]
   

  Objective: Lipoprotein metabolism in FIC I deficiency due to ATP8B1 mutations has never been studied sufficiently. This study was performed to investigate the detailed lipoprotein metabolism in benign recurrent intrahepatic cholestasis (BRIC) caused by FIC1 deficiency. Patients and Methods: Lipoprotein profile and major lipoprotein regulators such as lecithin: cholesterol acyltransferase (LCAT), hepatic triglyceride lipase (HTGL), lipoprotein lipase, and cholesteryl ester transfer protein in a Japanese patient with BRIC were serially examined during a bout of cholestasis. Liver expression of farnesoid X receptor (FXR), which suppresses high-density lipoprotein (HDL) generation., was also examined. Results: Hypercholesterolemia and lipoprotein X accumulation were never observed throughout this study. When the cholestasis was severe, triglyceride-rich low-density lipoprotem (LDL) accounted for most of the plasma lipoproteins whereas HDL was hardly detectable. Concurrently, activities of all regulators were decreased, together with decreases of the serum parameter for liver protein synthesis. In particular, suppressions of LCAT and HTGL activities were severe and greatly contributed to the appearance of triglyceride-rich LDL. As the cholestasis improved, this LDL gradually transformed into normal LDL with the recoveries of LCAT and HTGL activities. The activities of all regulators for the last I to 2 months were normal but HDL remained depleted. His liver showed low FXR expression compared with control livers. Conclusions: The present study showed an appearance of triglyceride-rich LDL due to suppressions of LCAT and HTGL activities and a depletion of HDL that is not able to be explained by lipoprotein regulators or FXR in our patient.
• Improved HPLC assay for lipid peroxides in human plasma using the internal standard of hydroperoxide

  SP Hui, T Murai, T Yoshimura, H Chiba, H Nagasaka, T Kurosawa

  LIPIDS 40 (5) 515 - 522 0024-4201 2005/05 [Refereed][Not invited]
   

  We have developed a sensitive reversed-phase chemiluminescence HPLC approach for simultaneous quantitative and qualitative analyses of hydroperoxides of cholesteryl ester and TG in human plasma. Standard hydroperoxides of cholesteryl ester and TG and a novel internal standard (1 -tetradecanyl 3-octadecenoyloxy-5 beta-cholan-24-oate monohydroperoxicle) (I.S.) were chemically synthesized and the standard curves confirmed to be linear throughout the calibration range (1-1000 pmol). Within-day and between-day CV were less than 7%, and the recoveries were within the range of 84-93%. With sample size minimized to 0.1 mL of plasma for each run, plasma cholesteryl ester hydroperoxide levels were 189 +/- 87 nM (mean SD) in healthy young (22-25 yr old; n = 15, male/female = 6:9) and 210 69 nM in healthy elderly (39-60 yr old; n = 6, male/female = 3:3). TG hydroperoxide was not detected in healthy subjects. In patients with advanced liver failure (36-67 yr old; n = 4, male/female = 2:2), hydroperoxide levels of plasma cholesteryl ester and TG were 11,903 9,553 nM and 3,318 1,590 nM, respectively, indicating an involvement of lipid oxidation. Sensitive and specific monitoring of plasma lipid peroxides using the present chemiluminescence HPLC approach with the synthesized I.S. may help our understanding of chemical and pathophysiological aspects of lipid peroxidation.
• Oxidized low-density lipoprotein upregulates GM2 activator protein gene expression

  Yanai H, Yoshida H, Fujiwara H, Yoshida S, Fuda H, Shu-Ping Hui, Nagasaka H, Tada N, Chiba H

  Am J Biochem Biotechnol 1 (2) 90 - 94 2005/05 [Refereed][Not invited]
  
• Simple chemical syntheses of TAG monohydroperoxides

  SP Hui, T Murai, T Yoshimura, H Chiba, T Kurosawa

  LIPIDS 38 (12) 1287 - 1292 0024-4201 2003/12 [Refereed][Not invited]
   

  For the purpose of synthesizing standards to be used in the quantification of TAG hydroperoxides, three TAG (1,2-dioleoyl-3-palmitoylglycerol, 1-oleoyl-2-linoleoyl-3-palmitoylglycerol, and triolein) monohydroperoxides were chemically synthesized as authentic specimens. TAG were prepared by using a simple condensation in pyridine of glycerol and the corresponding acid chlorides. These TAG were then converted into monohydroperoxides by a photosensitized peroxidation. The synthesized monohydroperoxides were analyzed by normal-phase and RP-HPLC. The results of normal-phase HPLC analysis showed that monohydroperoxides from a corresponding TAG were a mixture of regioisomers. In RP-HPLC, however, the regioisomers of monohydroperoxides were not separated and gave a single peak, which may improve the sensitivity for the detection of TAG monohydroperoxides. In this study TAG monohydroperoxide standards were synthesized; these will be useful for the study of yet unknown biological and pathological roles of TAG hydroperoxides.
• Unique character and metabolism of high. density lipoprotein (HDL) in fetus

  H Nagasaka, H Chiba, H Kikuta, H Akita, Y Takahashi, H Yanai, SP Hui, H Fuda, H Fujiwara, K Kobayashi

  ATHEROSCLEROSIS 161 (1) 215 - 223 0021-9150 2002/03 [Refereed][Not invited]
   

  Lipid and lipoprotein profiles, and enzymes for the lipid metabolism were compared between cord and adult blood. Consistent with previous reports, the major lipoprotein in cord blood was high-density lipoprotein (HDL), and that in adult blood was low-density lipoprotein (LDL). The level of apolipoprotein E (apo E) in cord blood was almost equivalent to that in adult blood, while other apolipoproteins and lipids were all lower than the adult level. In cord blood, apo E-rich HDL cholesterol represented more than 30% of total HDL cholesterol (around 11% in adult), and the concentration was about twice of that in adult blood. This apo E-rich HDL cholesterol was poorly esterified (E/T 56%) compared with that in adults (93%). The lecithin:cholesterol acyltransferase (LCAT) activity in cord blood was extremely low, while the activity and mass of cholesteryl ester transfer protein (CETP) were higher than those in adult blood. The apo E genotype did not show influences on total cholesterol, LDL cholesterol, total HDL cholesterol, and apo E rich HDL cholesterol levels in cord blood, as opposed to those in adult blood. The association of D442G mutation of the CETP gene with the increased HDL cholesterol in adult blood was not seen in cord blood. Rather, the mutation was associated with low total cholesterol and LDL cholesterol levels in cord blood. These results indicate that, in fetus, the character and metabolism of HDL, especially of apo E-rich HDL cholesterol, are distinct from those in adults. (C) 2002 Elsevier Science Ireland Ltd. All rights reserved.
• Metabolic changes in human CD36 deficiency displayed by glucose loading

  H Yanai, H Chiba, H Fujiwara, M Morimoto, Y Takahashi, SP Hui, H Fuda, H Akita, T Kurosawa, K Kobayashi, K Matsuno

  THROMBOSIS AND HAEMOSTASIS 86 (4) 995 - 999 0340-6245 2001/10 [Refereed][Not invited]
   

  Previous in vitro studies have shown that CD36 participates in cellular fatty acid (FA) uptake. In vivo evidence for a physiologic role of CD36 in this process is poor and mostly obtained in animals. To examine the metabolic role of human CD36, we performed a glucose loading, test for normals (n = 16) and subjects with CD36 deficiency, both Type I (n = 5) and Type Il (n = 16). After 30 min, FA levels had fallen by 60.1% in normals but by only 31.7% in Type II deficiency (P <0.01 vs. normals) and 16.5% in Type I deficiency which remained significantly higher than the other two groups out to 2 h. Further, changes in triglyceride and glucose metabolism were observed in the both types of CD36 deficiency. Impaired fast FA clearance by muscle and consequently increased hepatic FA uptake seem to underlie these changes. We conclude that human CD36 deficiency causes systemic metabolic changes.
• Phenotype-genotype correlation in CD36 deficiency types I and II

  H Yanai, H Chiba, H Fujiwara, M Morimoto, K Abe, S Yoshida, Y Takahashi, H Fuda, SP Hui, H Akita, K Kobayashi, K Matsuno

  THROMBOSIS AND HAEMOSTASIS 84 (3) 436 - 441 0340-6245 2000/09 [Refereed][Not invited]
   

  CD36 deficiency was studied with attention to the phenotype-genotype relationship. The diagnosis of CD36 deficiency was made when CD36 was negative on platelets (type II) or on both platelets and monocytes (type I). Among 827 apparently healthy Japanese volunteers, the type I and II deficiencies were found in 8 (1.0%) and 48 (5.8%): respectively. The T for C substitution at nt478 for Pro90Ser and the insertion of A at nt1159 constituted the major causes of type I and II deficiencies. The dinucleotide deletion at nt539 had a minor role. In two family studies, we found a previously unreported polymorphic site in the 5'-proximal flanking region and the 3'-untranslated region. Including these new polymorphisms, DNA sequence other than the three known mutations affecting CD36 expression was not observed in the CD36:gene, calling into question the previous hypothesis that a platelet-specific silent allele exists near or at the CD36 gene.
• Human CD36 deficiency is associated with elevation in low-density lipoprotein-cholesterol

  H Yanai, H Chiba, M Morimoto, K Abe, H Fujiwara, H Fuda, SP Hui, Y Takahashi, H Akita, GA Jamieson, K Kobayashi, K Matsuno

  AMERICAN JOURNAL OF MEDICAL GENETICS 93 (4) 299 - 304 0148-7299 2000/08 [Refereed][Not invited]
   

  To find out whether CD36 plays a role in the human lipoprotein metabolism, we studied lipoprotein profiles in subjects with CD36 deficiency. Apparently healthy Japanese volunteers (n = 790) were classified by flow cytometry into three groups of normal (platelet and monocyte CD36+, n = 741, 93.8%), type-II deficiency (platelet CD36-and monocyte CD36+, n = 45, 5.7%), and type-II deficiency (platelet and monocyte CD36-, it = 4, 0.5%). At least one of reported mutations in the CD36 gene was found in all four subjects with type-I deficiency and in 23 of the 45 subjects with type II. Among 779 subjects (731 normals, 44 type II, and four type I) with serum triglyceride levels of <400 mg/dL, serum total cholesterol and low-density lipoprotein (LDL) cholesterol were significantly elevated in type-II deficiency (P = 0.0095 and 0.0382 versus normal, respectively, Scheffe's F-test), while differences were not significant in triglyceride and high-density lipoprotein-cholesterol. Similar tendency was observed in type-I deficiency, although the differences were not statistically significant because of small sample size. We conclude that CD36 deficiency elevates LDL cholesterol, indicating a contribution of CD36 to LDL metabolism. (C) 2000 Wiley-Liss, Inc.
• Quantitative and compositional changes in high density lipoprotein subclasses in patients with various genotypes of cholesteryl ester transfer protein deficiency

  H Chiba, H Akita, K Tsuchihashi, SP Hui, Y Takahashi, H Fuda, H Suzuki, H Shibuya, M Tsuji, K Kobayashi

  JOURNAL OF LIPID RESEARCH 38 (6) 1204 - 1216 0022-2275 1997/06 [Refereed][Not invited]
   

  High density lipoprotein (HDL) with and without apolipoprotein (apo) E was quantified and characterized in subjects with three genotypes of cholesteryl ester transfer protein (CETP) deficiency: the nonsense mutation in intron 14 (10 homozygotes and 5 heterozygotes); the missense mutation in the exon 15 (3 homozygotes and 9 heterozygotes); and the Int14A/D442G in 6 compound heterozygotes. ApoE-poor and apoE-rich HDL-cholesterol levels were elevated significantly in all genotypic groups with the decrease in CETP activity, indicating that both types of HDL-cholesterol can be a substrate for CETP. However, an unchanged or only slightly increased serum apoA-II level in each genotype indicated that the HDL particles with apoA-II are relatively resistant to CETP-mediated lipid transfer. Serum apoE-rich HDL level was considerably higher in the Int14A homozygotes than in the compound heterozygotes, in spite of similar apoE-poor HDL-cholesterol levels, which may indicate that apoE-rich HDL is a better substrate for CETP than apoE-poor HDL. Although the apoE-rich and apoE-poor HDL subclasses were similar in the accumulation of cholesteryl ester and depletion of triglyceride, the accumulation of free cholesterol was unique to apoE-rich HDL, indicating inhibited cholesterol esterification on this lipoprotein. Clinical laboratories should be aware of the discrepancy in HDL-cholesterol measurements that comes from the different recoveries of apoE-rich HDL using commercial reagents. In conclusion, CETP deficiency causes considerable quantitative and compositional changes in HDL subclasses, reflecting a significant physiological role for CETP in HDL metabolism.
• Frequency and Effect on Plasma Lipoprotein Metabolism of a Mutation in the Cholesteryl Ester Transfer Protein Gene in the Chinese

  HUI Shu-Ping

  北海道醫學雜誌 = Acta medica Hokkaidonensia 72 (3) 319 - 327 0367-6102 1997/05/01 [Not refereed][Not invited]
  
• [Frequency and effect on plasma lipoprotein metabolism of a mutation in the cholesteryl ester transfer protein gene in the Chinese].

  Hui SP

  [Hokkaido igaku zasshi] The Hokkaido journal of medical science 3 72 (3) 319 - 327 0367-6102 1997/05 [Refereed][Not invited]
  
• Effects of triamcinolone on brain and cerebrospinal fluid apolipoprotein E levels in rats

  H Chiba, H Akita, SP Hui, Y Takahashi, H Nagasaka, H Fuda, K Kobayashi

  LIFE SCIENCES 60 (20) 1757 - 1761 0024-3205 1997/04 [Refereed][Not invited]
   

  The effects of the short term administration of triamcinolone (0.5 mg per 100 g body weight, 5 days) on apolipoprotein E and A-I concentrations in cerebrospinal fulid (CSF), brain extract and serum were studied in male Wistar rats using enzyme immunoassays. ApoE was significantly increased by triamcinolone in apoE-rich HDL(1) in serum; 40+/-13 (mean+/-SD) vs. 68+/-23 mg/dl(15 saline-treated rats vs. 11 triamcinolone-treated rats)(P<0.01), which was paralleled by an increase in serum apoA-I (76+/-21 vs. 184+/-24 mg/dl), while serum lipids also increased significantly. No significant difference was observed in the apoE concentrations in CSF (296+/-170 vs. 269+/-67 mu g/dl) or brain extract (5.0+/-1.6 vs. 5.7+/-1.8 mu g/g wet weight). The apoA-I concentrations found in CSF and brain extract were much lower than those for apoE and were not appreciably affected by triamcinolone: 7.7+/-5.5 vs. 4.5+/-3.1 mu g/dl for CSF and <0.5 vs. <0.5 mu g/g wet weight for brain extract. The apo E metabolism in the rat central nervous system appears to be refractory to the short term administration of triamcinolone and to changes in the serum lipoprotein metabolism. ApoA-I appears unlikely to play a significant role in the rat central nervous system.
• Lipoprotein(a), apolipoprotein(a) phenotypes, and nephropathy in NIDDM patients

  H Akita, H Chiba, M Noritake, Y Katsura, Y Udo, H Shibuya, SP Hui, Y Takahashi, H Nagasaka

  DIABETES CARE 19 (9) 1036 - 1037 0149-5992 1996/09 [Refereed][Not invited]
  
• Native lipoproteins inhibit platelet activation induced by oxidized lipoproteins

  Y Takahashi, H Chiba, K Matsuno, H Akita, SP Hui, H Nagasaka, H Nakamura, K Kobayashi, NN Tandon, GA Jamieson

  BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 222 (2) 453 - 459 0006-291X 1996/05 [Refereed][Not invited]
   

  Copper-catalyzed oxidation of low-density lipoproteins (LDL) (0.8 g protein/l LDL, 20 mu mol/l CuSO4, 37 degrees C) resulted in the formation of thiobarbituric reactive substances that was substantially complete at 24 hrs whereas their formation from high-density lipoproteins (HDL) plateaued at only 25% of that amount after 8 hrs. The oxidized lipoproteins induced aggregation and increases in [Ca2+]i in washed platelets, but not in platelet-rich plasma, and these activating effects were not inhibited by aspirin or EGTA but were inhibited by both of the native lipoproteins. These results show that oxidized HDL, like oxidized LDL, have platelet activating ability and suggest that the native lipoproteins may play a crucial role in preventing the oxidized lipoprotein-mediated platelet activation. (C) 1996 Academic Press, inc.
• EVALUATION OF G-TO-A SUBSTITUTION IN THE APOLIPOPROTEIN-A-I GENE PROMOTER AS A DETERMINANT OF HIGH-DENSITY-LIPOPROTEIN CHOLESTEROL LEVEL IN SUBJECTS WITH AND WITHOUT CHOLESTERYL ESTER TRANSFER PROTEIN-DEFICIENCY

  H AKITA, H CHIBA, M TSUJI, SP HUI, Y TAKAHASHI, K MATSUNO, K KOBAYASHI

  HUMAN GENETICS 96 (5) 521 - 526 0340-6717 1995/11 [Refereed][Not invited]
   

  The effect of a polymorphism, guanine (G) to adenine (A) substitution in the promoter of apolipoprotein A-I gene at a position 78 bp upstream of the transcription initiation site, on the serum high-density lipoprotein (HDL)-cholesterol level was studied in 168 Japanese subjects with HDL-cholesterol levels ranging from 26 to 171 mg/dl. Considering the significant effect of cholesteryl ester transfer protein (CETP) on the HDL-cholesterol level and the common occurrence of its deficiency, we per formed statistical analyses separately for two groups: one without CETP deficiency (n = 126) and the other with CETP deficiency (n = 42). In the group without CETP deficiency, in which the numbers of G/G, G/A, and A/A genotypes were 92 (73.0%), 28 (22.2%), and 6 (4.8%), respectively, the frequency of the A allele in the subjects with HDL-cholesterol levels of greater than or equal to 70 mg/dl did not differ from subjects with HDL-cholesterol levels of less than or equal to 69 mg/dl, irrespective of gender: 0.154 and 0.145 in males, and 0.182 and 0.174 in females, respectively, for the greater than or equal to 70 mg/dl and less than or equal to 69 mg/dl groups. Additionally, the HDL-cholesterol levels for the subjects with the G/G genotype did not differ from those for the subjects with the A allele: 64 +/- 22, 58 +/- 14, 77 +/- 14 and 62 +/- 16 mg/dl, respectively, for the G/G, G/A, A/A, and G/A + A/A in males, and 72 +/- 18, 74 +/- 24, 63 +/- 4, and 73 +/- 23 mg/dl in females. For the group with CETP deficiency, in which the numbers of G/G and G/A + A/A genotypes were 25 (59.5%) and 17 (40.5%), the HDL-cholesterol levels also did not differ: 98 +/- 24 mg/dl and 99 +/- 30 mg/dl, respectively, for the G/G and G/A + A/A genotypes. Thus, there is no evidence that the polymorphism has any effect on serum HDL-cholesterol levels regardless of CETP status. We conclude that the G-to-A substitution in the promoter of apolipoprotein A-I gene does not significantly alter serum HDL-cholesterol level.
• CHOLESTERYL ESTER TRANSFER PROTEIN-DEFICIENCY - IDENTIFICATION IN THE CHINESE

  H AKITA, H CHIBA, SP HUI, Y TAKAHASHI, K MATSUNO, K KOBAYASHI

  AMERICAN JOURNAL OF MEDICAL GENETICS 59 (3) 399 - 400 0148-7299 1995/11 [Refereed][Not invited]
  

Books etc

• バイオセンサの迅速・簡易・高機能化技術と課題解決書：過酸化脂質・酸化LDL測定のためのバイオセンサ

  武田 晴治, 惠 淑萍, 千葉 仁志 (Joint work)
  （株）技術情報協会 2014/04
• 医用質量分析ガイド 内分泌・代謝疾患，診断と治療

  千葉仁志, 惠 淑萍 (Joint work)
  診断と治療社 2013/12
• “パザパ”薬学演習シリーズ７

  黒澤隆夫, 豊田栄子, 村井毅, 惠 淑萍 (Joint work)
  京都廣川書店 2010/04

Conference Activities & Talks

• From the standpoint of a lipid researcher, strategic measures for pre-illness with an eye on the next era  [Invited]

  Shu-Ping Hui

  第27回 日本未病学会学術総会 シンポジウム８（次の時代を見据えた戦略的未病対策とは ）  2020/11
• Analyses of hydroperoxide lipids in human lipoprotein and cell by mass spectrometry  [Invited]

  SHU-PING HUI

  2018 Annual Medical Laboratory Forum (Taipei Medical University)  2018/10
• 異所性脂肪蓄積と酸化ストレス  [Not invited]

  惠 淑萍

  第31回日本小児脂質研究会学術集会 2017年度  2017/11
• 質量分析による分子レベルの脂質研究  [Not invited]

  惠 淑萍

  第56回日本臨床化学会年次学術集会（教育講演）  2016/12

MISC

• 非標的LC/MSを用いたナマコ中の健康機能性脂質の分析と同定

  南 雄介, ボメゴウダ・シッダバサーブゴウダ, デヴィヤヴァニ・ゴウダ, 千葉 仁志, 惠 淑萍  JSBMS Letters  48-  (Suppl.)  77  -77  2023/08
• 培養ヒト肝細胞から放出されるVLDL中のTG及びTG-OOHの解析

  益子 真明, 櫻井 俊宏, 山端 ありさ, 佐崎 生, 茂木 すみれ, 千葉 仁志, 惠 淑萍  JSBMS Letters  48-  (Suppl.)  81  -81  2023/08
• 心筋梗塞のバイオマーカー探索のためのリピドミクス

  南 雄介, ボメゴウダ・シッダバサーブゴウダ, 千葉 仁志, 惠 淑萍  JSBMS Letters  48-  (2)  20  -26  2023/06  

  心筋梗塞は心不全の最も一般的な原因であり,世界の主要な死因として知られている.種々の心血管疾患の病態生理学プロセスは多様であるが,脂質はいつも非常に重要な役割を担っている.近年,質量分析法の進歩によって,生体組織における複雑な脂質分子種の同定が可能になり,心筋梗塞マーカーの探索に用いられている.リピドミクスは,心血管イベントの根底にある複雑な病態の解明や治療法の開発に貢献することが期待されている.この総説では,リピドミクスによって明らかになった心筋梗塞や関連疾患のバイオマーカーに焦点をあてる.(著者抄録)
• メナキノン-4による腎尿細管細胞のミトコンドリア保護作用

  江口 美祈, 何 欣蓉, 青木 菜摘, 高明 晨, 鈴木 拓貴, 千葉 仁志, 惠 淑萍  ビタミン  97-  (4)  217  -217  2023/04
• フラジンはヒト尿細管HK-2細胞における脂肪酸を介した脂質毒性を抑制する(Flazin attenuates fatty acid mediated lipotoxicity in human tubular HK-2 cells)

  呉 訓智, 何 欣蓉, 陳 震, 陳 一凡, 沈 テン秋, 千葉 仁志, 惠 淑萍  日本栄養・食糧学会大会講演要旨集  77回-  173  -173  2023/03
• ビタミンKによる腎尿細管細胞のミトコンドリア保護効果

  江口 美祈, 何 欣蓉, 青木 菜摘, 高 明晨, 鈴木 拓貴, 千葉 仁志, 惠 淑萍  日本栄養・食糧学会大会講演要旨集  77回-  302  -302  2023/03
• グルコシルセラミドフリッパーゼ発現によるゴーシェ病細胞の異常改善の検討

  喜多 夏暉, 濱本 明日香, ボメゴウダ・シッダバサーブゴウダ, 高津 宏之, 中山 和久, 惠 淑萍, 申 惠媛  日本生化学会大会プログラム・講演要旨集  95回-  2T02a  -04  2022/11
• Investigating oxidized lipids in an omics way: Oxidative lipidomics in biological applications using mass spectrometry(タイトル和訳中)

  Chen Zhen, Wu Xunzhi, Chiba Hitoshi, Hui Shu-Ping  Medical Mass Spectrometry  6-  (2)  72  -84  2022/11
• グルコシルセラミドフリッパーゼ発現によるゴーシェ病細胞の異常改善の検討

  喜多 夏暉, 濱本 明日香, ボメゴウダ・シッダバサーブゴウダ, 高津 宏之, 中山 和久, 惠 淑萍, 申 惠媛  日本生化学会大会プログラム・講演要旨集  95回-  2T02a  -04  2022/11
• マガキ由来抗酸化物質DHMBAの腎臓尿細管細胞における抗酸化作用

  何 欣蓉, 関根 かりん, 青木 菜摘, 櫻井 俊宏, 千葉 仁志, 渡辺 秀明, 渡辺 貢, 惠 淑萍  日本未病学会学術総会抄録集  29回-  73  -73  2022/10
• 食事摂取した海藻の脂質に関する液体クロマトグラフィー/質量分析計を利用した探索的なデータ分析研究(Exploratory data analysis on dietary seaweeds lipids by liquid chromatography/mass spectrometry)

  Gowda Siddabasave Gowda B., Yifan Chen, Gowda Divyavani, Tsuboi Yui, Chiba Hitoshi, Hui Shu-Ping  日本未病学会学術総会抄録集  29回-  73  -73  2022/10
• マガキ由来抗酸化物質DHMBAの腎臓尿細管細胞における抗酸化作用

  何 欣蓉, 関根 かりん, 青木 菜摘, 櫻井 俊宏, 千葉 仁志, 渡辺 秀明, 渡辺 貢, 惠 淑萍  日本未病学会学術総会抄録集  29回-  73  -73  2022/10
• LC/MSによるリピドミクスを用いた海藻由来植物ステロールの神経細胞に対する脂質異常改善作用の検討

  竹内 美音, 陳 震, Shen Nianqiu, Wu Yue, Wu Xunzhi, 千葉 仁志, 惠 淑萍  JSBMS Letters  47-  (Suppl.)  82  -82  2022/08
• 酸化LDLの肝カルジオリピン生合成への影響

  佐崎 生, 櫻井 俊宏, 茂木 すみれ, 千葉 仁志, 惠 淑萍  JSBMS Letters  47-  (Suppl.)  83  -83  2022/08
• 脂肪肝モデル細胞におけるサケ白子抽出物のカルジオリピン代謝改善作用

  櫻井 俊宏, 吉貝 あみ, 茂木 すみれ, 山端 ありさ, 関島 将人, 佐藤 美涼, 佐藤 浩志, 千葉 仁志, 惠 淑萍  JSBMS Letters  47-  (Suppl.)  84  -84  2022/08
• 食品由来の肥満関連スフィンゴ脂質代謝阻害剤の発見(Discovery of Food Derived Inhibitor for Sphingolipid Metabolism Associated to Obesity)

  Minami Yusuke, Gowda Siddabasave Gowda B., Gowda Divyavani, Chiba Hitoshi, Hui Shu-Ping  JSBMS Letters  47-  (Suppl.)  80  -80  2022/08
• LC/MSおよびNanoESI-MSを用いて検討した腎細胞での脂質過負荷に対するFlazinの保護作用(Investigation on Protective Effect of Flazin Against Lipid Overloading in Renal Cells Using LC/MS and NanoESI-MS)

  Wu Xunzhi, Chen Zhen, Chen Yifan, Shen Nianqiu, Chiba Hitoshi, Hui Shu-Ping  JSBMS Letters  47-  (Suppl.)  81  -81  2022/08
• 食用魚11品目の未知の脂質に挑む Untargeted LC/MSによる組成分析(Tackling Unknown Lipids in Eleven Dietary Fish and Their Composition Analysis by Untargeted LC/MS)

  Gowda Siddabasave Gowda B., Minami Yusuke, Gowda Divyavani, Chiba Hitoshi, Hui Shu-Ping  JSBMS Letters  47-  (Suppl.)  85  -85  2022/08
• ヒドロキシ脂肪酸の短鎖脂肪酸エステル類の合成と簡便な分析法の開発(Synthesis and Development of a Facile Analytical Method for Short Chain Fatty Acid Esters of Hydroxy Fatty Acids)

  Hou Fengjue, Gowda Siddabasave Gowda B., Gowda Divyavani, Chiba Hitoshi, Hui Shu-Ping  JSBMS Letters  47-  (Suppl.)  86  -86  2022/08
• LC/MSによるリピドミクスを用いた海藻由来植物ステロールの神経細胞に対する脂質異常改善作用の検討

  竹内 美音, 陳 震, Shen Nianqiu, Wu Yue, Wu Xunzhi, 千葉 仁志, 惠 淑萍  JSBMS Letters  47-  (Suppl.)  82  -82  2022/08
• 酸化LDLの肝カルジオリピン生合成への影響

  佐崎 生, 櫻井 俊宏, 茂木 すみれ, 千葉 仁志, 惠 淑萍  JSBMS Letters  47-  (Suppl.)  83  -83  2022/08
• 脂肪肝モデル細胞におけるサケ白子抽出物のカルジオリピン代謝改善作用

  櫻井 俊宏, 吉貝 あみ, 茂木 すみれ, 山端 ありさ, 関島 将人, 佐藤 美涼, 佐藤 浩志, 千葉 仁志, 惠 淑萍  JSBMS Letters  47-  (Suppl.)  84  -84  2022/08
• 【質量分析法を活用したバイオマーカー分析研究】リゾホスファチジルエタノールアミンと肝疾患

  櫻井 俊宏, 惠 淑萍  臨床化学  51-  (3)  174  -178  2022/07
• 【質量分析法を活用したバイオマーカー分析研究】リゾホスファチジルエタノールアミンと肝疾患

  櫻井 俊宏, 惠 淑萍  臨床化学  51-  (3)  174  -178  2022/07
• 腎臓近位尿細管細胞におけるメナキノン-4の酸化ストレス軽減効果

  何 欣蓉, 青木 菜摘, 高 明晨, 鈴木 拓貴, 千葉 仁志, 惠 淑萍  ビタミン  96-  (4)  193  -193  2022/04
• 3,5-dihydroxy-4-methoxybenzyl alcohol(DHMBA)は心筋細胞の冷保存傷害を軽減する

  深井 原, 大谷 晋太郎, 千葉 仁志, 惠 淑萍, 坂本 聡大, 柴田 賢吾, 石川 隆壽, 川村 典生, 嶋村 剛, 武冨 紹信  日本外科学会定期学術集会抄録集  122回-  SF  -8  2022/04
• 乳牛の血漿遊離脂肪酸濃度上昇に伴う卵子エネルギー代謝関連脂質プロファイルの変化

  古川瑛理, 陳震, 窪友瑛, CHELENGA Madalitso, WU Yue, 千葉仁志, 柳川洋二郎, 片桐成二, 惠淑萍, 永野昌志  日本生殖医学会雑誌  67-  (3)  2022
• HK2細胞における異所性脂肪蓄積・酸化と、フラジンによる予防効果(Ectopic lipid accumulation and oxidation in HK2 cells and the prevention effects of flazin)

  惠 淑萍, Wu Xunzhi, Chen Zhen, Wu Yue, Chen Yifan, 千葉 仁志  日本未病学会学術総会抄録集  28回-  89  -89  2021/11
• 貯蔵環境の違いによる食品中ビニルエーテル結合型リン脂質の変化(Alterations of vinyl ether-linked phospholipids in foodstuffs under different storing conditions)

  惠 淑萍, Jia Jiaping, Chen Zhen, Wu Yue, 千葉 仁志  日本未病学会学術総会抄録集  28回-  118  -118  2021/11
• 日常的に摂取する食品に含まれるプラズマローゲン種の違いと比較(Variation and comparison of plasmalogen species in daily foodstuffs)

  惠 淑萍, Wu Yue, 陳 震, Jia Jiaping, 千葉 仁志  日本未病学会学術総会抄録集  28回-  120  -120  2021/11
• ホルスタイン種泌乳牛の分娩後日数及びボディコンディションスコアと卵子中トリアシルグリセロール量の関係

  古川 瑛理, 陳 震, 窪 友瑛, Madalitso Chelenga, Yue Wu, 千葉 仁志, 柳川 洋二郎, 片桐 成二, 惠 淑萍, 永野 昌志  日本獣医学会学術集会講演要旨集  164回-  [GO  -5]  2021/09
• ホルスタイン種泌乳牛の分娩後日数及びボディコンディションスコアと卵子中トリアシルグリセロール量の関係

  古川 瑛理, 陳 震, 窪 友瑛, Madalitso Chelenga, Yue Wu, 千葉 仁志, 柳川 洋二郎, 片桐 成二, 惠 淑萍, 永野 昌志  日本獣医学会学術集会講演要旨集  164回-  [GO  -5]  2021/09
• 酸化ストレス誘導下HK-2細胞におけるDHMBAによるカルジオリピンの組成変化に関する研究(Alteration of cardiolipin profile in HK-2 proximal tubule cells under oxidative stress and the reverse effects of 3,5-dihydroxy-4-methoxybenzyl alcohol(DHMBA))

  呉 亦柔, 何 欣蓉, 陳 震, 賈 佳萍, 青木 菜摘, 千葉 仁志, 渡辺 孝之, 渡辺 貢, 惠 淑萍  JSBMS Letters  46-  (Suppl.)  106  -106  2021/08
• 脂肪滴の蓄積(LDA)と酸化脂肪滴(oxLDs)を標的とする生物活性アルカロイドの発見(Discovery of bioactive alkaloids targeting the lipid droplet accumulation(LDA) and oxidized lipid droplets oxLDs)

  ディブエ・ディア・フィタ, 大場 早紀, 武石 にれ, ロジート・シュレスタ, 仁志 千葉, 惠 淑萍  日本薬学会年会要旨集  141年会-  29P01  -034  2021/03
• β-カルボリンアルカロイドのデザイン・合成と、その抗脂肪滴蓄積活性(Design and synthesis of β-carboline alkaloids and their anti-lipid droplet accumulation activity)

  武石 にれ, ディブエ・ディア・フィタ, 大場 早紀, 千葉 仁志, 惠 淑萍  日本薬学会年会要旨集  141年会-  29P01  -035S  2021/03
• Allium抽出物の直接誘導体化と、HepG2肝細胞を用いた遊離脂肪酸誘導脂肪滴集積に対する阻害効果(Direct derivatization of Allium extracts and their inhibitory effect on the free fatty acid-induced lipid droplet accumulation in HepG2 Hepatocyte)

  大場 早紀, ディブエ・ディア・フィタ, 武石 にれ, 千葉 仁志, 惠 淑萍  日本薬学会年会要旨集  141年会-  29P01  -036S  2021/03
• Investigation of the storage-induced changes of plasmalogens in fish by LC/MS

  JIA Jiaping, CHEN Zhen, WU Yue, CHIBA Hitoshi, HUI Shu-Ping  JSBMS Letters  46-  (Supplement)  2021
• Comprehensive analysis of lipid composition in the dietary Japanese fish using UHPLC/LTQ Orbitrap MS

  MINAMI Yusuke, BOMMEGOWDA Siddabasavegowda, GOWDA Divyavani, CHIBA Hitoshi, HUI Shu-Ping  JSBMS Letters  46-  (Supplement)  2021
• Lipid fingerprinting for lipid biomarker discovery of drug-resistance in leukemia

  MATSUHISA Chihiro, BOMMEGOWDA Siddabasavegowda, USUKI Seigo, SAKURAI Toshihiro, CHIBA Hitoshi, HUI Shu-Ping  JSBMS Letters  46-  (Supplement)  2021
• Effects of lysophosphatidylethanolamine on hepatic lipid and mitochondrial metabolism

  櫻井俊宏, 井上夏緒, 山本祐輔, CHEN Zhen, 千葉仁志, HUI Shu-Ping  JSBMS Letters  46-  (Supplement)  2021
• Lipids and lipid hydroperoxides in multiple tissues from type 2 diabetic model mice

  CHIN Shin, LIANG Qiangrong, WU Yue, CHIBA Hitoshi, HUI Shu-Ping  JSBMS Letters  46-  (Supplement)  2021
• One-step derivatization assisted targeted analysis of fatty acid hydroperoxides by LC/MS.

  LIANG Chongsheng, BOMMEGOWDA Siddabasavegowda, SAKURAI Toshihiro, CHIBA Hitoshi, HUI Shu-Ping  JSBMS Letters  46-  (Supplement)  2021
• Global lipid fingerprinting for early disease biomarker discovery by LC/MS

  BOMMEGOWDA Siddabasavegowda, GOWDA Divyavani, LIANG Chongsheng, CHIBA Hitoshi, HUI Shu-Ping  JSBMS Letters  46-  (Supplement)  2021
• Synthesis and Development of a Facile Analytical Method for Short Chain Fatty Acid Esters of Hydroxy Fatty Acids

  HOU Fengjue Hou, BOMMEGOWDA Siddabasavegowda, GOWDA Divyavani, CHIBA Hitoshi, HUI Shu-Ping  JSBMS Letters  46-  (Supplement)  2021
• 次の時代を見据えた戦略的未病対策とは 脂質研究者の立場から、次の時代を見据えた戦略的未病対策

  惠 淑萍  日本未病学会学術総会抄録集  27回-  72  -72  2020/10
• NASH鑑別マーカーとしての血中LDL-TG値の有用性

  櫻井 俊宏, 高橋 祐司, 藤井 佑樹, 能祖 一裕, 太田 素子, 伊藤 康樹, 千葉 仁志, 惠 淑萍  臨床化学  49-  (Suppl.1)  141  -141  2020/10
• 酸化HDLの肝脂質代謝及びミトコンドリアへの影響

  山端 ありさ, 櫻井 俊宏, 関島 将人, 上野 朱音, 千葉 仁志, 惠 淑萍  臨床化学  49-  (Suppl.1)  150  -150  2020/10
• 酸化HDLにより誘導された肝細胞過酸化脂質プロフィールの変動

  上野 朱音, 櫻井 俊宏, 関島 将人, 山端 ありさ, 陳 震, 千葉 仁志, 惠 淑萍  臨床化学  49-  (Suppl.1)  151  -151  2020/10
• サケ白子抽出物の培養ヒト肝細胞におけるミトコンドリア活性化作用

  関島 将人, 櫻井 俊宏, 佐藤 浩志, 何 欣蓉, 千葉 仁志, 惠 淑萍  臨床化学  49-  (Suppl.1)  152  -152  2020/10
• Nrf2介在抗酸化物質と考えられるn-3脂肪酸由来脂質の発見(Discovery of n-3 fatty acid derived lipids as potent Nrf2 mediated antioxidants)

  千葉 仁志, Siddabasavegowda B. Gowda, 布田 博敏, 津久井 隆行, 惠 淑萍  臨床化学  49-  (Suppl.1)  193  -193  2020/10
• 心筋梗塞におけるスフィンゴシン-1-リン酸塩シグナル伝達の役割(Role of Sphingosine-1-phosphate signalling in myocardial infarction)

  Siddabasavegowda B. Gowda, Gowda Divyavani, 千葉 仁志, Halade Ganesh, 惠 淑萍  臨床化学  49-  (Suppl.1)  213  -213  2020/10
• 健常者及び非アルコール性脂肪性肝疾患患者の血清中LysoPEプロファイル

  山本 祐輔, 櫻井 俊宏, 陳 震, Wu Yue, 藤井 佑樹, 能祖 一裕, 千葉 仁志, 惠 淑萍  臨床化学  49-  (Suppl.1)  213  -213  2020/10
• エーテル結合型リン脂質は、Keap1-Nrf2を介した酸化損傷から肝細胞を保護する(Ether-linked phospholipids protect hepatocytes from oxidative damage via Keap1-Nrf2 pathway)

  Wu Yue, 陳 震, 千葉 仁志, 惠 淑萍  臨床化学  49-  (Suppl.1)  215  -215  2020/10
• IVIG感受性川崎病患児と抵抗性患児との間にみられる脂質の特徴の違い(Differential lipid characteristics in Kawasaki disease between IVIG sensitive and resistant children)

  陳 震, 齋 秀二, Wu Yue, 千葉 仁志, 惠 淑萍  臨床化学  49-  (Suppl.1)  215  -215  2020/10
• LC/HR-MS/MSによる、2型糖尿病マウスの血清と組織におけるリピドームプロファイルの変化(Lipidomic profile variation in serum and tissues from type 2 diabetic mice by LC/HR-MS/MS)

  惠 淑萍, 陳 震, Liang Qiangrong, Wu Yue, 高 紫君, 梁 崇晟, 千葉 仁志  臨床化学  49-  (Suppl.1)  218  -218  2020/10
• one-step derivatization coupled targeted LC-MS/MSによる脂質ヒドロペルオキシドの簡易測定(Facile determination of lipid hydroperoxides by one-step derivatization coupled targeted LC-MS/MS)

  梁 崇晟, Siddabasavegowda B. Gowda, 千葉 仁志, 惠 淑萍  臨床化学  49-  (Suppl.1)  218  -218  2020/10
• NASHモデルマウスを用いた肝・腎内の脂質に関する網羅的解析(Comprehensive analysis of lipids in the liver and kidney from NASH model mice)

  惠 淑萍, 齋藤 捺希, 陳 震, 櫻井 俊宏, Wu Yue, 津久井 隆行, 布田 博敏, 千葉 仁志  日本未病学会学術総会抄録集  27回-  100  -100  2020/10
• LC-MS/MSによる血中脂肪酸同時測定法の開発と最適化(Method development and optimization of simultaneous measurement for fatty acids in blood by LC-MS/MS)

  惠 淑萍, 陳 震, 高 紫君, Wu Yue, Shrestha Rojeet, 千葉 仁志  日本未病学会学術総会抄録集  27回-  101  -101  2020/10
• 調理法による牛肉中プラズマローゲン含有量の変化(Changes of Plasmalogen content in beef induced by cooking way)

  惠 淑萍, Wu Yue, 陳 震, 千葉 仁志  日本未病学会学術総会抄録集  27回-  113  -113  2020/10
• COPDに対する中鎖脂肪酸-グレリン併用療法

  三木 啓資, 北田 清悟, 惠 淑萍, 三木 真理, Rojeet Shrestha, 辻野 和之, 香川 浩之, 押谷 洋平, 好村 研二, 木田 博, 前倉 亮治, 寒川 賢治  日本呼吸器学会誌  9-  (増刊)  274  -274  2020/08
• COPDに対する中鎖脂肪酸-グレリン併用療法

  三木 啓資, 北田 清悟, 惠 淑萍, 三木 真理, Rojeet Shrestha, 辻野 和之, 香川 浩之, 押谷 洋平, 好村 研二, 木田 博, 前倉 亮治, 寒川 賢治  日本呼吸器学会誌  9-  (増刊)  274  -274  2020/08
• リポ蛋白質の酸化脂質種 その意義と分析による考察(Oxidized lipid species in lipoproteins: Significance and analytical considerations)

  Shrestha Rojeet, Chiba Hitoshi, Hui Shu-Ping  Medical Mass Spectrometry  4-  (1)  2  -23  2020/06
• 質量分析を用いたカルジオリピンの分析及び生物医学への応用

  陳 震, 山本 祐輔, 千葉 仁志, 惠 淑萍  JSBMS Letters  45-  (2)  18  -24  2020/06  

  カルジオリピンはミトコンドリア内膜に局在するユニークなリン脂質であり、エネルギー代謝をはじめとする様々なミトコンドリア機能に重要な役割を果たしている。近年、質量分析によるカルジオリピンの分析法が開発され、生物医学的研究へ応用されている。そのことは、カルジオリピンの化学的プロファイル、リモデリング、酸化的変化の理解や、ミトコンドリア機能障害と代謝異常の関係の理解に貢献すると思われる。今後、カルジオリピン分子種のいずれかが健康障害・疾病の潜在的バイオマーカーとして見いだされる可能性もある。この総説では、質量分析により明らかとなったカルジオリピンの化学的プロファイリング、リモデリング、および酸化について焦点をあてる。(著者抄録)
• One-pot Pictet-Spengler反応とその生物学的活性を用いたβ-carboline alkaloidの設計と合成(Design and synthesis of β-carboline alkaloids using a one-pot Pictet-Spengler reaction and their biological activity)

  ディブエ・ディアフィタ, 布田 博敏, 山本 祐輔, シュレスタ・ロジート, 千葉 仁志, 惠 淑萍  日本薬学会年会要旨集  140年会-  28Q  -pm105  2020/03
• 【これからの臨床化学教育】大学院教育が臨床化学の基盤を築く

  千葉 仁志, 惠 淑萍  臨床化学  48-  (4)  321  -327  2019/10  [Not refereed][Not invited]
  
• ヒト近位尿細管上皮細胞の脂肪滴形成がミトコンドリア固有リン脂質への影響に関する検討

  惠 淑萍, 三浦 佑介, 櫻井 俊宏, 陳 震, 千葉 仁志  日本未病システム学会学術総会抄録集  26回-  108  -108  2019/10  [Not refereed][Not invited]
  
• LC/MSによる糖尿病性腎症患者の尿中コレステリルエステル定量分析

  惠淑萍, 三浦祐介, 辻川諒哉, 櫻井俊宏, 高田康徳, 千葉仁志  臨床病理  67-  (補冊)  260  -260  2019/10  [Not refereed][Not invited]
  
• 臓器脂質代謝異常へのアプローチ 糖尿病マウスの心臓における脂質組成異常とマイトファジー-リソソーム経路の機能障害(Altered lipidomic profile and dysfunctional mitophagy-lysosome pathway in the diabetic mouse heart)

  Liang Qiangrong, Chen Zhen, Wu Yue, Liang Chongsheng, Kobayashi Satoru, Patel Joy, Huang Yuan, Kobayashi Tamayo, Chiba Hitoshi, Hui Shu-Ping  臨床化学  48-  (Suppl.1)  181  -181  2019/08
• 糖化がLDLの物性に与える影響について(Effect of glycation on the physical properties of low density-lipoprotein)

  Takeda Seiji, Sakurai Toshihiro, Hui Shu-Ping, Chiba Hitoshi  生物物理  59-  (Suppl.1-2)  S310  -S310  2019/08
• 臓器脂質代謝異常へのアプローチ 細胞内脂肪滴の分析法の開発

  惠 淑萍  臨床化学  48-  (Suppl.1)  183  -183  2019/08  [Not refereed][Not invited]
  
• ヒト肝培養細胞における酸化HDLの線維化促進作用

  関島 将人, 櫻井 俊宏, 千葉 仁志, 惠 淑萍  臨床化学  48-  (Suppl.1)  205  -205  2019/08  [Not refereed][Not invited]
  
• ヒト腎近位尿細管上皮細胞を用いたマガキ由来抗酸化物質の抗酸化能

  布田 博敏, 窪田 航, 上甲 紗愛, 渡邉 貢, 武田 晴治, 惠 淑萍, 千葉 仁志  臨床化学  48-  (Suppl.1)  248  -248  2019/08  [Not refereed][Not invited]
  
• DDTはヒト肝臓(HepG2)細胞のレドックス恒常性とビタミンCとEの保護作用を変化させる(DDT alters redox homeostasis in human liver(HepG2) cells and protection with vitamins C and E)

  武石 にれ, Darwish Wageh, 千葉 仁志, 惠 淑萍  臨床化学  48-  (Suppl.1)  248  -248  2019/08  [Not refereed][Not invited]
  
• 酸化リポタンパク質はHepG2細胞の脂質滴中の脂質過酸化水素の蓄積を誘発する(Oxidized lipoproteins induce accumulation of lipid hydroperoxides in lipid droplets of HepG2 cells)

  Shrestha Rojeet, 陳 震, 桜井 俊宏, 千葉 仁志, 惠 淑萍  臨床化学  48-  (Suppl.1)  258  -258  2019/08  [Not refereed][Not invited]
  
• ヒト血漿中の脂肪酸に関する派生体化およびLC-MS/MSを用いた定量的組成分析(Quantitative profiling of fatty acids in human plasma by derivatization and LC-MS/MS)

  陳 震, 高 紫君, Wu Yue, ロジート・シュレスタ, 千葉 仁志, 惠 淑萍  臨床化学  48-  (Suppl.1)  259  -259  2019/08  [Not refereed][Not invited]
  
• 質量分析による生細胞中の単一脂肪滴の分析

  惠 淑萍, 趙 瑤瑤, 陳 震, 呉 げつ, 高 紫君, 張 新栄, 千葉 仁志  JSBMS Letters  44-  (Suppl.)  94  -94  2019/08  [Not refereed][Not invited]
  
• 鉛誘導したHepG2細胞によるプラズマローゲンの保護作用

  陳 震, Yue Wu, Wageh Darwish, 寺田 航, 千葉 仁志, 惠 淑萍  JSBMS Letters  44-  (Suppl.)  95  -95  2019/08  [Not refereed][Not invited]
  
• 質量分析を用いる肺サーファクタントリン脂質膜に対する環境濃度オゾンの影響に関する研究(Study on the Lipid Monolayer of Lung Surfactant in Low-level Ozone by LC/MS)

  喬 琳, 陳 震, 高田 俊智, 千葉 仁志, 叶 深, 惠 淑萍  JSBMS Letters  44-  (Suppl.)  101  -101  2019/08  [Not refereed][Not invited]
  
• 高炭水化物食由来脂肪肝マウスの肝臓中cardiolipin及びmonolysocardiolipinの組成解析

  櫻井俊宏, CHEN Zhen, 早坂孝宏, 千葉仁志, HUI Shu‐Ping  JSBMS Letters  44-  (Suppl.)  99  -99  2019/08  [Not refereed][Not invited]
  
• HK‐2細胞における脂肪滴とカルジオリピンの関連性についての検討

  三浦佑介, 櫻井俊宏, 陳震, 千葉仁志, 惠淑萍  臨床化学  48-  (Suppl.1)  256  -256  2019/08  [Not refereed][Not invited]
  
• 非アルコール性脂肪性肝炎発症に対する酸化HDLの関与

  櫻井俊宏, 関島将人, 田村宥人, 仲門菜月, 津久井隆行, 津久井隆行, 布田博敏, 千葉仁志, 惠淑萍  臨床化学  48-  (Suppl.1)  256  -256  2019/08  [Not refereed][Not invited]
  
• リゾホスファチジルエタノールアミンの定量系開発のための標準品および内部標準物質の合成

  山本祐輔, 櫻井俊宏, 三浦佑介, 陳震, 千葉仁志, 惠淑萍  臨床化学  48-  (Suppl.1)  259  -259  2019/08  [Not refereed][Not invited]
  
• 環境濃度オゾンによる肺サーファクタントリン脂質膜の酸化に関する研究

  喬 琳, 陳 震, 高田 俊智, 千葉 仁志, 叶 深, 惠 淑萍  脂質生化学研究  61-  224  -224  2019/06  [Not refereed][Not invited]
  
• 臨床栄養学 脂肪蓄積とマガキから抽出した抗酸化物質

  惠 淑萍, 布田 博敏, 千葉 仁志  医学のあゆみ  269-  (10)  814  -815  2019/06  [Not refereed][Not invited]
  
• 環境濃度オゾンによる肺サーファクタントリン脂質膜の酸化に関する研究

  喬 琳, 陳 震, 高田 俊智, 千葉 仁志, 叶 深, 惠 淑萍  脂質生化学研究  61-  224  -224  2019/06  [Not refereed][Not invited]
  
• 教育と研究の両立、ロールモデルから学ぶ 高度脂質分析ラボにおける教育研究 フロンティアへの挑戦

  惠 淑萍  臨床検査学教育  11-  (1)  49  -53  2019/03  [Not refereed][Not invited]
  
• 教育と研究の両立、ロールモデルから学ぶ 高度脂質分析ラボにおける教育研究 フロンティアへの挑戦

  惠 淑萍  臨床検査学教育  11-  (1)  49  -53  2019/03  [Not refereed][Not invited]
  
• Altered lipidomic profile and dysfunctional mitophagy-lysosome pathway in the diabetic mouse heart

  LIANG Qiangrong, CHEN Zhen, WU Yue, LIANG Chongsheng, KOBAYASHI Satoru, PATEL Joy, HUANG Yuan, KOBAYASHI Tamayo, CHIBA Hitoshi, CHIBA Hitoshi, HUI Shu-Ping  臨床化学  48-  2019
• Dysregulation of lipid metabolism in HepG2 cells exposed to a mixture of 4 kinds of PAHs

  GAO Zijun, DARWISH Wageh Sobhy, CHEN Zhen, CHIBA Hitoshi, HUI Shu-Ping  JSBMS Letters  44-  (Supplement)  2019
• 新たな創薬資源としてのゼブラフィッシュ・マイクロバイオーム

  島田康人, 島田康人, 島田康人, 岡崎文美, 岡崎文美, 岡崎文美, 臧黎清, 臧黎清, 中山寛子, 中山寛子, 三谷隆敦, 先沖陽貴, 恵淑萍, 西村訓弘, 西村訓弘  日本分子生物学会年会プログラム・要旨集(Web)  42nd-  2019
• 質量分析法による25(OH)D測定法の開発および免疫学的測定法との比較検討

  清水 力, 陳 震, 惠 淑萍, 千葉 仁志  日本内分泌学会雑誌  94-  (3)  732  -732  2018/12  [Not refereed][Not invited]
  
• 質量分析法による25(OH)D測定法の開発および免疫学的測定法との比較検討

  清水 力, 陳 震, 惠 淑萍, 千葉 仁志  日本内分泌学会雑誌  94-  (3)  732  -732  2018/12  [Not refereed][Not invited]
  
• 細胞内脂質および過酸化脂質に関する研究

  惠 淑萍, 李 泳翰, 陳 震, ダルウィッシュ・ワギ, 呉 Yue, 趙 瑤瑤, 高 紫君, 千葉 仁志  日本未病システム学会学術総会抄録集  25回-  141  -141  2018/10  [Not refereed][Not invited]
  
• 未病の可視化と臨床検査 質量分析による未病の可視化

  千葉 仁志, 惠 淑萍  日本未病システム学会学術総会抄録集  25回-  80  -80  2018/10  [Not refereed][Not invited]
  
• マススペクトロメトリーの検査応用 現状と今後の課題 質量分析による低分子の分析および臨床検体への応用

  惠 淑萍  臨床病理  66-  (9)  1001  -1001  2018/09  [Not refereed][Not invited]
  
• 液体クロマトグラフィー高分解能質量分析を用いた牛卵子中遊離脂肪酸およびトリアシルグリセロールの分析

  植芝 滉己, 呉 ユエ, 陳 震, 坂口 謙一郎, 柳川 洋二郎, 片桐 成二, 千葉 仁志, 惠 淑萍, 永野 昌志  日本獣医学会学術集会講演要旨集  161回-  405  -405  2018/08  [Not refereed][Not invited]
  
• 加齢・疾病対策のための酸化脂質研究の最前線 LC/MSを用いたカルジオリピン過酸化物の分析(Analysis of cardiolipin peroxidation products by LC/MS)

  陳 震, 呉 げつ, 馬 逸興, 小林 悠, 趙 瑤瑤, 三浦 佑介, 千葉 仁志, 惠 淑萍  JSBMS Letters  43-  (Suppl.)  51  -51  2018/08  [Not refereed][Not invited]
  
• 加齢・疾病対策のための酸化脂質研究の最前線 nanoESI-MSおよびLC/MSによる過酸化脂質の分析

  惠 淑萍  JSBMS Letters  43-  (Suppl.)  52  -52  2018/08  [Not refereed][Not invited]
  
• LC/MSによるヒト血清中における分子種別コレステリルエステルの定量分析

  三浦 佑介, 加藤 颯太, 櫻井 俊宏, 陳 震, 呉 げつ, 高 紫君, ロジート・シュレスタ, 中村 幸志, 鵜川 重和, 中川 貴史, 玉腰 暁子, 千葉 仁志, 惠 淑萍  JSBMS Letters  43-  (Suppl.)  82  -82  2018/08  [Not refereed][Not invited]
  
• 血中短鎖脂肪酸と肥満との関連

  山村 凌大, 陳 震, 呉 げつ, 高 紫君, 惠 淑萍, 千葉 仁志, 中川 貴史, 鵜川 重和, 中村 幸志, 玉腰 暁子  JSBMS Letters  43-  (Suppl.)  84  -84  2018/08  [Not refereed][Not invited]
  
• LC-MS/MSによる糖尿病性腎症患者の尿中コレステリルエステル分析

  辻川 諒哉, 三浦 佑介, 陳 震, 高田 康徳, 千葉 仁志, 惠 淑萍  JSBMS Letters  43-  (Suppl.)  86  -86  2018/08  [Not refereed][Not invited]
  
• HepG2細胞内の単一脂肪滴に関するnanoESI-MS直接脂質分析(Direct lipid analysis of single lipid droplets in HepG2 cells by nanoESI-MS)

  趙 瑤瑤, 呉 げつ, 陳 震, 高 紫君, 張 新榮, 千葉 仁志, 惠 淑萍  JSBMS Letters  43-  (Suppl.)  102  -102  2018/08  [Not refereed][Not invited]
  
• 単一脂質滴のトリグリセリド構成に及ぼす中鎖脂肪酸の影響(Effect of medium-chain fatty acid on triglyceride profile of single lipid droplets)

  趙 瑤瑤, 陳 震, 呉 げつ, 高 紫君, 張 新榮, 千葉 仁志, 惠 淑萍  JSBMS Letters  43-  (Suppl.)  103  -103  2018/08  [Not refereed][Not invited]
  
• LC/Orbitrap-MSを用いた牛卵子中脂質の網羅的解析(Lipidomic profiling in bovine oocytes by LC/Orbitrap-MS)

  呉 げつ, 植芝 滉己, 陳 震, 坂口 謙一郎, 柳川 洋二郎, 片桐 成二, 永野 昌志, 千葉 仁志, 惠 淑萍  JSBMS Letters  43-  (Suppl.)  105  -105  2018/08  [Not refereed][Not invited]
  
• LC/HR-MS/MSを用いた肝硬変ラット肝臓中脂質と過酸化脂質の定量分析(Quantitative profiling of lipids and their hydroperoxides in cirrhotic rats by LC/HR-MS/MS)

  陳 震, 玉井 康将, 江口 暁子, 岩佐 元雄, 呉 げつ, 竹井 謙之, 千葉 仁志, 惠 淑萍  JSBMS Letters  43-  (Suppl.)  106  -106  2018/08  [Not refereed][Not invited]
  
• 脂肪蓄積および酸化ストレス曝露による代謝変化

  津久井 隆行, 郭 先薈, 陳 震, 惠 淑萍, 千葉 仁志  JSBMS Letters  43-  (Suppl.)  107  -107  2018/08  [Not refereed][Not invited]
  
• LC-MS/MSによる短鎖、中鎖、長鎖、極長鎖脂肪酸の一斉定量分析(Simultaneous quantitation of short-, medium-, long-, and very long-chain fatty acids by LC-MS/MS)

  高 紫君, 陳 震, 呉 げつ, 三浦 祐介, 千葉 仁志, 惠 淑萍  JSBMS Letters  43-  (Suppl.)  136  -136  2018/08  [Not refereed][Not invited]
  
• 食餌由来脂肪肝モデルマウスにおいて鮭白子抽出物の摂取は肝機能障害の改善とミトコンドリアを活性化させる

  櫻井 俊宏, 早坂 孝宏, 関口 博太, 佐藤 浩志, 陳 震, 千葉 仁志, 惠 淑萍  JSBMS Letters  43-  (Suppl.)  147  -147  2018/08  [Not refereed][Not invited]
  
• 破骨細胞の分化を阻害する化合物(ursolic acid)の新しい標的タンパク質を発見(Nuclear export exportin -5 as a target for ursolic acid regulates bone resorption)

  譚 慧, 千葉 仁志, 惠 淑萍  JSBMS Letters  43-  (Suppl.)  148  -148  2018/08  [Not refereed][Not invited]
  
• 低濃度オゾンによる不飽和リン脂質膜の酸化に関する研究(Study on the Oxidation of the Unsaturated Phospholipid Monolayer in Low-level Ozone)

  喬 琳, 陳 震, 高田 俊智, 呉 げつ, 千葉 仁志, 叶 深, 惠 淑萍  JSBMS Letters  43-  (Suppl.)  151  -151  2018/08  [Not refereed][Not invited]
  
• 環境濃度オゾンによるHepG2細胞の酸化に関する研究(Study on the Oxidation of Ambient-level Ozone on HepG2 Cells)

  朱 子健, 喬 琳, 陳 震, 呉 げつ, 千葉 仁志, 惠 淑萍  JSBMS Letters  43-  (Suppl.)  152  -152  2018/08  [Not refereed][Not invited]
  
• 混合単分子膜の低濃度オゾンによる酸化に及ぼすコレステロール添加の影響

  渡部 勇樹, 喬 琳, 陳 震, 高田 俊智, 呉 げつ, 叶 深, 千葉 仁志, 惠 淑萍  JSBMS Letters  43-  (Suppl.)  153  -153  2018/08  [Not refereed][Not invited]
  
• 細胞ベース検定により検査したNrf2依存性抗酸化物質 抗酸化物質の特性分析(Nrf 2 dependent antioxidants screened by cell-based assay: characterization of antioxidant)

  Roan Yuning, Fuda Hirotoshi, Umetsu Satomi, Furukawa Takayuki, Joko Sae, Miyanaga Satoshi, Suzuki Hirotaka, Hui Shu-Ping, Chiba Hitoshi  臨床化学  47-  (Suppl.1)  252  -252  2018/07
• 細胞ベース検定により検査したNrf2依存性抗酸化物質 抗酸化物質の特性分析(Nrf 2 dependent antioxidants screened by cell-based assay: characterization of antioxidant)

  Roan Yuning, Fuda Hirotoshi, Umetsu Satomi, Furukawa Takayuki, Joko Sae, Miyanaga Satoshi, Suzuki Hirotaka, Hui Shu-Ping, Chiba Hitoshi  臨床化学  47-  (Suppl.1)  351  -351  2018/07
• 細胞ベース検定により検査したNrf2依存性抗酸化物質 抗酸化物質のスクリーニングと同定(Nrf 2 dependent antioxidants screened by cell-based assay: screening & identification of antioxidant)

  Fuda Hirotoshi, Roan Yuning, Umetsu Satomi, Furukawa Takayuki, Joko Sae, Miyanaga Satoshi, Suzuki Hirotaka, Hui Shu-Ping, Chiba Hitoshi  臨床化学  47-  (Suppl.1)  380  -380  2018/07
• 近位尿細管上皮細胞の脂肪酸負荷による脂質変化

  三浦 佑介, 櫻井 俊宏, 陳 震, 布田 博敏, 千葉 仁志, 惠 淑萍  臨床化学  47-  (Suppl.1)  378  -378  2018/07  [Not refereed][Not invited]
  
• リポータージーンアッセイを用いた食品由来抗酸化物質の抗酸化能評価

  鈴木 拓貴, 布田 博敏, 上甲 紗愛, 渡邊 貢, Roan Yuning, 武田 晴治, 古川 貴之, 惠 淑萍, 千葉 仁志  臨床化学  47-  (Suppl.1)  381  -381  2018/07  [Not refereed][Not invited]
  
• LDLの硬さと加齢について

  武田 晴治, 櫻井 俊宏, 惠 淑萍, 千葉 仁志  臨床化学  47-  (Suppl.1)  382  -382  2018/07  [Not refereed][Not invited]
  
• LDL-TG測定試薬の臨床検体への応用

  櫻井 俊宏, 高橋 祐司, 藤井 佑樹, 能祖 一裕, 太田 素子, 伊藤 康樹, 永坂 敦, 和田 典男, 千葉 仁志, 惠 淑萍  臨床化学  47-  (Suppl.1)  383  -383  2018/07  [Not refereed][Not invited]
  
• フッ素置換構造を鍵としたリゾリン脂質異性体の合成と物性に関する研究

  山本 祐輔, 古川 貴之, 樫田 紘之, 武田 晴治, 惠 淑萍  臨床化学  47-  (Suppl.1)  385  -385  2018/07  [Not refereed][Not invited]
  
• 質量分析の活用事例とピットフォール 健康科学研究におけるLC/MS分析法とピットフォール

  惠 淑萍  臨床化学  47-  (Suppl.1)  180  -180  2018/07  [Not refereed][Not invited]
  
• 臨床化学の未病対策への挑戦 脂肪蓄積と酸化ストレスに関する研究

  惠 淑萍  臨床化学  47-  (Suppl.1)  196  -196  2018/07  [Not refereed][Not invited]
  
• 臨床化学の食への挑戦 食品機能性への臨床化学的アプローチ

  千葉 仁志, 布田 博敏, 惠 淑萍  臨床化学  47-  (Suppl.1)  213  -213  2018/07  [Not refereed][Not invited]
  
• フッ素置換構造を鍵としたリゾリン脂質異性体の合成と物性に関する研究

  山本 祐輔, 古川 貴之, 樫田 紘之, 武田 晴治, 惠 淑萍  臨床化学  47-  (Suppl.1)  231  -231  2018/07  [Not refereed][Not invited]
  
• LDL-TG測定試薬の臨床検体への応用

  櫻井 俊宏, 高橋 祐司, 藤井 佑樹, 能祖 一裕, 太田 素子, 伊藤 康樹, 永坂 敦, 和田 典男, 千葉 仁志, 惠 淑萍  臨床化学  47-  (Suppl.1)  244  -244  2018/07  [Not refereed][Not invited]
  
• ヒト血清中のブタン酸とカプロン酸のLC-MS/MSを用いた定量法(Determination of butanoic and caproic acid in human serum by LC-MS/MS)

  陳 震, Yue Wu, シュレスタ・ロジート, 三浦 祐介, 趙 瑤瑤, 玉腰 暁子, 千葉 仁志, 惠 淑萍  臨床化学  47-  (Suppl.1)  377  -377  2018/07  [Not refereed][Not invited]
  
• 近位尿細管上皮細胞の脂肪酸負荷による脂質変化

  三浦 佑介, 櫻井 俊宏, 陳 震, 布田 博敏, 千葉 仁志, 惠 淑萍  臨床化学  47-  (Suppl.1)  378  -378  2018/07  [Not refereed][Not invited]
  
• リポータージーンアッセイを用いた食品由来抗酸化物質の抗酸化能評価

  鈴木 拓貴, 布田 博敏, 上甲 紗愛, 渡邊 貢, Roan Yuning, 武田 晴治, 古川 貴之, 惠 淑萍, 千葉 仁志  臨床化学  47-  (Suppl.1)  381  -381  2018/07  [Not refereed][Not invited]
  
• LDLの硬さと加齢について

  武田 晴治, 櫻井 俊宏, 惠 淑萍, 千葉 仁志  臨床化学  47-  (Suppl.1)  382  -382  2018/07  [Not refereed][Not invited]
  
• LDL-TG測定試薬の臨床検体への応用

  櫻井 俊宏, 高橋 祐司, 藤井 佑樹, 能祖 一裕, 太田 素子, 伊藤 康樹, 永坂 敦, 和田 典男, 千葉 仁志, 惠 淑萍  臨床化学  47-  (Suppl.1)  383  -383  2018/07  [Not refereed][Not invited]
  
• フッ素置換構造を鍵としたリゾリン脂質異性体の合成と物性に関する研究

  山本 祐輔, 古川 貴之, 樫田 紘之, 武田 晴治, 惠 淑萍  臨床化学  47-  (Suppl.1)  385  -385  2018/07  [Not refereed][Not invited]
  
• 脂質関連マーカーのUpdate 脂質・酸化脂質の分析と臨床応用

  惠 淑萍  臨床病理  66-  (4)  428  -428  2018/04  [Not refereed][Not invited]
  
• LC/MSによる腎疾患患者の血清・尿中の脂質分析

  惠淑萍, 三浦佑介, 寺田航, 櫻井俊宏, 陳震, 古川貴之, 小林美穂, 清水力, 千葉仁志  腎と脂質研究会プログラム・抄録集  30th-  43  2018  [Not refereed][Not invited]
  
• 脂質関連マーカーのUpdate 脂質・酸化脂質の分析と臨床応用

  惠 淑萍  臨床病理  65-  (補冊)  025  -025  2017/10  [Not refereed][Not invited]
  
• 地域医療におけるアンチエイジングを包括的に考える 臨床検査に時間軸を入れよう

  千葉 仁志, 惠 淑萍  日本未病システム学会学術総会抄録集  24回-  67  -67  2017/10  [Not refereed][Not invited]
  
• 腎疾患における尿中バイオマーカーとしてのコレステリルエステルの有用性

  三浦 佑介, 古川 貴之, 小林 美穂, 清水 力, 千葉 仁志, 惠 淑萍  日本未病システム学会学術総会抄録集  24回-  93  -93  2017/10  [Not refereed][Not invited]
  
• 直接及び間接抗酸化物質における分配係数、細胞毒性及び細胞保護能の比較

  布田 博敏, 上甲 紗愛, 渡邉 貢, 武田 晴治, 古川 貴之, Shrestha Rojeet, 惠 淑萍, 千葉 仁志  日本未病システム学会学術総会抄録集  24回-  98  -98  2017/10  [Not refereed][Not invited]
  
• ヒトリポ蛋白中の過酸化コレステリルエステル及びトリグリセリドの同定

  惠 淑萍, 三浦 佑介, 櫻井 俊宏, 武田 晴治, 布田 博敏, 千葉 仁志  日本未病システム学会学術総会抄録集  24回-  99  -99  2017/10  [Not refereed][Not invited]
  
• 脂質関連マーカーのUpdate 脂質・酸化脂質の分析と臨床応用

  惠 淑萍  臨床病理  65-  (補冊)  025  -025  2017/10  [Not refereed][Not invited]
  
• マススペクトロメトリーの検査応用 現状と今後の課題 質量分析による低分子の分析および臨床検体への応用

  惠 淑萍  臨床病理  65-  (補冊)  070  -070  2017/10  [Not refereed][Not invited]
  
• ヒト近位尿細管上皮細胞における蓄積脂質の分析

  三浦佑介, 櫻井俊宏, 津久井隆行, 陳震, 布田博敏, 千葉仁志, 惠淑萍  臨床化学  46-  315  2017/09/01  [Not refereed][Not invited]
  
• ヒト近位尿細管上皮細胞における蓄積脂質の分析

  三浦佑介, 櫻井俊宏, 津久井隆行, 陳震, 布田博敏, 千葉仁志, 惠淑萍  臨床化学  46-  203  2017/09/01  [Not refereed][Not invited]
  
• プラズマローゲンライブラリーの合成とLC/MSを用いた絶対定量系の開発

  寺田 航, 古川 貴之, 李 娜, 三浦 佑介, 千葉 仁志, 惠 淑萍  臨床化学  46-  (Suppl.1)  209  -209  2017/09  [Not refereed][Not invited]
  
• フッ素置換構造を鍵としたリゾリン脂質異性体の合成と物性に関する研究

  山本 祐輔, 古川 貴之, 武田 晴治, 千葉 仁志, 惠 淑萍  臨床化学  46-  (Suppl.1)  211  -211  2017/09  [Not refereed][Not invited]
  
• HepG2細胞におけるカルジオリピンとそのヒドロペルオキシド類のLC/MSによる組成分析(Profiling of Cardiolipins and Their Hydroperoxides in HepG2 Cells by LC/MS)

  陳 震, Wu Yue, 馬 逸興, 小林 悠, 趙 瑤瑤, 三浦 佑介, 千葉 仁志, 惠 淑萍  臨床化学  46-  (Suppl.1)  284  -284  2017/09  [Not refereed][Not invited]
  
• 高血糖ゼブラフィッシュにおけるリン脂質とそのヒドロペルオキシド類のLC/MSによる同時組成分析(Simultaneous profiling of phospholipids and their hydroperoxides in hyperglycemia zebrafish by LC/MS)

  Wu Yue, 陳 震, 臧 黎清, 趙 瑤瑤, 三浦 佑介, 西村 訓弘, 千葉 仁志, 惠 淑萍  臨床化学  46-  (Suppl.1)  284  -284  2017/09  [Not refereed][Not invited]
  
• 血漿カプリン酸を定量するための改良LC/MS検定法(An improved LC/MS assay for the measurement of plasma capric acid)

  冨田 優理, Shrestha Rojeet, 平野 賢一, 千葉 仁志, 惠 淑萍  臨床化学  46-  (Suppl.1)  285  -285  2017/09  [Not refereed][Not invited]
  
• リゾホスファチジルエタノールアミンの簡便合成法及び分析法の開発とNASHモデルマウスへの応用

  古川 貴之, 布田 博敏, 千葉 仁志, 惠 淑萍  臨床化学  46-  (Suppl.1)  286  -286  2017/09  [Not refereed][Not invited]
  
• 質量分析による血清コレステリルエステルの分析

  加藤 颯太, 三浦 佑介, 櫻井 俊宏, Shrestha Rojeet, 陳 震, 玉腰 暁子, 千葉 仁志, 惠 淑萍  臨床化学  46-  (Suppl.1)  287  -287  2017/09  [Not refereed][Not invited]
  
• オレイン酸負荷により細胞内に形成させた脂肪滴の酸化変動解析および抗酸化物質による酸化抑制の検討

  大浦 弘太郎, 津久井 隆行, 千葉 仁志, 惠 淑萍  臨床化学  46-  (Suppl.1)  288  -288  2017/09  [Not refereed][Not invited]
  
• 脂肪蓄積と酸化ストレス曝露による肝細胞の代謝変化と過酸化脂質放出

  千葉 仁志, 郭 先薈, 津久井 隆行, 馬 逸興, 陳 震, 布田 博敏, 惠 淑萍  臨床化学  46-  (Suppl.1)  288  -288  2017/09  [Not refereed][Not invited]
  
• マガキ由来抗酸化物質のKeap1-Nrf2経路活性化と抗酸化能

  布田 博敏, 上甲 紗愛, 渡邉 貢, 武田 晴治, 古川 貴之, 惠 淑萍, Shrestha Rojeet, 千葉 仁志  臨床化学  46-  (Suppl.1)  304  -304  2017/09  [Not refereed][Not invited]
  
• LDLの酸化および酵素処理が硬さに与える影響について

  武田 晴治, アグス・スバギョ, 布田 博敏, 惠 淑萍, 末岡 和久, 千葉 仁志  臨床化学  46-  (Suppl.1)  329  -329  2017/09  [Not refereed][Not invited]
  
• 健常日本人被験者における血清総脂肪酸および非エステル化脂肪酸の組成分析法(Serum total and non-esterified fatty acid profiling in healthy Japanese individuals)

  惠 淑萍, Shrestha Rojeet, 三浦 佑介, 陳 震, 玉腰 暁子, 千葉 仁志  臨床化学  46-  (Suppl.1)  329  -329  2017/09  [Not refereed][Not invited]
  
• ヒトリポタンパク質における非極性脂質の酸化に関する分析

  惠 淑萍, 三浦 佑介, Shrestha Rojeet, 櫻井 俊宏, 武田 晴治, 布田 博敏, 千葉 仁志  臨床化学  46-  (Suppl.1)  330  -330  2017/09  [Not refereed][Not invited]
  
• 高脂肪食および酸化LDL負荷マウスにおける腎臓への脂肪蓄積

  津久井 隆行, 布田 博敏, 惠 淑萍, 千葉 仁志  臨床化学  46-  (Suppl.1)  332  -332  2017/09  [Not refereed][Not invited]
  
• 酸化HDLが肝由来株化細胞C3Aに及ぼす影響

  仲門 菜月, 櫻井 俊宏, 岸田 佳倫, 布田 博敏, 津久井 隆行, 千葉 仁志, 惠 淑萍  臨床化学  46-  (Suppl.1)  333  -333  2017/09  [Not refereed][Not invited]
  
• プロテオミクスを用いる抗酸化酵素群の発現量解析

  岸田 佳倫, 櫻井 俊宏, 仲門 菜月, 布田 博敏, 古川 貴之, 千葉 仁志, 惠 淑萍  臨床化学  46-  (Suppl.1)  347  -347  2017/09  [Not refereed][Not invited]
  
• 化学先取りシンポジウム グライコブロッティング法によるリポタンパク質中の脂質アルデヒド分析

  古川 貴之, 比能 洋, 武田 晴治, 千葉 仁志, 西村 紳一郎, 惠 淑萍  臨床化学  46-  (Suppl.1)  173  -173  2017/09  [Not refereed][Not invited]
  
• 化学先取りシンポジウム 酸化還元電位および硬さ分布からLDLの酸化状態を評価

  武田 晴治, アグス・スバギョ, 布田 博敏, 惠 淑萍, 末岡 和久, 千葉 仁志  臨床化学  46-  (Suppl.1)  175  -175  2017/09  [Not refereed][Not invited]
  
• マススペクトロメトリーの医療への応用 体外受精から未病診断まで 体外受精における使用済み胚培養液の組成と胚発育の関係

  惠 淑萍  臨床化学  46-  (Suppl.1)  181  -181  2017/09  [Not refereed][Not invited]
  
• 冷保存肝の脂質の網羅的解析

  白澤 憲典, 早坂 孝宏, 深井 原, 梅本 浩平, 石川 隆壽, 櫻井 俊宏, 布田 博敏, 橋本 咲月, 大谷 晋太郎, 中藪 拓哉, 島田 慎吾, 嶋村 剛, 武冨 紹信, 千葉 仁志, 惠 淑萍  臨床化学  46-  (Suppl.1)  198  -198  2017/09  [Not refereed][Not invited]
  
• 酵素処理がLDLの物性に与える影響(Effect of enzymes treatment on physical properties of lowdensity lipoprotein)

  Takeda Seiji, Subagyo Agus, Hui Shu-Ping, Fuda Hirotoshi, Sueoka Kazuhisa, Chiba Hitoshi  生物物理  57-  (Suppl.1-2)  S278  -S278  2017/08
• LC/MS/MSを用いた過酸化カルジオリピン分子種の一斉分析(Simultaneous analysis of cardiolipin hydroperoxide molecular species by liquid chromatography/tandem mass spectrometry)

  陳 震, 呉 ゆえ, 馬 逸興, 小林 悠, 趙 瑤瑤, 三浦 佑介, 惠 淑萍, 千葉 仁志  JSBMS Letters  42-  (Suppl.)  67  -67  2017/08  [Not refereed][Not invited]
  
• エポキシ化反応及びMSによる不飽和脂肪酸の二重結合に関する分析と定量(Identification and quantitation of C=C location isomers of unsaturated fatty acids by epoxidation reaction and tandem mass spectrometry)

  趙 瑤瑤, 千葉 仁志, 惠 淑萍  JSBMS Letters  42-  (Suppl.)  73  -73  2017/08  [Not refereed][Not invited]
  
• グライコブロッティング法に基づくリポソーム及びリポたんぱく質中酸化脂質の迅速解析

  古川 貴之, 比能 洋, 武田 晴治, 千葉 仁志, 西村 紳一郎, 惠 淑萍  JSBMS Letters  42-  (Suppl.)  96  -96  2017/08  [Not refereed][Not invited]
  
• LC-MS/MSによる近位尿細管上皮細胞の脂質分析

  三浦 佑介, 櫻井 俊宏, 津久井 隆行, 陳 震, 布田 博敏, 千葉 仁志, 惠 淑萍  JSBMS Letters  42-  (Suppl.)  110  -110  2017/08  [Not refereed][Not invited]
  
• 中鎖脂肪酸トリグリセリドの短期経口投与中における血漿カプリン酸濃度(Plasma capric acid concentrations during a short-term oral administration of medium-chain triglycerides)

  Shrestha Rojeet, Hirano Kenichi, Suzuki Akira, Yamaguchi Satoshi, Chiba Hitoshi, Hui Shu-Ping  日本動脈硬化学会総会プログラム・抄録集  49th-  255 (WEB ONLY)  -255  2017/06/25  [Not refereed][Not invited]
  
• 質量分析イメージング法(IMS)阻血再灌流における新規予後予測マーカーの探索

  橋本 咲月, 梅本 浩平, 大谷 晋太郎, 中薮 拓哉, 三野 和宏, 武冨 紹信, 深井 原, 木村 太一, 早坂 孝宏, 惠 淑萍, 千葉 仁志, 嶋村 剛  北海道外科雑誌  62-  (1)  101  -101  2017/06  [Not refereed][Not invited]
  
• 中鎖脂肪酸療法により難治性狭心症が改善した特発性中性脂肪蓄積心筋血管症の一例

  平野 賢一, 安井 洋子, 東 将浩, 坂口 学, 鈴木 朗, 高木 敦子, Shrestha Rojeet, 橋本 千佳子, 李 銘, 原 康洋, 山口 知是, 池田 康行, 惠 淑萍, 内藤 博昭, 長坂 博範  日本動脈硬化学会総会プログラム・抄録集  49回-  241  -241  2017/06  [Not refereed][Not invited]
  
• 中鎖脂肪酸療法が著効を示した特発性中性脂肪蓄積心筋血管症の一例

  橋本 千佳子, 鈴木 朗, 高木 敦子, シュレスタ・ロジート, 中林 十士紀, 李 銘, 池田 康行, 惠 淑萍, 和田 晃, 平野 賢一  日本動脈硬化学会総会プログラム・抄録集  49回-  297  -297  2017/06  [Not refereed][Not invited]
  
• マガキ抽出物はNASHモデルマウスの肝臓において脂肪肝,炎症,アポトーシス,線維化を抑制する

  布田博敏, 渡邉貢, 岡部浩昭, 上甲紗愛, 三浦佑介, HUI Shu‐Ping, YI Min, 濱岡直裕, 三木恵美子, 千葉仁志  日本農芸化学会大会講演要旨集(Web)  2017-  ROMBUNNO.2B09p07 (WEB ONLY)  2017/03/05  [Not refereed][Not invited]
  
• イメージング質量分析法における超臨界洗浄法の開発

  松下 祥子, 正木 紀隆, 佐藤 浩平, 早坂 孝宏, Paxton Thanai, 杉山 栄二, 佐藤 太, 寺崎 真樹, 惠 淑萍, 千葉 仁志, 間瀬 暢之, 瀬藤 光利  日本薬学会年会要旨集  137年会-  (2)  297  -297  2017/03  [Not refereed][Not invited]
  
• 酸化と酵素切断がLDLの物性に与える影響(Physical properties of low-density lipoprotein after oxidation or proteolytic enzyme treatment)

  Takeda Seiji, Subagyo Agus, Hui Shu-Ping, Fuda Hirotoshi, Sueoka Kazuhisa, Chiba Hitoshi  生物物理  56-  (Suppl.1-2)  S290  -S290  2016/10
• 未病に役立つ栄養 マガキ抽出物による抗肥満作用、インスリン抵抗性改善作用、及び肝臓保護作用

  布田 博敏, 渡邉 貢, 惠 淑萍, 千葉 仁志  日本未病システム学会学術総会抄録集  23回-  68  -68  2016/10  [Not refereed][Not invited]
  
• NASHモデルマウスにおけるマガキ抽出物の肝臓保護作用

  布田 博敏, 渡邉 貢, 岡部 浩昭, 上甲 紗愛, 三浦 佑介, 惠 淑萍, 伊 敏, 濱岡 直裕, 三木 恵美子, 千葉 仁志  日本未病システム学会学術総会抄録集  23回-  115  -115  2016/10  [Not refereed][Not invited]
  
• 北海道における地方生活者と都市生活者の血中25-ヒドロキシビタミンD量の比較

  岡部 浩昭, 岡田 恵美子, 鵜川 重和, 中村 幸志, 中川 貴史, 玉腰 暁子, 菊地 玲, 南 昭子, 清水 力, 陳 震, 千葉 仁志, 惠 淑萍  日本未病システム学会学術総会抄録集  23回-  120  -120  2016/10  [Not refereed][Not invited]
  
• 質量分析による分子レベルの脂質研究

  惠 淑萍  臨床化学  45-  (Suppl.1)  89  -90  2016/10  [Not refereed][Not invited]
  
• リゾリン脂質の新たな可能性 リゾホスファチジルエタノールアミンの合成とフッ素アナログへの展開

  古川 貴之, 惠 淑萍, 山本 祐輔, 樫田 紘之, 小路 明日香, 布田 博敏, 千葉 仁志  臨床化学  45-  (Suppl.1)  129  -129  2016/10  [Not refereed][Not invited]
  
• 脂質アルデヒド類の新規分析法 グライコブロッティング法によるアプローチ 合成標品からリポソームまで

  古川 貴之, 惠 淑萍, 比能 洋, 西村 紳一郎, 千葉 仁志  臨床化学  45-  (Suppl.1)  167  -167  2016/10  [Not refereed][Not invited]
  
• 内部標準物質及び標品として用いるカルジオリピンの合成研究と定量分析へ向けた基礎的検討

  高橋 遼地, 古川 貴之, 三浦 佑介, 惠 淑萍, 千葉 仁志  臨床化学  45-  (Suppl.1)  181  -181  2016/10  [Not refereed][Not invited]
  
• 脂肪滴と酸化ストレスが骨格筋に及ぼす影響に関する研究

  小林 悠, 馬 逸興, 中島 進吾, 惠 淑萍, 千葉 仁志  臨床化学  45-  (Suppl.1)  219  -219  2016/10  [Not refereed][Not invited]
  
• LC/MSを用いたヒト胚培養後の培養液における脂肪酸分析

  惠 淑萍, 八木 亜希子, 宮永 賢, Shrestha Rojeet, 武田 晴治, 神谷 博文, 千葉 仁志  JSBMS Letters  41-  (Suppl.)  68  -68  2016/08  [Not refereed][Not invited]
  
• 質量分析による生体試料中のコレステリルエステルの分析

  三浦 佑介, 惠 淑萍, 高橋 遼地, 小林 美穂, 清水 力, 千葉 仁志  日本分析化学会講演要旨集  65年会-  308  -308  2016/08  [Not refereed][Not invited]
  
• 電気泳動による臨床検査・機能性食品の新たな展開 検査室でのApoE-rich HDLとの遭遇から診断薬開発まで

  千葉 仁志, 高橋 祐司, 伊藤 康樹, 惠 淑萍  電気泳動  60-  (Suppl.)  s5  -s5  2016/08  [Not refereed][Not invited]
  
• 質量分析手法を用いた次世代臨床検査の新展開 質量分析法による種々の脂質分子の定性及び定量分析

  惠 淑萍  臨床病理  64-  (6)  674  -674  2016/06  [Not refereed][Not invited]
  
• マガキ由来抗酸化物質によるNrf2標的遺伝子の発現

  布田 博敏, 上甲 紗愛, 渡邉 貢, 惠 淑萍, 武田 晴治, 渡邉 孝之, 千葉 仁志  脂質生化学研究  58-  50  -50  2016/05  [Not refereed][Not invited]
  
• LC-MS/MSによるコレステリルエステル定量法の開発

  三浦 佑介, 惠 淑萍, 高橋 遼地, 千葉 仁志  臨床化学  45-  (2)  127  -134  2016/04  [Not refereed][Not invited]
   

  脂質分子の多様性がもつ意義を解明することは重要である。複雑な生体試料の脂質分子種を正確に分析するためには、信頼できる標準品が必要となる。液体クロマトグラフィー/タンデム質量分析(LC-MS/MS)法の内標準物質には、安定同位元素標識体が最も適している。しかし、標識体は市場からの入手が難しく、その標識純度も定量分析には適さない。そこで我々は、LC-MS/MS法の内標準物質となる重水素標識コレステリルエステル(CE)の化学合成を行った。主要な反応として、(1)2α,3α-epoxy-5α-steroidsのLiAlD4による開環反応、(2)NaBD4を用いた3,6-diketo-5α-steroidsの立体選択的な(3α,6α-D2)-3β,6β-diolへの還元、(3)6β-hydroxy-5α-steroidにPOCl3を反応させることによる5位からの脱水、および1-(3-dimethylaminopropyl)-3-ethylcarbodiimideを触媒とした重水素標識コレステロールと脂肪酸のエステル化反応があげられる。合成した標識体を用いてCEのLC-MS/MS定量法を確立した。本法は様々な臨床研究に対し、高感度・高選択的なCE定量法として貢献することが期待される。(著者抄録)
• 抗酸化物質の概念の変化と新しい抗酸化食品開発戦略

  布田 博敏, 岡部 浩昭, 惠 淑萍, 千葉 仁志  JSBMS Letters  41-  (1)  7  -13  2016/03  [Not refereed][Not invited]
   

  酸素は生物にとって欠くことができず、絶えず活性酸素種(ROS)の生成が行われている。高度に分化し、かつ統合された一連の抗酸化酵素群がROSから細胞を保護しているが、年齢とともに低下することが報告されている、そのために食品中の抗酸化物質を取ることが健康維持のために有意義と考えられている。Dinkonva-Kostova、Talalayらが抗酸化物質をdirect antioxidantとindirect antioxidantの2つに分類して以来、抗酸化、抗酸化物質の概念は大きく変わり始めた。彼らはdirect antioxidantを「ROSに直接反応して、不活化する物質」、indirect antioxidantを「Keap1-Nrf2経路を活性化し、一連の抗酸化酵素群を発現することにより生体内のROSや異物を不活化・代謝する物質」と定義した。本稿では抗酸化物質の概念の変化について解説し、我々が考える新しい抗酸化食品開発戦略についても述べる。具体的な開発事例として、我々がマガキ抽出液から見いだした新規抗酸化物質の研究を紹介する。質量分析は抗酸化物質の探索、同定、定量分析において重要な役割を果たしている。(著者抄録)
• 多項目自動血球分析装置XNシリーズによる,原発性中性脂肪蓄積心筋血管症由来の好中球脂肪滴の検出

  和田淳, 鈴木朗, 鈴木朗, 山本志緒里, 海道雅子, 越智康浩, 惠淑萍, 千葉仁志, 平野賢一, 平野賢一  臨床病理  63-  250  2015/10/20  [Not refereed][Not invited]
  
• 質量分析法による種々の脂質分子の定性及び定量分析

  惠淑萍  臨床病理  63-  029  2015/10/20  [Not refereed][Not invited]
  
• LC‐MS/MSによるコレステリルエステル定量法の開発

  三浦佑介, 惠淑萍, 高橋遼地, 千葉仁志  臨床化学  44-  197  2015/10/07  [Not refereed][Not invited]
  
• グライコブロッティング法を用いた脂質アルデヒドの定量測定法の開発

  古川貴之, 惠淑萍, 比能洋, 西村紳一郎, 千葉仁志  臨床化学  44-  209  2015/10/07  [Not refereed][Not invited]
  
• 酸化ストレスによるヒト肝培養細胞の脂肪滴形成と脂質分泌に及ぼす影響

  梅津里美, 宮永賢, 布田博敏, 惠淑萍, 千葉仁志  臨床化学  44-  222  2015/10/07  [Not refereed][Not invited]
  
• 内部標準物質として用いる非天然型カルジオリピンの合成研究と定量分析へ向けた基礎的検討

  高橋遼地, 古川貴之, 惠淑萍, 千葉仁志  臨床化学  44-  218  2015/10/07  [Not refereed][Not invited]
  
• HILIC‐LC/MSによる体外受精用培養液におけるアミノ酸のメタボローム解析

  林沙紀, 宮永賢, 千葉仁志, 八木亜希子, 八木亜希子, 神谷博文, 惠淑萍  臨床化学  44-  210  2015/10/07  [Not refereed][Not invited]
  
• マガキ由来抗酸化物質,DHMBA,によるKeap1‐Nrf2経路活性化

  上甲紗愛, 布田博敏, 渡邉貢, 惠淑萍, 武田晴治, 渡邉孝之, 千葉仁志  臨床化学  44-  227  2015/10/07  [Not refereed][Not invited]
  
• LDL酸化に及ぼすアミノ酸の影響に関する研究

  橋本咲月, 武田晴治, 寺嶋駿, 布田博敏, 惠淑萍, 千葉仁志  臨床化学  44-  220  2015/10/07  [Not refereed][Not invited]
  
• 臨床化学の未来を拓く 臨床化学教育・研究における国際交流

  惠 淑萍  臨床化学  44-  (Suppl.1)  135  -136  2015/10  [Not refereed][Not invited]
  
• 一年を通しての貯蔵型ビタミンDと紫外線量との関連性

  菊地玲, 南昭子, 道又理恵, 山下直樹, 安田慶子, 澁谷斉, 加畑馨, 岡部浩昭, 惠淑萍, 千葉仁志, 清水力  日本未病システム学会学術総会抄録集  22nd-  165  2015/09/10  [Not refereed][Not invited]
  
• マガキ抽出物中の両親媒性抗酸化フェノールの定量

  岡部浩昭, 惠淑萍, 布田博敏, 大谷晋太郎, 渡辺孝之, 渡辺貢, 千葉仁志  日本未病システム学会学術総会抄録集  22nd-  175  2015/09/10  [Not refereed][Not invited]
  
• ラットC6グリア細胞株における脂肪滴形成とその影響

  中島進吾, 牧原圭佑, 惠淑萍, 千葉仁志  日本未病システム学会学術総会抄録集  22nd-  155  2015/09/10  [Not refereed][Not invited]
  
• 脂肪滴の分析技術

  惠淑萍  日本未病システム学会学術総会抄録集  22nd-  79  2015/09/10  [Not refereed][Not invited]
  
• メタボロミクスによる体外受精用培養液のアミノ酸の解析

  林沙紀, 宮永賢, 千葉仁志, 八木亜希子, 八木亜希子, 神谷博文, 惠淑萍  日本未病システム学会学術総会抄録集  22nd-  179  2015/09/10  [Not refereed][Not invited]
  
• 脂肪細胞に蓄えられた脂肪滴より過酸化トリグリセリドを検出した

  惠淑萍, 繁富(栗林)香織, 小林悠, 千葉仁志  日本未病システム学会学術総会抄録集  22nd-  158  2015/09/10  [Not refereed][Not invited]
  
• ブロッティング技術に基づく脂質アルデヒド類の網羅的定量解析

  古川貴之, 惠淑萍, 比能洋, 西村紳一郎, 千葉仁志  日本未病システム学会学術総会抄録集  22nd-  136  2015/09/10  [Not refereed][Not invited]
  
• マガキ由来の抗酸化物質における肝細胞保護作用

  布田博敏, 渡邉貢, 上甲紗愛, 惠淑萍, 武田晴治, 渡邉孝之, 千葉仁志  日本未病システム学会学術総会抄録集  22nd-  174  2015/09/10  [Not refereed][Not invited]
  
• 牛乳ビタミンD類の定量に向けた新規ラベル化試薬の合成と応用

  中藪拓哉, 岡部浩昭, 古川貴之, 惠淑萍, 千葉仁志  日本未病システム学会学術総会抄録集  22nd-  151  2015/09/10  [Not refereed][Not invited]
  
• 尿中コレステリルエステルは腎脂質代謝を反映する

  三浦佑介, 惠淑萍, 小林美穂, 清水力, 千葉仁志  日本未病システム学会学術総会抄録集  22nd-  152  2015/09/10  [Not refereed][Not invited]
  
• Glycoblotting法を用いた牛乳中遊離オリゴ糖の網羅的定量解析

  山本祐輔, 古川貴之, 惠淑萍, 比能洋, 西村紳一郎, 千葉仁志  日本未病システム学会学術総会抄録集  22nd-  134  2015/09/10  [Not refereed][Not invited]
  
• 酸化LDL由来の酸化分解物質の組成と性質

  寺嶋駿, 武田晴治, 宮永賢, 吉田繁, 惠淑萍, 千葉仁志  日本未病システム学会学術総会抄録集  22nd-  153  2015/09/10  [Not refereed][Not invited]
  
• マガキ由来抗酸化物質のKeap1‐Nrf2経路活性化による肝保護作用

  上甲紗愛, 布田博敏, 渡邉貢, 惠淑萍, 武田晴治, 渡邉孝之, 千葉仁志  日本未病システム学会学術総会抄録集  22nd-  116  2015/09/10  [Not refereed][Not invited]
  
• 酸化されたHepG2細胞による酸化トリグリセリド分泌と脂肪滴形成

  梅津里美, 宮永賢, 布田博敏, 惠淑萍, 千葉仁志  日本未病システム学会学術総会抄録集  22nd-  156  2015/09/10  [Not refereed][Not invited]
  
• LC‐MS/MSによるマガキ由来フェノール性抗酸化物質の測定法の開発とその応用

  大谷晋太郎, 惠淑萍, 岡部浩昭, 布田博敏, 渡辺貢, 千葉仁志  日本未病システム学会学術総会抄録集  22nd-  115  2015/09/10  [Not refereed][Not invited]
  
• システインはLDLを安定化させて酸化を抑制する

  橋本咲月, 武田晴治, 寺嶋駿, 布田博敏, 惠淑萍, 千葉仁志  日本未病システム学会学術総会抄録集  22nd-  117  2015/09/10  [Not refereed][Not invited]
  
• LC/MSを用いた、血漿中・triglyceride rich-lipoprotein中のphosphatidylcholine hydroperoxideの分析(Analysis of phosphatidylcholine hydroperoxides in plasma and triglyceride-rich lipoproteins by LC/MS)

  Shrestha Rojeet, 惠 淑萍, 武田 晴治, 布田 博敏, 千葉 仁志  日本未病システム学会学術総会抄録集  22回-  118  -118  2015/09  [Not refereed][Not invited]
  
• Glycoblotting法を用いた牛乳中遊離オリゴ糖の網羅的定量解析

  山本 祐輔, 古川 貴之, 惠 淑萍, 比能 洋, 西村 紳一郎, 千葉 仁志  日本未病システム学会学術総会抄録集  22回-  134  -134  2015/09  [Not refereed][Not invited]
  
• ブロッティング技術に基づく脂質アルデヒド類の網羅的定量解析

  古川 貴之, 惠 淑萍, 比能 洋, 西村 紳一郎, 千葉 仁志  日本未病システム学会学術総会抄録集  22回-  136  -136  2015/09  [Not refereed][Not invited]
  
• 牛乳ビタミンD類の定量に向けた新規ラベル化試薬の合成と応用

  中藪 拓哉, 岡部 浩昭, 古川 貴之, 惠 淑萍, 千葉 仁志  日本未病システム学会学術総会抄録集  22回-  151  -151  2015/09  [Not refereed][Not invited]
  
• 尿中コレステリルエステルは腎脂質代謝を反映する

  三浦 佑介, 惠 淑萍, 小林 美穂, 清水 力, 千葉 仁志  日本未病システム学会学術総会抄録集  22回-  152  -152  2015/09  [Not refereed][Not invited]
  
• 酸化LDL由来の酸化分解物質の組成と性質

  寺嶋 駿, 武田 晴治, 宮永 賢, 吉田 繁, 惠 淑萍, 千葉 仁志  日本未病システム学会学術総会抄録集  22回-  153  -153  2015/09  [Not refereed][Not invited]
  
• ラットC6グリア細胞株における脂肪滴形成とその影響

  中島 進吾, 牧原 圭佑, 惠 淑萍, 千葉 仁志  日本未病システム学会学術総会抄録集  22回-  155  -155  2015/09  [Not refereed][Not invited]
  
• 酸化されたHepG2細胞による酸化トリグリセリド分泌と脂肪滴形成

  梅津 里美, 宮永 賢, 布田 博敏, 惠 淑萍, 千葉 仁志  日本未病システム学会学術総会抄録集  22回-  156  -156  2015/09  [Not refereed][Not invited]
  
• 筋細胞において、DHMBAはミトコンドリアの機能を向上させ、脂肪酸の利用を促進する(DHMBA increases mitochondrial function and fatty acid utilization in muscle cells)

  馬 逸興, 惠 淑萍, 古川 貴之, 布田 博敏, 千葉 仁志  日本未病システム学会学術総会抄録集  22回-  158  -158  2015/09  [Not refereed][Not invited]
  
• 脂肪細胞に蓄えられた脂肪滴より過酸化トリグリセリドを検出した

  惠 淑萍, 繁富 香織, 林, 小林 悠, 千葉 仁志  日本未病システム学会学術総会抄録集  22回-  158  -158  2015/09  [Not refereed][Not invited]
  
• 一年を通しての貯蔵型ビタミンDと紫外線量との関連性

  菊地 玲, 南 昭子, 道又 理恵, 山下 直樹, 安田 慶子, 澁谷 斉, 加畑 馨, 岡部 浩昭, 惠 淑萍, 千葉 仁志, 清水 力  日本未病システム学会学術総会抄録集  22回-  165  -165  2015/09  [Not refereed][Not invited]
  
• マガキ由来の抗酸化物質における肝細胞保護作用

  布田 博敏, 渡邉 貢, 上甲 紗愛, 惠 淑萍, 武田 晴治, 渡邉 孝之, 千葉 仁志  日本未病システム学会学術総会抄録集  22回-  174  -174  2015/09  [Not refereed][Not invited]
  
• マガキ抽出物中の両親媒性抗酸化フェノールの定量

  岡部 浩昭, 惠 淑萍, 布田 博敏, 大谷 晋太郎, 渡辺 孝之, 渡辺 貢, 千葉 仁志  日本未病システム学会学術総会抄録集  22回-  175  -175  2015/09  [Not refereed][Not invited]
  
• メタボロミクスによる体外受精用培養液のアミノ酸の解析

  林 沙紀, 宮永 賢, 千葉 仁志, 八木 亜希子, 神谷 博文, 惠 淑萍  日本未病システム学会学術総会抄録集  22回-  179  -179  2015/09  [Not refereed][Not invited]
  
• 脂肪細胞の脂肪滴における過酸化トリグリセリドの分析

  惠 淑萍, 繁富 香織, 林, 小林 悠, 千葉 仁志  JSBMS Letters  40-  (Suppl.)  71  -71  2015/08  [Not refereed][Not invited]
  
• 脂肪細胞の脂肪滴における過酸化トリグリセリドの分析

  惠 淑萍, 繁富 香織, 林, 小林 悠, 千葉 仁志  JSBMS Letters  40-  (Suppl.)  71  -71  2015/08  [Not refereed][Not invited]
  
• 細胞折り紙を用いるHepG2・3T3細胞共培養のための3D微細構造形成(Formation of 3D microstructure for HepG2 and 3T3 cells co-culture using cell origami)

  何 倩, 惠 淑萍, 繁富 香織  北海道外科雑誌  60-  (1)  98  -98  2015/06  [Not refereed][Not invited]
  
• 中性脂肪蓄積心筋血管症におけるJordans’ anomalyを自動血球分析装置で検出する

  鈴木朗, 和田淳, 長坂博範, 越智康浩, 千葉仁志, 惠淑萍, 平野賢一  臨床病理  62-  126  2014/10/31  [Not refereed][Not invited]
  
• 他項目自動血球分析装置XE‐5000WBC/BASOチャンネルによる,中性脂肪蓄積心筋血管症判別原理の検証

  和田淳, 鈴木朗, 山本志緒里, 越智康浩, 惠淑萍, 千葉仁志, 平野賢一  臨床病理  62-  127  2014/10/31  [Not refereed][Not invited]
  
• 中鎖脂肪酸のHPLC法による定量分析

  惠淑萍, ろじーと しゅれすた, 平野賢一, 鈴木朗, 千葉仁志  臨床病理  62-  299  2014/10/31  [Not refereed][Not invited]
  
• 機能性脂肪酸に関する分析

  惠淑萍, SHRESTHA Rojeet, 平野賢一, 鈴木朗, 千葉仁志  日本未病システム学会学術総会抄録集  21st-  121  2014/10/14  [Not refereed][Not invited]
  
• Keap1‐Nrf2経路活性化によるマガキ由来の抗酸化物質の肝臓保護作用

  布田博敏, 渡邉貢, 上甲紗愛, 惠淑萍, 武田晴治, 渡邉孝之, 千葉仁志  日本未病システム学会学術総会抄録集  21st-  141  2014/10/14  [Not refereed][Not invited]
  
• マガキ抽出液中のフェノール性抗酸化物質の定量

  岡部浩昭, 惠淑萍, 布田博敏, 大谷晋太郎, 渡辺孝之, 渡辺貢, 千葉仁志  日本未病システム学会学術総会抄録集  21st-  142  2014/10/14  [Not refereed][Not invited]
  
• ミネラルオイルへの培養液溶出現象に関する検討

  八木亜希子, 惠淑萍, 小林清一, 千葉仁志, 神谷博文  日本生殖医学会雑誌  59-  (4)  396  2014/10/01  [Not refereed][Not invited]
  
• 機能性脂肪酸に関する分析

  惠 淑萍, Shrestha Rojeet, 平野 賢一, 鈴木 朗, 千葉 仁志  日本未病システム学会学術総会抄録集  21回-  121  -121  2014/10  [Not refereed][Not invited]
  
• Keap1-Nrf2経路活性化によるマガキ由来の抗酸化物質の肝臓保護作用

  布田 博敏, 渡邉 貢, 上甲 紗愛, 惠 淑萍, 武田 晴治, 渡邉 孝之, 千葉 仁志  日本未病システム学会学術総会抄録集  21回-  141  -141  2014/10  [Not refereed][Not invited]
  
• マガキ抽出液中のフェノール性抗酸化物質の定量

  岡部 浩昭, 惠 淑萍, 布田 博敏, 大谷 晋太郎, 渡辺 孝之, 渡辺 貢, 千葉 仁志  日本未病システム学会学術総会抄録集  21回-  142  -142  2014/10  [Not refereed][Not invited]
  
• ミネラルオイルへの培養液溶出現象に関する検討

  八木 亜希子, 惠 淑萍, 小林 清一, 千葉 仁志, 神谷 博文  日本生殖医学会雑誌  59-  (4)  396  -396  2014/10  [Not refereed][Not invited]
  
• 中性脂肪蓄積心筋血管症におけるJordans' anomalyを自動血球分析装置で検出する

  鈴木 朗, 和田 淳, 長坂 博範, 越智 康浩, 千葉 仁志, 惠 淑萍, 平野 賢一  臨床病理  62-  (補冊)  126  -126  2014/10  [Not refereed][Not invited]
  
• 他項目自動血球分析装置XE-5000 WBC/BASOチャンネルによる、中性脂肪蓄積心筋血管症判別原理の検証

  和田 淳, 鈴木 朗, 山本 志緒里, 越智 康浩, 惠 淑萍, 千葉 仁志, 平野 賢一  臨床病理  62-  (補冊)  127  -127  2014/10  [Not refereed][Not invited]
  
• 中鎖脂肪酸のHPLC法による定量分析

  惠 淑萍, しゅれすた・ろじーと, 平野 賢一, 鈴木 朗, 千葉 仁志  臨床病理  62-  (補冊)  299  -299  2014/10  [Not refereed][Not invited]
  
• LC/MSによる誘導体化脂肪酸の分析

  惠 淑萍, ロジート・シュレスタ, 平野 賢一, 鈴木 朗, 千葉 仁志  JSBMS Letters  39-  (Suppl.)  52  -52  2014/09  [Not refereed][Not invited]
  
• 質量分析イメージングを用いたNASHモデルマウス腎組織における脂質の可視化

  早坂 孝宏, 布田 博敏, 惠 淑萍, 千葉 仁志  JSBMS Letters  39-  (Suppl.)  107  -107  2014/09  [Not refereed][Not invited]
  
• 多重重水素標識コレステロールの合成

  三浦佑介, 惠淑萍, 池川繁男, 千葉仁志  臨床化学  43-  236  2014/08/31  [Not refereed][Not invited]
  
• LC‐MS/MSによるマガキ由来フェノール性抗酸化物質の定量

  岡部浩昭, 惠淑萍, 池川繁男, 比留間貴久, 布田博敏, 渡辺貢, 千葉仁志  臨床化学  43-  259  2014/08/31  [Not refereed][Not invited]
  
• DEVELOPMENT OF AN AUTOMATED ASSAY FOR APOE-CONTAINING HDL-CHOLESTEROL

  Y. Takahashi, T. Sakurai, M. Fujikawa, A. Nagasaka, S. Hui, S. Jin, S. Takeda, H. Fuda, Y. Ito, H. Chiba  ATHEROSCLEROSIS  235-  (2)  E102  -E102  2014/08  [Not refereed][Not invited]
  
• 質量分析技術の社会実装 地域・産業振興のための高度脂質分析ラボの立ち上げから将来構想まで

  惠 淑萍  臨床化学  43-  (Suppl.1)  119  -120  2014/08  [Not refereed][Not invited]
  
• 多重重水素標識コレステロールの合成

  三浦 佑介, 惠 淑萍, 池川 繁男, 千葉 仁志  臨床化学  43-  (Suppl.1)  236  -236  2014/08  [Not refereed][Not invited]
  
• LC-MS/MSによるマガキ由来フェノール性抗酸化物質の定量

  岡部 浩昭, 惠 淑萍, 池川 繁男, 比留間 貴久, 布田 博敏, 渡辺 貢, 千葉 仁志  臨床化学  43-  (Suppl.1)  259  -259  2014/08  [Not refereed][Not invited]
  
• 新しい脂質評価系と食品開発

  千葉仁志, 惠淑萍  日本栄養・食糧学会大会講演要旨集  68th-  107  2014/04/30  [Not refereed][Not invited]
  
• 新しい脂質評価系と食品開発

  千葉 仁志, 惠 淑萍  日本栄養・食糧学会大会講演要旨集  68回-  107  -107  2014/04  [Not refereed][Not invited]
  
• マガキ由来の抗酸化物質における培養肝細胞保護作用

  布田博敏, 渡邉貢, 上甲紗愛, 神繁樹, 川西範明, 吉田繁, HUI Shu‐Ping, 武田晴治, 櫻井俊宏, SHRESTHA Rojeet, 池川繁男, 三木恵美子, 渡邉孝之, 千葉仁志  日本農芸化学会大会講演要旨集(Web)  2014-  2B04A04 (WEB ONLY)  2014/03/05  [Not refereed][Not invited]
  
• タンデム型質量分析計による臨床サンプルの定量分析 ハイブリッドステロイド質量分析による原発性アルドステロン症の鑑別診断

  和田 典男, 神 繁樹, 惠 淑萍, 柳澤 克之, 黒澤 隆夫, 千葉 仁志  臨床病理  62-  (3)  276  -282  2014/03  [Not refereed][Not invited]
  
• タンデム型質量分析計による臨床サンプルの定量分析 LC/MSを用いた過酸化脂質および生理活性脂質の分析

  惠 淑萍  臨床病理  62-  (3)  283  -290  2014/03  [Not refereed][Not invited]
  
• Analyses of Lipid Hydroperoxides and Physiologically Active Lipids by Mass Spectrometry

  惠 淑萍  臨床病理 : 日本臨床検査医学会誌  62-  (3)  283  -290  2014/03  [Not refereed][Not invited]
  
• Differential Diagnosis of Primary Aldosteronism by Measurement of Hybrid Steroids Using Mass Spectrometry

  和田 典男, 神 繁樹, 惠 淑萍  臨床病理 : 日本臨床検査医学会誌  62-  (3)  276  -282  2014/03  [Not refereed][Not invited]
  
• マガキ由来の新規抗酸化物質における肝保護作用

  布田博敏, 渡邉貢, 神繁樹, 上甲紗愛, 惠淑萍, 武田晴治, 櫻井俊宏, 渡邉孝之, 千葉仁志  日本未病システム学会学術総会抄録集  20th-  173  2013/10/15  [Not refereed][Not invited]
  
• リポ蛋白における過酸化リン脂質と抗酸化脂質に関する研究

  惠淑萍, 武田晴治, 神繁樹, 布田博敏, 千葉仁志  日本未病システム学会学術総会抄録集  20th-  173  2013/10/15  [Not refereed][Not invited]
  
• ARTに用いる市販ミネラルオイルにおける過酸化脂質の分析

  八木亜希子, 惠淑萍, ROJEET Shrestha, 小林清一, 千葉仁志, 神谷博文  日本生殖医学会雑誌  58-  (4)  283  -283  2013/10/01  [Not refereed][Not invited]
  
• マガキ由来の新規抗酸化物質における肝保護作用

  布田 博敏, 渡邉 貢, 神 繁樹, 上甲 紗愛, 惠 淑萍, 武田 晴治, 櫻井 俊宏, 渡邉 孝之, 千葉 仁志  日本未病システム学会学術総会抄録集  20回-  173  -173  2013/10  [Not refereed][Not invited]
  
• リポ蛋白における過酸化リン脂質と抗酸化脂質に関する研究

  惠 淑萍, 武田 晴治, 神 繁樹, 布田 博敏, 千葉 仁志  日本未病システム学会学術総会抄録集  20回-  173  -173  2013/10  [Not refereed][Not invited]
  
• LC/MSを用いた過酸化脂質および生理活性脂質の分析

  惠淑萍  臨床病理  61-  17  2013/09/30  [Not refereed][Not invited]
  
• ハイブリッドステロイド質量分析による原発性アルドステロン症の鑑別診断

  和田典男, 神繁樹, 惠淑萍, 柳澤克之, 黒澤隆夫, 千葉仁志  臨床病理  61-  16  2013/09/30  [Not refereed][Not invited]
  
• 高度生殖医療に用いられるミネラルオイル中の過酸化脂質

  惠淑萍, 八木亜希子, SHRESTHA Rojeet, 小林清一, 千葉仁志, 神谷博文  質量分析総合討論会講演要旨集  61st-  217  2013/09/02  [Not refereed][Not invited]
  
• タンデム型質量分析計による臨床サンプルの定量分析 ハイブリッドステロイド質量分析による原発性アルドステロン症の鑑別診断

  和田 典男, 神 繁樹, 惠 淑萍, 柳澤 克之, 黒澤 隆夫, 千葉 仁志  臨床病理  61-  (補冊)  16  -16  2013/09  [Not refereed][Not invited]
  
• タンデム型質量分析計による臨床サンプルの定量分析 LC/MSを用いた過酸化脂質および生理活性脂質の分析

  惠 淑萍  臨床病理  61-  (補冊)  17  -17  2013/09  [Not refereed][Not invited]
  
• 血漿、VLDL、およびIDL中のコレステロールエステル過酸化水素のLC/MSによる定性分析(Qualitative analysis of cholesteryl ester hydroperoxides in plasma, VLDL and IDL by LC/MS)

  Shrestha Rojeet, 惠 淑萍, 櫻井 俊宏, 八木 亜希子, 高橋 祐司, 武田 晴治, 神 繁樹, 布田 博敏, 千葉 仁志  臨床病理  61-  (補冊)  126  -126  2013/09  [Not refereed][Not invited]
  
• A phenolic antioxidant from the Pacific oyster (Crassostrea gigas) inhibits oxidation of cultured human hapatocytes mediated by diphenyl-l-pyrenylphospline

  渡邉 貢, 布田 博敏, 神 繁樹, 櫻井 俊宏, 惠 淑萍, 武田 晴治, 渡邉 孝之, 小池 隆夫, 千葉 仁志  北海道醫學雜誌 = Acta medica Hokkaidonensia  88-  (4)  141  -142  2013/09/01  [Not refereed][Not invited]
  
• Defection of oxidized LDL using a carbon nanotube electrode

  武田 晴治, 惠 淑萍, 福田 慶介, 布田 博敏, 神 繁樹, 櫻井 俊宏, 石井 睦, 武笠 幸一, 末岡 和久, 千葉 仁志  北海道醫學雜誌 = Acta medica Hokkaidonensia  88-  (4)  151  -151  2013/09/01  [Not refereed][Not invited]
  
• Triglyceride-rich lipoproteinsの過酸化脂質の分析(Analyses of lipid hydroperoxides in triglyceride-rich lipoproteins)

  Shrestha Rojeet, 惠 淑萍, 八木 亜希子, 櫻井 俊宏, 高橋 祐司, 武田 晴治, 神 繁樹, 布田 博敏, 池川 繁男, 千葉 仁志  JSBMS Letters  38-  (Suppl.)  68  -68  2013/08  [Not refereed][Not invited]
  
• LC/MSによる高度生殖医療に用いられるミネラルオイル中のヒドロペルオキシドの検出

  八木 亜希子, 惠 淑萍, Shrestha Rojeet, 小林 清一, 千葉 仁志, 神谷 博文  JSBMS Letters  38-  (Suppl.)  69  -69  2013/08  [Not refereed][Not invited]
  
• TGCVの皮膚線維芽細胞におけるトリグリセリドのLC/MSによる分析

  惠 淑萍, 平野 賢一, Shrestha Rojeet, 千葉 仁志  JSBMS Letters  38-  (Suppl.)  70  -70  2013/08  [Not refereed][Not invited]
  
• 金属酸化LDLに由来する粒子の電気化学的検討

  寺嶋駿, 武田晴治, 山田理絵, 大川芙多葉, 恵淑萍, 布田博敏, 櫻井俊宏, 神繁樹, 千葉仁志  臨床化学  42-  184  2013/07/31  [Not refereed][Not invited]
  
• マガキ由来の新規抗酸化物質における肝保護作用

  上甲紗愛, 布田博敏, 渡邉貢, 神繁樹, 惠淑萍, 武田晴治, 櫻井俊宏, 渡邉孝之, 千葉仁志  臨床化学  42-  (Suppl.1)  229  -229  2013/07/31  [Not refereed][Not invited]
  
• グリセリンカプリン酸エステルの合成

  惠淑萍, 三浦佑介, 池川繁男, 千葉仁志  臨床化学  42-  (Suppl.1)  185  -185  2013/07/31  [Not refereed][Not invited]
  
• apoE‐containing HDL‐cholesterol直接法の開発

  高橋祐司, 櫻井俊宏, 惠淑萍, 神繁樹, 武田晴治, 布田博敏, 伊藤康樹, 千葉仁志  臨床化学  42-  (Suppl.1)  219  -219  2013/07/31  [Not refereed][Not invited]
  
• 原子間力顕微鏡によるLDL酸化状態評価

  武田晴治, 櫻井俊宏, 大川芙多葉, 惠淑萍, 神繁樹, 布田博敏, 千葉仁志  臨床化学  42-  (Suppl.1)  216  -216  2013/07/31  [Not refereed][Not invited]
  
• Qualitative Determination of Triacylglycerol Hydroperoxide in VLDL, Intermediate Density Lipoprotein and Human Plasma using Orbitrap Mass Spectrometer.

  Shrestha R, Hui SP, Sakurai T, Takahashi Y, Ohkawa F, Miyazaki R, Xiao N, Takeda S, Jin S, Fuda H, Chiba H  American Association for Clinical Chemistry  2013/07  [Not refereed][Not invited]
  
• Detection and characterization of lipid hydroperoxides in human lipoproteins by LC/MS.

  Hui SP, Shrestha R, Sakurai T, Takahashi Y, Xiao N, Takeda S, Jin S, Fuda H, Chiba H  American Association for Clinical Chemistry, Houston  2013/07  [Not refereed][Not invited]
  
• LC/MSによるVLDLとIDL中のトリアシルグリセロール過酸化水素の定性分析(Qualitative analysis of triacylglycerol hydroperoxide in VLDL and IDL by LC/MS)

  Shrestha Rojeet, 惠 淑萍, 櫻井 俊宏, 高橋 祐司, 武田 晴治, 神 繁樹, 布田 博敏, 千葉 仁志  臨床化学  42-  (Suppl.1)  185  -185  2013/07  [Not refereed][Not invited]
  
• 脂質酸化の新しい評価系の開発~未病の理解と予防を目指して~

  千葉仁志, 惠淑萍, 武田晴治  日本未病システム学会雑誌  19-  (1)  53  -59  2013/05/17  [Not refereed][Not invited]
  
• Analysis of Triacylglycerol Hydroperoxide in VLDL, Intermediate Density Lipoprotein and Human Plasma using LC/MS

  SHRESTHA Rojeet, HUI Shu-Ping, SAKURAI Toshihiro, TAKAHASHI Yuji, YAGI Akiko, TAKEDA Seiji, JIN Shigeki, FUDA Hirotoshi, CHIBA Hitoshi  バイオメディカル分析科学シンポジウム講演要旨集  26th-  2013
• 教育講演 脂質酸化の新しい評価系の開発 : 未病の理解と予防を目指して (第19回日本未病システム学会学術総会)

  千葉 仁志, 惠 淑萍, 武田 晴治  日本未病システム学会雑誌  19-  (1)  53  -59  2013  [Not refereed][Not invited]
  
• LC/MSによる酸化リポタンパク質の過酸化リン脂質の定量分析

  惠 淑萍, 田口 裕大, 横井 俊宏, 大川 芙多葉, 武田 晴治, 神 繁樹, 布田 博敏, 黒澤 隆夫, 千葉 仁志  北海道醫學雜誌 = Acta medica Hokkaidonensia  88-  (1)  39  -39  2013/01/01  [Not refereed][Not invited]
  
• リポタンパク質の金属酸化時のphosphatidyl choline hydroperoxides及びTBARSの経時変化

  惠淑萍, 櫻井俊宏, 櫻井俊宏, 神繁樹, 布田博敏, 武田晴治, 千葉仁志  臨床病理  60-  (補冊)  113  -113  2012/10/20  [Not refereed][Not invited]
  
• TBARSは酸化リポタンパク質中のphosphatidylcholine hydroperoxidesのレベルを反映していない

  HUI Shu‐Ping, 櫻井俊宏, 神繁樹, 布田博敏, 武田晴治, 黒澤隆夫, 千葉仁志  JSBMS Lett  37-  (Supplement)  58  -58  2012/09/10  [Not refereed][Not invited]
  
• 原子間力顕微鏡を用いた酸化LDLの電位変化測定

  武田晴治, 大川芙多葉, 櫻井俊宏, 神繁樹, 布田博敏, 惠淑萍, 末岡和久, 千葉仁志  臨床化学  41-  (Suppl.1)  226  -226  2012/07/31  [Not refereed][Not invited]
  
• 直接法によるHDL亜分画中のコレステロール同時測定法の開発

  高橋祐司, 櫻井俊宏, 櫻井俊宏, 永坂敦, 藤川正人, 惠淑萍, 神繁樹, 武田晴治, 布田博敏, 伊藤康樹, 千葉仁志  臨床化学  41-  (Suppl.1)  227  -227  2012/07/31  [Not refereed][Not invited]
  
• 酸化リポタンパク質における過酸化脂質とプラズマローゲンの濃度変化

  惠淑萍, 櫻井俊宏, 武田晴治, 神繁樹, 布田博敏, 黒澤隆夫, 千葉仁志  臨床化学  41-  (Suppl.1)  239  -239  2012/07/31  [Not refereed][Not invited]
  
• 過酸化脂質測定CNT電極センサーを用いた抗酸化能測定法

  大川芙多葉, 武田晴治, 櫻井俊宏, 櫻井俊宏, 惠淑萍, 布田博敏, 神繁樹, 千葉仁志  臨床化学  41-  (Suppl.1)  200  -200  2012/07/31  [Not refereed][Not invited]
  
• 脂質異常症患者および健常者における粒子サイズ別の酸化LDLの解析

  櫻井俊宏, 櫻井俊宏, 高橋祐司, 高橋祐司, 和田典男, 古川博之, 永坂敦, 惠淑萍, 神繁樹, 武田晴治, 布田博敏, 小林清一, 千葉仁志  臨床化学  41-  (Suppl.1)  226  -226  2012/07/31  [Not refereed][Not invited]
  
• カーボンナノチューブセンサーの臨床検査化学への応用

  武田晴治, 惠淑萍, 櫻井俊宏, 石井睦, 武笠幸一, 神繁樹, 布田博敏, 末岡和久, 千葉仁志  臨床病理  60-  (7)  19  -630  2012/07/25  [Not refereed][Not invited]
  
• 技術講座 過酸化脂質測定法

  惠 淑萍, 千葉 仁志  細胞  44-  (8)  370  -373  2012/07  [Not refereed][Not invited]
   

  過酸化脂質(脂質ヒドロペルオキシド)の測定法には、チオバルビタール酸(TBA)法、ヘモグロビン-メチレンブルー法、FOX2法、共役ジエン法、カーボンナノチューブセンサー法、高速液体クロマトグラフィー化学発光法、高速液体クロマトグラフィー質量分析法(LC-MS法)などがある。このうちTBA法は脂質ヒドロペルオキシドよりもアルデヒド類の貢献が大きい。LC-MS法は分子種が同定でき、感度も優れており、SRMスキャンモードの使用により試料中の夾雑ピークの影響を避けることが可能であることから、今後の発展が期待される。(著者抄録)
• 新しい臨床検査技術の開発 カーボンナノチューブセンサーの臨床検査化学への応用

  武田 晴治, 惠 淑萍, 櫻井 俊宏, 石井 睦, 武笠 幸一, 神 繁樹, 布田 博敏, 末岡 和久, 千葉 仁志  臨床病理  60-  (7)  630  -630  2012/07  [Not refereed][Not invited]
  
• LC‐MS/MSによる酸化リポタンパク質における過酸化リン脂質の定量分析

  惠淑萍, 櫻井俊宏, 武田晴治, 神繁樹, 布田博敏, 千葉仁志  日本酸化ストレス学会学術集会プログラム・抄録集  65th-  72  2012/06/05  [Not refereed][Not invited]
  
• Increase of oxidized lipoproteins in liver disease, detected by monoclonal antibody recognizing early lipoprotein change during copper-oxidation.

  Sakurai T, Ichikawa A, Furukawa F, Wada N, Nagasaka A, Takahashi Y, Fujikawa M, Ikuta A, Furumaki H, Shiga M, Shimizu C, Hui SP, Jin S, Takeda S, Fuda H, Nagasaka S, Kobayashi S, Chiba H  European Atherosclerosis Society, Milan  2012/05  [Not refereed][Not invited]
  
• 酸化低比重リポ蛋白質検出におけるCNT電極の特異性

  武田晴治, 惠淑萍, 櫻井俊宏, 櫻井俊宏, 石井睦, 武笠幸一, 神繁樹, 布田博敏, 末岡和久, 千葉仁志  電気化学会大会講演要旨集  79th-  38  2012/03/29  [Not refereed][Not invited]
  
• LC/MSによる酸化リポタンパク質におけるcholesterylester hydroperoxidesの分析

  惠淑萍, 田口裕大, 櫻井俊宏, 櫻井俊宏, 神繁樹, 布田博敏, 武田晴治, 黒澤隆夫, 千葉仁志  日本薬学会年会要旨集  132nd-  (4)  135  2012/03/05  [Not refereed][Not invited]
  
• LC/MSによるヒト血中コリン型プラズマローゲンに関する分析

  山木志展, 惠淑萍, 田口裕大, 千葉仁志  臨床病理  59-  (補冊)  206  -206  2011/10/15  [Not refereed][Not invited]
  
• 自製ELISAによる酸化リポ蛋白の測定とその有用性の検討

  櫻井俊宏, 櫻井俊宏, 生田知子, 古牧宏啓, 高橋祐司, 西端友香, 石川明彦, 古川博之, 和田典男, 永坂敦, 惠淑萍, 神繁樹, 武田晴治, 布田博敏, 小林清一, 千葉仁志  臨床病理  59-  (補冊)  204  -204  2011/10/15  [Not refereed][Not invited]
  
• LC/MSによるLDL及びHDLにおける過酸化トリグリセリドの分析

  惠淑萍, 田口裕大, 櫻井俊宏, 櫻井俊宏, 山木志展, 古牧宏啓, 神繁樹, 布田博敏, 武田晴治, 千葉仁志  臨床病理  59-  (補冊)  206  -206  2011/10/15  [Not refereed][Not invited]
  
• 食品由来の新規抗酸化物の同定

  布田博敏, 渡邉貢, 神繁樹, 櫻井俊宏, 櫻井俊宏, 惠淑萍, 武田晴治, 千葉仁志  臨床病理  59-  (補冊)  123  -123  2011/10/15  [Not refereed][Not invited]
  
• 新しい臨床検査技術の開発 カーボンナノチューブセンサーの臨床検査化学への応用

  武田 晴治, 惠 淑萍, 櫻井 俊宏, 石井 睦, 武笠 幸一, 神 繁樹, 布田 博敏, 末岡 和久, 千葉 仁志  臨床病理  59-  (補冊)  19  -19  2011/10  [Not refereed][Not invited]
  
• 各種の過酸化脂質分子種及びプラズマローゲンのLC/MSによる定性定量分析

  HUI Shu‐Ping, 田口裕大, 黒澤隆夫, 千葉仁志  JSBMS Lett  36-  (Supplement)  52  -52  2011/08/10  [Not refereed][Not invited]
  
• 原発性アルドステロン症鑑別診断における血清18‐ヒドロキシコルチゾール定量の有用性の検討

  神繁樹, 和田典男, HUI Shu‐Ping, 櫻井俊宏, 櫻井俊宏, 高橋祐司, 柳澤克之, 黒澤隆夫, 千葉仁志  JSBMS Lett  36-  (Supplement)  69  -69  2011/08/10  [Not refereed][Not invited]
  
• LC‐MS/MSを用いた血清18‐ヒドロキシコルチゾール定量法の開発

  神繁樹, HUI Shu‐Ping, 和田典男, 櫻井俊宏, 櫻井俊宏, 高橋祐司, 柳澤克之, 黒澤隆夫, 千葉仁志  JSBMS Lett  36-  (Supplement)  68  -68  2011/08/10  [Not refereed][Not invited]
  
• LC/MSによるLDL及びHDLにおけるtriglyceride hydroperoxidesの分析

  HUI Shu‐Ping, 田口裕大, 櫻井俊宏, 櫻井俊宏, 山木志展, 古牧宏啓, 神繁樹, 布田博敏, 武田晴治, 黒澤隆夫, 千葉仁志  JSBMS Lett  36-  (Supplement)  67  -67  2011/08/10  [Not refereed][Not invited]
  
• LC‐MS/MSによる健常者血中phosphatidyl choline monohydroperoxidesの分析

  惠淑萍, 田口裕大, 黒澤隆夫, 千葉仁志  臨床化学  40-  203  2011/07/31  [Not refereed][Not invited]
  
• LC‐MS/MSによるヒト血中プラズマローゲンの新規測定法の開発

  惠淑萍, 山木志展, 黒澤隆夫, 千葉仁志  臨床化学  40-  201  2011/07/31  [Not refereed][Not invited]
  
• LC‐MS/MSによる血清中18‐ヒドロキシコルチゾールの測定法の開発

  神繁樹, 惠淑萍, 和田典男, 櫻井俊宏, 櫻井俊宏, 柳澤克之, 黒澤隆夫, 千葉仁志  臨床化学  40-  229  2011/07/31  [Not refereed][Not invited]
  
• 2種類のHDL‐C直接法を用いたapoE‐containing HDL定量法

  高橋祐司, 櫻井俊宏, 櫻井俊宏, 永坂敦, 藤川正人, 惠淑萍, 神繁樹, 武田晴治, 布田博敏, 千葉仁志  臨床化学  40-  209  2011/07/31  [Not refereed][Not invited]
  
• 様々な修飾カーボンナノチューブ電極を利用した過酸化物質の測定

  武田晴治, 惠淑萍, 石井睦, 武笠幸一, 末岡和久, 千葉仁志  臨床化学  40-  220  2011/07/31  [Not refereed][Not invited]
  
• 原発性アルドステロン症の鑑別診断における血清18‐hydroxycortisolの有用性の検討

  和田典男, 神繁樹, 惠淑萍, 柳澤克之, 黒澤隆夫, 千葉仁志  臨床化学  40-  230  2011/07/31  [Not refereed][Not invited]
  
• カーボンナノチューブ電極を利用した酸化LDLの測定

  熊野綾子, 武田晴治, 惠淑萍, 石井睦, 武笠幸一, 末岡和久, 千葉仁志  臨床化学  40-  221  2011/07/31  [Not refereed][Not invited]
  
• Oxidized LDL and HDLにおけるコレステリルエステル過酸化脂質のLC/MSによる分析

  惠淑萍, 櫻井俊宏, 古牧宏啓, 黒澤隆夫, 千葉仁志  臨床化学  40-  202  2011/07/31  [Not refereed][Not invited]
  
• マガキ由来の新規抗酸化物質の抗酸化能

  布田博敏, 渡邉貢, 神繁樹, 櫻井俊宏, 櫻井俊宏, 惠淑萍, 武田晴治, 千葉仁志  日本酸化ストレス学会学術集会プログラム・抄録集  64th-  97  2011/06/27  [Not refereed][Not invited]
  
• 金属酸化により得られた酸化LDLと酸化HDLにおける過酸化トリグリセリドのLC/MS定性分析

  惠淑萍, 櫻井俊宏, 櫻井俊宏, 古牧宏啓, 山木志展, 田口裕大, 神繁樹, 布田博俊, 武田晴治, 黒澤隆夫, 千葉仁志  日本酸化ストレス学会学術集会プログラム・抄録集  64th-  87  2011/06/27  [Not refereed][Not invited]
  
• 新規測定法による酸化リポ蛋白の解析

  櫻井俊宏, 櫻井俊宏, 生田知子, 古牧宏啓, 高橋祐司, 西端友香, 石川明彦, 古川博之, 和田典男, 永坂敦, 惠淑萍, 神繁樹, 武田晴治, 布田博敏, 小林清一, 千葉仁志  日本酸化ストレス学会学術集会プログラム・抄録集  64th-  72  2011/06/27  [Not refereed][Not invited]
  
• CD36欠損者の長期フルクトース摂取による代謝変化

  鈴木純子, 千葉仁志, 櫻井俊宏, 高橋祐司, 神繁樹, 惠淑萍, 小池隆夫  糖尿病  54-  (Supplement 1)  S.359  -359  2011/04/25  [Not refereed][Not invited]
  
• LC‐MS/MSによる血清18‐hydroxycortisol測定法の開発

  和田典男, 神繁樹, 柳澤克之, 惠淑萍, 黒澤隆夫, 千葉仁志  日本内分泌学会雑誌  87-  (1)  286  -286  2011/04/01  [Not refereed][Not invited]
  
• 原発性アルドステロン症の鑑別診断における血清18‐hydroxycortisol測定の意義

  和田典男, 神繁樹, 柳澤克之, 惠淑萍, 黒澤隆夫, 千葉仁志  日本内分泌学会雑誌  87-  (1)  286  -286  2011/04/01  [Not refereed][Not invited]
  
• LC/MSによるプラズマローゲンの分析

  HUI Shuping, 千葉仁志, 黒澤隆夫  JSBMS Lett  35-  (Supplement)  46  -46  2010/08/03  [Not refereed][Not invited]
  
• Molecular analyses of choline and ethanolamine plasmalogens in the blood plasma of human and rats

  Megum Nishimukai, Yuya Yamazaki, Toru Nezu, Shu-Ping Hui, Hitoshi Chiba, Ryouta Maeba, Hiroshi Hara  CHEMISTRY AND PHYSICS OF LIPIDS  163-  S42  -S42  2010/08  [Not refereed][Not invited]
  
• リポ蛋白中プラズマローゲンの分布に関する研究

  生田知子, 西向めぐみ, 惠淑萍, 櫻井俊宏, 神繁樹, 原博, 千葉仁志  臨床化学  39-  (Suppl.1)  117  -117  2010/07/31  [Not refereed][Not invited]
  
• apoCIIIとapoEの両者を持つリポ蛋白粒子(Lp‐CIII‐E)の分布の検討

  古牧宏啓, 櫻井俊宏, 高橋祐司, 生田知子, 西端友香, 惠淑萍, 神繁樹, 布田博敏, 武田晴治, 千葉仁志  臨床化学  39-  (Suppl.1)  123  -123  2010/07/31  [Not refereed][Not invited]
  
• HDL‐C直接法における界面活性剤種による反応性変化について

  高橋祐司, 高橋祐司, 古牧宏啓, 櫻井俊宏, 永坂敦, 藤川正人, 惠淑萍, 神繁樹, 武田晴治, 布田博敏, 千葉仁志  臨床化学  39-  (Suppl.1)  121  -121  2010/07/31  [Not refereed][Not invited]
  
• カーボンナノチューブ電極を利用した過酸化脂質の測定

  武田晴治, 福田佳祐, 惠淑萍, 中村基訓, 石井睦, 武笠幸一, 末岡和久, 千葉仁志  臨床化学  39-  (Suppl.1)  135  -135  2010/07/31  [Not refereed][Not invited]
  
• LC‐MS/MSによる健常人と肝不全患者血中過酸化リン脂質の分析

  惠淑萍, 千葉仁志, 櫻井俊宏, 神繁樹, 黒澤隆夫  臨床化学  39-  (Suppl.1)  119  -119  2010/07/31  [Not refereed][Not invited]
  
• プラズマローゲンの基質となる物質の化学合成

  山木志展, 惠淑萍, 黒澤隆夫, 神繁樹, 千葉仁志  臨床化学  39-  (Suppl.1)  117  -117  2010/07/31  [Not refereed][Not invited]
  
• 酸化リポ蛋白の新規測定法の開発

  櫻井俊宏, 生田知子, 古牧宏啓, 高橋祐司, 西端友香, 古川博之, 和田典男, 永坂敦, 惠淑萍, 神繁樹, 武田晴治, 布田博敏, 小林清一, 千葉仁志  臨床化学  39-  (Suppl.1)  121  -121  2010/07/31  [Not refereed][Not invited]
  
• 抗apoE抗体と抗apoCIII抗体のsandwich‐ELISAによるレムナントリポ蛋白の検出について

  古牧宏啓, 櫻井俊宏, 高橋祐司, 生田知子, 西端友香, 惠淑萍, 神繁樹, 布田博敏, 千葉仁志  臨床病理  58-  (補冊)  96  -96  2010/07/30  [Not refereed][Not invited]
  
• 酸化リポ蛋白自己抗体の研究:臨床的検討

  櫻井俊宏, 西端友香, 高橋祐司, 古川博之, 和田典夫, 永坂敦, 惠淑萍, 生田知子, 古牧宏啓, 神繁樹, 武田晴治, 布田博敏, 小林清一, 千葉仁志  臨床病理  58-  (補冊)  94  -94  2010/07/30  [Not refereed][Not invited]
  
• 酸化リポ蛋白自己抗体の研究:モノクローナル抗体の作製

  西端友香, 櫻井俊宏, 高橋祐司, 古川博之, 和田典男, 永坂敦, 惠淑萍, 生田知子, 古牧宏啓, 神繁樹, 武田晴治, 布田博敏, 小林清一, 千葉仁志  臨床病理  58-  (補冊)  173  -173  2010/07/30  [Not refereed][Not invited]
  
• LC‐MS/MSによるヒト血中過酸化リン脂質の分析

  惠淑萍, 千葉仁志, 櫻井俊宏, 神繁樹, 黒澤隆夫  臨床病理  58-  (補冊)  162  -162  2010/07/30  [Not refereed][Not invited]
  
• LC‐MS/MSによるプラズマローゲンのリポ蛋白分布の検討

  生田知子, 西向めぐみ, 惠淑萍, 櫻井俊宏, 神繁樹, 原博, 千葉仁志  臨床病理  58-  (補冊)  92  -92  2010/07/30  [Not refereed][Not invited]
  
• 疾患と酸化ストレス:過酸化脂質分析

  惠淑萍, 千葉仁志, 黒澤隆夫  バイオメディカル分析科学シンポジウム講演要旨集  23rd-  56  -57  2010/07/21  [Not refereed][Not invited]
  
• 過酸化脂質測定法の進歩

  惠 淑萍  臨床化学  39-  (3)  268  -275  2010/07  [Not refereed][Not invited]
   

  不安定で多様な構造を持つ過酸化脂質測定では、標品と内部標準物質(IS)の有機合成のステップを踏むことが唯一の確かな道である。我々はコレステリルエステル、トリグリセリド、リン脂質、遊離脂肪酸の各脂質種について、光増感反応により過酸化脂質(脂質ヒドロペルオキシド)の標品を化学合成した。コレステリルエステルヒドロペルオキシド(CE-OOH)およびトリグリセリドヒドロペルオキシド(TG-OOH)の同時分析のためのISとして、リトコール酸ミリスチルアルコールエステルをオレイン酸エステルへと導き、光増感反応によりヒドロキシペルオキシドとした。これらを用いて、CE-OOHとTG-OOHの同時測定化学発光HPLC法を確立した。肝疾患患者ではCE-OOHとTG-OOHが著しく増加していた。過酸化リン脂質(PC-OOH)についても、標品と合成ISを用いる化学発光HPLC測定法を開発し、健常者の血漿濃度を決定した。また、PC-OOHの液体クロマトグラフ質量分析法(内部標準法)による定量も開発した。本論文では、我々の研究成果を中心に過酸化脂質分析法について概説するとともに、過酸化脂質測定の臨床的意義に関する研究を紹介する。(著者抄録)
• LC‐MS/MSによる健常者血中過酸化リン脂質の分析

  惠淑萍, 千葉仁志, 櫻井俊宏, 黒澤隆夫  日本酸化ストレス学会学術集会プログラム・抄録集  63rd-  83  2010/06/24  [Not refereed][Not invited]
  
• 酸化リポ蛋白に対する自己抗体の新規測定法の開発

  櫻井俊宏, 西端友香, 高橋祐司, 古川博之, 和田典男, 永坂敦, 惠淑萍, 生田知子, 古牧宏啓, 神繁樹, 武田晴治, 布田博敏, 小林清一, 千葉仁志  日本酸化ストレス学会学術集会プログラム・抄録集  63rd-  89  2010/06/24  [Not refereed][Not invited]
  
• Measurement of the Transport Activities of Bile Salt Export Pump Using Chemiluminescence Detection Method

  Kana Yamaguchi, Tsuyoshi Murai, Hikaru Yabuuchi, Shu-Ping Hui, Takao Kurosawa  YAKUGAKU ZASSHI-JOURNAL OF THE PHARMACEUTICAL SOCIETY OF JAPAN  130-  (5)  755  -761  2010/05  [Not refereed][Not invited]
   

  Monovalent bile acids, such as taurine- and glycine-conjugated bile acids, are excreted into bile by bile salt export pumps (BSEP, ABCB11). Human BSEP (hBSEP) is physiologically important because it was identified as the gene responsible for the genetic disease: progressive familial intrahepatic cholestasis type 2 (PFIC-2). The evaluation of the inhibitory effect of hBSEP transport activity provides significant information for predicting toxic potential in the early phase of drug development. The role and function of hBSEP have been investigated by the examination of the ATP-dependent transport of radioactive isotopically (RI) -labeled bile acid such as a tritium labeled taurocholic acid, in membrane vesicles obtained from hBSEP-expressing cells. The chemiluminescence detection method using 3 alpha-hydroxysteroid dehydrogenase (3 alpha-HSD) had been developed for a simple analysis of bile acids in human biological fluids. This method is extremely sensitive and it may be applicable for the measurements of bile acid transport activities by hBSEP vesicles without using RI-labeled bile acid. The present paper deals with an application of the chemiluminescence detection method using 3 alpha-HSD with enzyme cycling method to the measurement of ATP-dependent transport activities of taurocholic acid (T-CA) in membrane vesicles obtained from hBSEP-expressing Sf9 cells. Calibration curves for T-CA was linear over the range from 10 to 400 pmol/ml. The values of the kinetic parameters for hBSEP vesicles obtained by the chemiluminescence detection method were comparable with the values of that obtained by liquid chromatography-mass spectrometry method. This assay method was highly useful for the measurements of bile acid transport activities.
• ELISAとLC‐MS/MSによる原発性アルドステロン症における尿中18‐ヒドロキシコルチゾールの測定

  和田典男, 神繁樹, 千葉仁志, 惠淑萍, 黒澤隆夫, 柳澤克之  日本内分泌学会雑誌  86-  (1)  162  -162  2010/03/20  [Not refereed][Not invited]
  
• 【広範囲 血液・尿化学検査免疫学的検査[第7版] その数値をどう読むか】 生化学的検査 脂質関係 過酸化脂質

  惠 淑萍  日本臨床  68-  (増刊1 広範囲血液・尿化学検査 免疫学的検査(2))  120  -124  2010/01  [Not refereed][Not invited]
  
• 過酸化ホスファチジルコリンのLC‐MS分析

  HUI Shuping, 千葉仁志, 神繁樹, 黒澤隆夫  JSBMS Lett  34-  (Supplement)  83  -83  2009/08/15  [Not refereed][Not invited]
  
• LC-MS/MSを用いた尿中の18-ヒドロキシコルチゾールの定量

  神 繁樹, 和田 典男, 岩渕 亜美, 佐藤 祐輔, 櫻井 俊宏, 高橋 祐司, 柳澤 克之, 惠 淑萍, 黒澤 隆夫, 千葉 仁志  JSBMS Letters  34-  (Suppl.)  56  -56  2009/08  [Not refereed][Not invited]
  
• 欲速則不達:急がば回れの過酸化脂質研究の道

  惠淑萍  臨床化学  38-  (0)  46  -46  2009/07/31  [Not refereed][Not invited]
  
• 18‐ヒドロキシコルチゾール定量分析におけるLC‐MSの有用性の検討

  神繁樹, 岩渕亜美, 櫻井俊宏, 佐藤祐輔, 高橋祐司, 和田典男, 柳澤克之, 惠淑萍, 黒澤隆夫, 千葉仁志  臨床化学  38-  (Suppl.1)  115  -115  2009/07/31  [Not refereed][Not invited]
  
• apoE‐richHDLの分離とHDL測定試薬の反応性

  高橋祐司, 高橋祐司, 古牧宏啓, 櫻井俊宏, 神繁樹, 惠淑萍, 伊敏, 千葉仁志  臨床化学  38-  (Suppl.1)  147  -147  2009/07/31  [Not refereed][Not invited]
  
• プラズマローゲン過酸化物の合成と分析(2)

  惠淑萍, 千葉仁志, 神繁樹, 黒澤隆夫  臨床化学  38-  (Suppl.1)  147  -147  2009/07/31  [Not refereed][Not invited]
  
• 原発性アルドステロン症の鑑別診断における種々の採尿条件での尿中18‐hydroxycortisol測定の評価

  和田典男, 神繁樹, 岩渕亜美, 佐藤祐輔, 櫻井俊宏, 高橋祐司, 柳澤克之, 惠淑萍, 黒澤隆夫, 千葉仁志  臨床化学  38-  (Suppl.1)  114  -114  2009/07/31  [Not refereed][Not invited]
  
• 生理活性リン脂質プラズマローゲンの化学合成

  生田知子, 惠淑萍, 黒澤隆夫, 神繁樹, 千葉仁志  臨床化学  38-  (Suppl.1)  148  -148  2009/07/31  [Not refereed][Not invited]
  
• apoE‐rich HDLと各種沈殿試薬の反応性について

  古牧宏啓, 高橋祐司, 櫻井俊宏, 伊敏, 惠淑萍, 神繁樹, 千葉仁志  臨床化学  38-  (Suppl.1)  146  -146  2009/07/31  [Not refereed][Not invited]
  
• 動脈硬化惹起性リポ蛋白測定系の若年健常者における検討

  鈴木純子, 櫻井俊宏, 高橋祐司, 惠淑萍, 神繁樹, 伊敏, 千葉仁志  臨床化学  38-  (Suppl.1)  151  -151  2009/07/31  [Not refereed][Not invited]
  
• 酸化LDLによるCD36の発現変化

  伊敏, 櫻井俊宏, 松岡志保, 須藤祐, 古牧宏啓, 神繁樹, 恵淑萍, 千葉仁志  臨床病理  57-  175  2009/07/28  [Not refereed][Not invited]
  
• 尿中18‐hydroxycortisolを測定した副腎癌の1例

  和田典男, 神繁樹, 岩渕亜美, 佐藤祐輔, 櫻井俊宏, 高橋祐司, 柳澤克之, 惠淑萍, 黒沢隆夫, 千葉仁志  臨床病理  57-  (補冊)  273  -273  2009/07/28  [Not refereed][Not invited]
  
• 18‐ヒドロキシコルチゾールのLC‐MS/MS定量法の検討

  神繁樹, 岩渕亜美, 櫻井俊宏, 佐藤祐輔, 高橋祐司, 高橋祐司, 和田典男, 柳澤克之, 惠淑萍, 黒澤隆夫, 千葉仁志  臨床病理  57-  (補冊)  206  -206  2009/07/28  [Not refereed][Not invited]
  
• CD36欠損者の長期フルクトース摂取による代謝変化―脂肪負荷試験による検討―

  高橋祐司, 鈴木純子, 櫻井俊宏, 神繁樹, 惠淑萍, 伊敏, 千葉仁志  臨床病理  57-  (補冊)  176  -176  2009/07/28  [Not refereed][Not invited]
  
• CD36欠損者の長期フルクトース摂取による代謝変化―糖負荷試験による検討―

  櫻井俊宏, 鈴木純子, 高橋祐司, 神繁樹, 惠淑萍, 伊敏, 千葉仁志  臨床病理  57-  (補冊)  175  -175  2009/07/28  [Not refereed][Not invited]
  
• CD36欠損者の長期フルクトース摂取が,H‐MRS測定法による骨格筋細胞内脂質量に及ぼす影響

  鈴木純子, 山本徹, 辻真太郎, 櫻井俊宏, 高橋祐司, 神繁樹, 惠淑萍, 伊敏, 千葉仁志  臨床病理  57-  (補冊)  176  -176  2009/07/28  [Not refereed][Not invited]
  
• 過酸化遊離脂肪酸に関する研究(3)

  惠淑萍, 千葉仁志, 神繁樹, 黒澤隆夫  臨床病理  57-  (補冊)  177  -177  2009/07/28  [Not refereed][Not invited]
  
• 臨床化学の若い力で発信しつつある新たな検査、病態解析法 欲速則不達 急がば回れの過酸化脂質研究の道

  惠 淑萍  臨床化学  38-  (Suppl.1)  46  -46  2009/07  [Not refereed][Not invited]
  
• プラズマローゲン過酸化物の化学合成と分析(1)

  仲野真悟, 惠淑萍, 千葉仁志, 増山秀幸, 水野文音, 黒澤隆祐, 黒澤隆夫  日本薬学会年会要旨集  129th-  (4)  96  2009/03/05  [Not refereed][Not invited]
  
• リン脂質過酸化物のLC‐MS分析

  惠淑萍, 千葉仁志, 神繁樹, 仲野真悟, 増田秀幸, 水野文音, 黒澤龍祐, 黒澤隆夫  日本薬学会年会要旨集  129th-  (4)  99  2009/03/05  [Not refereed][Not invited]
  
• Quantitative analysis of curcuminoids in human plasma by means of HPLC with UV detection

  HUI Shuping, HAYASHI Chitose, SAKURAI Toshihiro, JIN Shigeki, ISHII Satomi, MURAI Tsuyoshi, UEBABA Kazuo, OGAWA Hiroko, CHIBA Hitoshi, KUROSAWA Takao  臨床化学  38-  (1)  59  -68  2009/01/31  [Not refereed][Not invited]
   

  血中クルクミノイドのUV検出高速液体クロマトグラフィー(HPLC)による分離定量法を確立し、ヒトでのクルクミノイドの血中濃度推移と血中の抱合様式を検討した。ターメリック粉3g(クルクミノイド60〜90mg含有)を服用した健常成人7例と市販ウコン錠(90%精製クルクミノイド480mg含有)を服用した健常成人7例を対象とした。ターメリック粉群では服用1時間後に最大血中濃度が142.9±63.7nmol/lに達した。ウコン錠群では服用8時間に最大血中濃度が26.9±17.7nmol/lとなり、クルクミノイドの精製度が消化管吸収速度に影響する可能性が示唆された。非還元型クルクミノイドは大部分がグルグロン酸抱合体かグルクロン酸と硫酸の二重抱合型として存在し、一部が遊離型として存在することが推測された。HPLC法は、ウコン食品の薬理学的研究に有用なツールとなることが示唆された。
• 111 Effects of thickness of the cerebrospinal fluid and skull layers on optical mapping

  WANG Shuping, SHIBAHARA Nanae, KURAMASHI Daishi, OKAWA Shinpei, KAKUTA Naoto, OKADA Eiji, MAKI Atsushi, YAMADA Yukio  The Proceedings of the Bioengineering Conference Annual Meeting of BED/JSME  2008.21-  21  -22  2009  [Not refereed]
  
• 過酸化遊離脂肪酸に関する研究(2)

  惠 淑萍, 千葉 仁志, 仲野 真悟, 生田 知子, 増田 秀幸, 水野 文音, 村井 毅, 黒澤 隆夫  臨床化学  37-  (Suppl.1)  103  -103  2008/08  [Not refereed][Not invited]
  
• LC‐ESI‐MSを用いたBSEP輸送活性の測定

  山口佳奈, 村井毅, 惠淑萍, 薮内光, 黒澤隆夫  日本薬学会年会要旨集  128th-  (4)  33  2008/03/05  [Not refereed][Not invited]
  
• B217 Effects of the anatomy of the head on the optical mapping of brain activities

  OOKI Mariko, SHIBAHARA Nanae, TOGASHI Ryo, WANG Shuping, OOKAWA Shinpei, HOSHI Youko, YAMADA Yukio  The Proceedings of the JSME Conference on Frontiers in Bioengineering  2008.19-  71  -72  2008  [Not refereed]
  
• 健常者の血中過酸化リン脂質濃度と他の脂質項目との関係

  惠淑萍, 千葉仁志, 桜井俊宏, 浅川千登勢, 村井毅, 黒澤隆夫  臨床病理  55-  (Suppl.2)  142  -142  2007/10/31  [Not refereed][Not invited]
  
• ターメリック摂取後のクルクミノイドの血中動態と存在様式

  浅川千登勢, 浅川千登勢, 惠淑萍, 山田小百合, 櫻井俊宏, 小川弘子, 許鳳浩, 上馬場和夫, 黒澤隆夫, 千葉仁志  臨床病理  55-  (Suppl.2)  209  -209  2007/10/31  [Not refereed][Not invited]
  
• 酸化ストレスに対する単球CD36のtranslocationの検討

  竹内淳, 三好秀明, 櫻井俊宏, 惠淑萍, 浅川千登勢, 清水力, 小池隆夫, 千葉仁志  臨床病理  55-  (Suppl.2)  141  -141  2007/10/31  [Not refereed][Not invited]
  
• クルクミン製剤の剤型によるバイオアベイラビリティと抗酸化能の違い

  上馬塲和夫, 小川弘子, 許鳳浩, 浅川千登勢, 惠淑萍, 千葉仁志  応用薬理  72-  (5)  177  -178  2007/08/15  [Not refereed][Not invited]
  
• ウコンの多機能性とその将来性展望 クルクミン製剤の剤型によるバイオアベイラビリティと抗酸化能の違い

  上馬塲 和夫, 小川 弘子, 許 鳳浩, 浅川 千登勢, 惠 淑萍, 千葉 仁志  応用薬理  72-  (5-6)  177  -178  2007/08  [Not refereed][Not invited]
   

  健常ヒトを対象とした無作為クロスオーバー試験によって、クルクミン濃縮製剤とターメリックの乾燥粉末とで、抗酸化能とクルクミンのバイオアベイラビリティの違いがあるか検討した。ヒト血漿やサンプルの抗酸化能測定は、PAO値を用い、製剤摂取後の血漿PAO値とクルクミンの動態の変化を、2種類の製剤間で比較した。試験管実験では、ターメリックの粉に比べ濃縮クルクミンの方が高い抗酸化能を示した。経口投与では、ターメリック投与群は投与後2〜8時間にわいて対照群より高い血中抗酸化能の傾向を示した。ターメリックの粉の方が、クルクミン濃縮製剤より、バイオアベイラビリティが高く、そのため抗酸化能も高くなることが示された。ターメリック中のクルクミン以外の成分が、腸管内でのクルクミンの吸収や代謝に影響を及ぼすことが示唆された。
• 透析患者の血中過酸化脂質の同定と定量

  竹内淳, 惠淑萍, 千葉栄市, 菅原剛太郎, 千葉仁志  日本透析医学会雑誌  40-  (Supplement 1)  375  -375  2007/05/15  [Not refereed][Not invited]
  
• 胎児性胆汁酸のBSEP輸送活性の測定

  山口佳奈, 村井毅, 惠淑萍, 竹村理明, 薮内光, 黒澤隆夫  日本薬学会年会要旨集  127th-  (3)  33  2007/03/05  [Not refereed][Not invited]
  
• 内部標準法による血中クルクミノイドのHPLC測定

  千葉仁志, 惠淑萍, 山田小百合, 黒澤隆夫  日本未病システム学会学術総会抄録集  13th-  48  2006/11/15  [Not refereed][Not invited]
  
• 血液透析における血漿過酸化脂質と単球CD36の変動

  惠淑萍, 竹内淳, 清水力, 村井毅, 黒澤隆夫, 千葉仁志, 千葉仁志  臨床化学  35-  55  2006/09/08  [Not refereed][Not invited]
  
• 血液透析による血漿過酸化脂質と単球CD36の変動

  惠 淑萍, 竹内 淳, 清水 力, 村井 毅, 黒澤 隆夫, 千葉 仁志  臨床化学  35-  (Suppl.2)  55  -55  2006/09  [Not refereed][Not invited]
  
• 脂肪酸CoAエステルの合成とβ‐酸化系酵素群の特異性の解析 (3)―3‐hydroxyacyl‐CoAの分析法の検討―

  土田史郎, 村井毅, 恵淑へい, 青木隆, 渡部博之, 黒沢隆夫  日本薬学会年会要旨集  126th-  (2)  48  2006/03/06  [Not refereed][Not invited]
  
• ジヒドロキシ胆汁酸生合成におけるβ‐酸化系酵素の基質特異性

  尾迫明日香, 土田史郎, 惠 淑萍, 村井毅, 黒沢隆夫  日本薬学会年会要旨集  126th-  (2)  49  -49  2006/03/06  [Not refereed][Not invited]
  
• トリアシルグリセロール過酸化物の1,3‐Specific lipaseを用いたGC/MSによる分析法の検討

  鈴木亜維, 恵淑へい, 村井毅, 黒沢隆夫  日本薬学会年会要旨集  126th-  (2)  57  2006/03/06  [Not refereed][Not invited]
  
• 胆汁酸による脂質抗酸化作用における代謝成績体の分析

  村井毅, 桜井岳史, 山口佳奈, HUI Shu-Ping, 黒澤隆夫  バイオメディカル分析科学シンポジウム講演要旨集  19th-  2006
• 高分解能GC/MS法によるモノヒドロキシ胆汁酸生合成中間体の分析

  山口佳奈, 村井毅, 桜井岳史, HUI Shu-Ping, 黒澤隆夫  バイオメディカル分析科学シンポジウム講演要旨集  19th-  2006
• エリートランナーにおける夏季のマラソン完走後の腎機能検査項目

  石井好二郎, 恵淑へい, 千葉仁志  臨床病理  53-  (Suppl.2)  169  -169  2005/10/31  [Not refereed][Not invited]
  
• エリートランナーの夏季マラソン完走後の血しょう過酸化脂質

  恵淑へい, 石井好二郎, 村井毅, 黒沢隆夫, 千葉仁志  臨床病理  53-  307  2005/10/31  [Not refereed][Not invited]
  
• エリートランナーの夏季マラソン完走後の血漿過酸化脂質

  惠 淑萍, 石井 好二郎, 村井 毅, 黒澤 隆夫, 千葉 仁志  臨床化学  34-  (Suppl.2)  307  -307  2005/10  [Not refereed][Not invited]
  
• 胆汁酸による脂質抗酸化作用について

  原健太郎, 村井毅, 恵淑へい, 黒沢隆夫  日本薬学会年会要旨集  125th-  (2)  61  -61  2005/03/05  [Not refereed][Not invited]
  
• 高分解能GC/MS法による胆汁アルコール側鎖水酸化活性の測定(2)

  前田純佳, 村井毅, 恵淑へい, 黒沢隆夫  日本薬学会年会要旨集  125th-  (2)  61  2005/03/05  [Not refereed][Not invited]
  
• モノヒドロキシ高級胆汁酸のβ‐酸化による側鎖切断反応の解析

  矢沢諭, 土田史郎, 尾迫明日香, 村井毅, 恵淑へい, 黒沢隆夫  日本薬学会年会要旨集  125th-  (2)  58  2005/03/05  [Not refereed][Not invited]
  
• 化学発光検出HPLCによる肝不全患者血中過酸化脂質の分析

  恵淑へい, 村井毅, 千葉仁志, 黒沢隆夫  臨床病理  52-  (Suppl.3)  221  -221  2004/07/31  [Not refereed][Not invited]
  
• 高分解能GC/MSを用いた胆汁アルコール側鎖水酸化反応の立体化学的研究

  前田純佳, 村井毅, 恵淑へい, 黒沢隆夫  分析化学討論会講演要旨集  65th-  287  2004/05/01  [Not refereed][Not invited]
  
• 過酸化脂質の合成と分析法に関する研究

  恵淑へい, 村井毅, 黒沢隆夫  分析化学討論会講演要旨集  65th-  318  2004/05/01  [Not refereed][Not invited]
  
• Simple chemical syntheses of MAG monohydroperoxides (vol 38, pg 1287, 2003)

  SP Hui, T Murai, T Yoshimura, H Chiba, T Kurosawa  LIPIDS  39-  (4)  403  -403  2004/04  [Not refereed][Not invited]
  
• モノヒドロキシ胆汁酸の生合成におけるβ‐酸化の検討

  矢沢諭, 土田史郎, 村井毅, 恵淑へい, 黒沢隆夫  日本薬学会年会要旨集  124th-  (3)  39  -39  2004/03/05  [Not refereed][Not invited]
  
• クルクミン誘導体のHPLCによる分析

  大倉亜矢子, 恵淑へい, 村井毅, 黒沢隆夫  日本薬学会年会要旨集  124th-  (3)  39  -39  2004/03/05  [Not refereed][Not invited]
  
• β‐酸化酵素SCP‐2/thiolaseの特異性の解析

  土田史郎, 村井毅, 矢沢諭, 恵淑へい, 黒沢隆夫  日本薬学会年会要旨集  124th-  (3)  39  2004/03/05  [Not refereed][Not invited]
  
• 過酸化脂質の合成と分析

  恵淑へい, 村井毅, 黒沢隆夫  日本薬学会年会要旨集  124th-  (3)  39  2004/03/05  [Not refereed][Not invited]
  
• 高分解能GC/MS法による胆汁アルコール側鎖水酸化活性の測定

  前田 純佳, 村井 毅, 惠 淑萍, 黒澤 隆夫  日本薬学会年会要旨集  124年会-  (3)  35  -35  2004/03  [Not refereed][Not invited]
  
• モノヒドロキシ高級胆汁酸のβ‐酸化による側鎖切断反応の解析

  矢沢諭, 土田史郎, 村井毅, 恵淑へい, 黒沢隆夫  胆汁酸研究会講演要旨集  26th-  6  2004  [Not refereed][Not invited]
  
• 25‐Hydroxylase pathwayにおける胆汁酸生合成中間体の高分解能GC/MS法による分析

  村井毅, 吉村昭毅, 恵淑へい, 黒沢隆夫  胆汁酸研究会講演要旨集  25th-  7  2003/11/01  [Not refereed][Not invited]
  
• 化学発光検出HPLCによる血中過酸化脂質の分析

  惠 淑萍, 千葉仁志, 村井毅, 衛藤雅昭, 斎藤美恵子, 石井好二郎, 小林邦彦, 黒沢隆夫  臨床化学  32-  (補冊)  127  -127  2003/09/25  [Not refereed][Not invited]
  
• ATB8B1及びABCB11遺伝子異常におけるリポ蛋白代謝異常

  長坂 博範, 嶋村 英貴, 千葉 仁志, 小林 邦彦, 惠 淑萍, 黒澤 隆夫  BIO Clinica  18-  (10)  920  -921  2003/09  [Not refereed][Not invited]
  
• 高分解能GC/MSによる重水素化胆汁アルコールを利用した酵素活性の測定

  村井 毅, 惠 淑萍, 黒澤 隆夫  BIO Clinica  18-  (10)  924  -925  2003/09  [Not refereed][Not invited]
  
• 25‐Hydroxylase pathwayによる5β‐cholestane類の側鎖開裂反応の解析 III 胆汁酸生合成中間体の分析

  山口佳奈, 村井毅, 吉村昭毅, 恵淑へい, 黒沢隆夫  日本薬学会年会要旨集  123rd-  (3)  44  2003/03/05  [Not refereed][Not invited]
  
• 脂肪酸コエンザイムAエステルの合成とβ‐酸化系酵素の特異性の解析 (2) β‐ヒドロキシαメチル脂肪酸の異性体の分離

  土田史郎, 村井毅, 恵淑へい, 黒沢隆夫  日本薬学会年会要旨集  123rd-  (3)  38  2003/03/05  [Not refereed][Not invited]
  
• 化学発光検出HPLCによる血中過酸化脂質の分析

  恵淑へい, 村井毅, 黒沢隆夫  日本薬学会年会要旨集  123rd-  (3)  38  -38  2003/03/05  [Not refereed][Not invited]
  
• ガスクロマトグラフィー/質量分析法による胆汁酸生合成中間体の定量法の開発

  山口佳奈, 村井毅, 吉村昭毅, 恵淑へい, 黒沢隆夫  日本分析化学会年会講演要旨集  51st-  268  -268  2002/09/05  [Not refereed][Not invited]
  
• ガスクロマトグラフィー/質量分析法を用いた胆汁酸生合成中間体の分析

  村井毅, 山口佳奈, 吉村昭毅, 恵淑へい, 黒沢隆夫  バイオメディカル分析科学シンポジウム講演要旨集  15th-  147  -148  2002/07/30  [Not refereed][Not invited]
  
• ガスクロマトグラフィー/質量分析法による胆汁酸生合成中間体の定量法の開発

  村井 毅, 山口 佳奈, 吉村 昭毅, 惠 淑萍, 黒澤 隆夫  分析化学  51-  (6)  443  -448  2002/06  [Not refereed][Not invited]
   

  モノヒドロキシ胆汁酸の生成機構と25-水銀酸による胆汁酸の生成経路の関連性を明らかにする為,モノヒドロキシ胆汁酸,生合成中間体と推測される各種胆汁アルコール及びヒドロキシコレステロールの,高分解能ガスクロマトグラフィー/質量分析法による一斉分析法を開発した.各種中間体標品をトリメチリシリルエーテル誘導体とした後,高分解能選択イオンモニタリング法によって分析した.得られた検量線は,どれも10〜250pmolの範囲で良好な直線性を示した.検出限界は50〜100fmolであった.逆相系固相抽出法を用いる前処理のみで,生体由来の妨害ピークは認められなかった.以上より本分析法は各種生体試料中の前記中間体と胆汁酸の分析法として,適応可能であることが明らかになった
• Development of an analytical method for intermediates in bile acid biosynthesis by GC/MS

  T Murai, K Yamaguchi, SP Hui, T Kurosawa  BUNSEKI KAGAKU  51-  (6)  443  -448  2002/06  [Not refereed][Not invited]
   

  A method for the analysis of bile alcohols and hydroxycholesterols as intermediates in monohydroxyl bile acid biosynthesis has been developed by gas chromatography/mass spectrometry (GC/MS) with selected ion monitoring in a high-resolution mode. The linearity of the calibration curves for the bile alcohols, hydroxycholesterol and bile acids ranged from 10 similar to 250 pmol (r > 0.999), and the detection limits were 50 similar to 100 fmol (S/N= 2 similar to 6). The recoveries of the bile alcohols, hydroxycholesterol and bile acids from the rat liver mitochondrial fraction ranged over 97.9 similar to 111%, respectively, of the added amounts of their standard samples. The present method was made available for the determination of bile alcohols, hydroxycholesterol and related monohydroxy bile acids in incubation mixtures after only a simple extraction procedure using a solid-phase extraction cartridge.
• 脂肪酸コエンザイムAエステルの合成とβ‐酸化系酵素の特異性の解析 (1) 脂肪酸CoAエステルの分析法の開発

  土田史郎, 藤原雅人, 村井毅, 恵淑へい, 黒沢隆夫  日本薬学会年会要旨集  122nd-  (3)  51  -51  2002/03/05  [Not refereed][Not invited]
  
• 25‐Hydroxylase pathwayによる5β‐cholestane類の側鎖開裂反応の解析 II モノヒドロキシ胆汁酸の生成機構について

  山口佳奈, 村井毅, 吉村昭毅, 藤原雅人, 恵淑へい, 黒沢隆夫  日本薬学会年会要旨集  122nd-  (3)  38  2002/03/05  [Not refereed][Not invited]
  
• 25-Hydroxylase pathwayによる5β-cholestane類の側鎖開裂反応の解析 モノヒドロキシ胆汁酸の生成機構について

  山口 佳奈, 村井 毅, 吉村 昭毅, 藤原 雅人, 惠 淑萍, 黒澤 隆夫  日本薬学会年会要旨集  122年会-  (3)  38  -38  2002/03  [Not refereed][Not invited]
  
• 化学発光検出HPLCによる血中過酸化脂質の分析

  惠 淑萍, 村井 毅, 黒澤 隆夫  日本薬学会年会要旨集  122年会-  (3)  51  -51  2002/03  [Not refereed][Not invited]
  
• 脂肪酸コエンザイムAエステルの合成とβ-酸化系酵素の特異性の解析 脂肪酸CoAエステルの分析法の開発

  土田 史郎, 藤原 雅人, 村井 毅, 惠 淑萍, 黒澤 隆夫  日本薬学会年会要旨集  122年会-  (3)  51  -51  2002/03  [Not refereed][Not invited]
  
• 胆汁酸生合成関連酵素Bifunctional Proteinの特異性について

  黒沢隆夫, 村井毅, 恵淑へい, 吉村昭毅, 藤原雅人  Bio Clin  16-  (10)  950  -950  2001/09/10  [Not refereed][Not invited]
  
• コレステロールエステル水酸化体の蛍光ラベル化による分析

  恵淑へい, 吉村昭毅, 村井毅, 黒沢隆夫  日本薬学会年会要旨集  121st-  (4)  122  -122  2001/03/05  [Not refereed][Not invited]
  
• 24‐Oxo‐5β‐cholestanoic acid CoAのSCPxによる開裂反応

  黒沢隆夫, 吉村昭毅, 恵淑へい, 村井毅  日本薬学会年会要旨集  121st-  (4)  122  2001/03/05  [Not refereed][Not invited]
  
• コレステロールエステル過酸化物の分析

  惠 淑萍, 黒澤隆夫, 吉村昭毅  北海道支部2001年冬季研究発表会  2001/02  [Not refereed][Not invited]
  
• コレステロールエステル過酸化物の分析

  恵淑へい, 吉村昭毅, 黒沢隆夫  日本化学会北海道支部研究発表会講演要旨集  2001-  110  2001/01/19  [Not refereed][Not invited]
  
• Sterol Carrier Protein xによる24‐oxo‐5β‐cholestanoic acid CoAの開裂反応

  黒沢隆夫, 吉村昭毅, 恵淑へい, 村井毅, 中野浩行  胆汁酸研究会講演要旨集  22nd-  19  2000/11/25  [Not refereed][Not invited]
  
• 脂肪酸コレステロールエステル過酸化物異性体の分析研究

  恵淑へい, 吉村昭毅, 黒沢隆夫  日本薬学会年会要旨集  120th-  (4)  176  -176  2000/03/05  [Not refereed][Not invited]
  
• Heterogenous reactivities in homogenous LDL-cholesterol assays of serum lipoproteins from patients with complete cholesteryl ester transfer protein deficiency.

  AKIZAWA KOJI, SHIMAMURA HIDEKI, AKITA HARUKUNI, MEGUMI YOSHIHEI, TAKAHASHI YUKIHIRO, YANAUCHI HIDEKATSU, FUJIWARA HIROMI, ISHIZUKA SHOJI, CHIBA HITOSHI  月刊医学と薬学  43-  (1)  137  -142  2000/01/25  [Not refereed][Not invited]
   

  CETP完全欠損患者の血清HDLは脂質組成の変化を伴うため,一部のLDLコレステロール自動測定試薬ではHDLコレステロールを部分的にLDLコレステロールとして回収する結果,正誤差を生じることが明らかとなった.脂質代謝に影響し得る様々な病態に日常的に遭遇する臨床の場では,リポ蛋白の質的変化に対して一定の安定性を有する試薬の使用が望ましく,その意味で市販試薬にも更に改良が必要と思われた
• Determination of regioisomeric hydroperoxides of fatty acid cholesterol esters produced by photosensitized peroxidation using HPLC

  Shu-Ping Hui, Teruki Yoshimura, Tsuyoshi Murai, Hitoshi Chiba, Takao Kurosawa  Analytical Sciences  16-  (10)  1023  -1028  2000  [Not refereed][Not invited]
   

  In order to examine the process of the lipid oxidation, the regioisomers of hydroperoxides produced by the photosensitized peroxidation of cholesteryl oleate, cholesteryl linoleate and cholesteryl linolenate by photosensitized peroxidation and the separation of each isomer by HPLC were investigated. The olefinic parts in fatty acid chains of the above cholesteryl esters received hydroperoxidation predominantly by photosensitized oxidation. Cholesteryl oleate was subjected to hydroperoxidation at the 9 and 10 position of the oleic acid chain. Cholesteryl linoleate and cholesteryl linolenate were also subjected to hydroperoxidation at the 9, 10, 12 and 13 positions of the linoleic acid chain and at the 9, 10, 12, 13, 15 and 16 positions of the linolenic acid chain, respectively. Hydroperoxidation on the cholesteryl ring was also observed, however, their formation was negligibly small compared with hydroperoxidation of the fatty acid chain. The elution order of isomeric peroxides in HPLC was determined using a normal-phase column (Mightysil 60) eluted with hexane/2-propanol by the detected wavelength at 210 and 233 nm.
• Significance of membrane glycoproteins in platelet interaction with oxidized low-density lipoprotein

  Y Takahashi, H Fuda, H Yanai, H Akita, SP Hui, H Chiba, K Matsuno  SEMINARS IN THROMBOSIS AND HEMOSTASIS  24-  (3)  251  -253  1998  [Not refereed][Not invited]
   

  The significance of three platelet membrane glycoproteins, (CD62p, CD63, and CD36) was studied in platelet interaction with native and copper-oxidized plasma low-density lipoproteins (LDL). Native LDL acquired platelet-activating ability only after oxidation, Flow-cytometric studies showed that CD62P and CD63 were rapidly expressed on platelets at a low concentration of oxidized LDL, indicating that they are more sensitive markers than platelet aggregation for detection of platelet activation by oxidized LDL. CD36 was found to be contributing to the binding and activation of platelets with oxidized LDL, but not to the binding of platelets with native LDL. Although the presence of oxidized LDL in plasma is known, its effect on platelet function remains to be studied.
• 中国人におけるCETP欠損症

  惠 淑萍  臨床病理  44-  (補冊)  110  -110  1996/10  [Not refereed][Not invited]
  

Industrial Property Rights

• 特開2023-022363:細胞保護用剤  2023/02/15

  惠 淑萍, 櫻井 俊宏, 千葉 仁志, 佐藤 浩志, 盛 孝男  国立大学法人北海道大学, 日生バイオ株式会社  202303011329545757
• 特許第7201164号:Keap1-Nrf2システムによる生体防御遺伝子発現の活性化用剤  

  惠 淑萍, 千葉 仁志, 布田 博敏, 陳 震, 佐藤 浩志  国立大学法人北海道大学, 日生バイオ株式会社  202303007910252313
• 特開2022-044259:非アルコール性脂肪性肝炎の診断方法  2022/03/17

  櫻井 俊宏, 惠 淑萍, 千葉 仁志  国立大学法人北海道大学  202203001827564731
• 特開2022-017829:ミトコンドリア機能活性化剤  2022/01/26

  惠 淑萍, 櫻井 俊宏, 千葉 仁志, 佐藤 浩志  国立大学法人北海道大学, 日生バイオ株式会社  202203001556225251
• 特許第6964290号:ATP産生促進用剤  

  惠 淑萍, 馬 逸興, 陳 震, 布田 博敏, 佐藤 浩志  国立大学法人北海道大学, 日生バイオ株式会社  202103002742300521
• 特願PCT/JP2021/004008:Method for aiding detection of nonalcoholic steatohepatitis  2021年/02/04

  櫻井俊宏, 水田 正弘, 惠淑萍, 能祖一裕, 岡田裕之, 佐藤 謙亨, 伊藤康樹
• 特許第6692089号:抗酸化能測定用リポソーム  

  武田晴治, 千葉仁志, 惠淑萍  国立大学法人北海道大学  202003011814405260
• 特願2020- 16820:非アルコール性脂肪肝炎の検出を補助する方法  2020年/02/04

  櫻井俊宏, 水田 正弘, 惠淑萍, 能祖一裕, 岡田裕之, 佐藤 謙亨, 伊藤康樹  北海道大学、岡山大学、デンカ生研株式会社
• 特開2020-002082:Keap1-Nrf2システムによる生体防御遺伝子発現の活性化用剤  2020/01/09

  惠 淑萍, 千葉 仁志, 布田 博敏, 陳 震, 佐藤 浩志  国立大学法人北海道大学, 日生バイオ株式会社  202003011226334241
• 特許第6628071号:低密度リポタンパク質酸化抑制剤  

  惠 淑萍, 武田晴治, 佐藤浩志  国立大学法人北海道大学, 日生バイオ株式会社, 株式会社エル・エスコーポレーション  202003004721253693
• 特開2019-073476:ATP産生促進用剤  2019/05/16

  惠 淑萍, 馬 逸興, 陳 震, 布田 博敏, 佐藤 浩志  国立大学法人北海道大学, 日生バイオ株式会社  201903010526815163
• 特開2017-116308:抗酸化能測定用リポソーム、抗酸化能測定方法及びリポソーム  2017/06/29

  武田 晴治, 千葉 仁志, 惠 淑萍  国立大学法人北海道大学  201703018950381340
• 特開2016-204340:低密度リポタンパク質酸化抑制剤  2016/12/08

  惠 淑萍, 武田 晴治, 佐藤 浩志  国立大学法人北海道大学, 日生バイオ株式会社, 株式会社エル・エスコーポレーション  201703002913011520
• 特開2015-087199:カプリン酸定量方法  2015/05/07

  惠 淑萍, 平野 賢一  国立大学法人北海道大学, 興和株式会社  201503084209484465
• 特許第5433839号:CNTセンサーによる過酸化物を電気的に測定する方法  2013/12/20

  千葉 仁志, 武田 晴治, 石井 睦, 中村 基訓, 武笠 幸一, 惠 淑萍, 黒澤 隆夫  国立大学法人北海道大学, 学校法人東日本学園  201403022970417426
• 特開2011-136926:プラズマローゲンの分析方法  2011/07/14

  西向 めぐみ, 千葉 仁志, 原 博, 惠 淑萍, 黒澤 隆夫  国立大学法人北海道大学, 惠 淑萍, 黒澤 隆夫  201103095521466523
• WO2011-007582:CNTセンサーによる過酸化物を電気的に測定する方法  2011/01/20

  千葉 仁志, 武田 晴治, 石井 睦, 中村 基訓, 武笠 幸一, 惠 淑萍, 黒澤 隆夫  国立大学法人北海道大学, 学校法人東日本学園・北海道医療大学  201303065579784108
• WO2011-007582:CNTセンサーによる過酸化物を電気的に測定する方法  2011/01/20

  千葉 仁志, 武田 晴治, 石井 睦, 中村 基訓, 武笠 幸一, 惠 淑萍, 黒澤 隆夫  国立大学法人北海道大学, 学校法人東日本学園・北海道医療大学  201303065579784108

Awards & Honors

• 2022/05 令和3年度 北海道大学保健科学研究院長賞
  
• 2021/09 from by the Lipoprotein and Vascular Disease Division of AACC, USA 2021 Zak Award
   

  受賞者: Shu-Ping Hui
• 2020/06 令和元年度 北海道大学保健科学研究院長賞
  
• 2018/05 平成29年度 北海道大学保健科学研究院長賞
   

  受賞者: 惠 淑萍
• 2014/11 第21回日本未病システム学会優秀賞
   

  受賞者: 惠 淑萍
• 2011/08 2011年度 日本臨床化学会 学術賞
   

  受賞者: 惠 淑萍

Research Grants & Projects

• Proton isotope effect as the method to repair marginal liver graft by extracorporeal perfusion

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research

  Date (from‐to) : 2023/04 -2026/03 

  Author : 嶋村 剛, 深井 原, 惠 淑萍, 暮地本 宙己, 亀田 浩之, 西川 祐司
• 新規抗酸化物質の糖尿病性腎症の有効性に関する研究

  日本学術振興会：科学研究費助成事業

  Date (from‐to) : 2023/04 -2026/03 

  Author : 惠 淑萍
• Development and application of novel technique for evaluation of mitochondrial function

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research

  Date (from‐to) : 2023/04 -2026/03 

  Author : 千葉 仁志, 櫻井 俊宏, 惠 淑萍
• 化学物質の次世代影響-出生から青年期の脂質代謝やエピゲノムの作用機序

  日本学術振興会：科学研究費助成事業

  Date (from‐to) : 2023/04 -2026/03 

  Author : 宮下 ちひろ, 池田 敦子, 田村 菜穂美, 山口 健史, 惠 淑萍
• Elucidation of oxidized HDL and NASH pathogenesis focusing on mitochondrial dysfunction

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research

  Date (from‐to) : 2022/04 -2025/03 

  Author : 櫻井 俊宏, 惠 淑萍
• Adverse health effects of environmental chemical exposure: elucidation of mechanisms via human and animal studies

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research

  Date (from‐to) : 2021/04 -2025/03 

  Author : 池田 敦子, 小島 弘幸, 池中 良徳, Goudarzi Houman, 山口 健史, 惠 淑萍, 中村 明枝, アイツバマイ ゆふ, 増田 秀幸, POUDEL KRITIKA

   

  疫学研究では、「北海道スタディ」に参加する9-11歳の前思春期児から収集した血漿を用いて、性ステロイドホルモン14物質の一斉分析、LH、FSH、Inhibin B、INSL3の測定を行った。曝露評価として尿中フタル酸エステル類代謝物、ノニルフェノールを測定した。フタル酸エステル類代替物質DINCH、DEHTP、DEHA代謝物分析に向けた条件検討を実施し、ピークが得られることを確認した。 曝露と健康を介在するバイオマーカーの測定に向けて、96穴プレートを用いて一度に数十種類のたんぱく質やペプチドを、20μLという非常に少量の検体を用いて分析できるBio-Plex200 multiplexアッセイシステムを購入した。機器メーカによるトレーニングに加えて、ヒトテストサンプルを用いて、Th1/Th2サイトカインにエオタキシンを加えたテストアッセイを実施し、手技や分析に問題がないことを確認し、子どもから収集した血漿を用いた測定に向けて準備を整えた。 子ども血中総IgE、好酸球、気道炎症マーカーFeNO値について、一般子ども集団のアレルギー有となしの子どもの比較により、成人のカットオフ値の妥当性について検討した。 動物・細胞実験分担研究においては、ヒト肝細胞に近い性質を示すHepaRG細胞株に0.1% DMSO（対照群）、MEHP 3 microM（低用量群）、MEHP 30 microM（高用量群）をそれぞれ24時間曝露してmRNAを採取した。GeneChip解析を用いた網羅的遺伝子発現解析を行い、核内受容体PPARalpha活性化に伴う遺伝子発現変化を認めた。さらに、妊娠ICRマウスに、0.1%DMSOを含む水（対照群）、DEHP 0.3 mg/kg（低用量群）及びDEHP 3 mg/kg（高用量群）を妊娠期間中経口曝露した。それぞれ出生後７日目の仔マウス肝臓からmRNAを採取した。
• プラスチックによる脂質代謝異常およびNASHや発癌との関連性に関する研究

  日本学術振興会：科学研究費助成事業 基盤研究(C)

  Date (from‐to) : 2020/04 -2023/03 

  Author : DARWISH WAGEH・SOBHY・ABDELRAHEM・ABDALLAH, 津久井 隆行, 惠 淑萍
• Effects of feeding management on bovine fertility and ovarian function: relationship between lipid profile and developmental competence of oocytes

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research

  Date (from‐to) : 2019/04 -2022/03 

  Author : Nagano Masashi

   

  Oocyte triacylglycerols (TAG) were significantly higher in cows in the early lactation stage than in heifers. Plasma free fatty acids (FFA) were high at 20-30 after days in milk (DIM). Oocyte FFA positively correlated with plasma FFA, but negatively correlated with the mean energy balance and 21 days before oocyte collection. Elevated oocyte FFA in cows in the early ostpartum period under intensive feeding management suggested that oocytes were at a high risk of FFA lipotoxicity. In addition, the effects modulated (5-20%) oxygen environments on the in vitro development of bovine oocyte-cumulus-granulosa cell complexes (OCGCs) cultured in the presence or absence of an antioxidant (astaxanthin: Ax) were evaluated by using gas permeable culture wells. The presence of Ax in the 5-20% O2 condition promoted oocyte nuclear maturation and development of OCGCS.
• 糖尿病性腎症発症メカニズムの解明と制御：コレステリルエステル蓄積からのアプローチ

  日本学術振興会：科学研究費助成事業 基盤研究(C)

  Date (from‐to) : 2019/04 -2022/03 

  Author : 惠 淑萍

   

  糖尿病性腎症は、先進国における末期腎臓病の主な原因であり、活性酸素種（ＲＯＳ）が極めて重要な役割を果たすことが知られている。本研究課題は、糖尿病性腎症を、近位尿細管細胞への脂肪酸流入とコレステリルエステル（ＣＥ）蓄積を発端に始まり、ミトコンドリアを巻き込んで進行する酸化的悪性サイクルとして理解し、それを制御する手段を提案することを目的とする。 ２０１９年度はまず、化学合成により得られた８種類ＣＥ標準物質およびそれぞれ８種類の重水素標識ＣＥの内部標準物質を用いて、液体クロマトグラフィー質量分析定量法（ＬＣ／ＭＳ）を確立した。また、ヒト腎近位尿細管上皮細胞由来であるＨＫ－２細胞に脂肪酸添加を行い、オレイン酸（ＦＡ１８：１），リノール酸（ＦＡ１８：２），アラキドン酸（ＦＡ２０：４），エイコサペンタエン酸（ＦＡ２０：５），およびドコサヘキサエン酸（ＦＡ２２：６）添加群において脂肪滴の形成を確認した。しかし，パルミチン酸（ＦＡ１６：０）添加群では脂肪滴は確認されなかった。なお、脂肪酸の添加による脂肪滴形成時のＣＥをＬＣ／ＭＳ定量法により分析した。同様に不飽和脂肪酸添加群においてＣＥの有意な増加が確認された。 さらに、ＣＥ代謝酵素とトリグリセリド（TG）の合成系酵素を評価した。HK-2細胞（健常成人腎臓由来の近位尿細管上皮細胞株）への脂肪酸の過剰流入により、Sterol regulatory element-binding proteinの発現が低下した。HK-2細胞への脂肪酸負荷により、Diacylglycerol acyltransferaseおよびAdipose triglyceride lipaseの変化はなかった。
• A community-based epidemiological study on the links between characteristics of low-density lipoprotein and lifestyles/metabolic disorders/arteriosclerosis.

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research

  Date (from‐to) : 2017/04 -2022/03 

  Author : Nakamura Koshi

   

  A community-based epidemiological study was conducted to examine the association between serum low-density lipoprotein (LDL) cholesterol concentration and arteriosclerosis in the general population, with consideration of characteristics of LDL. Indices of arteriosclerosis of interest were cardio ankle vascular index (CAVI) and ankle brachial pressure index (ABI). When the study population was stratified by the median values of LDL characteristics (i.e., particle size, zeta-potential, hardness, and height), there were no associations between serum LDL cholesterol concentration and CAVI/ABI in each stratum.
• 中性脂肪蓄積心筋血管症の診断に直結するエビデンス創出研究

  日本医療研究開発機構(AMED)：

  Date (from‐to) : 2020 -2022 

  Author : 惠 淑萍
• Study of Rapid Detection Technology for Oil Quality and Toxicological Evaluation of Frying Oil

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for JSPS Fellows

  Date (from‐to) : 2017/11 -2020/03 

  Author : 惠 淑萍, 張 函, MA JIN-KUI

   

  It was established that an effective and rapid detection method based on the odor fingerprint of frying oil for a real-time detection and monitoring of frying oil quality. The potential toxicities of frying oils' polar substances and oxidative products (hydroperoxides) to cells in vivo were investigated. The major primary oxidative products of the linoleic acid, 9 and 13-Hydroxyoctadecadienoic acid (9 and 13-HPODE), were synthesized using soybean lipoxygenase I-B. The 9 and 13-HPODE were also subjected to HepG 2 cells to evaluate its possible adverse effects (cell viability, changes in the enzyme expression, lipidomic analysis) . The research results of 2019 are summarized as follows: 1.The synthesized hydroperoxides have been identified by the Mass spectroscopy and NMR. Results showed that the purity of the synthesized 13-HPODE was around 95% and the yield rate was 70% above which implies the method is successful and was feasible for preparing 9 and 13-HPODE in a regular laboratory. 2.In the cells’ experiments, there were two parallel experiments carried out: one is for the PAHs and another is for the HPODEs using target cell line- HepG 2 cells. Results showed that the mRNA expression of GSTO1, SOD2, and HO-1 have increased when the cells were treated with high HPODEs concentration (100 uM), while the GPX mRNA expression was inhibited after the treatment. Results also indicated that the expression level of CYP1A1 mRNA has been significantly increased after the cells treated with high concentration of HPODEs.
• Study on the Oxidation and Inhibition of Unsaturated Lipid s and

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2017/04 -2020/03 

  Author : Ye Shen

   

  In the present study, we demonstrated the risk that the unsaturated lipids in the lung surfactants could be oxidized by the low-level ozone in the ambient environment. The oxidation products of the unsaturated lipid monolayers have been quantitatively characterized by surface isotherm, liquid chromatography-mass spectrometry (LC-MS), and heterodyne-detected sum frequency generation (HD-SFG) spectroscopy. It was found that the unsaturated lipid molecules of the lung surfactants were quickly oxidized from its C=C moiety to form the aldehyde and acid terminated lipid derivatives with a trace amount of secondary ozonide. Detailed reaction mechanisms and kinetics have been discussed based on the experimental results. Furthermore, oxidation inhibition effects by adding ascorbic acid or 3,5-dihydroxy-4-methoxybenzyl alcohol in the subphase have been partially confirmed.
• System development to realize transillumination and functional imaging of animal body using safe light

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2017/04 -2020/03 

  Author : Shimizu Koichi

   

  To realize 3D transillumination imaging of internal body structure and physiological function, we integrated our principles to establish a new technique, and developed a test system. Through this study, we achieved the following results. (1) An inverse scattering problem was solved to obtain internal structure in a turbid medium using scattered light from the medium. (2) Blurred transillumination image could be improved by the deconvolution with a point spread function. (3) The problem of image blur was resolved using the convolutional neural network obtained by deep learning. (4) The feasibility of 3D transillumination imaging was confirmed using a tissue-simulated model phantom. (5) A test system was designed and manufactured. (6) The feasibility of functional imaging was confirmed in the identification of vein and artery in a transillumination image. (7) 3D functional transillumination imaging was achieved in animal experiments.
• Novel biomarker for atherosclerosis: clinical significance of isoprostanes analyzed by LC-MS/MS

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

  Date (from‐to) : 2017/04 -2020/03 

  Author : Segawa Namiko

   

  Oxidative stress is linked with many diseases including atherosclerosis and Alzheimer disease. Urine 8-isoprostaglandin F2α (8-iso-PGF2α) was established as a biomarker of oxidative stress. However, reactive oxygen species (ROS) affects the polyunsaturated fatty acid (PUFA) arachidonic acid to produces many isoforms of 8-iso-PGF2α, the biological significance of which is unclear. To explore the diagnosis marker of atherosclerosis, the relation between atherosclerosis and 8-iso-PGF2αisoforms was examined using multivariate data analysis methods including principal component analysis (PCA).
• Exploration of graft conditioning methods during machine perfusion to augment mitochondrial function and antioxidant ability in steatotic liver graft

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2016/04 -2020/03 

  Author : Shimamura Tsuyoshi

   

  To establish the methods for pre-conditioning the steatotic graft by machine perfusion, and to find out the reliable marker to predict later severity of ischemia and reperfusion injury (IRI), we explored the following issues. Using steatotic rat livers and hepatocytes cell line treated by fatty acids-rich culture, they were subjected to cold storage and/or machine perfusion (or mimicking conditions) in various temperatures, preservation and perfusion solutions commercially available, with or without supplementation of antioxidant and other agents to augment some protective proteins. We found that pre-conditioning during machine perfusion with antioxidant and some agents, having property to induce expression of protective proteins and to activate protective transcription factors, effectively prevent progression of IRI. These results indicate that properly pre-conditioned steatotic graft could become a safe source of liver transplantation.
• 安全な光による生体透視と機能イメージングを実現するシステムの創成

  文部科学省：科学研究費助成事業 基盤研究(B)

  Date (from‐to) : 2017/04 -2020/03 

  Author : 分担者, 惠 淑萍
• 極低濃度オゾン曝露における不飽和脂質とヒトリポ蛋自の酸化及び抑制に関する研究

  文部科学省：科学研究費助成事業 基盤研究（B）

  Date (from‐to) : 2017/04 -2020/03 

  Author : 分担者, 惠 淑萍
• 血中低比重リポ蛋自の性質と生活習慣・代謝異常・動脈硬化に関する地域疫学研究

  文部科学省：科学研究費助成事業 基盤研究(B)

  Date (from‐to) : 2017/04 -2020/03 

  Author : 分担者, 惠 淑萍
• 脂肪肝グラフトのミトコンドリア機能と抗酸化能を増強する画期的な肝体外灌流法の開発

  文部科学省：科学研究費助成事業 基盤研究B

  Date (from‐to) : 2016/04 -2020/03 

  Author : 分担者, 惠 淑萍
• Impact of the fraction of serum fatty acids on health and regulatory role of foods and gastrointestinal microbiota on them

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research

  Date (from‐to) : 2016/04 -2018/03 

  Author : Tamakoshi Akiko

   

  We investigated the impact of fraction of serum fatty acids on body mass, body composition and hypertension and their relationship with food intake and gastrointestinal microbiota among the population in a town in Hokkaido, Japan. From previous studies, excessive intake of SFA and MUFA from foods increase the risk of various disease such as obesity and arteriosclerotic disease. In addition to these knowledge, this study newly revealed that the concentrations of some serum fatty acids were related to the risk of obesity and arteriosclerotic disease. Besides, the relationship between the fraction of serum fatty acids and a particular gastrointestinal bacterial species which affect host body mass and body composition was also observed. Further analysis is needed to examine these relations comprehensively
• Effect of dietary patterns and blood vitamin D levels on sarcopenia

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Young Scientists (B)

  Date (from‐to) : 2016/04 -2018/03 

  Author : Okada Emiko, TAMAKOSHI Akiko, NAKAMURA Koshi, UKAWA Shigekazu, SASAKI Sachiko, HUI Shu-ping

   

  This study aimed to investigate the effect of blood vitamin D levels and dietary patterns on sarcopenia among community-dwelling people enrolled the Dynamics of lifestyle and Neighborhood Community (DOSANCO) on Health Study. The serum 25-hydroxyvitamin D levels in participants was 24.8 ng/mL for males and 22.5 ng/mL for females. Three dietary patterns were identified from the results of factor analysis: vegetable, traditional Japanese, and low-confectionery. The higher vitamin D levels was significantly associated with the higher muscle strength. However, regarding three dietary patterns, no significant associations were observed with muscle mass, muscular strength, and sarcopenia. This study is limited to cross-sectional study. Further study is needed that follow-up of participants and investigate to effect of vitamin D levels and dietary patterns on the long-term care and mortality.
• 血中脂肪酸分画の健康影響ならびにその規定要因としての摂取食物と腸内細菌叢の関与

  文部科学省：科学研究費助成事業 挑戦的萌芽研究

  Date (from‐to) : 2016/04 -2018/03 

  Author : 分担者, 惠 淑萍
• 中性脂肪蓄積心筋血管症に対する中鎖脂肪酸を含有する医薬品の開発

  日本医療研究開発機構：難治性疾患実用化研究事業

  Date (from‐to) : 2015/04 -2018/03 

  Author : 分担者, 惠 淑萍
• Possibility of a new biomarker in the diagnosis of primary aldosteronism

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

  Date (from‐to) : 2013/04 -2017/03 

  Author : JIN SHIGEKI, CHIBA Hitoshi, HUI Shu-Ping, WADA Norio

   

  Primary aldosteronism is considered as approximately 10% of the causes of hypertension, so the diagnosis of its subtype is crucially important issue because cures are required on different kind of subtype. In this study, we found the substance which was detected more amount in the aldosterone-producing adenoma (APA) patient urine than that in the other subtypes of primary aldosteronism. We investigated the possibility of employing this substance as a new biomarker. The substance was detected as 1.5-2.0 fold amount in APA against essential hypertension and idiopathic hyperaldosteronism (IHA). However, its isolation and the structural analysis had not been executed until now.
• Comprehensive analyses and development of immunometric assay for plasmalogen

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

  Date (from‐to) : 2013/04 -2016/03 

  Author : Hui Shu-Ping

   

  Plasmalogens (Pls) are glycerophospholipids owning antioxidative effect, so it is conceivable that Pls may prevent arteriosclerosis and Alzheimer's disease. In our study, chemical synthesis method for choline Pls (PlsCho) has been improved; chemical synthesis method for ethanolamine Pls (PlsEtn) has been established. Comprehensively qualitative and quantitative analyses of Pls in human serum have been carried out by LC-MS/MS using the synthetic Pls standards. The concentration ofPlsEtn18:0/20:4shows higher than that of PlsCho18:0/20:4 in human serum. Some mouses were immunized with BSA-plasmalogens obtained from chemical synthesis.
• 地域イノベーション戦略支援プログラム（国際競争力強化地域）（機器共用化）

  科学技術振興機構：地域産学官連携科学技術振興事業費補助金・イノベーションシステム整備事業

  Date (from‐to) : 2013 -2016 

  Author : 惠 淑萍
• 地域イノベーション戦略支援プログラム（国際競争力強化地域）（研究者集積）

  科学技術振興機構：地域産学官連携科学技術振興事業費補助金・イノベーションシステム整備事業

  Date (from‐to) : 2013 -2016 

  Author : 惠 淑萍
• プラズマローゲンの網羅的解析と免疫測定法の開発

  文部科学省：科学研究費助成事業基盤研究C

  Date (from‐to) : 2013 -2016 

  Author : 惠 淑萍
• 中性脂肪蓄積心筋血管症に対する中鎖脂肪酸を含有する医薬品の開発(分担者）

  厚生労働省：難治性疾患克服研究経費・難治性疾患等克服研究事業

  Date (from‐to) : 2013 -2015/03 

  Author : 惠 淑萍
• 新規サケ白子加水分解物の機能性分析と血中動態

  北海道中小企業総合支援センター：中小企業競争力強化促進事業

  Date (from‐to) : 2013 -2014/03 

  Author : 惠 淑萍
• 食・運動・健康・医療をつなぐ知で家庭に拓く次世代健康生活創造の国際拠点

  科学技術振興機構：研究成果展開事業センター･オブ･イノベーション（COI）プログラム・COI-トライアル

  Date (from‐to) : 2013 -2014 

  Author : 惠 淑萍
• 中性脂肪蓄積心筋血管症に対する中鎖脂肪酸を含有する医薬品の開発 （分担者）

  平成 24 年度厚生労働科学研究費補助金（難治性疾患等克服研究事業）：

  Date (from‐to) : 2012/09 -2013/03 

  Author : 惠 淑萍
• 細胞内脂肪滴の酸化評価系の作製と食品開発への応用

  産学が連携した研究開発成果の展開 研究成果展開事業 研究成果最適展開支援プログラム(A-STEP) 探索タイプ

  Date (from‐to) : 2013 -2013 

  Author : 惠 淑萍

   

  針で採取した細胞の脂肪滴の量をモニターし、内標準法で脂質を定量できた時に、評価法の知財化と技術移転が可能になる。また、脂肪滴の酸化を阻止しミトコンドリア機能維持に役立つ物質が同定された時、最大の知財と技術移転の可能性が生じる。技術的課題、1)培養細胞に脂肪滴を形成、脂肪滴を定量的に評価、2)ミトコンドリア機能評価(膜電位測定)と脂肪滴との関係把握、3)脂肪滴採取と脂肪滴脂質(過酸化脂質を含む)の定量質量分析、以上の問題の解決に取り組み、1)、2)は実施できたが、3)は脂肪滴採取と内標準物質(安定同位体)合成までで時間切れとなった。達成率は90%。今後は現状の評価系での目的の食品成分の探索と、評価系の完成度向上に努め、両面で技術移転の可能性を追求する。
• Study of lipid hydroperoxides in metabolic syndrome

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research

  Date (from‐to) : 2010 -2012 

  Author : HUI Shu-ping

   

  Cholesterylester monohydroperoxides (CEOOH), triglyceride monohydroperoxides (TGOOH, and phosphatidylcholine monohydroperoxide (PCOOH) were chemically synthesized. oxLDL was prepared by incubating native LDL from human plasma with CuSO4 for up to 4 h. The PCOOH levels during oxidation were determined and showed time course consisted of three phases. We then immunized five mice using the synthetic PCOOH and oxLDL, and antibody titers were measured. For synthetic PCOOH, the antibody titer did not show increase, but significant increase for oxLDL.
• 地域ノベーションクラスタープログラム（分担者）

  地域産学官連携科学技術振興事業費補助金 イノベーションシステム整備事業：

  Date (from‐to) : 2010/04 -2011/03 

  Author : 惠 淑萍
• 特別研究員奨励費

  文部省科学研究補助基金：

  Date (from‐to) : 1999/01 -2001/03 

  Author : 惠 淑萍

Educational Activities

Teaching Experience