The 20th Symposium on Trace Nutrients Research(2003)
Special Session Kyoto, May 27, 2003

Theme	Reporter	Position	P
Functional Foods --View on the International Status of the Research	Soichi ARAI	Faculty of Applied Biological Science, Tokyo University of Agriculture	1-

Proceedings

NO	Theme	Reporter	Position	P
1	Pharmacokinetic study on gastrointestinal absorption and disposition in the blood of 48vanadium and 65zinc in healthy rats using radiotracer method	Hiroyuki Yasui(1),Yusuke Adachi(1),Jun Fugono(1),Hiromitsu Haba(2),Akihiro Nakayama(2),Rieko Hirunuma(2),Shuichi Enomoto(2)and Hiromu Sakurai(1)	(1) Department of Analytical and Bioinorganic Chemistry, Kyoto Pharmaceutical University (2) Cyclotron Center, RIKEN	5-
2	Effect of resupplying zinc in zinc-deficient rats	Makoto NODERA, Hiroyuki YANAGISAWA	Department of Hygiene and Preventive Medicine, Saitama Medical School	9-
3	Evaluation of in vivo acute thymus-toxicity of an inorganic arsenical using murine models	Teruaki SAKURAI, and Kitao FUZIWARA	Laboratory of Environmental Chemistry, School of Life Science, Tokyo University of Pharmacy and Life Science	13-
4	Are magnesium and calcium concentrations in bone of female rats higher than that in male rats? -A study on Spontaneously Obese Rat (Minko Rat) with abnormal lipid metabolism (17)	Ryuji Takeda(1,2), Takashi Nakamura(1),Madoka Ishida(2), Masayo Imanishi(2),Takahisa Takeda(2),Mieko Kimura(2)	(1) Graduate School of Medicine, Kyoto University (2) Takeda Research Institute of Life Science	17-
5	Digestibility of Zinc Contained in Oysters	Munehiro YOSHIDA(1)Natsumi SUMIDA(1), Yoshikazu MATSUDA(2),	(1)Laboratory of Food and Nutritional Sciences, Faculty of Engineering, Kansai University (2)Japan Clinic Co.	23-
6	Metal-induced histomorphological injury and the alterations in the amounts and proportion of unsaturated disaccharides of glycosaminoglycan derived from proteoglycan	Yasuaki Arakawa(1),Kanako Nishimura(1) and Toshio Imanari(2)	(1)Department of Hygiene & Preventive Medicine, Faculty of Health Science, The University of Shizuoka (2)Department of Analytical Chemistry, Faculty of Pharmaceutical Science, The University of Chiba	27-
7	A Polyphenol compound, Rutin suppresses Ultraviolet and Methylviologen-induced Oxidative Cellular Damage on Achlorophyllous Unicellular Eukaryote, Euglena gracilis SMZ	Helen Palmer, Mari Ohta, Masumi Watanabe and Tetsuya Suzuki	Laboratory of Food Wholesomeness, Department of Life Sciences, Graduate School of Fisheries Science, Hokkaido University	33-
8	Structure and Bioactivity of β-1,6-Glucan of Agaricus blazei	Akira MISAKI(1),Shinji MIYABE(2) and Mariko KAKUTA(3)	(1)Osaka City University (2)R&D,Taiai (3)Konan Women's University	39-
9	Inhibitory effects of taurine on platelet-derived growth factor-BB (PDGF-BB) signaling in rat vascular smooth muscle cells	Yu HOSOKAWA(1),Masaharu TERASHIMA(2),Yoshinori TANIGAWA(2),and Keisuke IMADA(3)	(1)Department of Food and Health Sciences, Jissen Women's University (2)Department of Biochemistry and Molecular Medicine, Shimane Medical University (3)Self Medication Laboratories, Taisho Pharmaceutical Co.	49-
10	Effects of Dietary Vitamin B12 Deficiency on Spermatogenesis during Early Developmental Period in Male Rats	Toshiaki WATANABE(1),Kunihiko MAEDA(2),Yoshitaka WATANABE(3),Syuhei EBARA(4) and Yoshihisa NAKANO(4)	(1)Department of Food Environment Analysis, School of Humanities for Environmental Policy and Technology, Himeji Institute of Technology (2)Departnient of Pathology, Yamagata University School of Medicine (3)Department of Chemistry, Ibaraki College of Technology (4)(5)Department of Applied Biological Chemistry, University of Osaka Prefectur	55-
11	Comparison of ascorbate content in hatchery-reared and wild black sea bream, Acanthopagrus schlegeli (Pisces)	Hong JI,Ahmad Daud OM, Tetsuya UMINO,Heisuke NAKAGAWA	Graduate School of Biosphere Science, Hiroshima University	61-
12	Role of Reducing Activity in the Antioxidant Action of Tocopherol	Keiko Murakami, Masae Itoh and Masataka Yoshino	Department of Biochemistry, Aichi Medical University School of Medicine	67-
13	Distribution of Accumulated Arsenic in the Seaweed Hijiki, Sargassum fusiforme (Harvey) Setchell (5)	Yohko SUGAWA-KATAYAMA(1),Masayuki KATAYAMA(2),Chie SAKIYAMA(1) and Yukihiro NAKANO(3)	(1) Department of Human Health Science, Graduate School of Human Environment Sciences, Fukuoka Women's University (2) Osaka Prefecture University (3) Research Reactor Institute, Kyoto University	73-
14	Pre-illness mineral intake tendency of Crohn disease	Naomasa Sakamoto(1),Takashi Shimoyama (1),Kenji Wakai (2),Keiko Saito(3),Torao Tanaka(3),Masakazu Takazoe(3), Suminori Kono (4),Yoshihiro Miyake(5),Kazusi Okamoto (6)Gen Kobasi (7),Masakazu Washio (8),Tetsuji Yokoyama (9),Chigusa Date (10),Satosi Sasaki(11),Heizo Tanaka(11),Yutaka Inaba (12)	(1) Hyogo College of Medicine (2) Aich Cancer research center (3) Social Insurance Central General Hospital (4) Kyushu University Graduate School of Medicine (5) Kinki University School of Medicine (6) Aichi Prefectural College of Nursing & Health (7) Hokkaido University Graduate School of Medicine (8) Sapporo Medical University (9) National Institute of Public Health (10) Mukogawa Women's University (11) National Institute of Health and Nutrition (12) Juntendo University School of Medicine	81-
15	Importance of Metal Binding to DNA on Regulation of Gene Expression	Riichi Tawa,Hiromu Sakurai	Department of Analytical and Bioinorganic Chemistry Kyoto Pharmaceutical University	85-
16	Changes in trace element balances in the Breath of Hypertensives	Yasuaki Arakawa(1),Takayoshi Endo(2),Sayoko Ohmori(3),Harunobu Nakashima(4),Takayuki Takeuchi(5)and Yukihiro Nakano(5)	(1)Department of Hygiene & Preventive Medicine, Faculty of Health Sciences, The University of Shizuoka. (2)Meditopia Numazu. (3)Ohzuma Women University. (4)Osaka Prefectural Institute of Public Health. (5)Research Reactor Institute, Kyoto University.	89-
17	Availability of plum-vinegar (umezu) calcium in rats	Satoshi Takasugi,Hideyuki Omori, Tohru Matsui and Hideo Yano	Division of Applied Biosciences, Graduate School of Agriculture, Kyoto University	97-
18	Insulinomimetic Zinc(II) complexes with maltol Derivatives	Yusuke Adachi1),Jiro Yoshida(2),Yukihiro Kodera(2),Akira Katoh(3),Hiromu Sakurai'(1)	(1)Department Analytical and Bioinorganic Chemistry, Kyoto Pharmaceutical University (2)Healthcare Institute, Wakunaga Pharmaceutical Co., Ltd. (3)Department of Applied Chemistry, faculty of Engineering, Seikei University	101-
19	Studies on Cyclic Feed Intake and Serotonin in Zinc Deficient Rats	Daisuke Sato, FumikoYano	School of Biology-Oriented Science and Technology, Kinki University	105-
20	Efficacy of NaFeEDTA for improving iron deficiency	Kaori IGARASHI(1),Yukiko NAKANISHI(2),Rieko HIRUNUMA(1),Shuichi ENOMOTO(1),Shuichi KIMURA(2)	(1) RIKEN (Institute of Physical and Chemical Research), (2) Graduate School of Human Life Sciences, Showa Women's University	109-
21	Reactivity to Reactive Oxygen Species, DNA-binding Activity, and DNA-cleaving Activity of Various Metal Containing Metallothionein I and II	Akihiro Nakayama(1),Rieko Hirunuma(1),Shuichi Enomoto(1),and Hiromu Sakurai(2)	(1) Cyclotron center, RIKEN (2) Department of Analytical and Bioinorganic Chemistry, Kyoto Pharmaceutical University	113-
22	Identification of Chemical Species of Selenium Contained in Dark Muscle of Tuna	Munehiro YOSHIDA,Satoru SUGIHARA,Yuko CHIHARA,Mariko KONDO	Department of Biotechnology, Faculty of Engineering, Kansai University	117-
23	D-Amino Acid in Food: Determination and Its possibility to micronutrient	Tadao Oikawa(1)(2),Takashi Yamada(1),Yuki Sugihara(1),Tomoko Miyata(1),Munehiro Yoshida(1)(2) and Kenji Soda(3)	(1)Department of Biotechnology Faculty of Engineering, Kansai University (2)Kansai University High Technology Research Center (3)Bioleaders,Japan	121-
24	Involvement of vanilloid receptor on capsaicin-induced ROS production in PC12	Shanlou QIAO,Weihua LI,Ryoko TSUBOUCHI,Keiko MURAKAMI and Masataka YOSHINO	Department of Biochemistry, Aichi Medical University School of Medicine	125-
25	Screening and partial purification of substances in some sea weeds which exert growth-suppressing action against mouse transformed fibroblasts	Thang Jun Hang(2),Mamiko inoue(1),Yuki Uzawa(1) and Yukio Kawamura(1)(2)	(1)Dept. of Food Science and Nutrition, School of Agriculture (2)Dept. of Applied Life Chemistry, Graduate School of Agriculture Kinki University	131-

SUMMARY 1

NO	20-1
Theme	Pharmacokinetic study on gastrointestinal absorption and disposition in the blood of 48vanadium and 65zinc in healthy rats using radiotracer method
Reporter	Hiroyuki Yasui(1), Yusuke Adachi(1), Jun Fugono(1), Hiromitsu Haba(2), Akihiro Nakayama(2),Rieko Hirunuma(2), Shuichi Enomoto(2) and Hiromu Sakurai(1)
Position	(1) Department of Analytical and Bioinorganic Chemistry, Kyoto Pharmaceutical University (2)Cyclotron Center, RIKEN
Volume	The 20th Symposium on Trace Nutrients Research(2003)
P	P.5-8
Summary	In recent years, vanadium and zinc compounds are demonstrated to exhibit insulinomimetic effects in in vitro and in vivo experiments, and to treat both insulin-dependent type1 and insulin-independent type 2 diabetes mellitus. We have reported that vanadyl complexes normalize the blood glucose level of streptozotocin-induced type 1 diabetic rats, and that zinc complexes decrease the blood glucose level of hereditary type 2 GK rats. To understand the action mechanism of vanadium and zinc treatment, in this study, we focused on investigating pharmacokinetics on gastrointestinal absorption and disposition in the blood of vanadium and zinc ions in healthy rats by administration of their radiotracers. Clearance curves of 48V in the blood of rats receiving intravenous injection of vanadyl (48VO2+) and vanadate (48VO43-) ions were not significantly different in spite of administration of the different oxidation state and chemical species of vanadium, because of rapid reduction of vanadate to vanadyl in the blood and organs of rats. On the other hand, absorption of 48VO43- was found to be 3 fold higher than that of 48VO2+, suggesting the chemical species-dependent mechanism on the gastrointestinal absorption of vanadium. Bioavailability of 65Zn was estimated to be 0.27, which coincides well with the result in rats given non-tracer ZnCl2, strongly indicating the need of developing zinc complexes with higher bioavailability to exhibit more hypoglycemic activity than zinc ion.

SUMMARY 2

NO	20-2
Theme	Effect of resupplying zinc in zinc-deficient rats
Reporter	Makoto NODERA, Hiroyuki YANAGISAWA
Position	Department of Hygiene and Preventive Medicine, Saitama Medical School
Volume	The 20th Symposium on Trace Nutrients Research(2003)
P	P.9-11
Summary	The effects of oral intake of zinc (Zn) were investigated using rats in a Zn deficient state. A Zn-deficient diet was fed to six-week-old male Sprague-Dawley rats for 60 days. Then, a diet containing 20 mg of Zn per 100 g of food was fed for the next 28 days. Abnormalities such as dermatitis, abnormal serum copper/zinc (Cu/Zn) ratio, and deformed thyroid improved. However, no germ or sperm cells were observed in the testis. These findings suggest that the effects of resupplying Zn are minimal in tissues affected by rapid cell death due to Zn deficiency.

SUMMARY 3

NO	20-3
Theme	Evaluation of in vivo acute thymus-toxicity of an inorganic arsenical using murine models
Reporter	Teruaki SAKURAI, and Kitao FUZIWARA
Position	Laboratory of Environmental Chemistry, School of Life Science, Tokyo University of Pharmacy and Life Science
Volume	The 20th Symposium on Trace Nutrients Research(2003)
P	P.13-15
Summary	It has been well known that inorganic arsenicals are potent toxicants and carcinogens in humans and most animals. However, immunotoxic effects of them have not been well studied. In this study, we observed in vivo acute immunotoxicity of an inorganic arsenical, sodium arsenite, using murine models. Sodium arsenite was administered intraperitoneally into male CDF1 mice aged 4 weeks at doses of 1.25 - 8.0 mg / kg (body weight) on day 0, and the effect of arsenite on thymus of these mice was assessed until day 8. As results, a half numbers of mice were died when 8 mg / kg of arsenite were administered. The mice body weights were decreased significantly by arsenite in a dose dependent manner at doses of 1.25 - 4.0 mg / kg on days 1 - 2, and it returned to the control level on day 4. Administered arsenite was distributed into thymus; the maximum arsenic concentrations were detected on day 1 and they dropped slowly to the control level during 8 days. The thymus weight was significantly decreased by arsenite at doses of 4 mg / kg on days 1 - 4, and they returned to the control levels on day 8. 4-mg / kg arsenite also decreased significantly the numbers of thymocytes of mice temporary on day 1. Furthermore, the thymocyte blatogenesis of these mice was significantly inhibited by 4 mg / kg arsenite on day 1. Taken together, arsenite has significant in vivo immunotoxicity both quantitatively and qualitatively on thymus, and the thymus is one of very sensitive immune organs to arsenic inhalations.

SUMMARY 4

NO	20-4
Theme	Are magnesium and calcium concentrations in bone of female rats higher than that in male rats? -A study on Spontaneously Obese Rat (Minko Rat) with abnormal lipid metabolism (17)
Reporter	Ryuji Takeda(1,2), Takashi Nakamura(1), Madoka Ishida(2), Masayo Imanishi(2), Takahisa Takeda(2), Mieko Kimura(2)
Position	(1)Graduate School of Medicine, Kyoto University (2)Takeda Research Institute of Life Science
Volume	The 20th Symposium on Trace Nutrients Research(2003)
P	P.17-21
Summary	Summary Osteoporosis is a major public health problem also in Japan. Although bone densitometry is often used as an indicator to evaluate bone fragility, bone mineral density isn't always refluxed bone strength. There are well known many factors affected bone strength. In this paper, the relationship between strength and mineral concentration in bone of spontaneously obese male and female rats (39 weeks old) named "Minko Rat" with abnormal lipid metabolism is studied. Mechanical study in bone indicated that the stiffness of male rats was significantly high compared to that of female rats. On the other hand, Mg, Ca, P, Na, Zn, Sr and Fe concentrations in male rats were significantly low compared to that of female rats, S and K concentrations in male rats were significantly high compared to that of female rats. These results suggested that bone strength isn't depending on only mineral concentrations such as Ca, Mg and P and sex-related variation in elemental concentrations of rats bone was found.

SUMMARY 5

NO	20-5
Theme	Digestibility of Zinc Contained in Oysters
Reporter	Munehiro YOSHIDA(1), Natsumi SUMIDA(1), Yoshikazu MATSUDA(2),
Position	(1)Laboratory of Food and Nutritional Sciences, Faculty of Engineering, Kansai University (2)Japan Clinic Co.
Volume	The 20th Symposium on Trace Nutrients Research(2003)
P	P.23-26
Summary	To obtain information on the luminal absorption of oyster (Crassostrea gigas) zinc, the zinc action during an in vitro protease digestion of oysters was examined. More than 90% of the zinc was solubilized at pH 1.3 or 3.0 irrespective of the pepsin digestion. The solubilized zinc was partially re-precipitated by neutralization and trypsin digestion did not solubilize the re-precipitated zinc. When the pepsin digestion was performed at pH 5.0, the ratio of soluble zinc in trypsin digest decreased. When the trypsin digest was fractionated by Sephadex G-25, zinc was eluted later than the peptide fragments. These results indicate that the pH of the stomach juice rather than peptides released by the digestion of oyster protein highly contributes to the solubilization of oyster zinc in small intestine.

SUMMARY 6

NO	20-6
Theme	Metal-induced histomorphological injury and the alterations in the amounts and proportion of unsaturated disaccharides of glycosaminoglycan derived from proteoglycan
Reporter	Yasuaki Arakawa(1), Kanako Nishimura(1) and Toshio Imanari(2)
Position	(1)Department of Hygiene & Preventive Medicine, Faculty of Health Science, The University of Shizuoka (2)Department of Analytical Chemistry, Faculty of Pharmaceutical Science, The University of Chiba
Volume	The 20th Symposium on Trace Nutrients Research(2003)
P	P.27-32
Summary	To determine the extent of the metal-induced histomorphological injury and degeneration of tissues, the relationship between organotin-induced histomorphological injury and the alterations in the amounts and proportion of unsaturated disaccharides of glycosaminoglycan derived from proteoglycan were analyzed by using a high performance liquid chromatography(HPLC). In this article, the alterations in the amounts and proportion of unsaturated disaccharides of glycosaminoglycan are discussed with a focus on the organotin-exposed hippocampus and thymus. The disaccharide compositions of glycosaminoglycan from the hippocampus and thymus were significantly altered by the exposure of organotin compounds. In the hippocampus, increases in the proportion of chondroitin(CS-0S) and dermatan(DS-0S) and decreases in the proportion of chondroitin 4-sulfate (CS-4S) and dermatan 4-sulfate (DS-4S) were seen in the one-week oral exposure of tributyltin. In the thymus, an increase in the proportion of unsaturated disaccharide units(ΔDi-diSB) derived from dermatan sulfate and a decrease in the proportion of unsaturated disaccharide units(ΔDi-0S) derived from chondroitin sulfate were seen in the one-week oral exposure of dibutyltin. An increase in the proportion of disaccharide units(ΔDi-0S) and a decrease in the proportion of disaccharide units(ΔDi-4S) were seen in the one-week oral exposure of tributyltin. These alterations in the thymus depend on an decrease in the absolute amounts of chondroitin sulfate derived from the atrophy of the thymus which contains lot amounts of chondroitin sulfate proteoglycan

SUMMARY 7

NO	20-7
Theme	A Polyphenol compound, Rutin suppresses Ultraviolet and Methylviologen-induced Oxidative Cellular Damage on Achlorophyllous Unicellular Eukaryote, Euglena gracilis SMZ
Reporter	Helen Palmer, Mari Ohta, Masumi Watanabe and Tetsuya Suzuki
Position	Laboratory of Food Wholesomeness, Department of Life Sciences, Graduate School of Fisheries Science, Hokkaido University
Volume	The 20th Symposium on Trace Nutrients Research(2003)
P	P.33-37
Summary	Antioxidative effect of the natural antioxidant rutin on the cellular damage induced by ultraviolet light radiation and methyl viologen (MV) in the unicellular flagellate achlorophyllous mutant of Euglena gracilis SMZ strain. UV-B irradiation and UV-B and MV combined exposure decreased phospholipid especially phosphatidylethanolamine contents, suggesting that combined exposure of UV and MV induced cell membrane damage under the chronic exposure conditions adapted in the present study. Exposure to UV-B and 200 μM MV induced the highest levels of TBARS, however, the supplementation of rutin suppressed the increase of TBARS.

SUMMARY 8

NO	20-8
Theme	Structure and Bioactivity of β-1,6-Glucan of Agaricus blazei
Reporter	Akira MISAKI(1),Shinji MIYABE(2)and Mariko KAKUTA(3)
Position	(1) Osaka City University (2)R&D,Taiaki (3)Konan Women's University
Volume	The 20th Symposium on Trace Nutrients Research(2003)
P	P.39-47
Summary	In a previous study fractionation of fruiting body of Agaricus blazei afforded an antioxidative substance (AOSA) from ethanol extract. Regarding β-glucans of major polysaccharide components of the fruiting body, we showed a diversity of β-1,3-linked glucans. In the present study, a careful fractionation of the mushroom revealed another molecular species of β-glucan, i.e. essentially linearβ-1,6-linked glucan, occurring in an alkali extract and hot-water extract., as presumed by Smith degradation analysis. The β-glucan, either from alkali-extract or hot water extract, was applied onto a Con A-affinity column and the pass-through fraction, free from glycogen, was collected. The glucan fraction contained some amount of protein but protease treatment followed by gel-filtration afforded an essentially protein-freeβ-1,6-glucan of Mw 20 - 25 x 104 and [α] -5.53°. Methylation analysis, Smith degradation, 13C -n.m.r spectrospecific analysis, and also hydrolysis by β-1,6-glucanase confirmed that this water-soluble glucan is an essentially linear glucan consisting solely of β-1,6-glucosidic linkages. Interactions with IgG against schizopyllan and β-1,6-specific banana lectin also supported above structure. The β-1,3-linked glucan, a major component of alkali-insoluble fraction, was converted to a soluble glucan polyol, which gave a linear β-1,3 glucan upon mild acid hydrolysis. Comparison in the growth inhibition activity of the mouse-transplanted sarcoma 180 revealed that the water-soluble, β-1,6-glucan, lack of a triple strand conformation, also showed a notable antitumor activity .

SUMMARY 9

NO	20-9
Theme	Inhibitory effects of taurine on platelet-derived growth factor-BB (PDGF-BB) signaling in rat vascular smooth muscle cells
Reporter	Yu HOSOKAWA(1), Masaharu TERASHIMA(2),Yoshinori TANIGAWA(2) and Keisuke IMADA(3)
Position	(1)Department of Food and Health Sciences, Jissen Women's University (2)Department of Biochemistry and Molecular Medicine, Shimane Medical University (3)Self Medication Laboratories, Taisho Pharmaceutical Co.
Volume	The 20th Symposium on Trace Nutrients Research(2003)
P	P.49-54
Summary	The effects of taurine on PDGF-BB-induced proliferation, immediate-early gene expression, and MAP kinase (MAPK) activation in rVSMCs were investigated. Taurine significantly inhibited both PDGF-BB (10 ng/ml)-induced proliferation and DNA synthesis on rat VSMCs in a concentration-dependent manner. Similarly, taurine significantly inhibited the mRNA expression of c-fos, c-jun, and c-myc induced by PDGF-BB in A7r5 VSMCs. In addition, taurine significantly inhibited PDGF-BB-induced phosphorylation of P44/P42 MAPK (ERK1/ERK2) in rVSMCs. These results suggest that taurine may inhibit PDGF-BB-induced rVSMC proliferation, and its activity may be mediated, at least in part, by down regulation of MAPK and immediate-early gene expression.

SUMMARY 10

NO	20-10
Theme	Effects of Dietary Vitamin B12 Deficiency on Spermatogenesis during Early Developmental Period in Male Rats
Reporter	Toshiaki WATANABE(1), Kunihiko MAEDA(2),Yoshitaka WATANABE(3),Syuhei EBARA(4),and Yoshihisa NAKANO(4)
Position	(1)Department of Food Environment Analysis, School of Humanities for Environmental Policy and Technology, Himeji Institute of Technology (2)Departnient of Pathology, Yamagata University School of Medicine (3)Department of Chemistry, Ibaraki College of Technology (4)Department of Applied Biological Chemistry, University of Osaka Prefecture
Volume	The 20th Symposium on Trace Nutrients Research(2003)
P	P.55-59
Summary	Vitamin B12 is an important nutrient for maintaining normal fertility in males. We have previously demonstrated that vitamin B12 deficiency during pregnancy may induce irreversible damage in the germ cells of rat embryos, and affect the maturation of spermatozoon. However, its mechanism and biological significance remain unclear. Therefore, to evaluate the role of cobalamin on spermatogenesis, the effect of dietary vitamin B12 deficiency on early spermatogenesis was examined in embryos and F1 males. There was no difference in the number of primordial germ cells and supporting cells in embryos and F1 males on 0 day of age between vitamin B12-deficient and control groups. However, on 21 days of age, numerous TUNEL positive cells were located in spermatocytes of the spermatogenic epithelium. The incidence of seminiferous tubules with apoptosis was 51.5% in the vitamin B12-deficient group. On 60 days of age, aplasia of spermatids and sperms were detected in the vitamin B12-deficient group. Spermatogenesis in F1 males was not sufficiently recovered in the vitamin B12-deficient group even if the control diet was given after weaning. From these findings, vitamin B12 deficiency during pregnant and lactating period may affect the germ cells, and especially damage spermatocytes in F1 male rats, which indicate that vitamin B12 may be an essential constituent in meiosis of spermatogenesis

SUMMARY 11

NO	20-11
Theme	Comparison of ascorbate content in hatchery-reared and wild black sea bream, Acanthopagrus schlegeli (Pisces)
Reporter	Hong JI, Ahmad Daud OM, Tetsuya UMINO, Heisuke NAKAGAWA
Position	Graduate School of Biosphere Science, Hiroshima University
Volume	The 20th Symposium on Trace Nutrients Research(2003)
P	P.61-65
Summary	Calcium ascorbate (CaA) has been mainly used as vitamin C source in fish feed in Japan. However CaA was instable in a composed diet and decomposed during storage. We compared ascorbate level in the brain, liver, muscle and eye between wild and hatchery-reared fish. Ascorbate was primarily accumulated in the brain. Hatchery-reared fish fed with ordinary composed diet incorporated less ascorbate than wild fish. However, ascorbate fortified diet elevated the level to that of wild fish. Although natural feed organisms ingested by wild fish contained low amounts of ascorbate, wild fish could effectively incorporate ascorbate in the organs. The low ascorbate in hatchery-reared fish might be explained by their poor ability to absorb dietary ascorbate and/or utilize the ascorbate inside their bodies. The current results suggested the importance of supplementation with a suitable amount of ascorbate in composed diet to exert the effects of vitamin C.

SUMMARY 12

NO	20-12
Theme	Role of Reducing Activity in the Antioxidant Action of Tocopherol
Reporter	Keiko Murakami, Masae Itoh and Masataka Yoshino
Position	Department of Biochemistry, Aichi Medical University School of Medicine
Volume	The 20th Symposium on Trace Nutrients Research(2003)
P	P.67-72
Summary	Antioxidant properties of tocopherol(Vitamin E) were analyzed in relation to its reducing activity. Tocopherol reduced ferric to ferrous ion but did not produce reactive oxygen species. Effect of tocopherol on the NADPH-generating system was examined under the in situ conditions of permeabilized yeast cells. NADP-isocitrate dehydrogenase, an NADPH-generating enzyme in mitochondria, was protected by tocopherol from the iron-mediated inactivation. Glutathione reductase, the regenerating reaction of reduced glutathione, was potently inhibited by cuprous ion; however, addition of tocopherol caused little or no inhibition by copper of the enzyme. Tocopherol may act as an antioxidant under the polar conditions, and can protect mitochondrial function against metal-induced oxidative damage

SUMMARY 13

NO	20-13
Theme	Distribution of Accumulated Arsenic in the Seaweed Hijiki, Sargassum fusiforme (Harvey) Setchell (5)
Reporter	Yohko SUGAWA-KATAYAMA(1), Masayuki KATAYAMA(2), Chie SAKIYAMA(1) and Yukihiro NAKANO(3)
Position	(1) Department of Human Health Science, Graduate School of Human Environment Sciences, Fukuoka Women's University, (2)Osaka Prefecture University, (3) Research Reactor Institute, Kyoto University
Volume	The 20th Symposium on Trace Nutrients Research(2003)
P	P.73-79
Summary	Summary In the preceding papers, we reported the distribution of arsenic accumulated by Hijiki plants harvested along the seashores of Tsushima Archipelago. To investigate them harvested in some other areas, we collected Hijiki plants along the seashores of Goto Archipelago, Japan, in May 2001. Hijiki plants were separated into several parts such as leaves, stems and roots. After washing them with distilled water, the samples were lyophilized. Arsenic was determined by neutron activation analysis in the nuclear reactor of Research Reactor Institute, Kyoto University. The non-uniformity of arsenic localization in individual plants along their stems was observed as shown previously when expressed in μg arsenic per g of wet or dry weight (ppm). Keywords: arsenic;Hizikia fusiforme Okam.; Sargassum fusiforme (Harvey) Setchell; s eaweeds; Goto Archipelago; neutron activation analysis

SUMMARY 14

NO	20-14
Theme	Pre-illness mineral intake tendency of Crohn disease
Reporter	Naomasa Sakamoto(1), Takashi Shimoyama (1), Kenji Wakai(2),Keiko Saito(2), Torao Tanaka(3), Masakazu Takazoe(3), Suminori Kono (4), Yoshihiro Miyake(5),Kazusi Okamoto (6), Gen Kobasi(7), Masakazu Washio (8),Tetsuji Yokoyama (9), Chigusa Date(10), Satosi Sasaki(11), Heizo Tanaka(11),Yutaka Inaba (12)
Position	(1)Hyogo College of Medicine (2)Aich Cancer research center (3)Social Insurance Central General Hospital (4)Kyushu University Graduate School of Medicine (5)Kinki University School of Medicine (6)Aichi Prefectural College of Nursing & Health (7)Hokkaido University Graduate School of Medicine (8)Sapporo Medical University (9)National Institute of Public Health (10)Mukogawa Women’s University (11)National Institute of Health and Nutrition (12)Juntendo University School of Medicine
Volume	The 20th Symposium on Trace Nutrients Research(2003)
P	P.81-84
Summary	Summary To evaluate the role of mineral intake tendency at the pre-illness periods in etiology of Crohn disease (CD), a hospital-based case-control study using a self-administered questionnaire was designed. Cases were patients aged 15-34 years with IBD (52 Male, 18 Female) within 3 years after diagnosis. One control was recruited for each case, matching by sex, age, in-, out-patient status and hospital. A semi-quantitative food frequency method is used to estimate average intakes of minerals five years ago. In Male, Fe and Na intake of the CD patients were significantly higher than that of controls, and those tendency was also observed in female. These findings suggested that dietary factors may be important for the incidence of IBD. Key words: iCrohn disease, case-control study, food frequency questionnaire, Fe,Na.

SUMMARY 15

NO	20-15
Theme	Importance of Metal Binding to DNA on Regulation of Gene Expression
Reporter	Riichi Tawa and Hiromu Sakurai
Position	Department of Analytical and Bioinorganic Chemistry Kyoto Pharmaceutical University
Volume	The 20th Symposium on Trace Nutrients Research(2003)
P	P.85-88
Summary	The biological trace elements contribute to many important cellular functions. It has been interested for their roles in regulation of gene expression. The metallothioneins (MTs) inductions in cells have been widely investigated as a cellular response for some exogeneous metals, and their inductions are mainly based on the transcriptional level which is regulated by some proteins reacting with metals. The upstream of the MT gene promoter region is located by the metal responsive elements (MRE), which react directly and characteristically with some metals, e.g., Cd, Zn, Ag, Cr or Cu. The many studies of finding and isolating the transcriptional factors as MT gene regulating protein have been started and some proteins, e.g., MTF-1 which reacts with zinc ion have been reported. Therefore, it is important to prove the secondary DNA structural changes by reaction with metal, especially those leading to study the metal effects to regulation of MT gene expression. In this study we measured the digestive inhibitions of some restriction endonucleases by Cu(II) ion in a relatively quantitative manner using the new constructed plasmid DNAs which contain the human MT-IIA promoter gene (pKB8) or the rat SOD-Cu chaperon cDNA (pRCuSD) in pUC19. It was also indicated that the complex formation between linearized pKB8 fragments by BssH II and the proteins involved in HeLa cell nuclear extract in the presence of Cu(II) ion.

SUMMARY 16

NO	20-16
Theme	Changes in trace element balances in the Breath of Hypertensives
Reporter	Yasuaki Arakawa(1),Takayoshi Endo(2),Sayoko Ohmori(3),Harunobu Nakashima(4),Takayuki Takeuchi(5),and Yukihiro Nakano(5)
Position	(1)Department of Hygiene & Preventive Medicine, Faculty of Health Sciences, The University of Shizuoka. (2)Meditopia Numazu. (3)Ohzuma Women University. (4)Osaka Prefectural Institute of Public Health. (5)Research Reactor Institute, Kyoto University.
Volume	The 20th Symposium on Trace Nutrients Research(2003)
P	P.89-95
Summary	To investigate the relationship between trace elements in the breath and morbid state, the element balances in the breath of various kind of patients were analyzed by using a thermal neutron activation analysis method. Of 22 elements, 16 elements such as Cl, Br, I, Na, K, Ca, Mg, Al, Mn, Zn, S, Fe, Co, Cr, V and Sc were found in the breath. In this article, trace element balances in the breath are discussed with a focus on a hypertension. In a complication of hypertension and lung disease, increases in percentage of K, and Br, and decreases in percentage of S, Al, Zn, and I were observed. In a complication of hypertension and diabetes, increases in percentage of K, and Mn and decreases in percentage of Al, and I were observed. In a complication of hypertension and heart disease, increases in percentage of K, and Mn and decreases in percentage of Al, and Zn were observed. In a essential hypertension, increases in percentage of K, and Mg and decreases in percentage of Na, Al, Cr, Se, and I were observed. Especially in a hypertension, an increase in K and a decrease in Na in the breath were observed. This indicates that the metabolism of Na/K in the lung is disorder. These results revealed that the excretion of trace elements in the breath changed significantly and specifically depending on morbid state. The excretion mechanism of the specific trace elements such as Na/K into the breath are now under investigation. key words ：Trace element balance, Breath, Hypertension

SUMMARY 17

NO	20-17
Theme	Availability of plum-vinegar (umezu) calcium in rats
Reporter	Satoshi Takasugi, Hideyuki Omori, Tohru Matsui and Hideo Yano
Position	Division of Applied Biosciences, Graduate School of Agriculture, Kyoto University
Volume	The 20th Symposium on Trace Nutrients Research(2003)
P	P.97-100
Summary	Plum-vinegar calcium is a byproduct of producing "plum wine". The present study was designed to compare the availability of calcium in plum-vinegar calcium (PV) with calcium carbonate (CAR) and calcium citrate (CIT). Rats were given diets containing one of 3 calcium sources at a level of 50% or 100% of calcium requirement for 4 weeks. Femur was collected and we measured defatted weight, density, ash weight, ash density and calcium content. All bone parameters increased with dietary calcium in rats given each calcium source. Femoral defatted weight, density, ash weight, and ash density were higher in PV group than in CAR group (p<0.05) when dietary calcium was low. These parameters did not differ between PV group and CIT group. When dietary calcium was high, femoral defatted weight was higher in CIT group than in the other groups (p<0.05) and the bone parameters did not differ between PV group and CAR group. We suggested that the availability of plum-vinegar calcium or calcium citrate was higher than calcium carbonate when intake of calcium source was low. Calcium citrate showed higher availability than the other calcium sources at high intake of calcium source but the advantage of plum-vinegar calcium disappeared. High content of sodium may interfere with calcium availability in plum-vinegar calcium when its intake is high.

SUMMARY 18

NO	20-18
Theme	Insulinomimetic Zinc(II) complexes with maltol Derivatives
Reporter	Yusuke Adachi(1),Jiro Yoshida(2),Yukihiro Kodera(2),Akira Katoh(3),Hiromu Sakurai(1)
Position	(1)Department Analytical and Bioinorganic Chemistry, Kyoto Pharmaceutical University (2)Healthcare Institute, Wakunaga Pharmaceutical Co., Ltd. (3)Department of Applied Chemistry, faculty of Engineering, Seikei University
Volume	The 20th Symposium on Trace Nutrients Research(2003)
P	P.97-100
Summary	Since the finding of insulinomimetic effect of zinc(II) ions, several insulinomimetic zinc(II) complexes have been proposed. Previous studies reported an extremely high inulinomimetic activities of zinc(II) complex with maltol ligand in In vitro and in vivo evaluations. Then we estimated insulinomimetic activities of zinc(II) complexes using several types of maltol ligand. In vitro insulinomimetic activity of zinc(II) complexes was estimated by inhibitory effect on free fatty acid (FFA)-release from the isolated rat adipocytes treated with epinephrine (adrenaline). Insulinomimetic activity of zinc(II) complex with allixin, [Zn(alx)2], was higher than those of ZnSO4 and other complexes. Moreover, Zn(alx)2 complex reduced the high blood glucose levels of type 2 diabetic KK-Ay mice. Based on these results, Zn(alx)2 complex is proposed to be a potent therapeutic agent to treat type 2 diabetes mellitus.

SUMMARY 19

NO	20-19
Theme	Studies on Cyclic Feed Intake and Serotonin in Zinc Deficient Rats
Reporter	Daisuke Sato, FumikoYano
Position	School of Biology-Oriented Science and Technology, Kinki University
Volume	The 20th Symposium on Trace Nutrients Research(2003)
P	P.105-108
Summary	It has been reported that change of serotonin(5HT) secretion in rat brain might induce a periodical change of feed intake in zinc deficient rats. To prove that 5HT is engaged in feeding cycle in Zn deficient rat, agonist and antagonist of 5HT receptors were administered into rat brains in this experiment. In experiment 1, 8-OH-DPAT was administered into lateral ventricle of zinc deficient rats for 14 days. The 8-OH-DPAT has been known to activate 5HT1A receptor and suppress 5HT secretion in brain. Periodical change of feed intake due to zinc deficiency didn't appear while the 8-OH-DPAT was administered, and the periodical change appeared after the 8-OH-DPAT dose ends. This result suggests that 5HT secretion rate in the brains participates in feeding cycle. In experiment 2, mianserine was administered into lateral ventricle of zinc deficient rats for 14 days since feeding depression in zinc deficiency might be controlled through the 5HT2C receptor, which has been shown to be involved in suppression of feeding frequency. However, periodical change of feeding rate, which appeared in zinc deficiency, was not disappeared by mianserine dose. Hence it　is suggested that 5HT2C receptor is not involved in regulations of periodical feeding pattern in zinc deficient rats.

SUMMARY 20

NO	20-20
Theme	Efficacy of NaFeEDTA for improving iron deficiency.
Reporter	Kaori IGARASHI(1), Yukiko NAKANISHI(2), Rieko HIRUNUMA(1), Shuichi ENOMOTO(1), Shuichi KIMURA(2)
Position	(1) RIKEN (Institute of Physical and Chemical Research), (2)Graduate School of Human Life Sciences, Showa Women’s University
Volume	The 20th Symposium on Trace Nutrients Research(2003)
P	P.109-112
Summary	Iron deficiency is one of the major nutritional problems in the developing countries, affecting primarily women of childbearing age, children and infants. The main causative factor is the poor absorption of iron from their foodstuffs. The bioavailability of dietary iron is extremely low because these inhibitors bind iron in the intestinal lumen. Iron fortification programs have been implemented to prevent iron deficiency in many developing countries. Sodium iron ethylenediaminetetraacetic acid (NaFeEDTA), a metal chelate, is one of the food fortificants. It is a flavor less and inert compound. In this study, to investigate the efficacy of NaFeEDTA for improving iron deficiency, the efficiency of NaFeEDTA for improving iron status compared with other iron fortificants, such as ferrous sulfate, ferrous citrate and emulsified ferric pyrophosphate in anemic rats was determined. Iron contents of liver at the end of experiments in rats fed the diet containing NaFeEDTA was higher than those in rats fed the diet containing ferrous citrate on rice-based diet. It is suggests that NaFeEDTA has high efficiency for improving iron status compared with ferrous citrate. Furthermore, to determine the interaction between NaFeEDTA and inhibitors, iron complexes in the solution of NaFeEDTA and phytic acid (PA) as an inhibitor was detected by using high performance liquid chromatography. In the eluent at pH 4.8, EDTA was bind to iron and keep it in the solution in the presence of PA. At pH 11, a little of iron was detected in the effluent of PA, suggesting that EDTA had strong chelating property of iron compared with phytic acid. It is assumed that EDTA prevents inhibitors binding iron in the gut. In this study, the results indicate that NaFeEDTA has high efficacy for improving iron deficiency in developing countries.

SUMMARY 21

NO	20-21
Theme	Reactivity to Reactive Oxygen Species, DNA-binding Activity, and DNA-cleaving Activity of Various Metal Containing Metallothionein I and II
Reporter	Akihiro Nakayama(1), Rieko Hirunuma(1), Shuichi Enomoto(1),and Hiromu Sakurai(2)
Position	(1)Cyclotron center, RIKEN (2)Department of Analytical and Bioinorganic Chemistry, Kyoto Pharmaceutical University
Volume	The 20th Symposium on Trace Nutrients Research(2003)
P	P.113-120
Summary	We previously reported that copper-containing metallothionein (MT) -I and MT-II react with superoxide anion radicals (·O2-) and generate hydroxyl radicals (·OH) in the different manner and the ·OH generation causes the development of hepatoma in the livers of Long-Evans rats with Cinnamon-like coat color (LEC rats) and human. Then, we investigated whether the other metal containing MT-I and MT-II (Zn- Cd-, Hg-, and Ag-MT-I and MT-II) react with reactive oxygen species (ROS). In addition, we investigated whether the metal containing MTs cleave DNA as an index of cellular damage. During the investigation, Cu containing MT binds to and cleaves DNA via generating ·OH by the contiguity site of DNA. Thus, we concluded that Cu-MT is one of the strong carcinogens.

SUMMARY 22

NO	20-22
Theme	Identification of Chemical Species of Selenium Contained in Dark Muscle of Tuna
Reporter	Munehiro YOSHIDA, Satoru SUGIHARA, Yuko CHIHARA, Mariko KONDO
Position	Department of Biotechnology, Faculty of Engineering, Kansai University
Volume	The 20th Symposium on Trace Nutrients Research(2003)
P	P.117-120
Summary	Chemical species of selenium (Se) contained in dark muscle of tuna were analyzed. About one-third of Se in the dark muscle were soluble in 0.2 N HCl. The acid extract was found to contain Se mainly as an unknown low molecular weight compound when analyzed using high-performance liquid chromatography-inductively coupled plasma mass spectrometry (HPLC-ICPMS). A proteolytic enzyme digestion released several amounts of selenocystine from the dark muscle. In a Sephadex G-25 gel chromatography of trypsin digest of the dark muscle, Se was eluted separately into several specific fractions. These results indicate that Se in dark muscle of tuna are present mainly as a protein-bound selenocysteine and an unknown low molecular-weight compound.

SUMMARY 23

NO	20-23
Theme	D-Amino Acid in Food: Determination and Its possibility to micronutrient
Reporter	Tadao Oikawa(1),(2), Takashi Yamada(1), Yuki Sugihara(1), Tomoko Miyata(1),Munehiro Yoshida(1),(2), and Kenji Soda(3)
Position	(1)Department of Biotechnology Faculty of Engineering, Kansai University (2)Kansai University High Technology Research Center (3)Bioleaders,Japan
Volume	The 20th Symposium on Trace Nutrients Research(2003)
P	P.121-124
Summary	D-Amino acids were once believed to be unnatural compounds. However, recently, a variety of D-amino acids were found to be both free and bound forms not only in bacteria, but in yeast, plants, insects, and animals. The recent development in analytical technique enables us to determine D-amino acids content exactly in organisms and tissues. We here describe the occurrence of D-amino acids in wine , sake, beer, tea, and coffee.

SUMMARY 24

NO	20-24
Theme	Involvement of vanilloid receptor on capsaicin-induced ROS production in PC12
Reporter	Shanlou QIAO, Weihua LI, Ryoko TSUBOUCHI, Keiko MURAKAMI, and Masataka YOSHINO
Position	Department of Biochemistry, Aichi Medical University School of Medicine
Volume	The 20th Symposium on Trace Nutrients Research(2003)
P	P.125-130
Summary	Capsaicin (CPS) is a major pungent principle present in hot peppers and is widely consumed as a food additive in the world. Capsaicin actions have traditionally been divided into specific (vanilloid receptor-mediated) and nonspecific effects. The vanilloid receptor 1 is a nonselective cation channel that is activated by pungent vanilloid compound, extra cellular protons or noxious heat. mRNA of VR1 and VRL1 were expressed in PC12 cells, and only VR1 mRNA was detected in glioma and A10 cell lines. VR1 protein was demonstrated in PC12 cells by immunocytochemistry and Western blotting. Capsaicin, the VR1 receptor agonist, led to an increase in intracellular calcium ion and this effect was blocked by pretreatment with VR1 receptor antagonist capsazepine (CPZ). Treatment of PC12 cells with low concentration of capsaicin (5-50 μM) increased reactive oxygen species production which was inhibited by capsazepine; by contrast, capsaicin inhibits the production of ROS at high dose, and capsazepine did not inhibit this effect. These findings suggest that PC12 cells can be used as a new model system for elucidating a molecular mechanism of the VR1-mediated signal pathway in neuron system, and differential effects of vanilloid receptor on capsaicin-induced ROS production in PC12.

SUMMARY 25

NO	20-25
Theme	マウスのガン化繊維芽細胞の増殖抑制を示す海藻成分の検索 Screening and partial purification of substances in some sea weeds which exert growth-suppressing action against mouse transformed fibroblasts
Reporter	Thang Jun Hang(2), Mamiko inoue(1), Yuki Uzawa(1) and Yukio Kawamura(1)(2)
Position	(1)Dept. of Food Science and Nutrition, School of Agriculture (2)Dept. of Applied Life Chemistry, Graduate School of Agriculture Kinki University
Volume	The 20th Symposium on Trace Nutrients Research(2003)
P	P.131-138
Summary	Anti-cancer action of the extracts of five kinds of sea weeds which were harvested from sea water and cultivated in the deep sea water was measured by using mouse fibroblast cells( wild type ) and the phenotype transformed with SV-40 virus. The ethanol extracts of the most sea weeds showed relatively high cytotoxic action to the fibroblast comparing with the aqueous extracts. The extracts of Ulva pertusa( or Ulva sp.) and Enteromorpha clathrta expressed high cytotoxic activity. Among them, the extracts from Enteromorpha clathrta cultivated in the deep sea water showed relatively selective cytotoxic action to the transformed cell. By gel filtration analysis, the cytotoxic activity was eluted in bimodal fractions with molecular weight around 2-3 kDa and 500Da.

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