d-Amino acids are widely detected in various organisms and foods and expected to have physiological functions for human. We measured all of the d- and L-amino acids in 48 kinds of brown rice for sake brewing using HPLC. We found that the almost all brown rice contained the d-amino acid forms of Ala, Arg, Asn, Asp, Glu, Gln, His, Ser, Val. Asp and Glu. d-Ala was detected only in brown rice produced in Hyogo, Hiroshima, Kochi, and Fukui Prefecture. The d-Ser concentration was relatively high in brown rice produced in Yamagata, and Tokushima Prefecture. d-Val was detected in brown rice produced in Hokkaido and Iwate Prefecture but not in Kochi and Kagawa Prefecture. d-Asp was detected only in brown rice but not in the 70%, 60%, and 50% polished rice. These results suggested that d-Asp existed around the center of the brown rice, and other d-amino acids existed near the surface of the brown rice. Accordingly, d-amino acid contents of brown rice for sake brewing are different significantly depend on the production area and the polishing ratio.

Sweet corn with husk (Zea mays L. var. saccharata f. Peter corn) was stored at 5 °C for 4 d under a photosynthetic photon flux density (PPFD) of 1 μmol m^-2s^-1 using red and blue LEDs. During storage, the sugar content (Brix) was measured consecutively for 4 d, and the amino acid content was analyzed after 4 d storage. Corn stored under blue LED light at 5 °C had higher sugar content than corn stored in a dark, cool condition. Similarly, the content of umami was increased by storage under the blue LED-cool condition, a result likely due to increases in aspartic acid and the major amino acid, glutamic acid. The red LED-cool condition was not as good as the blue LED-cool condition for preserving corn, but was much better than dark, room temperature storage. In contrast, the palatability of sweet corn kept at room temperature in the dark quickly declined. Blue LED-cool preservation appears to be a useful technique for storing sweet corn with husk.

It is generally considered that phylloquinone in forage along with menaquinones synthesized by intestinal microbes meet the requirement of horses. We investigated the concentrations of fecal and plasma menaquinone-7 (MK-7) and the effect of MK-7 supplementation on plasma vitamin K concentration in adult horses.

Fecal and plasma samples were collected from twenty Thoroughbred horses aged 9.0±2.9 yr before MK-7 supplementation, and MK-7 concentrations in feces and plasma were determined. Subsequently the horses were allocated to 4 groups (n = 5) and each group was given MK-7 at 20 μmol/d, 40 μmol/d, 60 μmol/d, or no MK-7 supplement for 7 d. Plasma samples were collected before feeding, and 2, 4, and 8 h after feeding on d 7, and plasma concentrations of menaquinone-7 were determined (Experiment 2). Although fecal MK-7 concentration can be measured in all horses without supplying vitamin K, plasma MK-7 concentration was extremely low and was under the measurement limit in some horses. Additionally, the relationship between plasma concentration and fecal excretion of MK-7 was not observed. We suggest that MK-7 produced by intestinal bacteria is not important for horses. Plasma MK-7 concentration was higher in the 40 μmol/d group (P &;lt 0.001) and the 60 μmol/d group (P < 0.001) than in the control group through the experiment. Further, plasma MK-7 concentration was not changed after feeding in the 40 μmol/d group; plasma MK-7 concentration was stably high in this group. These results suggest that MK-7 is a good source of vitamin K for bone health in horses because the supplementation of MK-7 increases its plasma concentration greatly and continuously.

Biotin belongs to the one of the water-soluble vitamin B groups, and it is a cofactor of carboxylase, such as acetyl-CoA carboxylase (ACC), pyruvate carboxylase (PC), propionyl-CoA carboxylase (PCC) and 3-methylcrotonyl-CoA carboxylase (MCC). 3-Hydroxyisovaleric acid (3-HIA) which is a metabolite of leucine catabolism is a kind of organic acid in urine. The leucine degradation pathway often works to create acetyl-CoA and acetoacetate. When biotin is deficient, an alternative metabolic pathway produces 3-HIA. In this study, when the activity of 3-methylcrotonyl-CoA carboxylase decreased, 3-HIA excretion increased in the urine. The urinary excretion of 3-HIA was determined by HPLC. Feeding an additional two-fold increase of leucine had no effect on urinary 3-HIA but urinary biotin was increased due to the leucine. In addition, leucine had effects on relative weight and biotin levels in some tissues. From these findings, although 3-HIA is a sensitive indicator of biotin deficiency, we have to pay attention when evaluating biotin deficiency.

Intake of microminerals (iron (Fe), zinc (Zn), copper (Cu), manganese (Mn), iodine, selenium (Se), chromium (Cr) and molybdenum (Mo)) from diets in hospitals and nursing home was determined by atomic absorption spectrometry or inductively coupled plasma mass spectrometry, and compared with those estimated by a calculation using the Standard Tables of Food Composition in Japan, 5th edition (Fe, Zn, Cu and Mn) or 2010 (iodine, Se, Cr and Mo). In the intake of Fe, Zn, Cu and Mn, correlation coefficients between the analyzed values and calculated values ranged from 0.6 to 0.9 and Y-intercepts of the regression equations were near zero; however, slopes of the equation ranged from 0.50 to 0.69. In the daily intake of Se and Mo, the estimated values using the Standard Tables, 2010 were almost equal to the analyzed values. On the other hand, the calculated iodine intake in some diets was markedly lower than the analyzed values. In addition, the calculated Cr intake was less than one-quarter of the analyzed values. These results indicate that considerable loss of Fe, Zn, Cu and Mn occurs in cooking and the estimated Se and Mo intake calculated using the Standard Tables of Food Composition in Japan, 2010 is reliable.

Deferiprone (3-hydroxy-1,2-dimethyl-4-pyridone), a hydroxyketone compound with pyridine ring, is a water soluble, orally active iron chelator used for the therapy of Friedreichʼs ataxia and -thalassemia. Toxic effect of deferiprone was analyzed in relation to the generation of reactive oxygen species. 1. Deferiprone/iron complex inactivated aconitase, the most sensitive enzyme to oxidative stress. The inactivation was dependent on sodium azide, an inhibitor of catalase, indicating that deferiprone/iron complex can generate hydrogen peroxide as a principal product. Deferiprone was more effective on aconitase than maltol or kojic acid, hydroxyketone compounds with pyrane ring. 2. Deferiprone, maltol and kojic acid stimulated the atuooxidation of Fe²⁺ suggesting that these compounds promote the activation of dioxygen molecule by reduced iron. When chelating agents are used for iron storage disease, attention should be paid to the transition metal-complex mediated generation of reactive oxygen species.

Capsaicin, the pungent secondary metabolite found in fruit of the genus Capsicum, and vanillylamine, a metabolic precursor to capsaicin, are used in the food science, pharmaceutical, medical, and forensic industries; however, a rapid method to evaluate fruit for the presence of these two compounds is lacking. The present research describes the in situ visualization of capsaicin and its precursor in Capsicum fruit by laser-induced fluorescence imaging and spectrometry. When excited by ultraviolet lasers at 266 nm, capsaicin and vanillylamine have peak fluorescence emissions at 310 nm. The fluorescence spectra of precursors of capsaicin and analogs, i.e., trans-caffeic acid, trans-ferulic acid, trans-para-coumaric acid, vanillyl alcohol, vanillin, vanillic acid, had different peaks than those of capsaicin and vanillylamine. The localization of capsaicin and its immediate precursor, vanillylamine, was imaged with an ultraviolet-sensitive camera after excitation with a laser at 266 nm. Fluorescence images detected at 310 nm showed the localization of capsaicinoid and/or vanillylamine on the surface of placenta and septa of Capsicum fruits. No fluorescence specific to capsaicin and vanillylamine was observed in seeds or pericarp. Both bell pepper and sweet pepper also showed 310 nm fluorescence on the placental surface, suggesting the accumulation of vanillylamine in the placenta. Laser-induced imaging shows considerable promise as a suitable technique for rapidly screening Capsicum fruit for their capsaicin and vanillylamine contents.

Oyster is an old worldwide food containing abundant nutritional elements including many kinds of minerals such as zinc, nucleic acid, glycogen, and amino acids. It is reported that Oyster Extract Powder exhibits several physiological activities including anti-platelet aggregation effect in human and experimental animals. Recently, possible interaction between medical drugs and health foods has attracted the public attention, and in some health foods, occurrence of drug interaction is reported by concomitant ingestion with medical drugs. This time, we investigated the safety and possible drug interaction of Oyster Extract Powder in rabbits and rats.

Safety assessment by a 90 days repeated oral dose toxicity study in rats revealed no significant toxic effects of Oyster Extract Powder. Possible drug interaction was investigated with anti-platelet aggregation drugs including aspirin and ticlopidine, and anti-blood coagulation drug, warfarin, and no interaction was observed with any drugs. From these results, it was confirmed that Oyster Extract Powder is a safety health food showing no possible drug interaction with anti-platelet aggregation and anti-blood coagulation drugs.

In addition, glutathione level in hippocampus was analyzed since it was reported that Oyster Extract Powder increased glutathione levels. Then, significant increase in total glutathione level in hippocampus was noticed by administration of Oyster Extract Powder.

To estimate the daily intake of total lipids, saturated fatty acids, n-6 fatty acids and n-3 fatty acids in Japanese woman vegans, 33 duplicate portion samples of daily diets were collected from 11 Japanese female vegans and their fatty acid compositions were determined by gas chromatography after methylation with sodium methoxide. Intake of total lipid and each fatty acid in vegan diets was as follows (mean±SD): total lipid (% energy), 20.5 ± 7.9; saturated fatty acids (% energy), 2.6 ± 1.0; n-6 fatty acids (% energy), 4.4 ± 1.8; n-3 fatty acids (g/d), 1.2 ± 0.8. Compared with the tentative dietary goal for preventing lifestyle-related diseases in female aged 30 to 49 years in the Dietary References Intakes for Japanese 2010, n-6 fatty acid intake is appropriate; however the intake of saturated fatty acids and n-3 fatty acids in vegan diets is quite low.

Aromatherapy and the plant-based essential oils are widely used for the complementary and alternative therapies of symptoms including anxiety and depression. Furthermore, it was reportedly effective for the care of patients of Alzheimerʼs disease. To investigate the pharmacological effects of essential oils, we have developed an in vitro assays system using GT1-7 cells (immortalized hypothalamic neurons). Among tested 21 essential oils, we found that the neuronal death induced by hydrogen peroxide was attenuated by lemon, black pepper, geranium, juniper berry, majoram, eucalyptus, lavender, vanilla, and clarysage. We also examined the radical scavenger activity using DPPH (1,1-diphenyl-2-picrylhydrazyl) and the SOD (super oxide dismutase)-like activity. Our developed in vitro assay system could be useful for the neuropharmacological study of essential oils.

In a cropland area near Lake Dian, large amount of the crop wastes, i.e. vegetable and flower residues, were fed to cows in dairy farms. Since several vegetables and flowers tend to accumulate nitrate, animals kept such situations were suspected to ingest large amount of nitrate. The objective of this study was to quantify the dietary nitrate loads on dairy cows in the area. Questionnaire surveys on feedstuffs fed to cows were conducted in the dairy farms from October 2009 to September 2010. Nitrate contents in the feedstuffs and methemoglobin concentrations in the blood of cows were measured. The feeding amounts of crop wastes from vegetables and flowers were 28.4 kg to 50.0 kg per head per day on a fresh weight basis. The contents of nitrate nitrogen in the residues of rape blossoms, parsley, stem lettuce and carnations were 0.19%DM to 0.37%DM, which were potentially toxic levels for ruminants. Those in the residues of celery and broccoli were 1.36%DM and 0.59%DM respectively, which were toxic levels. However, the nitrate nitrogen contents in the total diets (sum of feed ingredients) were 0.15%DM to 0.19%DM, which were below the allowable limit. The ratios of methemoglobin to the total hemoglobin in the blood were less than 10%, indicating that the dairy cows did not fall into acute nitrate poisoning.

Enzymes that catalyze the degradation of organohalogen compounds are useful in environmental technology and chemical industry. Burkholderia sp. WS is a Gram-negative bacterium that can utilize an aliphatic unsaturated organohalogen compound, 2-chloroacrylate (2-CAA), as its sole carbon source. The production of 2 proteins, CAA43 and CAA67_WS, is induced when the bacterium is grown in 2-CAA medium. CAA43 catalyzes the conversion of 2-CAA into (S)-2-chloropropionate, whereas the function of CAA67_WS remains unknown. Recently, a homolog of CAA67_WS from Pseudomonas sp. YL (CAA67_YL), subsequently named 2-haloacrylate hydratase, was shown to catalyze the FADH2-dependent hydration of 2-CAA to produce pyruvate. Our results suggest that CAA67_WS has a similar activity. Gene-disruption studies of CAA43 and CAA67_WS indicated that CAA67_WS is physiologically more important in the assimilation of 2-CAA in Burkholderia sp. WS. The enzyme CAA67_WS was purified from Burkholderia sp. WS and characterized. The UV-visible spectrum of the protein indicated the presence of bound flavin. CAA67_WS released chloride ions from 2-CAA in the presence of FAD and reducing agents such as NAD(P)H. CAA67_WS is similar to CAA67_YL in these respects. However, while the reduced form of flavin mononucleotide (FMN) served as a cofactor for CAA67_WS, it did not for CAA67_YL. CAA67_YL is a bifunctional enzyme that catalyzes the hydration of 2-CAA and the reduction of FAD by using NADH; CAA67_WS did not catalyze the reduction of FAD. Thus, comparative studies of these 2 proteins can provide valuable information on the structure-function relationship of these proteins.

Abbreviations: 2-CAA, 2-chloroacrylic acid (S)-2-CPA, (S)-2-chloropropionic acid ESI-MS, electrospray ionization mass spectrometry

We detected the d-amino acids forms of Ala, Asn, Asp, Arg, Glu, Gln, His, Ile, Leu, Lys, Ser, Tyr, Val, Phe, and Pro in 141 bottles of sakes using HPLC. We used three individual derivatization methods of HPLC: precolumn & postcolumn methods of o-phthalaldehyde and N-acetyl-L-cysteine and (+) -1-(9-fluorenyl)ethyl chloroformate/1-aminoadamantane method. The d-amino acid content of the sakes was increased by using deep-sea water “Kaiyoushinosousui”, “Kimoto yeast starter”, “Yamahaimoto”, and the long aging process “Choukijukusei”. After three months storage at 25°C, the sakes that were brewed with the adenine auxotroph of sake yeast (“Sekishoku seishu kobo”, Saccharomyces cerevisiae) without pasteurization (“Hiire”) contained large amount of d-Ala, d-Asp, and d-Glu.

Previously, we reported that there are no differences in biotin contents and expression levels of bio2 gene, which is a key enzyme in biotin biosynthesis pathway, in pea sprouts among various cultivation conditions. We also found that biotin required for the growth of pea sprout is sufficiently supplied from the seed. In this study, we investigated whether biotin biosynthesis is affected by cultivation conditions (temperature and insolation) using pea sprouts hydroponically cultivated without seeds.

Seeds were removed after their germinations, and then pea sprouts were grown with culture fluid without biotin under 5 conditions: Control (C), 25°C/12-h light; Small amount of insolation (SI), 25°C/4-h light; No insolation (NI), 25°C/0-h light; Moderate low temperature (MLT), 12°C/12-h light; Low temperature (LT), 4°C/12-h light.

Biotin contents in SI and NI were approximately 67% and 43% of C, and decreased in proportion to the amount of insolation (p &;lt 0.05 and p &;lt 0.001, respectively). Biotin content in LT was also lower than C (p &;lt 0.05).

Secondly, to clarify the seasonal differences of biotin contents in green vegetables, we measured biotin contents in garland chrysanthemum (shungiku), spinach, and potherb mustard (mizuna) harvested in summer and winter in Niigata, Japan.

Bitoin contents in shungiku and spinach in winter decreased to about 56% and 47% of those in summer. (p &;lt 0.001, respectively)

These results strongly suggest that biotin biosynthesis in plants fluctuates responding to the cultivation conditions, especially the amount of insolation. This may lead to seasonal differences of biotin contents in green vegetables.

Hijiki (Sargassum fusiforme) plants were collected during their growth period from the end of November through April at 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. Manganese (Mn) and zinc (Zn) in 1N HCl were determined with an atomic absorption spectrophotometer.

Mn and Zn accumulated in the respective tissues at concentrations of 10 to 50 μg/g dry weight of tissues at the beginning of growth, and their concentrations remained at similar levels until the beginning of April. These may, as discussed, suggest that the levels of Mn and Zn are their biochemically required concentrations in the tissues of Hijiki.

To estimate iodine and chromium intake in Japanese infants dependent on commercial baby food and human milk, 53 commercial baby food samples (24 samples for ≥ 7-month-old baby and 29 samples for ≥ 9-month-old baby) were collected and their iodine and chromium concentrations were determined by inductively coupled plasma mass spectrometry. The iodine concentrations were markedly elevated by the use of kombu or hijiki and their medians (25-75 percentile) in baby food for a ≥ 7-month-old baby and a ≥ 9-month-old baby were 30 (21-103) and 42 (27-1045) ng/g wet weight, respectively. Mean iodine intake by 6 to 8-month-old babies and 9 to 11-month-babies was estimated to be 144 and 691 μg/d, respectively although their medians were estimated to be 89 and 84 μg/d, respectively. On the other hand, chromium concentrations (median (25-75 percentile)) in baby foods for a ≥ 7-month-old baby and a ≥ 9-month-old baby were 12 (7-12) and 10 (7-16) ng/g wet weight, respectively. Mean chromium intake by 6 to 8-month-old babies and 9 to 11-month-babies was estimated to be 8 and 10 μg/d, which were about 10 times higher than the value shown as the Adequate Intake in the Dietary Reference Intakes for Japanese. These results indicate that intermittent high-iodine baby food ensures sufficient iodine intake, and chromium intake is increased in Japanese infants after beginning to eat baby food.

Iron and zinc are essential micronutrients for human health. Deficiencies in these 2 nutrients remain a global problem especially among childbearing-age women not only in developing countries but also developed countries. Iron-zinc interactions are increasingly important because deficiencies of these micronutrients often occur together. Subjects were classified as iron deficiency anemia (IDA) (Hb < 12 g/dl and serum ferritin < 20 ng/ml), iron deficiency without anemia (ID) (Hb ≥ 12 g/dl and serum ferritin < 20 ng/ml), and Normal (Hb ≥12 g/dl and serum ferritin ≥ 20 ng/ml). We examined hemoglobin (Hb), serum ferritin, serum iron, unsaturated iron binding capacity (UIBC), total iron binding capacity (TIBC) and transferrin saturation as iron status and serum zinc in young women (age 19 to 20). The mean (±SD) Hb, and ferritin, Fe, UIBC, TIBC, transferrin saturation and Zn concentrations in serum of the participants were 12.9 ± 0.8 g/dl, 12.3 ± 4.4 ng/ml, 90 ± 43 μg/dl, 312 ± 79 μg/dl, 408 ± 56 μg/dl, 22.8 ± 12.9% and 81.1 ± 12.1 μg/dl, respectively. Anthropometrics of participants were consistent with the National Health and Nutrition Survey 2004 in Japan. The interesting observation was found that serum zinc of ID (81.1 ± 12.1 μg/dl) was not significantly different compared to Normalʼs (87.8 ± 17.4 μg/ dl) regardless IDA (98.7 ± 15.9 μg/dl) was. It corresponds to the results of Yokoi et al. that zinc pool sizes declined in premenopausal American women with ID. Further investigations of the association of iron and zinc nutriture are expected among Japanese young women.

Iron (Fe), zinc (Zn) and copper (Cu) are essential trace metals for our lives. Recent advances in understanding of the molecules involved in their metabolism have allowed for elucidation of the mechanism of their absorption in the intestine. Fe-, Zn- and Cu-specific transporters operate in enterocytes. Transporters that locate at the apical surface mediates uptake of Fe, Zn and Cu in the diet into enterocytes across the apical membrane, while other transporters are involved in efflux of them from enterocytes across the basal membrane into circulation, although intracellular trafficking of these metals between apical and basal sides of the enterocytes has yet to be defined. Herein, we briefly review the molecular mechanism, focusing on the key players involved. Moreover, we discuss the possibility that dietary components may increase the absorption efficiency of Fe and Zn through transporters, because recent studies have indicated that a number of dietary components affect their bioavailability.