Proceedings The 10th Symposium on Trace Nutrients Research (1993)
Kyoto, June 23-24, 1993

NO	Theme	Reporter	Position	P
1	Inhibition of prostaglandin D synthesis by inorganic quadrivalant selenium compounds in vitro and in vivo	Osamu Hayaishi	Osaka Bioscience institute	1-
4	Influence of Oligopeptide to lipid metabolism	Sukehisa Izuta(1), Maki Hirose(1),Tadayoshi Fujita(1), Yoshikazu Matsuda(2),Takao Ohata(3), Masahiro NAkatsuka(3),and Yukiko Shibata(3)	(1)Agura Hospital (2)Japan Clinic Co., Ltd, Central Research Institute (3)Department of Biochemistry, Aichi Medical University	33-
5	Immunodeficiency and alopecia induced by zinc deficiency	Yasuaki Arakawa(1), Hiroaki Sakazaki(1), Takao Kurriyama(1), Harunobu Nakashima(2), Shinjiro Hori(2), Sayako Ohmori (3), Takayuki Takeuchi(4) , Yukihiro Nakano(4), Akira Morita(5), Toru Iwasaki(5), and Hideo Koishi(6)	(1)Department of Hygiene & Preventive Medicine, Faculty of Health Sciences, The University of Shizuoka (2)Osaka Prefectural Institute of Public Health (3)Ohzuma Women University (4)Research Reactor Institute, Kyoto University (5)Animal Experimental Center (6)Department of Human Nutrition, Faculty of Health Sciences, The University of Shizuoka	39-
6	Protection of D-Galactosamine Induced Hepatotoxicity by Zinc Metallothionein	Masaya Ikegawa, Mieko Kimura, Hirohide Tamura, and Yoschinori Itokawa	Kyoto University Graduate School of Medicine Department of Social Medicine	49-
7	Effect of Zinc deficiency on Brain Zinc Concentrations in Rats	Atsushi Toyoda(1), Takuma Shiraki(1), Fumiko Yano(2), Tohru Matsui(3), and Hideo Yano(3)	(1)Department of Neuroscience, Graduate School of Human and Environmental Studies, Kyoto University (2)Department of Biotechnological Science Faculty of Biology-oriented Science and Technology, Kinki University (3)Department of Animal Science, Faculty of Agriculture, Kyoto University	55-
8	Effect of Zinc Supplementation on Thyroid Hormone Status in Disable Patients with Zinc Deficiency	Sohroku Nishiyama(1), Tosiro Anakayama(1), Ichiro Matsuda(1) and Yoshio Sawada(2)	(1)Department of Pediatrics (2)Kumamoto Institute of Total Fitness	59-
9	Mechanism of Generation of Active Oxygen and Role pf Iron in Skeletal Muscle Atrophied by Immobilization	Hisao Kondo and Yoshinori Itokawa	Department of Social Medicine, Graduate School of Medicine, Kyoto University	63-
10	Behavior of Ascorbic Acid after long Term Drinking of Vanadate	Yukiko Nakanishi(1), Mieko Kawamura(1), Tatsuo Ido(2), Ren Iwata(2), and Syuichi Kimura(3)	(1)Faculty of Agriculture (2)Cyclotron and Radioisotope Center, Tohoku University (3)Showa Women's University, Graduate School, Course of Science for Living System	77-
11	Distribution and Physiological Roles of Vanadium	Hiromu Sakurai and Masaki Nakai	Department of Analytical Chemistry, Kyoto Pharmaceutical University	83-
12	Iron Overload Secondary to Selenium Deficiency in Rat	Nawarath Chareonpong-Kawamoto and Kyoden Yasumoto	Research Institute for Food Science, Kyoto University	87-
13	Intake and excretion of selenium and dietary selenium levels to maintain balance in adult females consuming ordinary Japanese foods	Ikuo Sato(1), Shiro Niizeki(2), and Kenji Yamaguichi	(1)Sanyo Gakuen Junior College (2)The National Institute of Health and Nutrition (3)Tokyo Kansai University	93-
14	A Longitudinal Study on the Relationship of Serum Selenium and Cholesterol Concentration in Japanese Rural Women	Yoji Deguchi	Department of Environment Health, Fukui Medical School	99-
15	Effect of Administration with Anti-thyroid Drug and Selenium Deficiency on Urinary Ketone Body Excretion in Rat	Munehiro Yoshida	Department of Public Health, Kansai Medical University	103-
16	Effects of methionine selenium deficiency on selenium and selenoenzyme in rats	Zongjian Zhu, Mieko Kimura and Yoshinori Itokawa	Department of Hygiene, faculty of Medicine, Kyoto University	107-
17	Glutathione peroxitase like reaction catalyzed by glutaselenone diastereomers.	Takashi Tamura, Hitoshi Nakazato, Nobuyoshi Esaki and Kenji Soda	Institute for Chemical Research	113-
18	The movement of calcium binding protein (D-28K) in Vitamin B6 deficient rat	Ryoko Tsubouchi(1),Tsunemasa Nonogaki(2), Fumio Takeuhi(1), Hideo Fuse(1), Shigehisa Aoki(2) and Shibata Yukio	(1)Department of Biochemistry, Aichi medical University (2)2nd Department of Pathology, Aichi medical University	117-
19	Vertebral Bone Mineral Density in Children	Takeaki Inomoto(1), Soroku Nishiyama(2), Shin-ichi Tomoeda(3), Nobutake Nakane(3), Hiroyuki Yonemitsu(3), Ichiro Matsuda(2), Yoshio Sawada(1)	(1)Kumamoto Institute of Total Fitness (2)Department of Pediatrics, Kumamoto University, medical School (3)Kumamoto Kinoh Hospital	121-
20	Effect of Caseinphosphopeptide on Mineral Metabolism in Rats	Miho Hirabayashi, Kin-ya Ashida, Tohru Matsui and Hideko Yano	Department of Animal Science Faculty of Agriculture, Kyoto University	127-
21	Effect of the Administration of Toxic Amounts of Copper with Other Chelating Compounds such as EDTA, Thiomolybdate, Ascorbate and Deferoxamine on Growth, Mortality and Hepatic Minerals of Chick Embryos	Tatsuo Hamada and Emiko Nakahama	National Institute of Animal Industry	131-
22	Effect of Aging on Trace Elements in Female Mice	Akemi Morita, Mieko Kimura and Yoshinori Itokawa	Department of Animal Social Medicine, Graduate School of Medicine, Kyoto University	139-
23	Tin Deficiency in Rats	Katsuhiko Yokoi, Mieko Kimura and Yoshinori Itokawa	Department of Animal Social Medicine, Graduate School of Medicine, Kyoto University	145-
24	A high Sensitive Determination of Glutathione and Glutathione disulfide by HPLC with Post-Label Fluorometric Detection	Kenji Fukunaga, Tetsuya Suzuki and Kozo Takayama	Department of Food Science and Technology, Faculty of Fisheries, Hokkaido University	151-
25	Primary Structure and Functional Expression of the Cysteine Dioxygenase (CDO) cDNA	Yu Hosokawa(1), Nobuyo Tsuboyama(2), Hitomi Tojo(3) and Masayuki Totani(1)	(1)The National Institute of Health and Nutrition (2)Kochi Medical University (3)Kanagawa Prefectural Junior College of Nutrition	155-
26	Effect of Vitamin A Deficiency on Fat Metabolism in Rats	Masayuki Kitagawa(1), Tadao Umeki(2), Yuko Kai(2), Takezo Nishino(2), Hideo Yano(1) and Ryoji Kawashima(1)	(1)Faculty of Agriculture, Kyoto University (2)Faculty of Agriculture, Yamaguchi University	161-
27	Effect of ethanol on rat brain gangliosides	Michiko Adachi(1), Akihiko fujii(1), Tatsuo Ido(1), Syuichi Kimura (2)	(1)Cyclotron and Radioisotope Center, Tohoku University (2)Course of Science for Living System, Showa Women's University	165-
28	Effect of caffeine on brain neurotransmitters	Yoshihiro Fujitani and Tatsuo Ido	Cyclotron and Radioisotope Center, Tohoku University	171-

SUMMARY 1

NO	10-1
Theme	Inhibition of prostaglandin D synthesis by inorganic quadrivalant selenium compounds in vitro and in vivo
Reporter	Osamu Hayashi
Position	Osaka Bioscience institute
Volume	The 10th Symposium on Trace Nutrients Research(1993)
P	P.1-11
Summary	During the last 10 years, we showed that prostaglandins (PG) D2 and E2 are the major sleep regulating substances in the brain, each promoting sleep and wakefulness. Recently we discovered that various inorganic quadrivalent selenium compounds are potent, specific and non-competitive inhibitors of brain prostaglandin D synthesis Ki is about 10 μM. Hexavalent compounds and organic selenium compounds are nor inhibitory. Other enzymes in the arachidonate cascade system are not inhibited. This inhibited is reversible and can be reversed by sulfhydryl compounds such as Glutathione or DTT. When one of these inorganic quadrivalent selenium compounds was infused into the third ventricle of a freely moving rat, sleep was inhibited time- and dose-dependently and reversibly, indicating that PGD synthesis is the key enzyme in sleep regulation and that PGD2 is the endogenous sleep-regulating substance in the rat under physiological conditions. Because the sleep pattern of rats is somewhat different from that of primates, we further extended these studies to the Macaca mulatta, that is, the Rhesus monkey. More recently, we found that intraventricular infusion of prostaglandin E2 reduces the amount of diurnal sleep of rats, ore increases the amount of wakefulness.

SUMMARY 4

NO	10-4
Theme	Influence of Oligopeptide to lipid metabolism
Reporter	Sukehisa Izuta(1), Maki Hirose(1), Tadayoshi Fujita(1), Yoshikazu Matsuda(2), TAkao Ohata(3), MAsahiro NAkatsuka(3), and Yukiko Shibata(3)
Position	(1)Agura Hospital (2)Japan Clinic Co., Ltd, Central Research Institute (3)Department of Biochemistry, Aichi Medical University
Volume	The 10th Symposium on Trace Nutrients Research(1993)
P	P.33-37
Summary	There are many reports that oligopeptides effect on the lipase activity or fatty acid synthesis. In this report, we studied about the influence of oligopeptides on the lipid metabolism and the total body fat by the continuous administration of oligopeptide to21 lipid pathobolism patients and healthy controls. Tablets of oligopeptide were administered orally to each group for 3 months (200mg oligopeptide par 1 tablet 10 tablets par day). Blood test, measurement of body weight and total body fat were carried out once a month. The body weight and total body gat decreased in lipid pathobolism patients and healthy control subjects. We conclude that the intake of oligopeptide might be available for the lipid metabolism.

SUMMARY 5

NO	10-5
Theme	Immunodeficiency and alopecia induced by zinc deficiency
Reporter	Yasuaki Arakawa(1), Hiroaki Sakazaki(1), Takao Kurriyama(1), Harunobu Nakashima(2), Shinjiro Hori(2), Sayako Ohmori (3), Takayuki Takeuchi(4) , Yukihiro Nakano(4), Akira Morita(5), Toru Iwasaki(5), and Hideo Koishi(6)
Position	(1)Department of Hygiene & Preventive Medicine, Faculty of Health Sciences, The University of Shizuoka (2)Osaka Prefectural Institute of Public Health (3)Ohzuma Women University (4)Research Reactor Institute, Kyoto University (5)Animal Experimental Center (6)Department of Human Nutrition, Faculty of Health Sciences, The University of Shizuoka
Volume	The 10th Symposium on Trace Nutrients Research(1993)
P	P.39-47
Summary	A Zinc deficiency induces a severe immunodeficiency and alopecia. The mechanism of zinc deficiency-induced immunodeficiency was examines by analyzing, the thymus atrophy and changes in the membrane surface antigens of T cells of zinc-deficient SPF Wistar-derives rats. Zinc deficiency induces severe and reversible thymus atrophy. The zinc-deficient thymocytes revealed significant increases in the percentage of CD4 antigen cells (rat T helper cells and macrophages), CD5 antigen cells(rat T suppressor/cytotoxic cells) and CD4 & CD8 double-negative cells, and percentage of CD4 & CD8 double-positive cells (undifferentiated cells) and Thy 1,1 antigen cells (total T cells). In addition, significant decreases in the percentage of The 1,1 antigen cells, slight decreases in the percentage of α/β (T cell receptor) antigen cells and CD2 antigen cells (E rosette firming cells), and a significant increase in the percentage of CD4 & CD8 double-positive cells were observed in the peripharal T cells of the zinc-deficient rats. Furthermore, thymulin activity deceased significantly in the serum of zinc-deficient rats. These findings indicate that the immunodeficiency induced by zinc deficiency is manifested as T cell dysfunction which is caused by not only quantitative changes of their membrane surface antigens, that is impairment of the process of differentiation and maturation of T cell in the thymus. The mechanism of zinc deficiency-induced alopecia was examined by analyzing the movement of trace elements in the hair and fatty acids in the skin of zinc-deficient SPF Wistar-derived rats. Zinc deficiency induced sever and reversible alopecia. With the progress of falling-out of the hair (or with the decrease in the concentration of zinc in the hair), a marked tendency for accumulation of manganese and calcium into the hair was seen, and a decrease in the concentration of linoleic acid and an increase in the concentration of palmitic acid in the skin were seen. The relations of manganese and unsaturated fatty acids such as linolenic acids to alopecia are now under investigation.

SUMMARY 6

NO	10-6
Theme	Protection of D-Galactosamine Induced Hepatotoxicity by Zinc mMetallothionein
Reporter	Masaya Ikegawa, Mieko Kimura, Hirohide Tamura, and Yoschinori Itokawa
Position	Kyoto university, Graduate School of Medicine, Department of Social Medicine
Volume	The 10th Symposium on Trace Nutrients Research(1993)
P	P.49-53
Summary	D-Galactosamine (D-GalN) induced hepatotoxicity was protected by a zinc (Zn) pretreatment in rats. Wistar rats, weighing about 200g, were divided into two groups; group 1 (G1) served as a control and group 2 (G2) received zinc-sulphate (10mg/kg Zn) intraperitoneally (i. p.). After 24 hours, various amounts of D-GalN (0-800mg/kg) was administered i.p. and 24 hours later, all rats were sacrificed. Plasma activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST) were dose-dependently elevated in G1, while marked reduction in AST and ALT was observed in G2. In both groups, plasma activity of alkaline phosphates (ALP) was increased without a difference significant. Zinc concentration in liver, kidney and tibia in G2 is significantly higher (p<0.01) than in G1. With an increasing dose of D-GalN administered, hepatic concentration of Zn was decreased dose-dependently. Zn in brain, heart, testis was preserved at ant dose of D-GalN and was not affected by a zinc pretreatment either. Hepatic concentration of metallothionein (MT), quantified by a high performance liquid chromatography method, revealed a large increase in G2 and D-GalN administration produced a marked decrease in hepatic concentration of MT in G1 and G2. In conclusion, we elucidate that Zn-Mt could be metabolized to extra a protective effect against D-GalN hepatotoxicity.

SUMMARY 7

NO	10-7
Theme	Effect of Zinc deficiency on Brain Zinc Concentrations in Rats
Reporter	Atsushi Toyoda(1), Takuma Shiraki(1), Fumiko Yano(2), Tohru Matsui(3), and Hideo Yano(3)
Position	(1)Department of Neuroscience, Graduate School of Human and Environmental Studies, Kyoto University (2)Department of Biotechnological Science Faculty of Biology-oriented Science and Technology, Kinki University (3)Department of Animal Science, Faculty of Agriculture, Kyoto University
Volume	The 10th Symposium on Trace Nutrients Research(1993)
P	P.55-58
Summary	There are three separate pools of brain zinc, i.e., free zinc, vesicular zinc, and protein-bound zinc. Each of them has physiological important roles. We suspected that behavioral disorders were induced by some change of brain zinc concentration during zinc deficiency. Twenty-seven male Wistar-strain rats (four weeks age) were divided into three groups: zinc deficient (ZD), pair fed (PF), and ad libtum (AD). Zinc concentrations in diets were 1mg/kg (ZD) and 30mg/kg (PF, AD). After three weeks of dietary period, the hippocampus, the cerebellum and the striatum were collected, and whole zinc concentrations in these tissues were measured by atomic absorption spectrophotometer. Moreover, homogenized hippocampus was divided two fractions ( >5kDa and <5kDa) by the dialectic filter, and zinc concentrations in these filtrate ( >5kDa) were measured in the same way. Zinc deficiency remarkably decreased the body weight gain and feed intake. In addition, plasma zinc concentration was lowered in the zinc concentration in brains and ultrafiltable zinc in the hippocampus were not changed.

SUMMARY 8

NO	10-8
Theme	Effect of Zinc Supplementation on Thyroid Hormone Status in Disable Patients with Zinc Deficiency
Reporter	Sohroku Nishiyama(1), Tosiro Anakayama(1), Ichiro Matsuda(1) and Yoshio Sawada(2)
Position	(1)Department of Pediatrics (2)Kumamoto Institute of Total Fitness
Volume	The 10th Symposium on Trace Nutrients Research(1993)
P	P.59-62
Summary	We examined zinc status in relation to thyroid hormone in disabled persons. Out of 134 patients, thirteen patients revealed low levels of serum free T3 and T3, elevated levels of serum reverse T3 and elevated reaction of TSH after TRH injection. The subjects also showed increase urinary zinc excretion, moderately zinc deficiency evaluated by total body zinc clearance and moderate to sever brain atrophy estimates by computes tomographic scan. After supplementation of zinc sulphate 5-10 mg per kg body weight to 13 disabled patients for 12 months, it was shown that levels of serum free T3 and T3 increased accompanied by decrease in serum reverse T3 and TSH injection. Serum concentration of selenium did not change in patients during zinc supplementation. The findings of the present study suggest that zinc may be related to the active mechanism of TypeII deiodinase which appears to carry out only 5'-deidination.

SUMMARY 9

NO	10-9
Theme	Mechanism of Generation of Active Oxygen and Role pf Iron in Skeletal Muscle Atrophied by Immobilization
Reporter	Hisao Kondo and Yoshinori Itokawa
Position	Department of Social Medicine, Graduate School of Medicine, Kyoto University
Volume	The 10th Symposium on Trace Nutrients Research(1993)
P	P.63-75
Summary	To clarify the mechanism of oxidative stress in skeletal muscle atrophied by immobilization, we measured the activities of antioxidant enzymes and xanthine oxidase (XOD) in a typical slow red muscle, the soleus. Male Wistar rats (14 wk old) whose one ankle joints were immobilized in the fully extended position were killed after 4, 8 and 12 days. The activities of Mn-containing superoxide dismutase (Mn-SOD), Cu, Zn-containing superoxide dismutase (Cu, Zn-SOD), Se-dependent glutathione peroxidase (Se-GSHPx), glutathione S-transferase, catalase, glutathione reductase were measured spectrophotometrically. The XOD activity and the concentrations of hypoxanthine, xanthine and urate were measured using a high performance liquid chromatography. Increased Cu, Zn-SOD and decreased Mn-SOD in atrophy might reflect increased generation of superoxide anions in the cytoplasm rather than in the mitochondria. The source of superoxide anions in the cytoplasm might be the increased superoxidase-producing XOD. Enhanced generation of superoxidase anions and increased Cu, Zn-SOD activity in atrophy suggested the enhanced generation of hydrogenvperoxide in the cytoplasm. Owing to the unchanged activity of Se-GSHPx and the unchanged or slightly increased activity of activity of catalase in atrophy, the ability to degrade hydrogen peroxidase might not increase so much. Hence, hydrogen peroxide is expected to be increased in atrophy. Because there is also an increase of iron in the microsome of atrophied muscles, the production of hydroxyl radicals, the most aggressive of radicals, might consequently be elevated. Key Words: muscle atrophy, oxidative stress, immobilization, iron, xanthine oxidase

SUMMARY 10

NO	10-10
Theme	Behavior of Ascorbic Acid after long Term Drinking of Vanadate
Reporter	Yukiko Nakanishi(1), Mieko Kawamura(1), Tatsuo Ido(2), Ren Iwata(2), and Syuichi Kimura(3)
Position	(1)Faculty of Agriculture (2)Cyclotron and Radioisotope Center, Tohoku University (3)Showa Women's University, Graduate School, Course of Science for Living System
Volume	The 10th Symposium on Trace Nutrients Research(1993)
P	P.77-81
Summary	We have been reported that the effects on neuroreceptor and metabolic analysis of vanadate for the correlation between neurological disorder and vanadium. This report is of biodistribution studies on the location of ascorbic acid in these animals. A solution of sodium ortovanadete (Na3 VO4, 100ppm) was administered into Wister rats as a drinking water for 10 months. In serum, liver, spleen, kidney and adrenal, the concentration of ascorbic acid were unchanged. The level of ascorbic acid was decreased in brain only with long-term drinking of vanadate. A reduction of ascorbic acid in each part of brain was different.

SUMMARY 11

NO	10-11
Theme	Distribution and Physiological Roles of Vanadium
Reporter	Hiromu Sakurai and Masaki Nakai
Position	Department of Analytical Chemistry, Kyoto Pharmaceutical University
Volume	The 10th Symposium on Trace Nutrients Research(1993)
P	P.83-86
Summary	As vanadium has a wide broad of redox potential, which depend on oxidation states and differ from the potentials of iron or copper ions, various physiological roles have been expected. For understanding the physiological and pharmacological roles of vanadium compounds, both distributions of vanadium, determined by neutron activation analysis (NAA) method and kinetics of vanadium, estimated by blood circulation monitoring electron spin resonance (BCM-ESR) method ware examined. When rats were treated with vanadyl sulfate (+4), vanadium was well incorporated in both normal and STD-diabetic rats, with vanadyl sulfate(+4) or sodium vanadate (+5), vanadium concentrations of nuclear fractions decreased and those of mitochondria and microsomal fractions increased, as the administered vanadium was elevated. Vanadium concentrations in nuclear fractions were higher in the rats treated with vanadyl sulfate than those with sodium vanadate. When rats were given vanadyl sulfate by i. v. injections, ESR spectra with a hyperfine structure of eight line signals due to vanadyl ions appeared immediately after the injection and decreased slowly. This BCM-ESR method can be applied for the real time analysis of not only blood vanadium concentrations but metal irons in the paramagnetic states.

SUMMARY 12

NO	10-12
Theme	Iron Overload Secondary to Selenium Deficiency in Rat
Reporter	Nawarath Chareonpong-Kawamoto and Kyoden Yasumoto
Position	Research Institute for Food Science, Kyoto University
Volume	The 10th Symposium on Trace Nutrients Research(1993)
P	P.87-92
Summary	The present study was aimed the effects of dietary of selenium (Se) and/or dietary excess of iron (Fe) on the iron levels in serum and various tissues of rats. The animals were fed either of following diets : Se-deficient/Fe-adequate (0.0035% Fe as ferric citrate) , Se-adequate (0.1ppm Se as sodium selenite)/Fe adequate, Se-deficient/Fe-excess (0.25% carbonyl Fe) and, Se-adequate /Fe-excess. Rats were killed after 30 weeks of feeding period and the levels of Fe and Se in serum, liver, kidney, spleen, heart and lung were determined by and most of analyzed tissues of Se-deficient rats than those of Se-adequate rats. The conclusion derived is that excess dietary Fe did not augment the conditions produced by Se deficiency. These results confirm the previously held notion that Se deficiency increases Fe levels in serum and other tissues because of Fe liberated by promoted hemolysis.

SUMMARY 13

NO	10-13
Theme	Intake and excretion of selenium and dietary selenium levels to maintain balance in adult females consuming ordinary Japanese foods
Reporter	Ikuo Sato(1), Shiro Niizeki(2), and Kenji Yamaguichi(3)
Position	(1)Sanyo Gakuen Junior College (2)The National Institute of Health and Nutrition (3)Tokyo Kansai University
Volume	The 10th Symposium on Trace Nutrients Research(1993)
P	P.93-98
Summary	To assess the selenium (Se) status of Japanese, ten 5-days balance study for Se were carried out in 60 healthy female subject aged 20-22 years. Experimental diets composed of ordinary foods and were typical of Japanese usual dietary habits. The same menu was used in each experiment during the 7 experimental days. Each experimental diet was characterized by protein source in the diets. Bread in the diets in Exp. 1-9 was substituted for rice in the diet in Exp. 10. The overall average with SD of dietary Se intake was 89±25 g/day, ranged from 4 to 141 μg/day. Apparent absorption, expressed as ratio of fecal excretion to intake, was 74±11% Se balance showed positive except Exp. 10. Se retention was 18±23μg/day. Se intake was significantly and positively correlated with urinary excretion (r=0.376), urinary and fecal excretion (r=0.556), and Se balance (r=0.834), Regression of Se balance on dietary Se intake indicated that these subject needed 52μ of dietary. Se per day to stay in balance. When balance and intake were adjusted for body weight, they needed 1μg of selenium per kg of body weight to maintain balance.

SUMMARY 14

NO	10-14
Theme	A Longitudinal Study on the Relationship of Serum Selenium and Cholesterol Concentration in Japanese Rural Women
Reporter	Yoji Deguchi
Position	Department of Environment Health, Fukui Medical School
Volume	The 10th Symposium on Trace Nutrients Research(1993)
P	P.99-102
Summary	Serum samples were collected from 167 Japanese rural women (age 35-81 years) in 1990 and from the same subject in 1992. Atherogenic index [= (total cholesterol - HDL -cholesterol)/HDL cholesterol] was a significant increasing factor of the serum selenium (Se) concentration in 1990 for women aged above 60 years by multiple regression analyses, which was consistent with our preliminary findings for coastal inhabitants of Fukui Pref. in 1986. Furthermore, the change in atherogenic cholesterol concentrations (total cholesterol - HDL-cholesterol/HDL-cholesterol) between the sampling years was a significant increasing factor of the changes in serum Se concentrations for all subjects. Thus our results suggest that serum Se concentration can increase as atherogenic cholesterol concentration increases for Japanese women and such a positive relationship may reflect some role of serum selenium in protecting atherogenic lipoproteins from oxidation.

SUMMARY 15

NO	10-15
Theme	Effect of Administration with Anti-thyroid Drug and Selenium Deficiency on Urinary Ketone Body Excretion in Rats
Reporter	Munehiro Yoshida
Position	Department of Public Health, Kansai Medical University
Volume	The 10th Symposium on Trace Nutrients Research(1993)
P	P.103-106
Summary	Effect of administration with 6-prople-2-thiouracil (PTU) to selenium (Se)-deficient or-adequate rats on the urinary ketone body excretion was studied. Rats (50 to 60g) were fed on Torula yeast-based Se deficient diet (Se content, <0.01μg/g) or the diet supplemented with sodium selenite (0.01μg Se/g). The rats were given 0.05%RTU solution or demonized water as a drinking water. After the feeding for 6 weeks, the PTU -loading made a severe inhibition in the growth. The rats feed the Se-deficient diet showed remarkably low hepatic Se contents and Glutathione peroxidase activities compared to the Se-adequate rats irrespective of the PTU-loading. Serum thyroid hormone levels were remarkably decreased in the PTU-received rats. In the rats without PTU, the Se deficiency made significantly higher serum T4 and lower T3 levels. Urinary ketone body excretion during the fasting was significantly higher in the Se-deficient rats than in the adequate rats irrespective of serum thyroid hormone levels. These results indicate that the increase of urinary ketone body excretion in the Se-deficient rats is independent to the serum thyroid hormone levels.

SUMMARY 16

NO	10-16
Theme	Effects of methionine selenium deficiency on selenium and selenoenzyme in rats
Reporter	Zongjian Zhu, Mieko Kimura and Yoshinori Itokawa
Position	Department of Hygiene, faculty of Medicine, Kyoto University
Volume	The 10th Symposium on Trace Nutrients Research(1993)
P	P.107-111
Summary	Studies were performed to clarify the effects of methionine and/or selenium deficiency and different forms of selenium (sodium selenate and selenomethionine) on selenium metabolism and the enzymatic activities of type I iodothyronine 5' deiodinase and Glutathione peroxidase. Forty-two weanling male Wister rats were divided into six groups and pairing fed the respective diets for 4 wk. The results showed that methionine-deficiency interfered with selenate absorption which induced lower selenium concentration, type I 5' deiodinase and glutathione peroxidase activities in liver whereas did not affect selenomethionine absorption. Glutathione of liver was decreased in methione-deficiency which may account type I 5' deiodinase and Glutathione peroxidase were decreased in selenium-deficiency. It is suggested that methionine-deficiency diminishes selenoenzymatic activities not only glutathione peroxidase but type I 5' deiodinase which may be due to the bad absorption or availability of selenium.

SUMMARY 17

NO	10-17
Theme	Glutathione peroxitase like reaction catalyzed by glutaselenone diastereomers
Reporter	Takashi Tamura, Hitoshi Nakazato, Nobuyoshi Esaki and Kenji Soda
Position	Institute for Chemical Research
Volume	The 10th Symposium on Trace Nutrients Research(1993)
P	P.113-116
Summary	Selenium analogue of glutathione (glutaselenone) catalyzes a glutathione peroxidase-like reaction. We found that glutaselenone and glutathione from a selenosulfide conjugate intermediate in the presence of the selenosulfide in the presence of glutaselenone diselenide and seleninic acid: they were converted to the selenosulfide in the presence of glutathione and hydrogen peroxide. Therefore, it appeared that the selenosulfide is essentially the catalyst in reduction of peroxide and oxidation of glutathione. Glutathione peroxidase-like activity is usually determined by the glutathione reductase method (GR-method). Four diastereomers of glutaselenone were different in catalytic activity from one another, when the activity was determined by GR-method. However, their activities were essentially the same when determined by the o-phthalaldehyde method (OPA-method). GR acts on glutaselenone diselenide and selenosulufide, and the discrepancy can be attributed to the action of GR among these selenium containing peptides.

SUMMARY 18

NO	10-18
Theme	The movement of calcium binding protein (D-28K) in Vitamin B
Reporter	Ryoko Tsubouchi(1),Tsunemasa Nonogaki(2), Fumio Takeuhi(1), Hideo Fuse(1), Shigehisa Aoki(2) and Shibata Yukio(1)
Position	(1)Department of Biochemistry, Aichi medical University (2)2nd Department of Pathology, Aichi medical University
Volume	The 10th Symposium on Trace Nutrients Research(1993)
P	P.117-120
Summary	The calcium-binding protein (CaBP) has been purified from rat kidney. The purified CaBP was about 28000 molecular weight and was detected in kidney and brain. The tissue distribution and the content of CaBP were determined in normal and vitamin B6 (B6) deficient rats. In rat kidney, CaBP was present as the distal tubules and located in nuclei and cytoplasm. There was no difference in CaBP distribution between two groups. However, the content of CaBP in B6deficient rats was significantly lower than that in normal rats. The content of serum calcium (Ca) in Ca deficient rats was the former was significantly higher than that in Ca administrated rats. On the other hand, the content of renal CaBP in the former was significantly higher than that in the latter. Thus the content of renal CaBP was inversely proportional to the content of serum Ca. We inferred that the decrease of the content of the renal CaBP was at least one factor in the formation of renal calculi observed in B6deficiency rats.

SUMMARY 19

NO	10-19
Theme	Vertebral Bone Mineral Density in Children
Reporter	Takeaki Inomoto(1), Soroku Nishiyama(2), Shin-ichi Tomoeda(3), Nobutake Nakane(3), Hiroyuki Yonemitsu(3), Ichiro Matsuda(2), Yoshio Sawada(1)
Position	(1)Kumamoto Institute of Total Fitness (2)Department of Pediatrics, Kumamoto University, medical School (3)Kumamoto Kinoh Hospital
Volume	The 10th Symposium on Trace Nutrients Research(1993)
P	P.121-125
Summary	The relationship between the physical performance and the vertebral bone mineral density was studied in 138(70 boys and 68 girls) normal children aged six to thirteen. The physical performance evaluated by the Sports activity and Bone mineral density of the lumbar vertebrae (L2-4BMD) was determined in the anteroposterior direction by the dual energy X-ray absorptiometry method (DEXA, QDR-10000, Hologic). The reported incidence of fracture varies; in general, it is highest among 12-year-old children, followed by 11-year-old children. Bone mineral density of the lumber vertebrae increased with height, weight, and age. The grip strength correlated significantly with the vertebral BMD. In the age of 11 and 12 years, vertebral BMD was significant hither in girls than in boys (P<0.05). The deleterious effects if immobilization and inactivity on bone are well known. Our data suggest that a positive relationship between physical activity and bone mineral density in lumber vertebrae.

SUMMARY 20

NO	10-20
Theme	Effect of Caseinphosphopeptide on Mineral Metabolism in Rats
Reporter	Miho Hirabayashi, Kin-ya Ashida, Tohru Matsui and Hideko Yano
Position	Department of Animal Science Faculty of Agriculture, Kyoto University
Volume	The 10th Symposium on Trace Nutrients Research(1993)
P	P.127-130
Summary	Effect of caseinphosphopeptide (CPP) on mineral metabolism was investigated in rats. Animals were fed a control diet (CPP%) or CPP diet (CPP0, 1, 0.3, 0.5%). All diets contained the same amounts of calcium (0.4%) , phosphorus (0.8%) and zinc (10.5mg/kg). Though an increase in the calcium retention was not observed, the deposition of calcium in a femur was greater in the CPP diet groups than those in the control diet group. Apparent absorption and retention of zinc were greater in two of the CPP diet groups (0.3,0.5%). Apparent absorption and retention of iron were grater in the all CPP diet groups. These results suggest that CPP supplementation may have a significant effect on the absorption of trace elements.

SUMMARY 21

NO	10-21
Theme	Effect of the Administration of Toxic Amounts of Copper with Other Chelating Compounds such sa EDTA, Thiomolybdate, Ascorbate and Deferoxamine on Growth, Mortality and Hepatic Minerals of Chick Embryos
Reporter	Tatsuo Hamada and Emiko Nakahama
Position	National Institute of Animal Industry
Volume	The 10th Symposium on Trace Nutrients Research(1993)
P	P.131-137
Summary	We administered copper and other chelating or reducing compounds directly into the air sacs of 14-day-old fertile eggs and after 5 days of incubation we examined embryonic growth, mortality and hepatic minerals of chick embryos. Administration of EDTA-2Na with copper (II) stimulated copper absorption and hepatic copper accumulation while that of tetrathiomolybdate with copper completely suppressed them. Although other agents such as deferoxamine mesylate, bathophenanthroline sulfonic acid, L-ascorbic acid and D-penicillamine could alleviate the copper toxicity to some extent, they were not so effective as EDTA or thiomolybdate in either stimulatory or suppressive ways for the hepatic copper accumulation. Examination of the hepatic minerals (Cu, Zn, Fe) revealed that the treatment group which had a higher death rate from excess copper absorption showed significantly higher tissue iron deposition probably due to the occurrence of hemolysis. These results suggest that excess copper absorption and that both free copper and free iron contribute to produce harmful radical species for membrane lipid peroxidation.

SUMMARY 22

NO	10-22
Theme	Effect of Aging on Trace Elements in Female Mice
Reporter	Akemi Morita, Mieko Kimura and Yoshinori Itokawa
Position	Department of Animal Social Medicine, Graduate School of Medicine, Kyoto University
Volume	The 10th Symposium on Trace Nutrients Research(1993)
P	P.139-143
Summary	To clarify the effect of aging on mineral status in female mice, mineral concentrations in tissues were determined. Five female mice (2, 6, 10 months old) were fed on commercial diet. Iron, zinc, copper, calcium and magnesium concentrations in blood, liver, kidney, heart, brain, lung and spleen of these mice were determined by a flame atomic absorption spectrophotometer. Iron concentrations in liver, kidney, heart, brain, and spleen were increased with age. Zinc concentrations in hearts and lung were decreased with age. It is noteworthy that the copper concentration in brain of 10 months old mice was markedly higher as compared with younger mice. Calcium concentration in kidney of 10 months ole mice was higher than 2 and 6 months old.

SUMMARY 23

NO	10-23
Theme	Tin Deficiency in Rats
Reporter	Katsuhiko Yokoi, Mieko Kimura and Yoshinori Itokawa
Position	Department of Animal Social Medicine, Graduate School of Medicine, Kyoto University
Volume	The 10th Symposium on Trace Nutrients Research(1993)
P	P.145-149
Summary	The effect of tin deficiency on the growth, iron and copper concentrations in tissues, and lipid parameters of plasma was studied. Tin, iron and copper concentrations in tissues, and triglycerides, total cholesterol and total phosupholipids if plasma were measured in rats fed a tin -adequate diet (2 mg tin/kg diet) or a tin-deficient diet (0.02 mg diet). Dietary tin deficiency decreased body weight gain and food efficiency. When rats fed a tin-deficient diet were compared with rats fed a tin-adequate diet, iron concentrations were higher in spleen and kidney, but lower in muscle; copper concentrations were lower in heart and tibia; triglycerides in plasma were higher, but total cholesterol and total phsuopholipids in plasma were lower. These results suggest that tin deficiency evokes ineffective utilization of iron in muscle, decreased copper retention and abnormalities in plasma lipid parameters.

SUMMARY 24

NO	10-24
Theme	A high Sensitive Determination of Glutathione and Glutathione disulfide by HPLC with Post-Label Fluorometric Detection
Reporter	Kenji Fukunaga, Tetsuya Suzuki and Kozo Takayama
Position	Department of Food Science and Technology, Faculty of Fisheries, Hokkaido University
Volume	The 10th Symposium on Trace Nutrients Research(1993)
P	P.151-154
Summary	A high sensitive high-performance liquid chromatographic method for determination of pmol levels glutathione (GSH) and glutathione disulfide (GSSG) in biological samples is described. GSH and GSSG were separated isocratically on a reversed-phase column (ODS column). The mobile phase consisted of 50mM sodium phosphate buffer (pH 3.0). After chromatographic separation, GSH and GSSG were converted to fluorescent derivatives by post-label reaction with o-phthalaldehyde under the alkaline condition. A 0.5pmol quantity of GSH and GSSG can be determined and the analytical recoveries were found to be almost 100% . The procedure is rapid, completed within 15 min, and it has been successfully applied to various biological samples.

SUMMARY 25

NO	10-25
Theme	Primary Structure and Functional Expression of the Cysteine Dioxygenase (CDO) cDNA
Reporter	Yu Hosokawa(1), Nobuyo Tsuboyama(2), Hitomi Tojo(3) and Masayuki Totani(1)
Position	(1)The National Institute of Health and Nutrition (2)Kochi Medical University (3)Kanagawa Prefectural Junior College of Nutrition
Volume	The 10th Symposium on Trace Nutrients Research(1993)
P	P.155-160
Summary	cDNA clones, rCDO-39 and hCDO-1, were isolates and characterized. rCDO-39, encoding the rat liver CDO contained a 1,458 b insert. HCDO-1, encoding the human liver CDO contained a 1,516 b insert. Both cDNAs encoded the polypeptides of 200 amino acids. Sequence comparison revealed a 91% homology in the amino acids between rat and human CDO. Significant expression of CDO gene was observed in liver, kidney, lung and brain but not in heart and skeletal muscle.

SUMMARY 26

NO	10-26
Theme	Effect of Vitamin A Deficiency on Fat Metabolism in Rats
Reporter	Masayuki Kitagawa(1), Tadao Umeki(2), Yuko Kai(2), Takezo Nishino(2), Hideo Yano(1) and Ryoji Kawashima(1)
Position	(1)Faculty of Agriculture, Kyoto University (2)Faculty of Agriculture, Yamaguchi University
Volume	The 10th Symposium on Trace Nutrients Research(1993)
P	P.161-164
Summary	Recently, farmers are likely to feed low vitamin A diets to beef cattle in Japan, because of the improvement of carcass quality. However, the relationship between vitamin A status and fat accumulation in animal is unknown. This study was conducted to elucidate the effect of vitamin A deficiency on fat metabolism in rats. Forty-mine female rats of Wister strain (4 weeks of age) were utilized in this study. They were fed vitamin A deficient diet during 40 weeks after the feeding of vitamin A adequate diet up to 10 weeks of age. They were sacrificed at 30,42 and 50 weeks of age in the fed state. Blood serum, liver and abdominal gat were collected for the determination of lipid levels and fatty acid composition. Rats fed vitamin A deficient diet exhibited an increase in serum and hepatic TG levels. Serum total cholesterol, free fatty acid (FFA) and glucose levers were found to be reduced during the experimental period, although not so severely as serum TG level. In contrast to the findings in serum and liver, lipid characteristics of abdominal gat in rats given vitamin A deficient diet were not different from control with the exception of FFA content at 50 weeks of age. These results suggest that TG rich lipoprotein secretion from liver is depressed when hepatic vitamin A store is completely exhausted in the deficient animals.

SUMMARY 27

NO	10-27
Theme	Effect of ethanol on rat brain gangliosides
Reporter	Michiko Adachi(1), Akihiko fujii(1), Tatsuo Ido(1), Syuichi Kimura (2)
Position	(1)Cyclotron and Radioisotope Center, Tohoku University (2)Course of Science for Living System, Showa Women's University
Volume	The 10th Symposium on Trace Nutrients Research(1993)
P	P.165-170
Summary	The effect of acute on rat brain gengliosides were investigated. 3g/kg of ethanol was administered intraperitoneally and the total amounts of N-acetylneuramic acid (NANA) decreased only in serum, but no in brain regions at two hours after injection. Time course of amounts of total and gangliosidic NANA were determined, and the decrease o f total NANA were only at 1 hr in cerebral cortex and returned to basal level at 4 hours. The amounts of gangliosidic NANA did not alter significantly. We investigated cleavage of NANA by using 3H labeled precursor of it, N-acetylmannosamine,-[N-mannosamine6H (N)]- . 48 hours after microinjection of precursor, dialysates were collected by microdialysis technique and were separated by HPLC system, and the liberation of NANA fraction were measured by liquid scintillation counter. Acute ethanol increased the liberation of NANA radioactivity's by neuraminidase in striatum. Therefore, we speculated that the effect of acute ethanol is a conformational change of gengliosides or their surrounding areas.

SUMMARY 28

NO	10-28
Theme	Effect of caffeine on brain neurotransmitters
Reporter	Yoshihiro Fujitani and Tatsuo Ido
Position	Cyclotron and Radioisotope Center, Tohoku University
Volume	The 10th Symposium on Trace Nutrients Research(1993)
P	P.171-176
Summary	We reported that acute and chronic caffeine administration affected neurotransmitters in tat brain striatum. We have measured extra cellular acetvlcholine (ACh) to investigate acute caffeine in rat hippocampus and chromic effect of caffeine by changing chronic period in rat striatum. The concentration of ACh in the rat hippocampus was increased was increased by the acute administration of caffeine (50 or 100mg/kg; i.p.) decreased the concentration of ACh. The similar effect was observed to the precious study. We have synthesized 11C-labeled caffeine's and examined the detail brain distribution of caffeine. Although the 11C-labeled caffeines distributed in the whole brain, considerably higher accumulation in the thalamus and hypothalamus was observed. The difference in the distribution of 11C-labeled caffeine's was due to their higher specific activity than commercially available caffeines which give a uniform distribution. These results suggest that the acute and chronic administrations of caffeine cause contrast, effect on in vivo extra cellular ACh concentration.

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