Calcium chloride or readily absorbable calcium preparation was administered to the experimental anim- als, and determining the change in blood levels of parathyroid hormore and calcitonine in the body. Male Wister rat were used as the experimental animals, and 3% or 5% of either type of Ca was administered orally with feed for 6 weeks, and then blood levels of parathyroid hormone and calcitonine were deter- mind, and any difference in Ca absorption was examined.

The field intervention in China has been done to establish the fact that Keshan Disease is related to the poor selenium (Se) of the local soils and can be prevented by the supplementation of Se. However, there also have certain seleniferous areas in china where chronic selenosis in the local inhabitants still persisted. Since 1982, we have been carrying out the determination on human Se req_uirements in the areas with different Se levels. The minimum physiological requirement and the range of daily safety in” take for adults were 50 and 50-250μg/ day, respectively.

We have attempted to synthesize the Neurospora crassαcopper metallothionein (MT) gene, clone it in the β-galactosidase gene, and express the MT-β-galactosidase fused gene in Escherichiαcoli cells. The fused protein in which the amino and carboxy termini of MT are linked to the β-galactosidase through methionine residues was purified by a single step, the immunoaffintiy chromatography with an anti-β-galactosidase antibody. The MT was prepared by cyanogen bromide cleavage of the fused protein followed by a reverse-phase HPLC separation. The spectral features of the MT and its copper complex were similar to those of the corresponding native MTs.

Superoxide anion production of PMN induced by FMLP or PMA was increased by magnesium deficiency in rats. The dose dependent increase of superoxide anion production was observed up to 10^-7M FMLP in control and 10^-6M FMLP in magnesium deficient group, respectively. Cytosolic free calcium concentrations after stimulation with 10^-7M FMLP in magnesium deficient group were higher than those in control. The inhibition of H-7, a protein kinase C agonist, on FMLP induced superoxide anion production in magnesium deficient group was less than that in control. These results indicate that cytosolic calcium and protein kinase C are involved in increase of FMLP induced superoxide anion production observed in magnesium deficient rats.

Growth Response of animal cells in culture to selenium (Se) are less well established than those for other nutrients. We added 0.01 to 1 ppm of selenite, selenomethionine (SeMet), and Ebselen (2-pheny-1, 2-benzoisoselenazol-( 2H) -one) to the culture medium and investigated their effects on proliferation and glutathion peroxidase ( GSHPx) activitiy of rat hepatocyte in primary culture ( HPC) and of human hepatoblastoma ( HeP G2) cells in culture. Among the Se compounds tested, selenite increased most significantly GSHPx activity in HPC from Se-deficient rats and maintained the enzyme activity in HPC from Se-sufficient rats. The proliferation of HeP G2 cells was stimulated by all of the Se comounds tested. GSHPx activities increased by the addition of selenite and Se-Met, but not by Ebselen addition. Ebselen and selenite showed an additive effect on the proliferation of Hep G2 cells.

In 1952, Kotake et al. have reported that Xanthurenic Acid ( XA) show the diabetogenic action in V.B6 deficient rats. From that time, we have experimented about the serum Ca and Mg value in Trp-sodium butyrate administered rats, V.B₆ deficient rats, alloxan diabetic rats and D.M. patients. And we detected the low Ca and slightly high Mg value in such an experimental rats serum.

In this report, we will report about the effect of Mg²⁺ andZn²⁺ related to Trp metabolism.

(1) In Mg deficient rats, only anthra凶 icacid (AA) excretion decreased. This effect is in accordance with the change of kynureninase activity.

(2) On the other hand, in Zn deficient rats, only XA excretion increased.

(3) From the experiment of kynureninase enzyme activity, -SH group existed in this apo enzyme, is important for this enzyme activity.

And it was inhibited by the p-OH-phenylpyruvate that is the metabolite of Tyr. From these data, the kynureninase reaction is very important for the regulation of 3-OH Kynurenine metabolism. And also it might be important for the metabolic regulation of D.M. patients in nutritional point.

In vivo studies on the oral treatment of organotin compounds with weanling rats, dibutyltin dichloride (Bu₂SnCl₂)and dioctyltin dichloride ( Oc₂SnCl₂) induced a severe thymus atrophy. This atrophy was reversible and its degree correlated to the concentration of Bu₂SnCl₂ in the thymus. At histopathological examination, there was marked depletion of lymphocytes in the thymic cortex rather than in the thymic medulla. Further, cortisone-pretreatment studies revealed that immature cells in the thymic cortex were far more sensitive to organotin compounds than mature cells. DNA synthesis of the thymic lymphocytes was significantly inhibited by Bu₂SnCl₂ and Oc₂SnCl₂ at the concentration of 10^-7M at which cell viability was not yet impaired. Moreover, a parallelism between dose-response curves of Bu₂SnCl₂ for DNA synthesis and cell viability was found. These results indicate that organotins primarily induces an inhibition of cell proliferation and secondarily causes cell death. Further studies on the movement of metals in the thymus of rats fed diets containing same molar concentration of individual organotin compound revealed that the degree of thymus atrophy correlated to the concentration of manganese in the thymus.

However, manganese was unevenly distributed to the epithelial connective tissues containing reticular cells and it was found in extremely small quantities in the thymic lymphocytes, depletion of which induces a severe thymus atrophy. Therefore, a relative increase of manganese concentration in the atrophic thymus depends on the immobility of manganese concentration in the epithelial connective tissues.

As previously reported, biodistribution studies with titanium-45 ascorbate (⁴⁵Ti-AsA)has shown that the effects of AsA on biodistribution of Ti was remarkably characteristic in the kidney¹⁾. In this paper, using ⁴⁵Ti-AsA, functional diagnosis of the kidney has been actively studied by animal models for renal disorder. As a technical requirements, autoradiography was selected. This work reveals the following results: 1) In Wistar rat which had made glomerulonephritis by injecting puromycin aminonucleoside²⁾, the highest concentration of ⁴⁵Ti was found in the cortex. 2) In stroke-prone spontaneously hypertensive rat ( SHRSP) fed a low protein and high sodium diet for four months which has the damage of renal tubule³⁾ , the highest uptake of ⁴⁵Ti was found in the medulla.

In conclusion, ⁴⁵Ti was localized the damaged region of the kidney in each inflammation rat. ⁴⁵Ti-AsA may be useful as a positron emmiting radiophrmaceutical or renal diagnosis.

As it becomes to clear that the incidence of hypogeusia is related to the deficiency of zinc or other transition element near to it in the periodic table, we feel the necessity to draw attention to the metals which are used for the restorative therapy of teeth. Among the metals the solder metals for dental use sometimes contain a large quantity of zinc or copper. In order to act these essential metals efficiently to our individual body should be dissolved at first. This study is concerned in the dissolution due to corrosion and potential difference.

If we consider the cases in which zinc or copper improve the taste acuity, the case will occur when the proliferation of Lactobacillus acidophilus advances and when the concentration of citric acid goes up after ingestion of citrus fruits, and also when acetic acid is elevated after taking of pickles.

Forty-nine cases of the "Pattern of Deficiency" were studied including 35 male cases of the "Pattern of Yin Deficiency" and 14 male cases of the "Pattern of Yon Deficiency". Cu, Zn, Pb, Cd, Mo levels in plasma were determined by atomic absorption spectrometry. Se level in plasma was determined fluorometrically.

Cu level in plasma increased singnificantly in cases of the "Pattern of Yin Deficiency" ( 1.03μg/ml in "Yin Deficiency", 0.75μg/ml in control), and decreased significantly in cases of the "Pattern of Yon Deficiency" ( 0.34μg/ ml in "Yon Deficiency"', 0.75μg/ml in control).

Se level in plasma was decreased significantly in cases of the "Pattern of Yin Deficiency" ( At 50 year group, 0.080μg/ml in "Yin Deficiency", 0.137μg/ml in control. At 60 year group, 0.075μg/ml in "Yin Deficiency", 0.102μg/ ml in control. At 70 year group, 0.071μg/ml in"Yin Deficiency", 0.100μg/ml in control). It was considered that "Yin Deficiency" was in the early aged state.

Vanadium has been shown to be as an essential trace element in experimental animals and proposed to be a possible etiological factor in manic depressive psychosis. To investigate the effects of vanadate on neuroreceptors, the binding properties of muscarir山 acetylcholinereceptor (mACh-R) and dopamine receptor ( D-2) were assayed in the brain of the rat treated under chronic administration of vanadate. Both of receptors were not changed in concentration (B max) but the binding affinity of mACh-R was decreased. The behavior of vanadium in vivo was investigated by use of cyclo-tron produced V-48 as a radioactive tracer. The major of vanadate in blood was reduced to vanady (4+) and bond to transferrin. The small part of unchanged vanadate was transferrd into brain through the anion channel. In cell fractionation of the brain, the cytosol had transferrin-vanadyl, free vanadate and no free vanadyl. The other cell component had about 60% of vanadium in the brain. The inhibition of Na⁺ ,K⁺-ATPase and the promotion of adenyl cyclase by vanadate may be concerned with the denature of mACh-R.

Vanadium has been known to be an essential trace elements for mammalians. Many physiological and biological functions of this element have been proposed; inhibition of Na⁺, K⁺-ATPase, anti-cancer effects, the presence in the active center of bromoperoxidase and insulin-mimetic effects for diabetic rats.

In the studies of insulin-mimetic effects of vanadium in rats, sodium vanadate (+5 oxidation state) in the drinking water has exclusively used. However, we used vanadyl sulfate (+4 oxidation state), which is reported to be less toxic than sodium vanadate in rats.

Vanadyl sulfate showed excellent insulin-mimetic effects for STZ-diabetic rats. Glucose tolerance test indicated that the STZ-diabetes was improved by vanadyl-administration, however, the serum insulin levels were not increased to the normal levels. Vanadium sulfate stimulated glucose incorpolation into adipocytes, suggesting that the effect of vanadium is due to the enhancement of glucose uptake in the periperal tissues. Determination of total vanadium and vanadyl species in organs indicated that most (~90%) vanadium ions present as a vanadyl form and coordinate with oxygen donors. On the basis of these results, the active form of vanadium for insulin-mimetic effects in rats was proposed to be vanadyl form as VO²⁺.

Feed oil have been widely employed as a feed additive for the purpose of accelerating growth and sparing dietary protein in ayu culture. However, the feed oil was not always useful for improvement of feed conversion efficiency nor protein efficiency ratio. In addition, lipid was excessively accumulated in muscle and intraperitoneal fat body with the increase of feed oil level.

When a combination of the feed oil and Chlorella-extract was supplemented tb commercial diet, the Chlorella-extract would cancel the evil effects of dietary lipid and might induce essential effects of dietary lipid such as sparing effect of dietary protein. As a result, Chlorella-extract improved feed conversion efficiency and protein efficiency ratio accompanied by suppression of lipid accumulation in muscle and intraperitoneal fat body. The phenomena implied the possibility of the improvement of fish quality of cultured ayu by the Chlorella-extract supplementation.

Mineral status of cattle in Java in Indonesia was investigated by evaluating mineral concentrations in forage and blood of cattle during both rainy and dry seasons in 1988. Investigations were conducted in Jonggol (West Java), Malang (East Java) and Mojokerto (East Java).

Low P content in Forage averaging 0.11% on dry matter basis, high Ca: P ratio in forage in rainy season (7.7:1) and low plasma Pi concentrations averaging 4.71mg/dl were observed in Jonggol. Although the Se content in forage from Jonggol in dry season, Malang and Mojokerto ranging from 0.05 to 0.08ppm was marginal the Se concentration in whole blood ranging from 0.11 to 0.21μg/ml was much higher than the critical level.

In Jonggol, the Cu content in forage averaging 8.2ppm was marginal and the plasma Cu concentration averaging 0.22μg/ml was much lower than the critical level. The Fe content in forage from three places was much higher than 1,000 ppm which is the maximal tolerable level of Fe for cattle.

Thirty samples of 22 species of useful plants in Thailand were analyzed by AAS method for Na, K, Ca and Mg and by ICP method for 23 trace elements. It was found that the logarithm of concentration ratio of each element relative to seawater was inversely proportional to the logarithm of the oceanic residence time.

A method of data analysis based on this relationship was applied to the analytical results. The values of statistical treatment on Thai plants were compared with those on Japanese seaweeds and other marine organisms.

A simple and sensitive liquid-chromatographic method has been developed for the separation and determination of vitamin B₆ derivatives in rat blood. The analyses were carried out with an ODS reversed phase column and fluorescence spectrometer ( ex. 298 nm, em. 392 nm) using a solution of 0.1M sodium perchloric acid in 0.1M potassium dihydrogen phosphate buffer (pH 2.5) as mobile phase. A standard solution of vitamin B₆ derivatives was eluted in the following order; PMP, PM, PLP, PL, PN and 4-PA. Deproteinized blood samples were applied to the HPLC system. PLP was the predominant derivative of the vitamin in rat blood.

Neonatal vitamin K deficiency causes hemorrhagic disease of the newborn. There have been some suggestions as to how to prevent babies from vitamin K deficiency. It is not yet clear which drug is more eff ecitve, vitamin K₁ or K₂ ( MK-n). It is recommended that prophylactic K₁ be administered parenterally to all newborn infants at a dose of 0.5 to 1.0mg by the American Academy of Pediatrics. In Japan, K₂ (MK-4) is generally administered with injection or syrup for babies or capsule for adults.

In this study, to find out the form of vitamin K present in fetus. We measured the concentrations of vitamin K in vernix caseosa of human neonate, because vitamin K is fat-soluble and may be contained abundantly in it. This is the first report of the determination of endogenous vitamin K in human vernix caseosa. Only MK-4 was found in vernix caseosa (Mean±S.D. 1.93ng/g±0.83),and K1 was not detected ( <0.05ng/g). As compared with the mean concentrations in maternal and umbilical cord plasma, in vernix caseosa there was high concentration of MK-4.

There is little vernix caseosa on the skin of neonates who are born before 32 weeks gestation or after 40 weeks. During 32 to 40 weeks gestation, vernix caseosa migth be stored up on the skin of fetus. From where vernix caseosa dose come? We suppose that it comes from the amniotic membrane. Fetus may get nutrition from placenta through the umbilical cord, and in addition to that we suppose he might get nutrition from amniotic membrane through the amniotic fluid and vernix caseosa.

Protein-calorie malnutrition is still a serious problem in developing countries. It is well-known that low-protein diet causes deficiency of trace nutrients. But the influence of low-protein diet on vitamin E status is not completely clear yet.

We studied the influence of low-protein diet on vitamin E status using male Wistar 3 week-old rats. The rats were devided into 3 groups: group 1; 5% casein, group 2; 10% casein, group 3 ( control group); 20% casein. They were fed ad libitum for 6 weeks. Tocopherols in plasma, liver, kidney, brain, spleen and testis were measured by HPLC-fluorometric method.

α-Tocopherol concentration in liver, kidney and plasma of rats fed 5% casein diet were significantly low compared with those of rats fed 20% casein diet. α-Tocopherol in liver and kidney of rats fed 10% casein diet were also significantly low compared with those of rats fed 20% casein diet. A uniform relationship between β, γ-Tocopherols concentrations in tissues and protein concentration in diet was not found.

The results in this study suggest that protein-mulnutrition at growing age influences vitamin E status.

To clarify the differences of trace elements distribution in red and white muscles, we analyzed the concentrations of Mn, Cu and Al among other elements ( Zn, Fe, Ca, Mg, P and S) in two types of muscie fiber. The extensor digitorum longus muscle (EDL) and soleus muscls (SOL) were taken as the white and red muscles respectively. The Mn and Cu concentrations were determined with a flameless atomic absorption spectrophotometer with a graphite furnace; the Al and other elements concentrations were determined with inductively coupled plasma atomic emisson spectrometer. The Mn, Al and Cu concentrations in SOL were significantly different from those in EDL and were two to three times higher than those in EDL. The Mg and P concentrations in SOL were significantly low and the Zn, Fe and S concentrations in SOL were significantly high compared with those in EDL.

A peptide which inhibits angiotensin converting enzyme was isolated from peptic hydrolyzates of water soluble protein of sardine muscle. The preparation procedure contains SP-Sephadex C-25 and Sephadex G-25 chromatography followed by twice high performance liquid chormatography. The purified peptide, a hexapeptide had the sequence of HQAAGW with the half maximum inhibition (IC₅₀) of 60μM. A peptide sequence with 66% homology to the purified peptide was found in some proteins such as enolase I, ferredoxin II, VB₁₂ receptor precursor, apo-lipoprotein C-III precursor, β-amylase, and reverse transcriptase.

Requirement of essential amino acid for growing rats was examined on the basis of hepatic cysteine dioxygenase ( CDO) activity and urinary ta urine. Young male rats were fed on partiallypurified diets of 0.45% sulfur amino acids ( Met : Cys=7 : 5) and different level of isoleucine, valine, leucine, phenylalanine and tyrosine for 18 days. Levels of isoleucine, leucine, phenylalanine + tyrosine in each experimental diet were adjusted to five levels, respectively. Minimum requirements of isoleucine, leucine, valine and phenylalanine+ tyrosine were resulted in 0.411, 0.508, 0.675 and 0.661 % in CDO activity and 0.392, 0.477, 0.622 and 0.627% in urinary taurine.

Contents of iron and arsenic in the different parts of Kombu blade (Laminaria Japonica) at different growth stages collected off the seashore of Aomori and Hokkaido prefectures in different seasons were determined by atomic absorption spectrometry. The iron content at the apex from the annual blades was higher than that of the base. However, the reverse tendency was recognized in the iron content in the samples obtained from the biennial blades: iron distribution in blades showed remarkable differences from other inorganic constituents. The arsenic content at the base, corresponding to the blade meristem, was higher than that of the apex at any stages examined. Although aresenic content varied with the season, and was highest in June, which was proportional to the increase in the size of blade length, it did not show any remarkable distribution change accompanied with growth of blade as recognized in iron.