We measured all of the d-amino acids in 37 samples taken in 3 kinds of sake brewing processes (Kimoto, Kimoto adding starter latctic acid bacteria, and Sokujomoto) and 4 samples of sake rice using high-performance liquid chromatography. We found that d-Asp, d-Glu, d-Ala, and d-Val were produced in all three sake brewing proceses, but d-His, d-Arg, and d-Pro were detected only in Kimoto. d-Ile and d-Phe were detected only in Kimoto adding starter lactic acid bacteria. d-Leu was detected in Kimoto adding starter latctic acid bacteria and Sokujomoto but not in Kimoto. Most of these d-amino acid concentrations increased from Fukuremae to Modosi period. The d-amino acid concentrations observed were different in each brewing process, and the highest amount of d-Asp (32.1 μM), d-Glu (25.3 μM), and d-Ala (160.8 μM) were contained in Kimoto at Jousou period. In contrast, sake rice contained d-Asp, d-Ala, d-Val, d-Glu, and d-Ser, but all of their concentrations were under 1 μM. Lactic acid bacteria type culture strains (Lacobacillus sakei NBRC 15893 and Leuconostoc mesenteroides NBRC 102480) isolated from Kimoto produced d-amino acids (for L. sakei: d-Ala, d-Glu, and d-Asp; for L. mesenteroides: d-Ala, d-Glu, and d-Lys). We found that all of the gene products of the amino acid racemase homologue genes from L. sakei and L. mesenteroides showed alanine, glutamate, aspartate, lysine, or histidine racemase activity. Accordingly, Kimoto is one of effective methods to increase d-amino acid contents in sake. The D-amino acids in sake were produced by lactic acid bacteria in Kimoto, and the amino acid racemases of the lactic acid bacteria probably catalyze the synthesis of various d-amino acids in the organisms.

Oyster contains much nutrients such as taurin, glycogen and zinc as called “marine milk”. A supplementary food containing oyster extract is reported to possess several physiological effects including anti-oxidative activity and supplementation of zinc. It is also known that “mental disorder” is caused by zinc deficiency, therefore oyster extract containing abundant zinc is expected to show some physiological effects on nervous systems.

Then, we assessed anxiolytic-like effect of Oyster Extract (prepared by Japan Clinic Co., Ltd.) in naïve mice and psychological stressed mice using Hole-Board test method, commonly used in the evaluation studies of anxiolytic activities of chemicals.

As a result, head-dipping behavior in naïve mice was significantly increased by oral administration of Oyster Extract for 28 days. On the other hand, head-dipping behavior in psychological stressed mice was significantly decreased from naïve mice. Oral administration with Oyster Extract significantly increased the number of head-dipping behavior in psychological stressed mice. Psychological stress induced anxiety-like behavior was suppressed by administration of oyster extract.

These results suggest that Oyster Extract have a potential of anxiolytic-like effect.

We focuses physiological activities of oyster extract and its possible indication to liver functional impairment, and investigated the effect of oyster extract on hepatic function in 20 male and female candidates with mild alcoholic liver damage by 12 weeks ingestion of oyster extract (Japan Clinic Co., Ltd.) prepared from Crassostrea gigas. Throughout the study period, there was no ethical issues and intentional or accidental medical accident.

The subjects with mild alcoholic liver damage ingested 4 pills (containing 230 mg of Oyster Extract Powder in 1 pill), tid, for 12 weeks. Physical examination, urinalysis, hematology, blood chemistry and clinical examinations by doctor were conducted at 0, 4, 8 and 12 weeks.

As the results, significant reduction was observed for ALT and ALP in male and female subjects and for γ-GTP in male subject at 12 weeks. A tendency of reduction to the reference ranges was observed for AST at 12 weeks.

As shown above, it was suggested that the hepatic functions of the subjects with mild alcoholic liver damage will be improved by 12 weeks ingestion of oyster extract.

To clarify the possibility of using an oyster-extract by-product as a health maintaining and promoting food material or supplement, this study investigated the effects of oyster-extract by-product powder (OEBP) on serum and liver lipid contents in rats. OEBP was prepared from washed and freeze-dried oyster-extract by-product, which contained 58.3% protein, 19.7% fat, and 19.7% carbohydrate. Male, 5-week-old Wistar rats were divided into 2 dietary groups of 7 rats each, one group receiving a control AIN-93G diet and the other receiving a 5% OEBP-containing diet. After the rats had been fed on their respective diets for 31 days, their serum and liver lipids contents and fecal cholesterol, bile acid, and nitrogen excretion were measured. The results demonstrated that OEBP decreased liver cholesterol content, in part owing to the enhancement of fecal cholesterol and bile acids excretion; however, serum cholesterol content in the OEBP diet did not decrease. In addition, the OEBP group exhibited lower protein digestibility during in vitro digestion analyses compared with the control diet. Therefore, in the OEBP group, the excretion of fecal cholesterol and bile acids was influenced by the low digestibility of OEBP protein. However, in order to develop OEBP as a lipid-lowering functional food and supplement, we concluded that OEBP required improvements in its formulation and further concentration of the bioactive components because OEBP has not the lowering effect of serum lipid contents.

Pyrimidine base is a constituent of nucleotides and pyrazine is a component of food and antibiotics. Biological effects of these compounds were analyzed in relation to the generation of reactive oxygen species. Pyrazine 2-carboxylate, pyrazine 2,3-dicarboxylate and orotic acid/iron complex inactivated aconitase, the most sensitive enzyme to oxidative stress. The inactivation was dependent on cyanide, an inhibitor of Cu/Zn SOD, indicating that these compounds can generate superoxide anion radical as a principal product. Pyrimidine and pyrazine carboxylate compounds were less effective than pyridine carboxylates on aconitase. Orotic acid, a pyrimidine carboxylate derivative stimulated the autooxidation of Fe²⁺ suggesting that this compound promoted the activation of dioxygen molecule by reduced iron. Pyrazine carboxylates did not stimulate the autooxidation, suggesting that these compounds produce perferryl ion. Injuries of tissue by pyrazine compounds can be partially explained by their transition metal complex-mediated generation of reactive oxygen species.

Vegetarian diets provide relatively large amounts of vegetable, cereals, nuts and fruits. Thus those diets have relatively low intakes of protein, saturated fatty acid, retinol and vitamin B₁₂. The plant foods do not contain vitamin B₁₂; in particularly, vegans may have low intakes of vitamin B₁₂. To estimate the daily intake of vitamin B₁₂ in Japanese woman vegans, 33 duplicate portion samples of daily diets were collected from 11 Japanese adult female vegans and their vitamin B₁₂ were determined by microbiological assay using Lactobacillus delbrueckii subsp. lactis ATCC7830. Content of vitamin B₁₂ in Japanese woman vegan diets were as follows: range, 0-2.1μg/day; median; 0.14μg/day. Compared with estimated average requirement and recommended dietary allowance in female aged 30 to 49 years in the Dietary References Intakes for Japanese 2010, vitamin B₁₂ in vegan diets is quite low. This study suggested that Japanese woman vegans should have the laver and permitted to have the vitamin B₁₂ supplement or vitamin B₁₂-fortified vegetable for preventing a vitamin B₁₂ deficiency.

Folic acid is a water soluble B vitamin which functions in single-carbon transfer reactions. The folate difining leads to the abnormal hematological changes such as megaloblastic anemia. Also the evidence is strong that the sick of having a fetus with NTD (Nutral Tube Defts) deseases with increasing intake of folate during gestation. However the folate intake of many young women is not sufficient to prevent the induction of NTD in Japan. Therefore aimed to evaluate the relationship between the folate intake and serum acid of folate in 17 young women, in Japan. The dietary folate intake was 208±104 μg/day for the annual average, although the RDA(Recommended Dietary Allowance) for adults in 240 μg/day of diet folates equivalunts. The serum concentration in folic acid 2.3±1.0 ng/ml for the annual average, which was below the lower limit of the normal range. The high end of serum folate can be partly explained by dieting folate intake. Therefore it is recommended that much more green and yellow vegetable should be intaken in young women.

Peroxisome proliferator-activated receptor γ (PPAR γ) is a ligand-dependent transcription factor that directly activates the expression of adipocyte-specific genes. PPAR γ is mainly distributed in adipose tissues, and regulates lipid and glucose metabolism. Thiazolidinediones (TZDs), as synthetic PPAR γ ligands and antidiabetic drugs, are able to stimulate adipogenesis and enhance insulin sensitivity by stimulating the transcriptional activity of PPAR γ. Through screening for natural PPAR γ activator in various plant seeds using the PPAR γ luciferase reporter assay, the methanol extracts of Wisteria floribunda seeds activated PPAR γ. To identify the active compound that activated PPAR γ, the methanol extracts of Wisteria floribunda seeds was partitioned between hexane, ethyl acetate and H₂O soluble fractions. Among these, treatment with the ethyl acetate extracts of Wisteria floribunda seeds enhanced a marker of adipocyte differentiation such as triglyceride accumulation in 3T3-L1 cells. These results suggest that Wisteria floribunda seeds possess PPAR γ activators. The ethyl acetate soluble fraction of Wisteria floribunda seeds was further separated by silica gel and ODS column chromatography. Based on NMR, ESI-MS, and luciferase reporter assay, we confirmed that blumenol A, wistin, and isomucronuratol were identified as PPAR γ activators in Wisteria floribunda seeds.

Fluoride (F) widely exists in nature and is contained in food and water. One of the target organs of F is the kidney. High IgA (HIGA) mice have been used as a model of IgA nephropathy. The objective of this study was to get information for the alterations in the renal function of HIGA mice exposed to F. The HIGA and BALB/c mice were exposed to F at 0, 50, 100, and 150 ppm in their drinking water for 4 weeks. For the protein levels in the urine, the mean of the 100-ppm group of HIGA mice after 3 weeks from the beginning of the exposure was significantly higher than those in the 0- and 50-ppm groups. For BALB/c mice, the mean protein level in the 100-ppm group was significantly higher than those in the 0- and 50-ppm groups after 2 weeks. The mean creatinine in the urine in the 50-, 100-, and 150-ppm groups of HIGA mice were significantly lower than that in the 0-ppm group after 4 weeks. For BALB/c mice, the mean creatinine in the 50-, 100-, and 150-ppm groups were significantly lower than that in the 0-ppm group after 2 weeks. The exposure to F at 100 ppm may induce temporal toxicity to the kidneys of mice, but no differences of toxic effects of F between the 11- to 12-week-old HIGA and BALB/c mice were observed.

Molybdenum (Mo) is a constituent of Mo cofactor (MoCo) that is indispensable for the activity of molybdoenzyme such as xanthine oxidase (XO). This molybdoenzyme plays an important role on purine-base metabolism in the liver where Mo mainly exists as MoCo. Our previous experiment indicated that magnesium (Mg) deficiency increased Mo concentration in rat liver. We hypothesized that the increase in Mo reflects up-regulating expression of molybdoenzyme such as XO in the liver of Mg-deficient rats. We investigated the effect of Mg deficiency on the metabolism of purinebase in rats. Twelve growing male rats were given a control diet or an Mg-deficient diet for 4 weeks. Urine was collected every day during the last 5 days in the feeding trial. At the end of feeding trial, liver was collected, and the hepatic XO activity and the concentration of purine-base metabolites such as hypoxanthine, xanthine, uric acid, and allantoin were determined. Additionally, urinary excretion of purine-base metabolites was determined. Mg deficiency did not affect the XO activity and the concentration of purine-base metabolites in the liver. One the other hand, urinary excretion of uric acid and allantoin was significantly decreased by Mg deficiency. These results suggested that the increase in hepatic Mo concentration does not affect purine-base metabolism in the liver of Mg-deficient rat, but that Mg deficiency decreases urinary excretion of purine-base metabolites.

A facultative halophyte Mesembryanthemum crystallinum L. changes its carbon fixation mechanism from C3 to crassulacean acid metabolism (CAM) in response to environmental stress such as drought and salinity. Induction of CAM also results in the accumulation of metabolites like malate, betacyanin, proline, and pinitol in the plant with mechanisms that are unknown as yet. In order to explore the factors that lead to the formation of these metabolites in this plant, we treated M. crystallinum L. with culture supernatants of Aspergillus oryzae, Nectria gracilipes, and Rhizopus sp. Among the fungi tested, culture supernatant from A. oryzae induced typical CAM phenotypes in this halophyte species even in the absence of salt. Endogenous malate levels in the treated plants increased 5-fold in comparison with those in the control plants. We investigated the expression patterns of genes like CAM-specific Ppck and Ppc1. Our analysis of expression patterns of these genes showed no significant increase in their expression levels in the A. oryzae-treated plant, indicating that the accumulation of malate was independent of CAM-photosynthesis. When the plant was treated simultaneously with the culture supernatant and 400 mM NaCl, circadian regulation of Ppck was disturbed. Expression patterns of a pinitol biosynthesis gene (Imt) in the treated plants also showed anomalies in the circadian rhythm. On the basis of these results, we suggest that Ppck and Imt genes may be regulated by a novel mechanism independent of CAM.

Insulin secretion is regulated by a number of factors, including a rise in a ATP/ADP ratio, cAMP production and plasma membrane depolarization, leading to a increase in intracellular Ca²⁺ ([Ca²⁺]i). Arginine, like glucose, is a well-known insulin secretagogue. It is recognized that arginine stimulates insulin secretion by a membrane depolarization with their positive charge. As another possible mechanism, it is proposed that arginine enhances a nitric oxide (NO) productions as its metabolites, which can activate several pathways and increase [Ca²⁺]i. Our previous study indicates that biotin directly acts in the glucose metabolic pathway for ATP production and modulates glucose-induced insulin secretion. But, it remains to clear whether biotin acts as a regulator of arginine-induced insulin secretion. We studied the effect of biotin deficiency on arginine-induced insulin secretion by isolated pancreas perfusion of rats. Insulin secretion was stimulated with l-arginine, n-nitro-l-arginine-methyl ester (l-NAME) which does not work as an NO-donor for NO production, and d-arginine which is not metabolized in the mammalian cells. In the moderate biotin deficient rats, the level of insulin response was significantly lower than that in the control rats only when insulin was stimulated with d-arginine. In the sever biotin deficient rats, insulin secretions stimulated with each of every secretagogues markedly diminished. These results indicate that arginine-induced insulin secretion was actually impaired by biotin deficiency, but biotin did not directly act as a regulator of arginine-induced insulin secretion. It may be caused by a histologic dysfunction in pancreatic β cells by biotin deficiency.

We used high-performance liquid chromatography to measure the concentration of d- and l-amino acids in 7 bottles of black vinegar produced by traditional field fermentation with an Aman pot in Fukuyama-Cho, Kagoshima, Japan. The data for the traditional black vinegar were compared with the data for bottles of black vinegar and rice vinegar produced by a modern fermentation method in a plant as well as black vinegar #001 that had been used to pickle 9 different fruits, such as blueberries, mandarin oranges, and apples, Japanese plums, green umes, persimmons, strawberries, acerolas, and pineapples. We found that d-Asn, d-Thr, d-Arg, d-Met, d-Trp, d-Ile, d-Lys, and L-Gln were specifically contained in black vinegar. The d- and L-amino acid concentrations in black vinegar were generally higher than the concentrations in rice vinegar. The d-amino acids with the highest concentrations in the analyzed traditional black vinegar samples produced by Fukuyama Kurozu Co. Ltd. were Glu, Ser, His, Ala, Val, and Pro. The use of black vinegar to pickle fruits generally increased the concentrations of d-amino acids of the vinegar, with Japanese plums providing the largest effect.

Twenty-five duplicate diet samples of homemade baby foods prepared for infants and toddlers aged 8 to 16 months were collected from their mothers living in an urban area of Hyôgo Prefecture in Japan and their mineral (sodium, potassium, calcium, magnesium, phosphorus, iron, zinc, copper, manganese, iodine, selenium, chromium and molybdenum) contents were determined. Mineral contents obtained were expressed as values per 1000 kcal and compared with the adequate intake (AI) for infants aged 6 to 11 months or the estimated average requirement (EAR) for toddlers aged 1 to 2 years described in Dietary Reference Intakes for Japanese, 2010 (DRI-J).

All mineral contents in the homemade baby foods were almost met the reference values in DRI-J. In particular, contents of magnesium, phosphorus, manganese, selenium, chromium and molybdenum in baby foods prepared for 6 to 11 months babies were markedly higher than the AI. Iodine contents in the baby foods were remarkably varied from near 0 to more than 1000 μg/1000 kcal. These results indicate that 1) mineral contents in the homemade baby foods collected fell within the suitable range, 2) intakes of magnesium, phosphorus, manganese, selenium, chromium and molybdenum increase with the progress of weaning, and 3) an intermittent high iodine intake is important to satisfy the iodine requirement in infants and toddlers.

A study reported that, during pregnancy, folic acid was involved in the continuation of pregnancy and fetal growth. However, there is no scientific evidence regarding optimal folic acid consumption in Japan. Folic acid deficiency results in the onset of neural tube defects such as spina bifida and anencephaly. We measured the blood level of folic acid in the first, second, and third trimesters of pregnancy in 28 pregnant women, and examined folic acid intake. The blood levels of folic acid in the first, second, and third trimesters of pregnancy were 0.27, 1.09, and 0.92 ng/mL, respectively. The mean folic acid intake was 235μg, being lower than that recommended in the Reference Intakes in Japanese [2010] (480μg). Trace nutrient consumption during pregnancy, including an additive volume, must be reviewed before pregnancy.

This study assessed bone mineral density (BMD) using dual-energy x-ray absorptiometry (DXA), the alkaline phosphatase (ALP) and tartrate-resistant acid phosphatase (TRAP) activity in the femur, and the excretion of calcium (Ca) and phosphate (P) in 15-h urine in zinc-deficient rats.

Male SD rats (150±10g) were used. They were kept in individual cages and fed zinc-containing (10 mg/100g diet; Pf group) or zinc-deficient (ZnD group) diets for 4 weeks. After 4 weeks, the BMD of the femur assessed by DXA was significantly lower in the ZnD group compared with the Pf group. The ALP level, a measure of osteoblastic activity in the femur, was significantly lower in the ZnD rats. The TRAP level, a measure of osteoclastic activity, was the same in both groups. The urine volume collected in a metabolic cage for 15 h was significantly lower in the ZnD group at 4 weeks. The urine Ca concentration did not differ between the two groups, but the mean urine P concentration was 1.535 and 0.025 mg/dl/mg creatinine in the ZnD and Pf groups, respectively. The urinary P excretion was significantly higher in the ZnD rats (p=0.0049). The results suggest that reduced BMD in zinc-deficient rats is related to increased urinary P excretion and decreased osteoblastic activity.

Food folates exist mainly as pteroylpolyglutamate, which is degraded to a monoglutamyl form by glutamate carboxypeptidase II (GCPII) in the intestine before absorption. Green tea is a traditional and widely consumed drink all over the world and one of the major sources of food folates. In this study, we examined the influence of food, including green tea, on porcine GCPII activity in vitro. Several foods were selected and their extracts were prepared in order to examine the influence on GCPII activity. The food extracts were included in the reaction mixture for the analysis of GCPII activity at the level of 10%. Little inhibition of GCPII activity was exhibited in extracts of cabbage, lettuce and spinach. In contrast, this activity was significantly inhibited by extracts of orange juice (68.6% of no addition), tomatoes (66.9%) and green tea (35.6%). The activity was inhibited in a concentration-dependent manner by the extract of green tea. These results indicate that green tea may partially influence folate bioavailability through the inhibition of GCPII.

A psychrotrophic bacterium, Shewanella livingstonensis Ac10, produces eicosapentaenoic acid (EPA) at 4°C, which plays a beneficial role in the cold adaptation of the bacterium. The gene cluster containing orf2, orf5, orf6, orf7, and orf8 is involved in EPA biosynthesis. Here, we investigated the function of orf2, which is thought to code for a phosphopantetheinyl transferase (PPTase). The western blot analysis using an anti-Orf2 antibody demonstrates that Orf2 is induced at 4°C and localized to the cytoplasm. PPTase catalyzes the transfer of a phosphopantetheine moiety from CoA to the serine residue of acyl carrier proteins (ACPs). To examine the phosphopantetheinylation of 5 conserved repeated ACP domains of Orf5 by Orf2, we established an in vitro phosphopantetheinylation system using the purified recombinant Orf2 as well as ACP domains of Orf5 as its putative substrates. When each of the ACP domains was incubated with Orf2 and CoA at 4°C, the phosphopantetheinylation of individual ACP domains was observed by HPLC and MALDI-TOF/MS analyses. Moreover, the in vitro modification did not occur when Ser-less ACP mutants were used, indicating that Orf2 catalyzes the phosphopantetheinylation of the serine residue in ACP domains of Orf5, which may probably be an initial step of EPA biosynthesis in S. livingstonensis Ac10.

Shewanella livingstonensis Ac10, a psychrotrophic bacterium, produces the omega-3 polyunsaturated fatty acid eicosapentaenoic acid (EPA), as a fatty acyl chain of phospholipids at low temperatures. EPA is incorporated into the sn-2 position of phospholipids. 1-Acyl-sn-glycerol-3-phosphate acyltransferase (PlsC) catalyzes the acylation at the sn-2 position of 1-acyl-sn-glycerol-3-phosphate to form phosphatidic acid (PA). We found that 5 genes code for proteins homologous to Escherichia coli PlsC (named PlsC1 through PlsC5), suggesting that these PlsCs are involved in the synthesis of EPA-containing phospholipids. To examine the role of these putative PlsCs, we constructed the knockout mutants of each plsC gene (∆plsC1 to ∆plsC5). In the mutant ∆plsC1, the amount of phospholipids containing EPA was less. Functional expression studies in a temperature-sensitive mutant of PlsC, E. coli JC201, showed that PlsC1 has a PlsC activity with a broad acyl-coenzyme A (acyl-CoA) specificity including EPA-CoA. These results indicate that PlsC1 is a key enzyme in the synthesis of EPA-containing PA in S. livingstonensis Ac10.

Hijiki (Sargassum fusiforme) plants were collected during their growth period from the end of November through April on the sea coast of Kushimoto, Wakayama, Japan. The samples were brought back to the laboratory under ice-cold conditions and washed thoroughly with artificial sea-water and distilled water successively. The plants were cut into pieces of a 10 cm length along the stalk, and separated into leaves and stalks. The samples were lyophilized, and decomposed in a mixture of conc H₂SO₄ and conc HNO₃ (3 to 1, v/v) on an electric heater. Iron (Fe) and magnesium (Mg) in 1N HCl were determined with an atomic absorption spectrophotometer.

Fe accumulated in the respective tissues at concentrations of 70 to 110μg/g dry weight of tissues at the beginning of growth and Mg, at concentrations of 7 to 9 mg/g dry weight of tissues. Fe accumulated at 20 to 300μg/g dry weight through the growing period until the beginning of April. Mg accumulated at 5 to 30 mg/g dry weight of tissues through the growing period. Mg accumulation seemed to be more than enough for its biochemically required concentrations in the tissues of Hijiki.

In our previous study, we reported the enhancement of glutathione in rat hippocampus with combined administration of oyster extract and warfarin. However, the effect of sole administration of oyster extract on glutathione in rat hippocampus was not investigated. Therefore, the principal aim of the present study was to determine if oyster extract increases glutathione in rat hippocampus and various organs. Moreover, the effect of oyster extract on glutathione S-transferase (GST) activity was investigated.

In the first experiment, after 7-week-old male Sprague-Dawley rats were administered oyster extract (200 or 500 mg/kg body weight P.O.) (high-volume group or low-volume group) or vehicle (control group) for 1 week, these various organs were isolated. The concentration of glutathione in these organs was determined. The concentrations of glutathione in hippocampus of the high-volume group and the low-volume group were twofold higher than that of the control group, with significant differences.

In the second experiment, after 7-week-old male Sprague-Dawley rats were administered oyster extract (500 mg/kg body weight P.O.) (oyster extract group) or vehicle (control group) for 1 week, these various organs were isolated. The concentration of glutathione and GST activity in these organs were determined. The concentration of glutathione in kidney in the treated group was significantly higher than that in the control group. GST activity in the hippocampus in the oyster extract group was significantly higher than that in the control group.

The present results suggest that oyster extract produces anti-oxidant effects in hippocampus and kidney.

To investigate the differences in the trace mineral intake of atherosclerosis patients at different stages. , the nutrition survey was made in 80 atherosclerosis patients and 184 healthy individuals. Studentʼs t-tests were performed to determine the differences in the mineral intake between atherosclerosis patients and healthy individuals, and the differences among the different stages of the disease. All subjects were divided into 2 groups according to body mass index above 25 (obese group) and below 25 (non-obese group). The Studentʼs t-test was performed for groups comprising only obese group and only non-obese group too.

In Serum measurements, the subjects included 16 male atherosclerosis patients and 35 healthy men. The differences between the average concentrations of each mineral were determined between the obese and non-obese groups; the atherosclerosis and non-obese groups; and the atherosclerosis and healthy groups. Studentʼs t-test was used to determine significance.

The results revealed that a significantly higher value of Fe intake was observed for the atherosclerosis group compared to the healthy group. Also Zn intake of the obese individuals of the non-atherosclerosis group was higher than that of the atherosclerosis group. The serum Zn level in the healthy and obese groups was higher than that in the atherosclerosis group.

The quantitative analysis of sulfhydryl group of Metallothionein was developed in the human serum for the biomarker of the stresses toward the environments. The serum was treated by diluted hydrochloric acid and the reducing agent under the precise conditions, and analyzed by HPLC with N-(9-acridinyl) maleimide fluorometry. Sulfhydryl group was detected in pico (10-12) mole level easily and sensitively.

Angiogenesis is a complex process of sprouting new capillaries from the pre-existing blood vessels. It happens in normal physiological processes, such as embryonic development, the female menstrual cycle, bone remodeling and wound healing. On the other hand, angiogenesis also plays a crucial role in many pathological conditions, including tumor growth, diabetic retinopathy, psoriasis, rheumatoid arthritis and atherosclerosis. Here the antiangiogenic effect of rice bran tocotrienol (T3) and its mechanism from the fundamental data of in vitro cell studies, DNA chip analysis, and in vivo egg and animal models are described.