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 2991 mg (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), Mn 6 .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.

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 concentrat_ion 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 is important for the prevention of osteoporosis.

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, an 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⁺/Calmodulin-dependent protein kinase II ( CaM kinase II) were seen. In case of zinc deficiency, a remarkable decrease of PTH and a little decreases of cAMP, IP₃ were seen in the olfactory tissues, and a 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 increasεof cAMP, and an excessive influx of extracellular 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 or apoptosis in vivo or toxic cell death in vitro.

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. Autoradiography of ²⁸Mg concentrations in the heart was also performed. ²⁸Mg was injected 10 min. after the administration of ouabain. The mice were sacrificed 10 min. after the 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.

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 overintake 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 Obtained results thus suggested that dietary magnesium deficiency gives rise to retrogressive changes in some organs such as heart, and concurrent calcium overintake synergistically enhances the myocardial injury due to magnesium deficiency.

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 extarct ( 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 treatment and partly prevented by the higher intake of cili extract. These results suggest that compared to 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.

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 selectivities of the ions were controlled by EDTA, Cl^-, pH and ion strength. Hg⁺, Hg²⁺ and CH₃Hg were selectivelly determined. The other ions were completely masked by the EDTA solution.

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, stepwize 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.

The present study was designed to examine if zinc (Zn) affects the development of unilateral ureteral obstruction (UUO)-induced tutbulointerstitial 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 or a high Zn diet, rats fed a Zn deficiency diet showed a more influx of leukocytes into the interstitium and m 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 mRN A in the kidney.

Insulin-dependent diabetes mellitus (IDDM) is controled 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 (FF A) 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 FF A release from adipocytes are essential to obtain a good effect of the complex on oral administration.

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 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-readion. NO has been shown 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.

In the present study, we demonstrated the cytotoxic effects of inorganic arsenicals, arsenite and arsenate, and organic arsenic compounds, monomethylarsonic acid (MAA), dimethylarsinic acid (DMAA) 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 (IC₅₀) 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 caused by inorganic arsenicals.

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-chains. 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. Based 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 aforementioned isoamylolysis and HP AEC 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.

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, Mn, 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 cells. It may also be considered that glassy surface ceramic treatment should be explained by other mechanisms.

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 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 cell 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, Chelex-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.

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 hem in 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 hemeoxygnase 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.

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 β₁ antagonist, metoprolol (3×10^-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^-5M), histamine (H₁) antagonist, markedly inhibited the PIE of OE, whereas cimetidine (10^-4M) , 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.

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⁺, andfurther 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⁺ ionfrom 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 complex, which cannot react with oxygen. Oxidation reduction properties of iron was applied to analyze the extract of oyster, Crassostrea 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.

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 synthesis by His. Decarboxylase in diabetic rats were also discussed.

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 DNA 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 splernocytes. 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.

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 (TW80-0CT) 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.

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 the depression of hepatic lipogenesis,but not involving the alteration in the activities of intestinal enzymes. These results also suggest that practical level of dietary phytate may affect hepatic lipid metabolism in animals fed on sucrose.

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.

Effects of acidic fibroblast growth factor (aFGF) on the hypothalamo pituitary-adrenocortical axis were examined in anethetized 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.

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 II - 8 medicines for eczema, urticaria, pruritus, and allergy, Group III - 3control medicines for non-skin disease. Biotin assay was performed by the agar plate method using Lαctobαcillus plαntarum ATCC 8014. The average content of free biotin in Group I, II and III was 118.0±4.6, 97.8±48.6 and 30.3±19.6 ng/g, respectively. Also total biotin content in each group was 313.3±116.0, 222.0±132.5 and 47.7±28.3 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 Kam po medi- cines contain a large amount of free biotin, which can be easily absorbed by the small intestine.