Proceedings The 15th Symposium on Trace Nutrients Research (1998)
Kyoto, May 26, 1998

NO	Theme	Reporter	Position	P
2	Studies of bio-trace elements using the multitracer	Fumitoshi Ambe	�����w������	3-
3	Extended All Present Theory of the Elements and Biological Trace Elements	Hiroki Haraguchi, Kazumi Inagaki, Akiko Hokura and Hirotaka Matsuura	Department of Applied Chemistry, Graduate School of Engineering, Nagoya University	11-
4	Study of antioxydative activity and estrogenic function of some plants extracts	Tomoko Iguchi(1), Masayuki Kasahara(2), Shoji Masiko(3), Kenichi Nakatsugawa(1), Shuichi Kimura(1)	(1) Showa Women's University (2)The Yamagata Prefectural Institute of Public Health (3)Utsunomiya University	23-
5	Participation of functional mineral-encaged zeolite treated water species in restoration of cell motility of organotin-intoxicated Euglena gracilis Z	Mari Ohta(1), Kaoru Nakamura(2), Yasuo Kurihara(3), Kozo Takama(1) and Tetsuya Suzuki(1)	(1)Laboratory of Food Wholesomeness, Department of Marine Biosources Chemistry, Faculty of Fisheries Hokkaido University (2)Institute of Chemical Research, Kyoto University (3)Central Laboratory, Sinanen Zeomic Corp., Ltd.	33-
6	Oxidative Stress on Liver Tissue: Damage in Liver Chromatin by Oxygen Free Radicals Generated from Reaction of Hydrogen Peroxide with Copper or Iron Ions	Riichi Tawa and Hiromu Sakurai	Department of Analytical and bioinorganic Chemistry, Kyoto Pharmaceutical University	41-
7	Effect of zinc and vanadium on 3T3-L1 adipocyte differentiation	Eiji Takahashi, Shin-ichiro Torii, Tohru Matsui and Hideo Yano	Laboratory of Animal Nutrition, Graduated School of Agriculture, Kyoto University	47-
8	Studies on Anti-cancer Substances in Food Stuffs by Using Transformed Cells Bearing the Oncogene-Oncosuppressor System.	Yukio Kawamura,Masaru Ishikawa, Manabu Muramoto, Kazumi Kitta and Mariko Manabe	Protein Science Laboratory, National Food Research Institute	53-
9	Biodistribution of sialoglycoconjugates and their transference into the brain	Mieko Kawamura(1), Tatsuo Ido(2), Noritaka Matsubara(3), Yoshihiro Ikeuchi(3) and Shuichi Yanahira(3)	(1)Faculty of Agriculture, Tohoku University (2)Cyclotron and Radioisotope Center, Tohoku University (3)Snow Brand Milk Products Co., Ltd.	59-
10	Effect of zinc deficiency on the expression of endothelin-1 in glomeruli of rats with unilateral ureteral obstruction	Hiroyuki Yanagisawa, Makoto Nodera and Osamu Wada	Department of Hygiene and Preventive Medicine, Faculty of Medicine, Saitama Medical School	65-
12	Effects of Oyster Extract on Teratogenicity and Embryonic Development in Zinc-Deficient Mice Oy-Ex Discussion	Toshiaki Watanabe(1), Yoshikazu Matsuda(2), Takao Ohta(2) and Yukio Shibata(2)	(1)Department of Hygiene and Preventive Medicine, Yamagata University School of Medicine (2)Central Research Institute, Japan Clinic, Co., Ltd.	73-
14	Biodynamics of radiactive trace elements in Se-deficient rats: application of the multitracer technique	Rieko Hirunurma, Shuichi Enomoto, Shizuko Ambe and Fumitoshi Ambe	The Institute of Physical and Chemical Research (RIKEN)	89-
15	Variation of Trace Element Contents in Black Tea Infusion	Tatsuhiko Ando(1), Hiroshi Tachi(1), Munehiro Yoshida(2)	(1) Department of Brewing and Fermentation, Junior College of Tokyo University of Agriculture (2)Department of Biotechnology, Faculty of Engineering, Kansai University	95-
16	Effects of Metal and Fatty Acids on Oxidative Degradation of Cholesterol	Kimiko Ohtani, Ayumi Kataoka and Takahisa Minamibe	Faculty of Human Environmental Science, Kyoto Prefectural University	101-
17	Application of Copper catalyzed Oxidative Inactivation of AMP Deaminase to Analysis of Antioxidant Action of Polyphenolic Compounds	Keiko Murakami, Toshitaka Mori, Mayuko Nagata, Masae Itoh and Masataka Yoshino	Department of Biochemistry, Aichi Medical University	107-
18	On the Kynureninase -Medico-historical aspect of tryptophan metabolism with emphasis on the action of taurin and Zn2+ to diabetic rats-	Yukio Shibata(1),Fumio Takeuchi(1),Yoshikazu Matsuda(2),Eiko Takaya(2), Masako Yamaguchi(2), Takao Ohta(3), Masahiro Nakatsuka(3), and Kiyoko Kawabata(3)	(1)Aichi Medical University (2)Japan Clinic Co., Ltd. (3)Japan Food Research and Development Institute Co., Ltd.	115-
19	The Effects of Oyster (Crassostrea gigas) Extract ob Cell Proliferation and Antitumor Activity Oy-Ex Discussion	Yoshikazu Matsuda(1), Eiko Takaya(1), Masako Yamaguchi(1), Takao Ohta(1), Mariko Manabe(2), Yukio Kawamura(2), Shingo Hiroishi(3) and Yukio Shibata(1)	(1) Japan Clinic Co., Ltd., Central Research Institute (2)National Food Research Institute (3)Fukui Prefectural University	121-
20	Effect of oyster extracts on exercise -Glucide and lipid metabolism- Oy-Ex Discussion	Eiko Takaya(1),Yoshikazu Matsuda(1),Masako Yamaguchi(1),Takao Ohta(1),Yuko Harada(2),Mika Tanaka(2),Teruko Nakasa(2),Yasushi Okinaka(2)and Yukio Shibata(1)	(1)Japan Clinic Co., Ltd., Central Research Institute (2)Nutritional Biochemistry, Doshisha Women's College	129-
21	Accelerated apoptosis in the zinc-defidient rat	Makoto Nodera,Hiroyuki Yanagisawa and Osamu Wada	Department of Hygiene and Preventive Medicine, Faculty of Medicine, Saitama Medical School	137-
22	Comparison of Copper-metallotionein in the livers of LEC rats and human hepatoma	Akihiro Nakayama(1),Hiroyuki Fukuda(2),Masaaki Ebara(2),Kenji Suzuki(3),Katsuyuki Nakajima(4) and Hiromu Sakurai	(1)Dep. of Analytical and Bioinorganic Chemistry, Kyoto Pharmaceutical Univ. (2)First Dep. of Medicine, School of Medicine, Chiba Univ. (3)Dep. of Pathology, College of Medical Care and Technology, Gumma Univ. (4)Otsuka Pharm. Co.	141-
23	Pharmacokinetic analysis of insulin-mimetics vanadium complexes in rats -metallokinetic analysis by BCM-ESR method-	Hiroyuki Yasui,Tae Fujisawa,Riichi Tawa and Hiromu Sakurai	Department of Analytical and Bioinorganic Chemistry, Kyoto Pharmaceutical University	147-
24	Study for in vitro Cytotoxicity of Arsenocholine, an Organic Arsenic Compound in seafood	Teruaki Sakurai,Toshikazu Kaise and Chiyo Matsubara	Laboratory of Environmental Chemistry, School of Life Science, Tokyo University of Pharmacy and Live Science	155-
25	Vitamin E status of prepartum and postpartum dairy cows and newborn calves	Hajime Kumagai and Yot Chaipan	Graduate School for International Development and Cooperation, Hiroshima University	161-
26	The Role of Trace Elements in Kampo Medicines for the Treatment of Anemic Women During Pregnancy	Minoru Tanaka(1),Mieko Kimura(2),Yoshinori Itokawa(2)	(1)Department of Obstetrics and Gynecology, Kyoto National Hospital (2)Department of Environmental Health, Kyoto University Graduate School of Medicine	167-
27	Anemia and food intake in female professional volleyball players	Kenji Fukunaga(1), Toshiko Ono(1), Takashi Kimura(2) and Munehiro Yoshida(3)	(1)Department of Public Health Kansai Medical University (2)Awazu Clinic (3)Laboratory of Food and Nutritional Sciences, Department Biotechnology, Faculty of Engineering, Kansai University	173-
28	Fluorometric Determination of Low Molecular Thiol (SH) and Disulfide (SS) Groups in Some Seafood	Eiko Hatakeyama, Tsuyoshi Hazu and Hiroshi Meguro	Laboratory of Food Hygiene, Tohoku Fukushi University	177-

SUMMARY 2

NO	15-2
Theme	Studies of bio-trace elements using the multitracer
Reporter	Fumitoshi Ambe
Position	RIKEN (The Institute of Physical and Chemical Research)
Volume	The 15th Symposium on Trace Nutrients Research(1998)
P	P.3-10
Summary	We have developed the multitracer technique using the RIKEN Ring Cyclotron, which can accelerate heavy ions up to 135MeV/nucleon. Irradiation with such heavy ions generates a wide rage of radioisotopes in targets mainly due to nuclear fragmentation. Our idea was to use these radioisotopes together as a multitracer in order to trace the behavior of many elements simultaneously in various systems. Radiochemical procedures for preparation of carrier- and salt-free multitracers were established. So far, more than fifty elements can be traced using this technique. The multitracers are now used to investigate the behavior of various elements in chemistry, biochemistry and biology in cooperation with laboratories in universities and research institute. Especially, uptake transfer and metabolism of trace elements in animal bodies are now extensively studied using the multitracer technique. Several examples of such studies are presented.

SUMMARY 3

NO	15-3
Theme	Extended All Present Theory of the Elements and Biological Trace Elements
Reporter	Hiroki Haraguchi, Kazumi Inagaki, Akiko Hokura and Hirotaka Matsuura
Position	Department of Applied Chemistry, Graduate School of Engineering, Nagoya University
Volume	The 15th Symposium on Trace Nutrients Research(1998)
P	P.11-22
Summary	In 1936, Ida Noddack proposed a hypothesis called "All Present Theory of the Elements", in which she predicted the presence of all elements in geological samples (minerals and rocks). Based on the Noddack's hypotheses, the present authors are proposing a mew concept of "Extended All Present Theory of the Elements", which indicates that the All Present Theory should be extended to all the biological systems including human beings. In order to elucidate or prove the Extended All Present Theory of the Elements, the experimental results for the determination of all rare earth elements in human blood serum, and multi-element determination of major-to-ultratrace elements in black and green tea leaves were introduced together with the chemical speciation of trace elements in black-tea infusion. Finally, the correlation of elemental concentrations between human blood serum and seawater was discussed from the viewpoints of chemical and biological evolution of the life in seawater. It should be stressed here that the recent research for trace elements has been supported by the development of high-sensitive and multi-element analytical methods, especially ICP-MS (inductively coupled plasma mass spectrometry).

SUMMARY 4

NO	15-4
Theme	Study of antioxydative activity and estrogenic function of some plants extracts
Reporter	Tomoko Iguchi(1), Masayuki Kasahara(2), Shoji Masiko(3), Kenichi Nakatsugawa(1), Shuichi Kimura(1)
Position	(1) Showa Women's University (2)The Yamagata Prefectural Institute of Public Health (3)Utsunomiya University
Volume	The 15th Symposium on Trace Nutrients Research(1998)
P	P.23-32
Summary	Finally, the antioxidative activities of the ten kinds of plants extracts were evaluated by three methods. The conker (Aesculus turbinata Blume.), the barilla plant (Salsola Komarovi Iijin.) and the purslane (Potulaca oleracea L.) extract was found to be a strong antioxidant when tested by the TBA method using deoxyribose as the substrate, whereas the Colorimetric assay using Methyl indole as a reagent and linolenic acid as the substrate indicated that the activity of others. The seed vessel of a conker, the barilla plant (Salsola Komarovi Iijin.) and the purslane (Potulaca oleracea L.) extracts were showed extremely strong ·OH radical scavenging activity when tested by the ESR method. Secondly, the effect of Salsola Komarovi Iijin. and Potulaca oleracea L. extracts studied in the overiectmized rat model of osteoporosis. Twelve weeks old female SD rats were randomly assigned to following four groups, i.e., sham operated (Sham); overectimized (OVX); OVX and fed a diet containing 1.5% Salsola Komarovi Iijin. extracts (OVX + SK); OVX and fed a diet containing 1.5% Potulaca oleracea L. extracts (OVX + PO). In conclusion, the OVX and OVX+SK groups were to be sthenia both deossification or osteogenesis.

SUMMARY 5

NO	15-5
Theme	Participation of functional mineral-encaged zeolite treated water species in restoration of cell motility of organotin-intoxicated Euglena gracilis Z
Reporter	Mari Ohta(1), Kaoru Nakamura(2), Yasuo Kurihara(3), Kozo Takama(1), and Tetsuya Suzuki(1)
Position	(1)Laboratory of Food Wholesomeness, Department of Marine Biosources Chemistry, Faculty of Fisheries Hokkaido University (2)Institute of Chemical Research, Kyoto University (3)Central Laboratory, Sinanen Zeomic Corp., Ltd.
Volume	The 15th Symposium on Trace Nutrients Research(1998)
P	P.33-39
Summary	Series of research in our laboratory has shown that restoration of morphology and motility of TBTCl-intoxicated Euglena gracilis Z should be resulting from trace minerals in the processed aqueous solution. To examine the detoxication effect of mineral-encaged zeolite on the impaired cells, we used the bioassay system using Euglena gracilis Z as the model organism and TBTCl as the model xenobiotic. Under the polluted environment, Euglena cells changes their shape to spherical form from spindle form. By taking advantage of this unique character, we examined the different types of zeolites which encaged different minerals. TBTCl-intoxicated Euglena cells were separately washed with the water species processed by zeolite, then incubated in the processed water for up to 3 hours .the restoration of motility of the cell was estimated by observing the motile cell number using a video-microscopy system. A remarkable recovery of cell motility was observed with the incubation system using Fe, Mn and Zn encaging zeolite. However they did not show any recovery effect when they were treated with a chelator, Chelex-100.

SUMMARY 6

NO	15-6
Theme	Oxidative Stress on Liver Tissue: Damage in Liver Chromatin by Oxygen Free Radicals Generated from Reaction of Hydrogen Peroxide with Copper or Iron Ions
Reporter	Riichi Tawa and Hiromu Sakurai
Position	Department of Analytical and bioinorganic Chemistry, Kyoto Pharmaceutical University
Volume	The 15th Symposium on Trace Nutrients Research(1998)
P	P.41-46
Summary	We examined the susceptibility of liver chromatin of mice at prenatally 16-days (fetus) and postnatally 19-23-weeks (adult) for hydroxyl radical (·OH). DNA strand breakages in both fetal and adult liver nuclei by Cu(II)-H2O2 system were shown to be more extensive that by Fe (III) -H2O2 system under the same experimental conditions. DNA damage in nuclei also showed higher rate and extent in fetal liver than that of adult, especially for oxidation by Cu (II) -H2O2 system. The reduced glutathione (GSH), N-acetylcysteine (NAC) and ascorbic acid (ASC) showed the promotive effects on CAN damages by ·OH, whereas cysteine (CYS) showed the antioxidative effect. In fetal liver, 5-methylcytosine (5mC) contents in chromatin were less than that of adult, suggesting the active chromatin structure in fetal liver nuclei. The increase of DNA damage by ·OH in fetal liver nuclei which accompanied the expansion of chromatin, indicated the importance of chromatin compaction for the oxygen free radical injury. The results also suggested that Cu (II) and Fe (III) ions act at different nuclei loci, and that Cu (II) ion binds to unmethylated sites on DNA constructing the nuclear matrix.

SUMMARY 7

NO	15-7
Theme	Effect of zinc and vanadium on 3T3-L1 adipocyte differentiation
Reporter	Eiji Takahashi, Shin-ichiro Torii, Tohru Matsui and Hideo Yano
Position	Laboratory of Animal Nutrition, Graduated School of Agriculture, Kyoto University
Volume	The 15th Symposium on Trace Nutrients Research(1998)
P	P.47-51
Summary	Zinc and vanadium have been reported to possess insulin-mimetic activity on various types of cells. 3T3-L1 preadipocyte which is derived from mouse can differentiate into adipocyte by several stimulators including insulin. We examined the stimulative effects of zinc and vanadium on the differentiation of 3T3-L1 preadipocyte. The supplementation of zinc during day 0-2 after confluence increased GPDH activity (a parameter of adipocyte differentiation) on day 8. The addition of zinc to 3T3-L1 cells for day 2-8 also enhanced GPDH activity. However no effect was observed when zinc was supplemented to fully differentiated 3T3-L1 adipocytes. Vanadium also increased GPDH activity in dose dependent manner when added for day 2-8.

SUMMARY 8

NO	15-8
Theme	Studies on Anti-cancer Substances in Food Stuffs by Using Transformed Cells Bearing the Oncogene-Oncosuppressor System.
Reporter	Yukio Kawamura, Masaru Ishikawa, Manabu Muramoto, Kazumi Kitta and Mariko Manabe
Position	Protein Science Laboratory, National Food Research Institute
Volume	The 15th Symposium on Trace Nutrients Research(1998)
P	P.53-58
Summary	By using cultured mouse fibroblast cells transformed with Simian virus 40 which bears the putative oncoprotein-oncosuppressor system, the rice bran was shown to contain anti-cancer substances which are selectively cytotoxic to the SV40-transformed cells but not to the molecular weights of 500 and 1,000 dalton, as revealed by molecular sieve chromatography on a Bio-gel P-2 column. LD50 and LD90 value of the active fraction was 1.0mg/ml and 3.0mg/ml to SV40-transformed cell, respectively, whereas at these concentrations nearly no Cytotoxicity was observed to nontransformed cells.

SUMMARY 9

NO	15-9
Theme	Biodistribution of sialoglycoconjugates and their transference into the brain
Reporter	Mieko Kawamura(1), Tatsuo Ido(2), Noritaka Matsubara(3), Yoshihiro Ikeuchi(3) and Shuichi Yanahira(3)
Position	(1)Faculty of Agriculture, Tohoku University (2)Cyclotron and Radioisotope Center, Tohoku University (3)Snow Brand Milk Products Co., Ltd.
Volume	The 15th Symposium on Trace Nutrients Research(1998)
P	P.59-63
Summary	The Biodistribution of the labeled sialoglycoconjugates, 14C-sialic acid, 14C-sialyllactose and 14C-ganglioside(GM3) and their transference into the brain of rats were studied. The intravenously administrated 14C-sialic acid and 14C-sialyllactose were excreted immediately from the kidney whereas 14C-ganglioside (GM3) was accumulated in the liver. By oral administration, 14C-sialyllactose and 14C-ganglioside (GM3) were accumulated in the salivary gland, spleen and brain. The oral administered 14C-sialyllactose was shown higher transference in to the brain than the case of intravenous administration. In the case of 14C-ganglioside (GM3), some was gradually metabolized to GM2, GD3 and Gd2 the remained in brain. It was clear that the orally administered ganglioside (GM3) was transferred into the brain, and was accumulated itself there and some metabolized.

SUMMARY 10

NO	15-10
Theme	Effect of zinc deficiency on the expression of endothelin-1 in glomeruli of rats with unilateral ureteral obstruction
Reporter	Hiroyuki Yanagisawa, Makoto Nodera, and Osamu Wada
Position	Department of Hygiene and Preventive Medicine, Faculty of Medicine, Saitama Medical School
Volume	The 15th Symposium on Trace Nutrients Research(1998)
P	P.65-68
Summary	In the present study, we examined the expression of endothelin (ET)-1 and a role of angiotensin IIin glomeruli from rats with unilateral ureteral obstruction (UUO) fed a zinc (Zn) deficiency or a standard from rats with UUO fed a Zn deficiency v.s. a standard diet. No expression of ET-1, however, was observed in glomeruli of the contralateral, non-obstructed kidney from rats with UUO fed a Zn deficiency or a standard diet. Indeed, the rennin-angiotensin system (RAS) is known to be activated after the onset of ureteral obstruction. These findings suggest that Zn deficiency intensifies the expression of ET-1 in glomeruli of the OK from rats with UUO by further enhancing the RAS upregulated after the onset of ureteral obstruction.

SUMMARY 12

NO	15-12
Theme	Effects of Oyster Extract on Teratogenicity and Embryonic Development in Zinc-Deficient Mice
Reporter	Toshiaki Watanabe(1), Yoshikazu Matsuda(2), Takao Ohta(2) and Yukio Shibata(2)
Position	(1)Department of Hygiene and Preventive Medicine, Yamagata University School of Medicine (2)Central Research Institute, Japan Clinic, Co., Ltd.
Volume	The 15th Symposium on Trace Nutrients Research(1998)
P	P.73-79
Summary	In the present study, we examined the expression of endothelin (ET)-1 and a role of angiotensin II in glomeruli from rats with unilateral ureteral obstruction (UUO) fed a zinc (Zn) deficiency or a standard from rats with UUO fed a Zn deficiency v.s. a standard diet. No expression of ET-1, however, was observed in glomeruli of the contralateral, non-obstructed kidney from rats with UUO fed a Zn deficiency or a standard diet. Indeed, the rennin-angiotensin system (RAS) is known to be activated after the onset of ureteral obstruction. These findings suggest that Zn deficiency intensifies the expression of ET-1 in glomeruli of the OK from rats with UUO by further enhancing the RAS upregulated after the onset of ureteral obstruction.

SUMMARY 14

NO	15-14
Theme	Biodynamics of radioactive trace elements in Se-deficient rats: application of the multitracer technique
Reporter	Reiko Hirunuma, Shuichi Enomoto, Shizuko Ambe, and Fumitoshi Ambe
Position	The Institute of Physical and Chemical Research (RIKEN)
Volume	The 15th Symposium on Trace Nutrients Research(1998)
P	P.89-93
Summary	The uptake and the distribution of radioactive trace elements in Se-deficient rats were examined by the multitracer technique, which can be used to evaluate the behavior of many elements under the same experimental condition. The uptake of Se was larger in the brain, spleen, and testicles of the Se-deficient rats than in those of the normal ones. The uptake of As, Fe, and Sc was larger in the liver of Se-deficient normal ones. Selenium is known to be in a competitive or synergetic relationship with several metals. From he present result on Sc and Zn, it was newly elucidated that there is also some interaction between those elements and Se.

SUMMARY 15

NO	15-15
Theme	Variation of Trace Element Contents in Black Tea Infusion
Reporter	Tatsuhiko Ando(1), Hiroshi Tachi(1), Munehiro Yoshida(2)
Position	(1)Department of Brewing and Fermentation, Junior College of Tokyo University of Agriculture (2)Department of Biotechnology, Faculty of Engineering, Kansai University
Volume	The 15th Symposium on Trace Nutrients Research(1998)
P	P.95-99
Summary	Contents of 12 trace elements (Na, Fe, Al, Zn, Mn, Cu, Ni, Cr, Se, As, Hg, Pb) in 30 black tea infusion samples were measured by inductively coupled plasma spectrometry or inductively coupled plasma mass spectrometry. The 30 tea samples consisted of Indian tea (9 samples), Ceylon tea (5 samples), Chinese tea (6 samples), Taiwan tea (3 samples), African tea (4 samples) and others (3 samples). The content of each element was as follows (mean ± SD); Na, 2.19 ± 2.33 ppm; Fe, 0.077 ± 0.036 ppm; Mn, 3.83 ± 2.31 ppm; Zn, 0.249 ± 0.113 ppm; Al, 3.44 ± 2.18 ppm; Ni, 0.040 ± 0.020 ppm; Cr, 0.002 ± 0.003 ppm; Cu, 0.042 ± 0.023 ppm; As, 1.52 ± 2.31 ppb; Hg, 0.027 ± 0.021 ppb; Se, 0.55 ± 0.51 ppb; Pb, 3.38 ± 3.26 ppb. A regional variation was observed in Na and As contents; African tea showed higher Na contents and Chinese tea showed higher As values than other samples. Factorial analysis showed that the 12 elements were classified 3 groups; elements to belong to the same group showed a similar variation in their concentration in the tea infusion each other. One group consisted of Mn, Zn, Al, Ni, Cr and Cu, and another group consisted of Fe, Se, As, Hg and Pb. Variation of Na was independent of those of other elements.

SUMMARY 16

NO	15-16
Theme	Effects of Metal and Fatty Acids on Oxidative Degradation of Cholesterol
Reporter	Kimiko Ohtani, Ayumi Kataoka and Takahisa Minamibe
Position	Faculty of Human Environmental Science, Kyoto Prefectural University
Volume	The 15th Symposium on Trace Nutrients Research(1998)
P	P.101-105
Summary	Cholesterol was heated at various temperature for various periods of time with or without metal salt and/or fatty acid. Cholesterol was degraded depending on the temperature and heating periods of time, but the amounts of cholesterol oxides produced were very little. In addition, the species and the molar ratio of cholesterol oxides produced were almost the same pattern. Then almost cholesterol might be degraded into much more small molecular substances. Fatty acids, especially polyunsaturated fatty acids, accelerated the degradation of cholesterol. However, large amounts of polyunsaturated fatty acids depressed the oxidative degradation of cholesterol. A small amount of copper salt, CuCl and CuCl2, accelerated the degradation of cholesterol especially in the presence of fatty acid, but large amounts of copper salts depressed the oxidation. In the case of iron salts, FeCl2 and FeCl3, oxidative degradation of cholesterol was accelerated depending on the amounts of iron salts added to the reaction mixture.

SUMMARY 17

NO	15-17
Theme	Application of Copper catalyzed Oxidative Inactivation of AMP Deaminase to Analysis of Antioxidant Action of Polyphenolic Compounds
Reporter	Keiko Murakami, Toshitaka Mori, Mayuko Nagata, Masae Itoh and Masataka Yoshino
Position	Department of Biochemistry, Aichi Medical University
Volume	The 15th Symposium on Trace Nutrients Research(1998)
P	P.107-113
Summary	Metal-catalyzed oxidative inactivation of AMP deaminase was applied to the structure-activity relationship studies of antioxidants in permeabilized yeast cells. AMP deaminase was readily inactivated by hydrogen peroxide plus reduced copper, and the inactivation may be due to the hydroxyl radical generated through the Fenton reaction at the copper-binding sites of the enzyme. Flavonol and flavone with both with both 2.3-double bond and 4-carbonyl group showed a protective effect on the copper-mediated inactivation of AMP deaminase. Baicalein representative of flavone without hydroxyl group at 3-position showed the most potent protective effect on the AMP deaminase. Flavonols, which has hydroxyl group at 3-position, also protected AMP deaminase to a lesser extent, but glycosylation of 3-hydroxyl group of flavonol nullified this protection. Flavanone and flavanol with saturated 2,3-bond and isoflavone with phenol group at 3-position showed little or no protection of the enzyme. Protective effect of Flavonoids on the oxidative inactivation of AMP deaminase was closely correlated with the inhibition of the formation of thiobarbituric acid-reactive substances as an index of lipid peroxidation. Antioxidant action of flavonoid is mainly depends on the 2,3-carbon-carbon double bond in conjugation with a 4-oxo group, which participates in the scavenging oxygen radicals and in the chelation of transition metals. This method may be useful for evaluating the antioxidant action of biological materials.

SUMMARY 18

NO	15-18
Theme	On the Kynureninase -Medico-historical aspect of tryptophan metabolism with emphasis on the action of taurin and Zn2+ to diabetic rats-
Reporter	Yukio Shibata(1), Fumio Takeuchi(1), Yoshikazu Matsuda(2), Eiko Takaya(2), Masako Yamaguchi(2), Takao Ohta(3), Masahiro Nakatsuka(3), and Kiyoko Kawabata(3)
Position	(1)Aichi Medical University (2)Japan Clinic Co., Ltd. (3)Japan Food Research and Development Institute Co., Ltd.
Volume	The 15th Symposium on Trace Nutrients Research(1998)
P	P.115-120
Summary	In 1925, Kynurenin has discovered by Matsuoka and Yoshimatsu and was named as Kynurenin by Prof. Yashiro Kotake. Since then, these metabolites have experimented by Yashiro Kotake, Yahito Kotake, Yazo Kotake, K Ichihara and Osamu Hayaishi et al. In this report, we display the action of taurin in diabetic rats through Kynureninase activity of rat liver. And we discuss about the interrelationship between the structure of Kynureninase protein and cystathionine synthase or lyase, and also about the difference against decarboxylase (5-OH Trp decarboxylase, DOPA decarboxylase and His decarboxylase related Zn2+ ion).

SUMMARY 19

NO	15-19
Theme	The Effects of Oyster (Crassostrea gigas) Extract ob Cell Proliferation and Antitumor Activity
Reporter	Yoshikazu Matsuda(1), Eiko Takaya(1), Masako Yamaguchi(1), Takao Ohta(1), Mariko Manabe(2), Yukio Kawamura(2), Shingo Hiroishi(3) and Yukio Shibata(1)
Position	(1) Japan Clinic Co., Ltd., Central Research Institute (2)National Food Research Institute (3)Fukui Prefectural University
Volume	The 15th Symposium on Trace Nutrients Research(1998)
P	P.121-127
Summary	The effects of oyster extract on tumor cells were studied using three cell lines (mice myeloma cell and A31 and SV-T2 cells originated from Balb/c 3T3 cell) and mice inoculated with P388 leukemia. Hot water extract was cytostatic against A31 cell and SV-T2 cell, but not against myeloma cell. And the high molecular weight fraction inhibited slightly the proliferation of myeloma cell and SV-T2 cell but not of A31 cell. The Low molecular weight fraction were cytostatic against these three cells. On mice with P388 leukemia, the high and low molecular weight fractions were more effective in prolonging life than hot water extract.

SUMMARY 20

NO	15-20
Theme	Effect of oyster extracts on exercise -Glucide and lipid metabolism-
Reporter	Eiko Takaya(1), Yoshikazu Matsuda(1), Masako Yamaguchi(1), Takao Ohta(1), Yuko Harada(2), Mika Tanaka(2), Teruko Nakasa(2), Yasushi Okinaka(2) and Yukio Shibata(1)
Position	(1)Japan Clinic Co., Ltd., Central Research Institute (2)Nutritional Biochemistry, Doshisha Women's College
Volume	The 15th Symposium on Trace Nutrients Research(1998)
P	P.129-135
Summary	In this study, we investigated about the effects of the oyster extract on the metabolism of the Glucide and lipid in exercised rats. The oyster extract was prepared from Oyster (Crassostrea gigas). Wistar male rats of 4 weeks age were divided into 3 groups: (1) the normal diet with exercise (T-group), (2) the normal diet without exercise (C-group), (3) the oyster extract added diet without exercise (O-group). After 4 weeks under these conditions, each group rats were forced to exercise. After the exercise, we analyzed triglyceride (TG), total cholesterol (TC), blood sugar level, lactate dehydrogenase (LDH), lactic acid, and free fatty acid in plasma, and also glycogen, TG, TC in liver. The change of blood sugar level LDH and lactic acid in T-group was less than those of the other groups. And the change of blood sugar level, LDH and lactic acid in O-group was less than that of D-group.

SUMMARY 21

NO	15-21
Theme	Accelerated apoptosis in the zinc-deficient rat
Reporter	Makoto Nodera, Hiroyuki Yanagisawa and Osamu Wada
Position	Department of Hygiene and Preventive Medicine, Faculty of Medicine, Saitama Medical School
Volume	The 15th Symposium on Trace Nutrients Research(1998)
P	P.137-140
Summary	To examine the effect of zinc (Zn) deficiency, we investigate the acceleration of apoptosis in Zn-deficient rat. Six-week-old male Sprague-Dawley (SD) rats were fed on diets supplemented 0or 0.02% Zn. After 90 days from administration of the Zn diet, detection of apoptotic cells skin, thymus and kidney was carried out using TUNEL (TdT-mediated dUTP-biotin nick end labeling) method. When compared to rat on 0.02% Zn diet. Furthermore, TINEL-positive cells were detected in the renal cortex on the 0% Zn diet, however, none were detected in the rat on the 0.02% Zn diet. These findings suggest that Zn deficiency may affect the cell cycle.

SUMMARY 22

NO	15-22
Theme	Comparison of Copper-metallotionein in the livers of LEC rats and human hepatoma
Reporter	Akihiro Nakayama(1), Hiroyuki Fukuda(2), Masaaki Ebara(2), Kenji Suzuki(3), Katsuyuki Nakajima(4) and Hiromu Sakurai
Position	(1)Dep. Of Analytical and Bioinorganic Chemistry, Kyoto Pharmaceutical Univ. (2)First Dep. Of Medicine, School of Medicine, Chiba Univ. (3)Dep. Of Pathology, College of Medical Care and Technology, Gumma Univ. (4)Otsuka Pharm. Co.
Volume	The 15th Symposium on Trace Nutrients Research(1998)
P	P.141-145
Summary	Long-Evans Cinnamon (LEC) rats develop spontaneous hepatic injury. We obtained evidence which shows unusual accumulation of copper (Cu) in the livers of LEC rats, followed by the finding of copper-metallothionein (Cu-MT) induction. On the other hand, Ebara et al. have found that copper is accumulated in the human liver suffered from hepatoma. Thus, we measured MT and 4 metals (Cu, Zn, Fe, and Mn) and compared the Cu in the LEC rat liver remarkably increased with age. The MT levels in human liver were evaluated as logarithmic concentration ratio [log((concentration in tumor site)/(concentration in juxta-tumor site))] was decreased with the tumor size, but no changes in the Cu levels were found. The, we separated the MT in human livers by gel filtration on Superose12. In tumor site of the human liver, MT was determined as Cu-MT, whereas in juxta-tumor site, the presence of Cu, Zn-MT was suggested, these results indicate that MT is induced as Cu-MT in the early stage of hepatoma, but it decreases in the development of hepatoma. Thus, se conclude that the development of hepatoma in LEC rats and human might be understood with respect of the induction of Cu-MT but both rates for the formation and degradation of MT are different between LEC rat and human hepatoma.

SUMMARY 23

NO	15-23
Theme	Pharmacokinetic analysis of insulin-mimetics vanadium complexes in rats -metallokinetic analysis by BCM-ESR method-
Reporter	Hiroyuki Yasui, Tae Fujisawa, Riichi Tawa and Hiromu Sakurai
Position	Department of Analytical and Bioinorganic Chemistry, Kyoto Pharmaceutical University
Volume	The 15th Symposium on Trace Nutrients Research(1998)
P	P.147-153
Summary	Biological functions of vanadium have been investigated by many researchers. Especially, the insulin-like effect of vanadium is the most remarkable. Recently, vanadyl ion (+4 oxidation state of vanadium) and its complexes have been shown to normalize the blood glucose levels of streptozotocin-induced diabetic rats (STZ-rats). We have examined to find more effective vanadyl complexes with insulin-mimetic activity and prepared less toxic complexes with several types of coordination mode. Among them, vanadyl-methylpicolinate complexes (VO-MPA) was found to exhibit higher insulin-mimetic activities and less toxic than other complexes as evaluated by both in vitro and in vivo experiments. Electron spin resonance (ESR) is available to measure the paramagnetic compound in biological samples. We have developed the in vivo blood circulation monitoring-electron spin resonance (BCM-ESR) method to analyze the ESR signals due to paramagnetic metal ions and stable radicals in real time. In the present pharmacokinetic investigation to elucidate the relation ship between the blood glucose normalizing effect and global disposition of vanadium, we report first the experiments, in which the BCM-ESR method was applied to evaluate the disposition of vanadyl states in circulating blood after intravenous administration to male Wistar rats. EST spectra due to vanadyl state were obtained in circulating blood, and pharmacokinetic, parameters for them were estimated by using the compartment model. The results of the experiments indicated that vanadium considerably distributed to peripheral tissues and eliminated from the body through urine, and the accumulation of vanadium in the issue was enhanced by VO-MPA treatment. The long-acting property of VO-MPA was suggested by the incorporation and accumulation of vanadium in the bone. We conclude that the tissue distribution of vanadium is closely related with the antidiabetic activity of the vanadyl complex.

SUMMARY 24

NO	15-24
Theme	Study for in vitro Cytotoxicity of Arsenocholine, an Organic Arsenic Compound in seafood
Reporter	Teruaki Sakurai, Toshikazu Kaise and Chiyo Matsubara
Position	Laboratory of Environmental Chemistry, School of Life Science, Tokyo University of Pharmacy and Live Science
Volume	The 15th Symposium on Trace Nutrients Research(1998)
P	P.155-160
Summary	In this study, we examined the in vitro cytotoxic effects of an organic arsenic compound containing in seafood, trimethyl (2-hydroxyethyl)-arsonium cation, namely Arsenocholine (AsCho), on marine splenocytes, thymocytes and bone marrow (BM) cells comparing with those of an inorganic arsenical, sodium arsenite using synthesized pure material. Sodium arsenite showed strong Cytotoxicity on all of these cells, and it's IC50 was 5-6 μM. In contrast, AsCho had no Cytotoxicity on these cells even at concentrations over 10mM, and it slightly enhanced the viability of BM cells at doses over 100 μ. It is interesting that this unique biological effect was found in AsCho, an organic arsenic compound containing in some marine animals which are daily ingested as seafood in many countries.

SUMMARY 25

NO	15-25
Theme	Vitamin E status of prepartum and postpartum dairy cows and newborn calves
Reporter	Hajime Kumagai and Yot Chaipan
Position	Graduate School for International Development and Cooperation, Hiroshima University
Volume	The 15th Symposium on Trace Nutrients Research(1998)
P	P.161-165
Summary	Eleven primiparous and ten multiparous Holstein Friesian cows and their calves were studied to evaluate vitamin E status of prepartum and postpartum period. Plasma α-tocopherol concentrations of multiparous cows were higher than those of primiparous cows. Multiparous cows had larger decrease and increase in plasma α-tocopherol and total lipid concentrations round about the time of parturition than primiparous cows. Alpha-tocopherol concentrations in colostrums of multiparous cows were twice high as those of primiparous cows. Plasma α-tocopherol concentrations of calves from multiparous cows were higher than those of primiparous cows on 5 days of the age. Plasma α-tocopherol concentrations of calves started to decrease on 5 or 15 days of the age and showed 23μg/dl on 90 days of the age. The results of this study indicated that higher plasma α-tocopherollevels of multiparous cows than primiparous cows afford their calves higher plasma α-tocopherol levels immediately after birth because of higher α-tocopherol concentrations in the colostrums. Plasma α-tocopherol concentrations of calves decrease as the calves become dependent upon calf starter with low α-tocopherol concentration.

SUMMARY 26

NO	15-26
Theme	The Role of Trace Elements in Kampo Medicines for the Treatment of Anemic Women During Pregnancy
Reporter	Minoru Tanaka(1), Mieko Kimura(2), Yoshinori Itokawa(2)
Position	(1)Department of Obstetrics and Gynecology, Kyoto National Hospital (2)Department of Environmental Health, Kyoto University Graduate School of Medicine
Volume	The 15th Symposium on Trace Nutrients Research(1998)
P	P.167-172
Summary	For the use of Kampo medicines in treating anemia during pregnancy, a total of 90 patients participated in this investigation. The patients were divided into Group A, B, and C. Group A had 38 patients taking Toki-shakuyaku-san and kojin, which contained relatively high Cu, Zn, and Ca for a moderate constitution, Group B had 17 patients taking Ryutan-shakan-to, which contained relatively high Cu, Fe and Mg concentrations for a robust constitution, and Group C had 35 patients taking Kami-kihi-to, which contained relatively low minerals for a deficient constitution. For the Hb increases in Group A following medications the increases in Group A were the most, and the Group B were the next, with a relatively lesser for the Group C. Kampo medicines, which were effective for the treatment of anemia during pregnancy, were found to contain relatively high Cu, Zn, Fe and Ca.

SUMMARY 27

NO	15-27
Theme	Anemia and food intake in female professional volleyball players
Reporter	Kenji Fukunaga(1), Toshiko Ono(1), Takashi Kimura(2) and Munehiro Yoshida(3)
Position	(1)Department of Public Health Kansai Medical University (2)Awazu Clinic (3)Laboratory of Food and Nutritional Sciences, Department Biotechnology, Faculty of Engineering, Kansai University
Volume	The 15th Symposium on Trace Nutrients Research(1998)
P	P.173-176
Summary	Anemia and food intake were examined on 14 female professional volleyball players (22.1±2.1 years). Daily total energy intake of the players was estimated to be more than 3600kcal. Because of no change of body weight for the fast 1 year, this high energy intake would equilibrate their energy consumption. Intake of all other nutrients were remarkably higher in the players than the student. Adjusted value of protein intake of the players was comparable to that of the students and exceeded the recommended dietary allowance. Higher value of adjusted iron intake was observed in the players than the students. Compared to the students, the players showed Serum haptoglobin was significantly lower in the players than the students, it suggested the hemolysis was cause by exercise. These results suggest that strenuous exercise such as professional volleyball play can result in a decrease of Hb and iron status.

SUMMARY 28

NO	15-28
Theme	Fluorometric Determination of Low Molecular Thiol (SH) and Disulfide (SS) Groups in Some Seafood
Reporter	Eiko Hatakeyama, Tsuyoshi Hazu and Hiroshi Meguro
Position	Laboratory of Food Hygiene, Tohoku Fukushi University
Volume	The 15th Symposium on Trace Nutrients Research(1998)
P	P.177-182
Summary	N- (9-Acridinyl) maleimide (NAM) fluorometry was applied to low molecular thiol (SH) and disulfide (SS) groups in the pacific saury (Coloalabis saira) and the oyster (Crassostrea gigas). The low molecular SH was measured in buffer (pH 8.80) at room temperature after standing for one hour. The method was preliminary applied to check the change of low molecular SH and low molecular SS in the pacific saury and the oyster during the preservation at 4°C and the storage refrigeration.

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