Proceedings The 14th Symposium on Trace Nutrients Research(1997)
Kyoto, May, 20 1997

NO	Theme	Reporter	Position	P
2	Mineral (Na, K, P, Ca, Mg, Fe, Cu, Zn, Mn, Se) intake in the residents of Jiangsu Province, China	Mieko Kimura(1),Masaya Ikegawa(1),H.J. Chen(1),Z.H.Zhang(1),Shinichi Nakagawa(1),Naoki Hatsuda(1), Yoshinori Itokawa(2)	(1)Kyoto University Graduate School of Medicine, Department of Preventive and Social Medicine, Social Medicine (2)Kyoto University Graduate School of Medicine, Department of Preventive and Social Medicine, International Health	7-
3	Effects of protein intake on intestinal absorption and bone mineral metabolism in growing rats	Toyoko Okuda(1), Tomomi Maekawa(2) and Noriko Ogawa(2)	(1)Osaka kyoiku University (2)Osaka City University	13-
4	Olfactory Disorders and Calcium Localization	Yasuaki Arakawa	Department of Hygiene & Preventive Medicine, Faculty of Health Sciences, The University of Shizuoka	19-
5	Effect of Ouabain on the Behavior of 28Mg in the Mice Myocardium	Tatsuo Ido(1),Ren Iwata(1) and Mieko Kawamura(2)	(1)Radiopharmaceutical Chemistry, Cyclotron and Radioisotope Center, Tohoku University (2)Department of Applied Biochemistry, Faculty of Agriculture, Tohoku University	29-
6	Effect of dietary magnesium deficiency in the rat -Histological and ultra structural examination	Takako Ikeda(1),Takayoshi Imazawa(1,2) and Shuichi Kimura(1)	(1)Showa Women's University (2)National Institute of Health Sciences	33-
7	Cili Extract Prevents Bone Loss in an Ovariectomized Rat Model of Osteoporosis	Yoko Kitade, Tomoya Yamada, Tohru Matsuiand Hideo Yano	Division of Applied Bioscience, Graduate School of Agriculture, Kyoto University	39-
8	Determination of Heavy Metal Ions Using Glutathion (GSH) Fixation	Eiko Hatakeyama,Hiroshi Meguro	Laboratory of Food Hygiene, Tohoku Fukushi University	45-
9	Effects of Serum Calcium, Iron and a Bone Metabolic Marker on Broadband Ultrasound Attenuation (BUA), and Speed of Sound (SOS) of os calcis of the Female College Students	Nobuko Hagiwara(1),Tatsuki Inoue(1), Eiko Kitamura(1) and Hideo Koishi(2)	(1)Department of Health Science & Physical Education, Notre Dame Women's College (2)Honoral Professor of Osaka City University	51-
10	Zinc and its action	Hiroyuki Yanagisawa, Makoto Nodera and Osamu Wada	Department of Hygiene & Preventive Medicine, Faculty of Medicine, Saitama Medical School	59-
11	Insulin-mimetic effects of vanadyl complexes and distribution of total vanadium in STZ-rats treated with vanadyl complexes	Yae Fujisawa, Seiki Fujimoto and Hiromu Sakurai	Department of Analytical and Bioinorganic Chemistry, Kyoto Pharmaceutical University	65-
12	Reaction of NO (nitric oxide) and Copper-Metallothionein in the liver of LEC rats	Mayumi Okita,Akihiro Tsuji and Hiromu Sakurai	Department of Analytical and Bioinorganic Chemistry, Kyoto Pharmaceutical University	73-
13	Cytotoxicity of Inorganic and Methylated Arsenic Compounds	Teruaki Sakurai, Toshikazu Kaise and Chiyo Matsubara	Laboratory of Environmental Chemistry, School of Life Science, Tokyo University of Pharmacy and Life Science	79-
14	Comparative studies for unit chain distributions of different glycogens and evidence of multiple branching of oyster glycogen.	Motoko Matsui(1),Mariko Kakuta(2) and Akira Misaki(3)	(1)Ichimura Gakuin Junior College (2)Konan Women's University (3)Osaka City University	85-
15	Participation of minerals in various functionalized aqueous solution or "Function Water", in exertion of detoxification effect on organotin intoxidated Euglea gracilis	Mari Ohta(1),Kaoru Nakamura(2),Kozo Takama(1)and Tetsuya Suzuki(1)	(1)Laboratory of Food Wholesomeness, Department of Marine Bioresources Chemistry, Faculty of Fisheries, Hokkaido University (2)Institute of Chemical Research, Kyoto University	95-
16	Effect of oyster extracts on the recovery of cell motility of TBTCl-intoxicated Euglena gracilis Z	Tetsuya Suzuki(1),Mari ohta(1), Kaoru Nakamura(2) and Kozo Takama(1)	(1)Laboratory of Food Wholesomeness, Department, of Marine Bioresoueces Chemistry, Faculty of Fisheries, Hokkaido University (2)Institute of Chemical Research, Kyoto University	103-
17	Studies on Interaction of Minerals in Intestinal Epithelial Cells by Using The Metal Stress Response as An Marker	Chieko Uno,Yukiko Nakanishi,Sakiyo Koseki (Yamaoka) and Kyoden Yasumoto	Research Institute for Food Science Kyoto University	131-
19	Effect of Oyster Extract on Guinea-Pig Papillary Muscle Oy-Ex Discussion	Michio Yajima(1),Yoshikazu Matsuda(2),Masahiro Nakatsuka(2),Takao Ohta(2)	(1)Dept. of Pharmacology, Aichi Medical University (2)Central Research Institute, Japan Clinic Co., Ltd.	123-
20	Mechanism of Antioxidant Action of Flavonoids and Polyphenolics	Keiko Murakami(1),Yoshikazu Matsuda(2),Masahiro Nakatsuka(2)and Masataka Yoshino(1)	(1)Department of Biochemistry, Aichi Medical University (2)Central Laboratory, Japan Clinic Co., Ltd.	129-
21	Importance of Kynureninase	Yukio Shibata(1), Masahiro Nakatsuka(1),Takao Ohta(1),Yoshikazu Masuda(1),Tomoyuki Shindo(1), Eiko Takaya(1),Hiroshi Ishizu(2),Taizo Yamada(3),Sukehisa Ishizu	(1)Japan Clinic Co., Ltd., Central Research Institute (2)Ueki Hospital (3)Tamai Orthopedics and Internal Medicine hospital (4)Akura Hospital	137-
22	Introduction of Apoptosis of HL60 Cells by Gallic Acid Derivatives	HLA HLA HTAY,Ryoko Tsubouchi,Miyako Haneda,Keiko Murakamiand Masataka Yoshino	Department of Biochemistry, Aichi Medical University School of Medicine, Nagakute, Aichi 480-1195,Japan	143-
23	Absorption in the Body of Inclusion Complex of γ-Cyclodexytrin with Octacosanol after Oral Administration in Mice	Mieko Kawamura(1),Tatsuo Ido(2),Tadashi Funada(3),Itaru Miyamoto(3)and Tatsuya Sugawara(3)	(1)Department of Applied Biochemistry (2)Cyclotron and Radioisotope Center, Tohoku University (3)Nippon Oil and Fats Co., Ltd.	149-
24	Effect of dietary level of phytic acid on lipid metabolism in rats fed on sucrose.	Shoko Onomi,Tetsuyuki Katayama	Laboratory of Nutritional Science, Faculty of Education, Hiroshima University	155-
25	Effect of strenuous exercise on blood constituent II	Kenji Fukunaga,Munehiro Yoshida,Toshiko Ono and Naoki Nakazono	Department of Public Health Kansai Medical University	161-
26	Acidic fibroblast growth factor activates hypothalamo-pituitary-adrenocortical axis	Kazuo Sasaki(1), Itsuro Matsumoto(2),Yutaka Ohmura(3)and Akira Niijima(4)	(1)Fac. of Eng., Toyama Univ. (2)Fac. of Med., Nagasaki Univ. (3)Inst. of Bio-Active Sci., Nippon Zoki Pharmaceu. Co. (4)Fac. of Med., Niigata Univ.	167-
27	Biotin Content in Kampo Medicines for Skin Disease	Toru Fukui(1) and Toshiaki Watanabe(2)	(1)Clinical Laboratory, Byotai-Seiri Laboratory (2)Department of Hygiene and Preventive Medicine, Yamagata University School of Medicine	171-

SUMMARY 2

NO	14-2
Theme	Mineral (Na, K, P, Ca, Mg, Fe, Cu, Zn, Mn, Se) intake in the residents of Jiangsu Province, China
Reporter	Mieko Kimura(1), Masaya Ikegawa(1), H.J. Chen(1), Z.H. Zhang(1), Shinichi Nakagawa(1), Naoki Hatsuda(1), Yoshinori Itokawa(2)
Position	(1)Kyoto University Graduate School of Medicine, Department of Preventive and Social Medicine, Social Medicine (2)Kyoto University Graduate School of Medicine, Department of Preventive and Social Medicine, International Health
Volume	The 14th Symposium on Trace Nutrients Research(1997)
P	P.7-11
Summary	A total of 107 women from a region in urban China in Jiangsu Province were selected to examine an effect of mineral intake on their health status. This study includes questionnaires, physical examinations and biochemistries of blood and urine samples. The daily intake was measured using 24 h dietary recall as well as seven-day food frequency record. Consequently, Chinese women exceed in body weight and height than age adjusted Japanese women with a lower incidence of obesity. An extremely low prevalence of hypertension, diabetes mellitus and liver dysfunction among Chinese population also shows an overall superiority in their health status. A daily total caloric intake of Chinese women are almost the same level with the Japanese women, while their mean fat intake exceeds 10g per day more than that of Japanese women. The majority of vitamin intake of Chinese women are sufficient to meet their physiological requirements. Daily mineral intake is as follows; Na 2991mg (3193 mg), K 1539 mg (856 mg), P 890 mg, Ca 392 mg (220 mg), Mg 270 mg (149 mg), Fe 20 mg (15 mg), Cu 2.0 mg (1.1 mg), Zn 10 mg (7.9 mg), Mn6.1 mg (2.7 mg), Se 42 mg (24 h recall determination). As a result, Chinese women take more magnesium, iron and copper than Japanese women. The role of these dietary pattern in their health status might be elucidated by extending these nutritional survey to another part of China, as we already performed.

SUMMARY 3

NO	14-3
Theme	Effects of protein intake on intestinal absorption and bone mineral metabolism in growing rats
Reporter	Toyoko Okuda(1), Tomomi Maekawa(2) and Noriko Ogawa(2)
Position	(1)Osaka kyoiku University (2)Osaka City University
Volume	The 14th Symposium on Trace Nutrients Research(1997)
P	P.13-17
Summary	Osteoporosis is a disease characterized by low bone mass, excessive loss of minerals and protein contents in bone. The effects of protein intake on mineral utilization in the body and bone metabolism in the femur were investigated in growing male rats. The apparent intestinal absorption of calcium, phosphorus and magnesium and their retention in the body and weight of femur and humerus increased linearly with dietary concentrations of 5, 10, 20, and 30% casein diet in growing male rats. Femur on 5% casein diet was significantly higher in concentration of calcium, was significantly lower in calcium content and breaking forth than those of other groups. Hence, dietary protein deprivation resulted in slow bone growth and weak bone strength. These findings indicated that protein intake during the crucial years of peak bone mass development in important for the prevention of osteoporosis.

SUMMARY 4

NO	14-4
Theme	Olfactory Disorders and Calcium Localization
Reporter	Yasuaki Arakawa
Position	Department of Hygiene & Preventive Medicine, Faculty of Health Sciences, The University of Shizuoka
Volume	The 14th Symposium on Trace Nutrients Research(1997)
P	P.19-28
Summary	Deficiency and excesses of trace elements induce various kinds of brain lesion. In this study, organotin exposure- and zinc deficiency- induced olfactory lesions were kinetically analyzed from the viewpoints of the movement of trace elements, and inhibition of signal transduction and toxic cell death (necrosis or apoptosis) in the olfactory systems. Remarkable results were that organotin exposure and zinc deficiency induced each excessive accumulation of calcium in the olfactory system such as olfactory epithelium and olfactory bulb. This calcium localization depended upon the number and species of ligand of organotin compounds in case of organotin exposure and was parallel with the degree of manifestation of deficiency symptom in case of zinc deficiency. In case of organotin exposure, excessive increases of parathyroid hormone (PTH), cAMP and excessive decreases of inositol triphosphate (IP3), Ca2+/Calnodulin-dependent protein kinase II(CaM kinase II) were seen. In case of zinc deficiency, a remarkable decrease of OTH and a little decreases of cAMP. IP3 were seen in the olfactory tissues, and remarkable decrease of calcium and a remarkable increase of PTH were seen in the cerebrospinal fluid. Putting all results together, the calcium localization in the olfactory system seems to be due to the processes such as the activation of adenylcyclase by an excessive increase of PTH in the special regions of the brain, excessive increase in the formation of cAMP by the activation of adenylcyclase, disorder of cAMP-activated channels by the excessive increase of cAMP, and an excessive influx of extra cellular Ca2+by the disorder of the channels. Moreover this rapid and excessive increase in the concentration of intracellular Ca2+ seems to inhibit CaM kinase II functions followed by leading to necrosis of apoptosis in vivo or toxic cell death in vitro.

SUMMARY 5

NO	14-5
Theme	Effect of Ouabain on the Behavior of 28Mg in the Mice Myocardium
Reporter	Tatsuo Ido(1), Ren Iwata(1) and Mieko Kawamura(2)
Position	(1)Radiopharmaceutical Chemistry, Cyclotron and Radioisotope Center, Tohoku University (2)Department of Applied Biochemistry, Faculty of Agriculture, Tohoku University
Volume	The 14th Symposium on Trace Nutrients Research(1997)
P	P.29-32
Summary	Magnesium is essential for maintaining ATPase reaction which is responsible for hydrolyzing ATP to ADP. Ouabain, a specific pump inhibitor, is known to inhibits the ATPase activity. The purpose of this study was to observe the effect of Ouabain on magnesium behavior in the mouse myocardium. Ouabain (0.147mg/mouse) was administered intraperitonealy to ddY male mice (6 weeks old, 25-30g). After the administration of Ouabain at intervals of 5, 10, 30, 60, and 120 min., each experimental mouse received a venous injection of 28Mg via the tail. After 10 min. the mice were sacrificed. The uptake of 28Mg in the blood, heart, liver, small intestine, muscle, and bone were then measured. Autradiography of 28Mg concentrations on the heart was also performed. 28Mg was injected 10 min. after the administration of ouabain. The mice were sacrificed 10 min, after injection of 28Mg and uptake levels were measured. Results indicated that the administration of Ouabain affected the uptake of 28Mg in the heart and small intestine. Compared to the control, the accumulation of 28Mg showed a substantial decrease within 30 min. after administration. However, after 120 min., the effect of Ouabain could not be observed and the accumulation of 28Mg in the heart among the experimental group showed no significant differences. Autoradiographical analysis revealed a low accumulation of 28Mg in the Ouabain treated mouse myocardium.

SUMMARY 6

NO	14-6
Theme	Effect of dietary magnesium deficiency in the rat -Histological and ultra structural examination-
Reporter	Takako Ikeda(1), Takayoshi Imazawa(1,2) and Shuichi Kimura(1)
Position	(1)Showa Women's University (2)National Institute of Health Sciences
Volume	The 14th Symposium on Trace Nutrients Research(1997)
P	P.33-38
Summary	Epidemiologically, it has been suggested that dietary magnesium/calcium imbalance is associated with the risk of heart diseases. In the present study, the effects of magnesium deficiency and /or calcium over intake were investigated in rats. Male Sprague-Dawley rats were divided into 4 groups, and respectively fed basal diet (AIN-76) alone (Group 1), calcium-doubled AIN-76 diet (Group 2), magnesium-deficient AIN-76 diet (Group 3) and magnesium-deficient/calcium-doubled AIN-76 diet (Group 4) for 19 days. Obtained results thus suggested that dietary magnesium deficiency gives rise to retrogressive changes in some organs such as heart, and concurrent calcium over intake synergistically enhances the myocardial injury due to magnesium deficiency.

SUMMARY 7

NO	14-7
Theme	Cili Extract Prevents Bone Loss in an Ovariectomized Rat Model of Osteoporosis
Reporter	Yoko Kitade, Tomoya Yamada, Tohru Matsui and Hideo Yano
Position	Division of Applied Bioscience, Graduate School of Agriculture, Kyoto University
Volume	The 14th Symposium on Trace Nutrients Research(1997)
P	P.39-44
Summary	Firstly, the effects of cili (Rosa roxburghii) extract on bone metabolism and uterotrophic activity were studied in an ovariectomized rat model of osteoporosis. Secondly, cili extract was examined for the effects on osteogenic activity in UMR106 cells, a clonal osteoblastic line derived from rat osteosarcoma. Thirteen-week old female Wistar rats were randomly assigned to following five groups, i.e., shamoperated (Sham); Ovariectomized (Ovx); Ovx and fed a diet containing 4.5% ciliextract (Ovx + LC); Ovx and fed a diet containing 9.0% cili extract (Ovx + HC); Ovx and injected daily with 5 μg of 17 β-etradiol/kg body wt./day (Ovx+E2). The Ovx group showed significantly lower bone mineral density of the right femur than did the Sham group. Bone mineral density did not differ among the other groups from the Ovx group. These results suggest that cili extract is effective in preventing bone loss induced by ovarian hormone deficiency. Ovariectomy caused atrophy of the uterus. The atrophy was completely prevented by the E2, cili extract has similar action on bone metabolism but less uterotrophic activity. Thus cili extract might be more suitable drug than estrogen for prevention of osteoporosis because estrogen administration is accompanied by severe adverse effects such as vaginal bleeding and carcinogenesis. In in vitro study, alkaline phosphatase activity was significantly increased in UMR106 cells by the additions of E2 or cili extract. This implies that cili extract directly stimulates osteoblastic activity, in conclusion, cili extract contains a phytoestrogen which is effective in preventing bone loss due to ovarian hormone deficiency.

SUMMARY 8

NO	14-8
Theme	Determination of Heavy Metal Ions Using Glutathion (GSH) Fixation
Reporter	Eiko Hatakeyama, Hiroshi Meguro
Position	Laboratory of Food Hygiene, Tohoku Fukushi University
Volume	The 14th Symposium on Trace Nutrients Research(1997)
P	P.45-49
Summary	A new method was developed to determine heavy metal ions taking advantage of the selective ion fixations. The principle is to measure the decrease of GSH by the fixation. GSH were measured by NAM fluorometry in the concentration range between 10-9 - 10-12 mol/ml. The decrease of the fluorescence was proportional to the metal ion concentrations. The selectivity's of the ions were controlled by EDTA, Cl-, pH and ion strength. Hg+,Hg2+ and CH3Hg were selectively determined. The other ions were completely masked by the EDTA solution.

SUMMARY 9

NO	14-9
Theme	Effects of Serum Calcium, Iron and a Bone Metabolic Marker on Broadband Ultrasound Attenuation (BUA), and Speed of Sound (SOS) of os calcis of the Female College Students
Reporter	Nobuko Hagiwara(1), Tatsuki Inoue(1), Eiko Kitamura(1) and Hideo Koishi(2)
Position	(1)Department of Health Science & Physical Education, Notre Dame Women's College (2)Honoral Professor of Osaka City University
Volume	The 14th Symposium on Trace Nutrients Research(1997)
P	P.51-57
Summary	Broadband ultrasound attenuation (BUA), which demonstrates structural quality of bone tissue (quality), and speed of sound (SOS), which shows the volume of bone minerals (quantity), of os calcis, were measured on 358 healthy female students by means of ultrasound bone densitometry. General data on the students were also obtained: 1) physical characteristics - height, weight, lean body mass (LBM), body fat percentage (%Fat); 2) serum calcium (Ca), iron (Fe) and alkaliphosphatase (ALP); 3) menstruation cycle, any special dietary details, nutritional balance of food intake. Finally, stepwise multiple regression analysis of BUA in relation to the students' physical characteristics, serum minerals and ALP was carried out. Results were as follows: positive correlations existed between BUA and SOS (r=+0.523, p<0.001), and BUA and all physical characteristics (height: r=+0.230, weight: r=+0.384, LBM: r=+0.368, %Fat: r=+0.265, p<0.001). In the next step, The sample was divided into four groups according to BUA and SOS values: Group 1 (185 subjects) under the mean BUA value, Group 2 (163 subjects) over the mean BUA value, Group 3 (194 subjects) under the mean SOS value, and Group 4 (154 subjects) over the mean SOS value. It was only in Group 1 that a positive correlation between BUA and all physical characteristics (p<0.001), and BUA and SOS (p<0.05) existed. The multiple regression analysis showed that SOS, LBM and %Fat contributed significantly and positively to variations in BUA, while serum ALP and Fe contributed negatively. However, variations in height, weight, and serum Ca were not found to be significant. In conclusion, positive correlations existed between bone quality and quantity. Above all, the former has a closer relation to general physical characteristics. In the immature bone quality group (Group 1), the students with more developed physical characteristics also showed a higher level of bone quality. We assume Group 1 will attain the final stage of both physical development and bone growth.

SUMMARY 10

NO	14-10
Theme	Zinc and its action
Reporter	Hiroyuki Yanagisawa, Makoto Nodera and Osamu Wada
Position	Department of Hygiene & Preventive Medicine, Faculty of Medicine, Saitama Medical School
Volume	The 14th Symposium on Trace Nutrients Research(1997)
P	P.59-63
Summary	The present study was designed to examine if zinc (Zn) affects the development of unilateral ureteral obstruction (UUO)-induced tubulointerstitial nephropathy and the expression of the proto-oncogene, c-fos mRNA relating to the induction of apotosis and carcinogenesis. Compared to rats fed a standard of a high Zn diet, rats fed a Zn deficiency diet showed a more influx of leukocytes into the interstitium and in part had the structural damage of glomeruli in the cortex of the obstructed kidney at day 3 following UUO. These histological changes, however, were ameliorated by administration of enalapril, an angiotensin (ANG) I converting enzyme inhibitor. Again, the expression of c-fos mRNA was observed only in the kidney of rats fed a Zn deficiency diet. Thus, it is suggested that Zn deficiency not only aggravates UUO-induced tubulointerstitial nephropathy via an increase in the action of ANG II but also enhances the expression of c-fos mRNA in the kidney.

SUMMARY 11

NO	14-11
Theme	Insulin-mimetic effects of vanadyl complexes and distribution of total vanadium in STZ-rats treated with vanadyl complexes
Reporter	Yae Fujisawa, Seiki Fujimoto and Hiromu Sakurai
Position	Department of Analytical and Bioinorganic Chemistry, Kyoto Pharmaceutical University
Volume	The 14th Symposium on Trace Nutrients Research(1997)
P	P.65-71
Summary	Insulin-dependent diabetes mellitus (IDDM) is controlled by daily injection of insulin. Therefore, the study of development of insulin replacements upon oral administration is important. Vanadate, vanadyl ion and their complexes have been reported to have insulin-mimetic activities by in vitro and in vivo experiments. During investigations of insulin-mimetic vanadyl complexes, we have found a vanadyl-picolinate (VO-PA) complex, which was orally active and low toxic. Then we have synthesized vanadyl-pyridinecarboxaldehyde (VO-PCA) and vanadyl-quinalginate (VO-QA) as related complexes of VO-PA. We tested first the insulin-mimetic activity of the complexes in inhibition of the release of free fatty acid (FFA) from isolated rat adipocytes treated with epinephrine. Both VO-PCA and VO-QA exhibited strong inhibitions, and thus we evaluated their insulin-mimetic effects on STZ-rats. In STZ-rats received i.p. injection of the complexes, normalization of blood glucose level was observed, but in STZ-rats received p.o.administration no normalization of blood glucose level was seen. We determined total vanadium concentration in organs of STZ-rats received i.p. injection of VO-PCA by a neutron activation analysis method and found that vanadium was accumulated in bone and kidney. In conclusion, both solubility of the complex and strong inhibition of FFA release from adipocytes are essential to obtain a good effect of the complex on oral administration.

SUMMARY 12

NO	14-12
Theme	Reaction of NO (nitric oxide) and Copper-Metallothionein in the liver of LEC rats
Reporter	Mayumi Okita, Akihiro Tsuji and Hiromu Sakurai
Position	Department of Analytical and Bioinorganic Chemistry, Kyoto Pharmaceutical University
Volume	The 14th Symposium on Trace Nutrients Research(1997)
P	P.73-77
Summary	LEC (Long-Evans cinnamon) rats develop spontaneous hepatic injury associated with severe jaundice about 4 months after birth. Recently, we obtained evidence which shows an unusual accumulation of copper (Cu) in the liver of LEC rats, followed by finding of the copper-metallothionein (Cu-MT) induction. We suggest the mechanism for the developments of the development of the hepatitis in LEC rats in relation to the accumulated copper and induced Cu-MT. On the other hand, nitric oxide (¥NO) is known to play a wide variety of physiological role such as control of the blood pressure and immuno-reaction. NO has been sown to react with free SH-groups of proteins yielding nitrosothiols. In the present study, we examined the age- and organ-dependent changes of NO in Lec rats. Then we studied the reaction of NO and Cu-MT in the liver of LEC rats.

SUMMARY 13

NO	14-13
Theme	Cytotoxicity of Inorganic and Methylated Arsenic Compounds
Reporter	Teruaki Sakurai, Toshikazu Kaise and Chiyo Matsubara
Position	Laboratory of Environmental Chemistry, School of Life Science, Tokyo University of Pharmacy and Life Science
Volume	The 14th Symposium on Trace Nutrients Research(1997)
P	P.79-83
Summary	In the present study, we demonstrated the cytotoxic effects of inorganic arsenicals, arsenite and arsenate, and organic arsenic compounds, monomethylarsonic acid (MAA), dimethylarsinic acid (CMAA) and trimethylarsine oxide (TMAO), which are metabolites of inorganic arsenicals in mammalian bodies, using murine peritoneal macrophages in vitro. Inorganic arsenicals, both arsenite and arsenate, were strongly toxic to macrophages, and their concentrations that decreased the number of surviving cells to 50% of that in untreated controls (ICsub>50) was 5 or 500 μM, respectively. These inorganic arsenicals mainly caused necrosis. In contrast, the cytotoxic effects of Methylated arsenic compounds were much lower than those of inorganic arsenicals. The IC50 of DMAA was about 5 mM, and MAA and TMAO had no toxicity even at concentrations over 10 mM, and DMAA mainly induced apoptotic cell death. These data imply that methylation of inorganic arsenicals in mammalia play an important role to suppress both severe immunosuppression and inflammatory responses cause by inorganic arsenicals.

SUMMARY 14

NO	14-14
Theme	Comparative studies for unit chain distributions of different glycogens and evidence of multiple branching of oyster glycogen.
Reporter	Motoko Matsui(1), Mariko Kakuta(2) and Akira Misaki(3)
Position	(1)Ichimura Gakuin Junior College (2)Konan Women's University (3)Osaka City University
Volume	The 14th Symposium on Trace Nutrients Research(1997)
P	P.85-94
Summary	In a series of investigation on properties of oyster glycogen, the fine structural features of glycogen was elucidated by a new analytical strategy. Thus, the multi-branched oyster glycogen molecule (A: B-chain, 0.7: 1) involved 5 to 6 times interlinkings of B-chain. More information on the glycogen structure was investigated by peeling action by Klebsiella pullulanase. The first treatment liberated short unit-chains (dp2-5), but it was tended to produce longer unit chains (dp3-17), as the pullulanase actions proceeded. Interestingly, after the 7th treatments small proportions of longer unit-chains were still present, suggesting the presence of longer tires are in inner molecule. Cased on these data, we proposed a computerized three-dimensional model of oyster glycogen. The oyster glycogen was acted with human pancreatic amylase. The analysis by HPAEC indicated production of various maltosaccharides including double branched oligosaccharides. The afore mentioned isoamylolysis and HPAEC analysis was applied to the unit chain distributions of glycogens from different biological origins. It was also successfully applied for clarification of the glycogen of human glycogen storage disease.

SUMMARY 15

NO	14-15
Theme	Participation of minerals in various functionalized aqueous solution or "Function Water", in exertion of detoxification effect on organotin intoxidated Euglea gracilis
Reporter	Mari Ohta(1), Kaoru Nakamura(2), Kozo Takama(1) and Tetsuya Suzuki(1)
Position	(1)Laboratory of Food Wholesomeness, Department of Marine Bioresources Chemistry, Faculty of Fisheries, Hokkaido University (2)Institute of Chemical Research, Kyoto University
Volume	The 14th Symposium on Trace Nutrients Research(1997)
P	P.95-102
Summary	Processing water with various mineral materials, electrostatic field, magnetic field gives functionalized aqueous solution, so-called "Function Water" in Japan. Series of research in our laboratory has revealed that restoration of morphology and motility of Tributyltin-chloride (TBTCl) intoxicated protozoa, Euglena gracilis should be due to Ca and Mg ions in the processed aqueous solution, in order to examine whether the detoxification effect could be observed commonly in the so-called "Functional Materials", we compared the extent of detoxification effect of various commercially available materials by evaluating restoration of motility of TBTCl-intoxicated Euglena gracilis. Materials examined in the present study were reduced-state mineral concentrate, glassy surface ceramic, and zeolite materials partly substituting Na with Fe, Mg, Zn, and Cu separately. Processed aqueous solutions were prepared by dissolving mineral concentrate in different concentrations, or immersing the materials for a period of time. TBTCl-intoxicated Euglena gracilis cells were separately washed with the processed aqueous solutions, then incubated in the solutions for up to 3 hrs. The restoration of morphology and motility was evaluated by observing the motile cell number under the video-microscope. Remarkable restoration was observed in the 250 folds diluted reduced mineral concentrate and solutions treated with zeolites encaging Fe, Mn, and Zn, respectively. However, those solutions did not show any restoration effect when they were treated with a chelator, Chelex-100. These results suggest that minerals should take part as the critical role in detoxification of TBTCl in Euglena cell. It may also be considered that glassy surface ceramic treatment should be explained by other mechanisms.

SUMMARY 16

NO	14-16
Theme	Effect of oyster extracts on the recovery of cell motility of TBTCl-intoxicated Euglena gracilis Z
Reporter	Tetsuya Suzuki(1), Mari ohta(1), Kaoru Nakamura(2), and Kozo Takama(1)
Position	(1)Laboratory of Food Wholesomeness, Department, of Marine Bioresoueces Chemistry, Faculty of Fisheries, Hokkaido University (2)Institute of Chemical Research, Kyoto University
Volume	The 14th Symposium on Trace Nutrients Research(1997)
P	P.103-112
Summary	Since oyster is an excellent source of minerals and taurine, it has been regarded as a health food. The authors have studied the biological role of role trace minerals on the detoxification of xenobiotics by using Tributyltin chloride (TBTCl)-intoxicated Euglena gracilis Z. In the present study, we examined detoxification effects by evaluating the recovery of cell motility for different kinds of oyster extracts; i.e., hot water extract of whole oyster flesh, extract rich in low molecular wt. Fraction, extract rich in high molecular wt. Fraction, and antioxidant rich fraction. After intoxication with 50 μM TBTCl, E. Gracilis Z cells were washed with aqueous solutions containing 4 different oyster extracts separately at the dilution ratio of 100-100,000-fold, then incubated for 0-180 minutes at 28°C under illumination (2800 lx). Recovery of cells motility was examined under the video microscope. Results showed that hot water extract at 1,000-fold dilution and the extract rich in the high molecular wt. Fraction at 10,000 to 100,000-fold dilution was most effective on the recovery of cell motility. Since the treatment with a chelator, Cheles-100, suppressed the recovery of cell motility by trapping Ca, Mg and Fe in the solution, these minerals seem to have participated in the detoxification of TBTCl and/or the recovery of cell motility.

SUMMARY 17

NO	14-17
Theme	Studies on Interaction of Minerals in Intestinal Epithelial Cells by Using The Metal Stress Response as An Marker
Reporter	Chieko Uno, Yukiko Nakanishi, Sakiyo Koseki (Yamaoka) and Kyoden Yasumoto
Position	Research Institute for Food Science Kyoto University
Volume	The 14th Symposium on Trace Nutrients Research(1997)
P	P.113-117
Summary	Hemeoxygenase, the rate-limiting enzyme in the heme-degradative pathway, is induced by addition of several metal ions. Hemeoxygenase is considered to function in the cellular defense mechanism against oxidative stress. We studied the induction of hemeoxygenase in intestinal epithelial cells (Caco-2 cell line) exposed to various types of minerals and the possible role of this enzyme in preventing the oxidative damage. The level of induced hemeoxygenase was found to depend on the species of minerals used (NaAsO2, hemin, and CuCl2). Both NaAsO2 and hemin strongly induced hemeoxygenase, but CuCl2 did not. Coexistence of NaAsO2 and hemin furthered the induction of hemeoxygenase as compared with that observed when the compound was added separately. However, the simultaneous addition of CuCl2 suppressed the hemeoxygenase induced in the presence of either NaAsO2 or hemin. Furthermore, in any case, the oxidized proteins accumulated gradually as the induction of hemeoxygenase was reduced. These results indicate that minerals directly affected the induction of hemeoxygenase and suggest the possibility that dietary minerals may regulate the induction of hemeoxygenase and the generation of oxidized protein in intestinal cells.

SUMMARY 19

NO	14-19
Theme	Effect of Oyster Extract on Guinea-Pig Papillary Muscle
Reporter	Michio Yajima(1), Yoshikazu Matsuda(2), Masahiro Nakatsuka(2), Takao Ohta(2)
Position	(1)Dept. of Pharmacology, Aichi Medical University (2)Central Research Institute, Japan Clinic Co., Ltd.
Volume	The 14th Symposium on Trace Nutrients Research(1997)
P	P.123-127
Summary	The effect of oyster extract(s) (OE) on the electrically-driven (force of) contraction in guinea-pig papillary muscles was studied. OE (1mg/ml) produced a transient negative inotropic effect followed by continuous positive inotropic effect (PIE). Increases in PIE was 6%, 20% and 33% at the 0.1, 0.3 and 1mg/ml of OE concentrations, respectively. Pretreatment of catecholamine β1antagonist, metoprolol (3 x 10-7M), and catecholamine α1 antagonist, prazosin (10-7M), did not inhibit the PIE of OE (1mg/ml). However, calcium channel antagonist, nicardipine (10-7M), partially inhibit the PIE of OE. Diphenhydramine (10-7M), histamine (H1) antagonist, markedly inhibited the PIE of OE, whereas cimetidine (10-7M), histamine (H2) antagonist, had no effect. These findings suggest that the PIE of OE was mediated mainly by the stimulation of histamine (H1) receptors and partially by activation of L-type calcium channels.

SUMMARY 20

NO	14-20
Theme	Mechanism of Antioxidant Action of Flavonoids and Polyphenolics
Reporter	Keiko Murakami(1), Yoshikazu Matsuda(2), Masahiro Nakatsuka(2) and Masataka Yoshino(1)
Position	(1)Department of Biochemistry, Aichi Medical University (2)Central Laboratory, Japan Clinic Co., Ltd.
Volume	The 14th Symposium on Trace Nutrients Research(1997)
P	P.129-136
Summary	Antioxidant action of flavonoids and Polyphenolics was analyzed. Flavonoids including baicalein, baicalin, quercetin and rutin with little or no reducing activity enhanced the autooxidation of Fe2+, and further inhibited the ascorbate-mediated reduction of iron. On the contrary, Polyphenolics belonging to non-flavonoids such as protocatechuic acid and chlorogenic acid showed a potent iron-reducing ability, and prevented Fe2+ ion from autooxidation completely. Both Flavonoids and non-flavonoids effectively inhibited the formation of thiobarbituric acid-reactive substances as a marker of lipid peroxidation of microsomes from rat liver. Flavonoids acted as antioxidants, which inhibit the formation of superoxide and hydroxyl radical by oxidation of Fe2+ ion acting as prooxidant. On the other hand, antioxidant properties of non-flavonoids can be explained by the formation of inactive Fe2+-polyphenolic complexes, which cannot react with oxygen. Oxidation-reduction properties of iron was applied to analyze the extract of oyster, Crassostera gigas. The oyster extract protected the Fe2+ from autooxidation, but showed inhibition of the ascorbate-induced reduction of iron. Oyster extract showed a potent inhibitory effect on lipid peroxidation of microsomes. These results suggest that two types of antioxidant, that is, flavonoid and non-flavonoid types play a central role in antioxidant action of oyster extract.

SUMMARY 21

NO	14-21
Theme	Importance of Kynureninase
Reporter	Yukio Shibata(1), Masahiro Nakatsuka(1), Takao Ohta(1), Yoshikazu Masuda(1), Tomoyuki Shindo(1), Eiko Takaya(1), Hiroshi Ishizu(2), Taizo Yamada(3), Sukehisa Ishizu
Position	(1)Japan Clinic Co., Ltd., Central Research Institute (2)Ueki Hospital (3)Tamai Orthopedics and Internal Medicine hospital (4)Akura Hospital
Volume	The 14th Symposium on Trace Nutrients Research(1997)
P	P.137-142
Summary	It was reported about the chemical structure of Kynureninase (KYN) and other B6 relating enzyme for example decarboxylases of amino acid metabolism. 1.The amino acid sequence of rat liver KYN and AST (GOT) have similarity in amino acid sequence. 2.And the amino acid sequence of active center in KYN, CGL and CGS have also similar sequence of amino acid. 3.5-OH-Trp. Decarboxylase and DOPA decarboxylase activity decreased in Zn2+ administrated rats. 4.Metabolism of Lys. and Trp. were interrelated through 3-amino adipate aminotransferase. 5.Zn2+ content increased in brain-stem especially in Corpus Striatum using VB6 deficient rats. 6.And finally, histamine syntheses by His. Decarboxylase in diabetic rats were also discussed.

SUMMARY 22

NO	14-22
Theme	The effect of sulfur amino acid intake on the proliferation response of splenocytes in growing mice
Reporter	Yu Hosokawa(1), Nobuyo Tsuboyama(1), Ritsuko Masuyama(2), Tomiko Yoshihara(3), Masayuki Totani(1)
Position	(1)Division of Maternal and Child Health Science, National Institute of Health and Nutrition (2)Department of Nutrition, Faculty of Agriculture, Tokyo University of Agriculture (3)School of Home Economics, Tokyo Kasei University
Volume	The 14th Symposium on Trace Nutrients Research(1997)
P	P.143-147
Summary	The effects of dietary proteins containing different levels of sulfur amino acid on the responsiveness of splenocytes to concanavalin A (Con A) were examined. The Con A-induced DNA synthesis of splenocytes in mice fed a purified egg protein (PEP) diet was significantly higher than those in mice fed a soy protein isolate (SPI) or casein diet. Reduced glutathione (GSH) and 2-mercaptoethanol (2-ME) markedly stimulated Con A-induced CAN synthesis in vitro, and the stimulatory effects of 2-Me were more marked in SPI and casein diet groups compared to the PEP diet group. In contrast, L-buthionine- (S, R)-sulfoximine (BSO) markedly inhibited Con A-induced DNA synthesis in splenocytes. The degree of inhibition was greater in the order SPI, casein and PEP diet groups. These results suggest that the sulfur amino acids concentration in the dietary protein may play a key role in the regulation of lymphocyte responsiveness to Con A in growing mice.

SUMMARY 23

NO	14-23
Theme	Absorption in the Body of Inclusion Complex of γ-Cyclodexytrin with Octacosanol after Oral Administration in Mice
Reporter	Mieko Kawamura(1), Tatsuo Ido(2), Tadashi Funada(3), Itaru Miyamoto(3) and Tatsuya Sugawara(3)
Position	(1)Department of Applied Biochemistry (2)Cyclotron and Radioisotope Center, Tohoku University (3)Nippon Oil and Fats Co., Ltd.
Volume	The 14th Symposium on Trace Nutrients Research(1997)
P	P.149-153
Summary	Octacosanol (CH3 (CH2) 26-CH2OH) is a natural substance which was contained the rice, the wheat with germs, the sugarcane, and so on. Although the increased motor endurance and physical performance from octacosanol were reported, their results did not coincide reason why that octacosanol absorption in the body was very slight. This present studies clarified with radioactive tracer (3H-octacosanol) that absorption in the body of inclusion complex of γ-cyclodexytrin (CD) with octacosanol (CD-OCT) was 2 times higher in blood and each organ than that of liposome (Lip-OCT) or Tween-80 (TW80-OCT) complex in conventional mice after 24 hrs oral administration. In germ free mice, however, the absorption in the body of CD-OCT was not higher than that of Lip-OCT or TW80-OCT after 24 hrs oral administration. Octacosanol in the form of CD inclusion complex was absorbed markedly, this result suggest the participation of intestinal flora on the process of intestinal absorption.

SUMMARY 24

NO	14-24
Theme	Effect of dietary level of phytic acid on lipid metabolism in rats fed on sucrose.
Reporter	Shoko Onomi, Tetsuyuki Katayama
Position	Laboratory of Nutritional Science, Faculty of Education, Hiroshima University
Volume	The 14th Symposium on Trace Nutrients Research(1997)
P	P.155-159
Summary	Sucrose-fed rats elevate hepatic concentrations of total lipids or triglyceride and hepatic activities of lipogenetic enzymes in comparison with corn starch-fed rats. In general, the elevations that had been caused by sucrose feeding were gradually suppressed by the increasing dietary phytate. Activities of intestinal enzymes in sucrose-fed rats were unaffected by dietary phytate. The curative effect of dietary phytate on fatty liver in rats caused by sucrose feeding may be mediated through enzymes. These results also suggest that practical level of dietary phytate may affect hepatic lipid metabolism in animals fed on sucrose.

SUMMARY 25

NO	14-25
Theme	Effect of strenuous exercise on blood constituent Π
Reporter	Kenji Fukunaga, Munehiro, Yoshida, Toshiko Ono and Naoki Nakazono
Position	Department of Public Health Kansai Medical University
Volume	The 14th Symposium on Trace Nutrients Research(1997)
P	P.161-165
Summary	The effect of strenuous exercise on blood constituent was examined. Studies were made on healthy 13 members (18 to 27 years) of the canoe club of a medical university who participated in summer training camp for 7 days. Situation of food intake on before and during training camp were also evaluated. Intake of total energy, protein, and carbohydrate during training camp were increased when compared with that of before training camp, but intake of fat as not changed. After training camp, serum total protein, albumin, and total electrolyte (Na+, K+, Ca2+) were not affected. Serum GOT, GPT, LDH, and CPK activities were significantly increased. RBC, Hb, Ht and serum haptoglobin were significantly decreased and reticulocyte was increased after the camp. Serum Zn level were significantly decreased but Cu and Fe levels were not changed. Serum antioxidative substance (ascorbic acid and α-tocopherol) levels were significantly decreased, while lipid peroxide level (malondialdehyde) was significantly increased. These data suggest that strenuous exercise can result in a decrease of RBC, Hb, and Ht with oxidative damage.

SUMMARY 26

NO	14-26
Theme	Acidic fibroblast growth factor activates hypothalamo-pituitary-adrenocortical axis
Reporter	Kazuo Sasaki(1), Itsuro Matsumoto(2), Yutaka Ohmura(3) and Akira Niijima(4)
Position	(1)Fac. Of Eng., Toyama Univ. (2)Fac. Of Med., Nagasaki Univ. (3)Inst. Of Bio-Active Sci., Nippon Zoki Pharmaceu. Co. (4)Fac. Of Med., Niigata Univ.
Volume	The 14th Symposium on Trace Nutrients Research(1997)
P	P.167-170
Summary	Effects of acidic fibroblast growth factor (aFGF) on the hypothalamo-pituitary-adrenocortical axis were examined in anesthetized rats. Intracerebroventricular (i.c.v.) and intravenous (i.v.) injections of aFGF increased the level of plasma corticosterone. Pretreatment of i.c.v. anti-CRF antibody abolished the increase of plasma corticosterone level induced by i.c.v. aFGF injection, but had no effect on that elicited by i.v. aFGF injection. The plasma ACTH level was increased by i.v. aFGF injection. The results indicate that centrally and peripherally applied aFGF activates the adrenocortical system via the CRF release and the ACTH release, respectively.

SUMMARY 27

NO	14-27
Theme	Biotin Content in Kampo Medicines for Skin Disease
Reporter	Toru Fukui(1) and Toshiaki Watanabe(2)
Position	(1)Clinical Laboratory, Byotai-Seiri Laboratory (2)Department of Hygiene and Preventive Medicine, Yamagata University School of Medicine
Volume	The 14th Symposium on Trace Nutrients Research(1997)
P	P.171-176
Summary	We studied the relation between biotin and Kampo medicines (traditional Chinese herbal medicines) for skin disease. The subjects consisted 14 Kampo medicines and their 24 crude powder materials. Kampo medicines were classified into three categories: Group I - 3 medicines which are generally prescribed for atopic dermatitis, eczema and pruritus, Group Π-8 medicines for eczema, urticaria, pruritus, and allergy, Group III - 3 control medicines for non-skin disease. Biotin assay was performed by the agar plate method using Lactobacillus plantarum ATCC 8014. The average content of free biotin in GroupI,Π and III was 118.0&plasmn;4.6, 97.8&plasmn;48.6 and 30.3&plasmn;19.6 ng/g on average, respectively. On the other hand, the free biotin in 11 crude powder materials was higher than 100 ng/g, and these materials were frequently used for making Kampo medicines for skin disease. It is known that these Kampo medicines have been effective against skin disease from ancient time. In this study, it is obvious that Kampo medicines contain a large amount of free biotin, which can be easily absorbed by the small intestine.

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