Authors recently found that olfactory stimulation with scents of grapefruit oil (GFO) and lavender oil (LVO) inversely affect activities of autonomic nerves innervating the white adipose tissue, brown adipose tissue, adrenal gland, kidney and stomach, which was measured by electrophysiological method under urethane-anesthesia, then cause changes in physiological functions such as lipolysis, heat production (energy expenditure), blood pressure, blood glucose and appetite in rats and mice. Moreover, authors obtained evidence that the biological clock located in the hypothalamic suprachiasmatic nucleus (SCN) and histaminergic neurons are involved in these changes elicited by these oils. That is, olfactory stimulation with scent of GFO elevated activities of sympathetic nerves innervating the white and brown adipose tissues, adrenal gland and kidney, lowered the gastric parasympathetic (vagus) nerve activity and increased lipolysis, heat production (thus body temperature), blood pressure and blood glucose, and suppressed appetite. While, olfactory stimulation with scent of LVO lowered activities of sympathetic nerves innervating the white and brown adipose tissues, adrenal gland and kidney, elevated the gastric parasympathetic nerve activity, and decreased lipolysis, heat production (thus body temperature), blood pressure and blood glucose, and increased appetite. Moreover, all of these changes, which induced by GFO and LVO, were eliminated by either electrolytic lesion of the SCN or histaminergic blockers. Changes in auotnomic nerve activities and physiological functions were also induced by intestinal administration of a kind of lactobacillus and music. Intraduodenal injection of Lactobacillus johnsonii La1 suppressed activities of sympathetic nerves innervating the adrenal gland and kidney and reduced the blood pressure and blood glucose. Repeated sound application of the music, Traeumerai composed by Schuman, to ears caused suppression of the renal sympathetic nerve and reduced the blood pressure. These changes were also eliminated by the SCN-lesions or histaminergic blockers. Furthermore, authors and others found evidence that the SCN is a control center of autonomic nerves using pseudorabies virus (PRV), which is reptrogradely and polysynaptically transported into the brain.These findings suggest that the SCN and histaminergic neurons are involved in the mechanisms of effects of GFO, LVO, the lactobacillus and music on autonomic nerve activities, thus physiological functions.

Recently, it was reported that alkaline phosphatase (ALP) reduces the toxicity of lipopolysaccharide (LPS), a toxin of Gram-negative bacteria. Therefore, we examined the change in ALP activity and induction of tumor necrosis factor-α (TNF-α) in the duodenum of zinc-deficient rats. Fifty-six Sprague-Dawley rats were divided into two groups fed diets containing 0 % (zinc-deficient group) or 0.01 % (control group) zinc. Blood and duodenal mucosa samples (5 cm below the stomach) were obtained every week for four weeks, for ALP and TNF-α analyses. At one week, the serum ALP and intestinal ALP activities in the zinc-deficient group were significantly lower than in the control group, and decreased further over four weeks. TNF-α was detected in the duodenum of the zinc-deficient rats at four weeks, but not in the control group. This study shows that dietary zinc deficiency induces the expression of TNF-α protein in the duodenal mucosa; one cause of this response is a marked reduction in the detoxification of LPS by intestinal ALP.

The nutritional adequacy of diets in Japanese prison was assessed in a total of 11 penal institutions. Menu of prison diet was analyzed based on the Standard Tables of Food Composition in Japan (5th. edition). The energy, protein, fat, and cholesterol content was broadly in line with current dietary recommendations of Correction Bureau, Ministry of Justice, and it also meets national RDA (30-49 years old man). Although vitamin and mineral provision was almost sufficient, however, marginally deprived supply at some prison was noticeable for calcium, Vitamin B₁, B₂, B₁₂ and D. It is worth noticing that Vitamin D content at some institutions was insufficient, which was particularly important for those with limited exposure to sunlight. Sodium content was high at all institution, which was not improved from the previous survey. Food choices made by prison kitchen resulted in a wide variation in dietary content. Milks and fruits should be fortified and seasonings, beverages and confectionary were successfully regulated in most of the institutions. Insufficient provision of mineral and vitamin in some institutions should be improved by learning from the menu of successful prison diet.

In a course of research on chemical properties and functions of Japanese traditional plant foods, our interests were drawn to “Mukago”, bulbils of Japanese yam, Dioscorea japonica Thumb., since our preliminary study indicated that it contains α-galactose-recognizing lectin (α-Gal-Lec) beside unique amylolytic enzyme(s).

Fresh Mukago was homogenized in PBS at cold. After removal of starch, protein was precipitated with ammonium sulfate(0.6 sat.), which was fractionated using affinity chromatographic techniques, involving β-cyclodextrin (CD) for α-amylolytic enzyme, and α-Galactose-conjugate column for a specific lectin, respectively. The purified α-Gal-Lec, showing a single band on SDS PAGE(38 kDa) was a dimeric molecule. This lectin interacted with various α-Gal-containing polysaccharides, e.g.plant galactomannans and also human blood type B substance, but not type A. Thus, chemical and binding properties of α-Gal-Lec appeared resemble to that of arrow-head tuber. The amylolytic enzyme, purified through β-CD was homogenous on SDS PAGE(31 kDa). Its action pattern on starch was appeared an intermediate manner of salivary (or bacterial) and funji (Takaamylase) types. It readily hydrolyzes Mukago starch, which might contain rather short α-1, 4 linked branches.

The effects of biotin deficiency on maternal metabolism and embryonic development in pregnant mouse dams were studied. In the biotin-deficient group, the inhibition of embryonic development and external malformations such as cleft palate (100 %), micrognathia (100 %) and micromelia (91.4 %) were detected in embryos. Biotin excretion in urine was decreased on day 4 of gestation (dg) and subsequently not detected, and the urinary concentration of 3-hydroxyisovaleric acid (3-HIA) was increased after dg 12. In contrast, the biotin concentration in urine was significantly increased on dgs 4, 8 and 12 in the biotin-supplemented group, and decreased on dg 16 in biotin-supplemented and biotin-control groups. Pyruvic acid in the biotin-deficient group was higher than that in the biotin-supplemented group throughout gestation. These findings demonstrated that the requirement of biotin increases at conception and/or during embryonic development, and a large amount of biotin is necessary for embryonic development in the late stage of gestation.

A labile selenium donor compound, selenophosphate, is synthesized from selenide and ATP by selenophosphate synthetase (SPS). Selenophosphate is required by several bacteria and by mammals for the specific incorporation of selenium into selenoproteins and modified seleno-tRNAs. Although free selenide can be used in vitro for synthesis of selenophosphate, the physiological selenium substrate has not been identified. To identify the selenite-reducing system in E. coli, we constructed a gor mutant strain and a trxB mutant strain of E. coli MC4100. Active FDHH can be detected by monitoring benzyl viologen reduction. In the cells growing under fermentative condition on low-salt-medium in the absence formate at pH 7.5, the activity of FDHH was optimum. The wild-type strain MC4100 and the gor mutant strain were able to produce an active FDHH, but the trxB mutant strain failed to show FDHH activity. When ΔselD mutant strains were complemented by human lung Sps2Cys, the trxB mutant strain couldn’t generate active FDHH. We suggest that reduction of selenite in vivo is not dependent upon the glutathione-reducing system, but instead on the thioredoxin-reducing system.

1. Pyridine compounds with 2-carboxylic group produced reactive oxygen species in the presence of transition metals, iron and copper. Aconitase, the most sensitive enzyme to oxidative stress was inactivated markedly by dipicolinic acid (pyridine 2, 6-dicarboxylic acid) and less potently by picolinic acid (pyridine 2-carboxylic acid) in the presence of ferrous sulfate. The inactivation was dependent on KCN, an inhibitor of Cu/Zn-SOD, suggesting that pyridine carboxylate/iron complexes can generate superoxide radical as a principal product. Introduction of carboxylic groups to 3, 4 or 5 in the pyridine ring decreased the inactivating effect. 2. Aconitase was also inactivated by copper/ascorbate complex, particularly in the presence of pyridine 2, 4-dicarboxylic acid (lutidinic acid). The inactivation was dependent on sodium azide, an inhibitor of catalase, suggesting that Cu/ascorbate produces hydrogen peroxide. 3. Dipicolinic acid and lutidinic acid at lower concentrations enhanced the Cu/ascorbate-dependent formation of 8-hydroxy-2’-deoxyguanosine in DNA, indicating generation of hydroxyl radical. Prooxidant action of pyridine carboxylic acids may explain the bactericidal or apoptotic action of these compounds reported previously.

Digestion is the process in which the bulk of ingested nutrients are broken down to oligomers or monomers in the mouth, stomach and intestine before they are absorbed and made available to all the cells of the body. Gastric juice is characterized by the presence of HCl, pepsin, mucus and an intrinsic factor. It is well established that the secretion of gastric juice is promoted by a type of amino acid and peptide. Oyster is a shellfish classified as Pelecypoda and is a nutritious food containing a great deal of zinc and taurine. Taurine has been reported to reduce myocardial damage and to have a beneficial effect on blood glucose and lipid levels. Thus, we investigated the effect of oyster extract on the gastric secretion in rats. Pylorus-ligated rats were sacrificed 6 h after the administration of oyster extract or taurine, and the gastric juice was collected and analyzed for volume, pH, acidity and pepsin activity. The results indicate that oyster extract may decrease pepsin activity and pepsin output; however, oyster extract significantly raised the acidity. Furthermore, it is suggested that the composition of oyster extract, excluding the taurine, may promote the secretion of HCl and organic acid because oyster extract more significantly raised acidity than taurine.

Students in agrarian villages of rural Cambodia are supposed to suffer high rates of iron-deficiency anemia in the context of a monotonous diet. To compare the efficacy and safety of daily consumption of 10 mg of iron in form of FeSO₄⁺ citric acid (Fe+CA group) and NaFeEDTA (Fe-EDTA group), each mixed in 10 mL of Khmer fish sauce as compared to a fish sauce placebo control with no iron added (Control group). A total of 140 students aged 6 to 21 years were enrolled in double blind, placebo-controlled intervention trial. They were randomly allocated to one of three treatment groups, and followed for 21 weeks including 12 national holidays (6 days/week). During this time 104 meals with 10 mL of fish sauce added were offered to each participant. Initial and final measurements of body weight and spinal length, of the concentrations of hemoglobin (Hb), ferritin (Fr) and C-reactive protein were obtained. The two different iron-fortified fish sauce increased Hb and Fr significantly as compared to the control group. No significant differences were observed between Fe+CA and Fe-EDTA fortification, neither regarding mitigation of iron-deficiency anemia nor regarding side-effects. Fe+CA and Fe-EDTA showed equivalent efficacy and safety during the intervention study. National launch of iron fortified fish sauce is one of the promising strategies to combat iron deficiency anemia in Cambodia.

To estimate the vanadium intake in the Japanese population, the vanadium content in various food and drinking water samples (3 Japanese tap water samples, 19 Japanese mineral water samples. 21 European mineral water samples and 6 North American mineral water samples) was determined using inductively coupled plasma mass spectrometry. All the tap water samples showed values of less than 5 ng/mL as vanadium concentration. Among 46 mineral water samples, 31 samples (67.4 %) showed less than 5 ng/mL and only 3 samples showed more than 50 ng/mL. Among the food samples analyzed, the highest vanadium content (> 1000 ng/g dry weight basis) was observed in several algae and shellfish samples. The moderate vanadium content (100-1000 ng/g dry weight basis) was observed in leaf vegetables, dairy products and white table bread while the low vanadium content (< 50 ng/g) was observed in cereals, soybean, potatoes, fruits, meats, eggs and fishes. Based on the present quantification of vanadium in foods and drinking water and the recent National Nutrition Survey in Japan, the average vanadium intake of Japanese population was preliminarily estimated as about 30 μg/d/capita. The principal vanadium source in the Japanese diet was thought to be wheat products, vegetables and dairy products.

In Japan, colorectal cancer demonstrates the third highest cancer mortality following lung cancer and gastric cancer. Western dietary habits symbolized by a high fat/low fiber diet are regarded as one of the risk factors on an epidemiological survey. The present study evaluated the preventive effects of oyster extract on the formation of chemically induced colonic aberrant crypt foci (ACF). Mice fed an AIN93G modified diet containing oyster extract (0.2 %, 1.0 % and 5.0 %) were subcutaneously injected with 1, 2-dimetylhydrazine (DMH) (15 mg/kg body wt) once weekly for 6 weeks (Experiment 1). Mice that were were first administered DMH once weekly for 6 weeks, were then fed an AIN93G modified diet-containing oyster extract (Experiment 2). The number of ACF was evaluated after 0.02 % methylene blue staining of the colon and was scored under a light microscope at 40 times magnification.

Experiment 1 demonstrated weight loss of 10-15 % in the Control group beginning one week after dosing with DMH. In the group receiving oyster extract powder, weight loss was slight. The mice fed the oyster extract diet showed a decreased number of total colonic ACF with a multiplicity of 4⁺ aberrant crypts compared with that in Control. In experiment 2, the mice fed an oyster extract diet did not show a change, total colonic ACF or a multiplicity of 4⁺ aberrant crypts compared with those in Controls. Oyster extract did not diminish or delay development depression after precancerous manifestation, however we could expect suppression of the initiator action of the carcinogen routine consumption.

The usage of enriched stable isotopes in nutritional sciences is increasing and includes measurements of kinetic parameters for micronutrients and amino acids, determination of true absorption rates based on single-tracer and double-tracer techniques, and quantification of endogenous losses. The suitable compartment model and required precision in the isotope ratio measurements by mass spectrometry for human zinc metabolism are discussed. The 500 data sets of the simulated plasma zinc disappearance obtained from giving 2 % normal random noise to Miller et al’s polyexponential model were analyzed by nonlinear regression. The three-compartment open mammillary model was found appropriate for the 9-day observation period; the closed model was found appropriate for the 24-hour observation. Approximately one percent overall precision in isotope ratio measurements is necessary and feasible in kinetic analyses of zinc metabolism over 24-hour to 9-day observation period.

Iron (Fe) and zinc (Zn) are suspected to be easily depleted in humans at the same time, because dietary sources such as meat and major absorption inhibitors such as calcium and phytate are common for Fe and Zn. To determine the effects of Fe and/or Zn deficiencies on physiological parameters in rats, forty 4-week-old male Sprague-Dawley rats were assigned into 4 dietary treatment groups: severely Fe-deficient (FD), moderately Zn-deficient (ZD), severely Fe-deficient/moderately Zn-deficient (FZD), and control (Cont). Body temperature of FD and ZD were decreased, but that of FZD was not changed. TIBC was decreased by Fe deficiency. Hemoglobin and hematocrit were decreased by Fe deficiency and increased by Zn deficiency. Total body water per body weight was increased in FZD and intracellular water/extracellular water ratio was increased in ZD. Plasma erythropoietin concentration was markedly decreased by Zn deficiency.

Organotin compounds are toxic to the immune system. Alkyltin compounds such as dibutyltin (DBT) and tributyltin (TBT) cause severe thymus atrophy. At histopathological examination, there is marked depletion of lymphocytes and the depletion is due to the cell death. In this study, the mechanism on the cell death induced by DBT or TBT was investigated from the angle of apoptosis in T-lymphocytes.

The results revealed that DBT induced caspase-independent cell death (necrosis), whereas TBT induced caspase dependent cell death (apoptosis). Furthermore, we demonstrated the mechanism on TBT-induced apoptosis. Bid cleavage by activation of caspase-8 produced tBid and released cytochrome c from mitochondria. Release of cytochrome c induced the activation of caspase-3 through caspase-9. CAD activated by caspase-3 induced DNA fragmentation. That is to say, it was elucidated that TBT induced caspase-dependent and mitochondria-mediated cell death.

The Japanese eel, Anguilla japonica, is the traditional staple food in summer in Japan. We have previously reported that both carnosine and an aqueous extract from eel muscle prevent zinc-induced GT1-7 cell death, which is associated with neuronal death after transient global ischemia. In this study, we found that these protective activities were exhibited by the heat-treated extracts from both eel muscle and skin. We developed a high-performance liquid chromatography system for quantifying carnosine; this system demonstrated that the muscle extract contained carnosine in an amount sufficient for preventing zinc-induced GT1-7 cell death, which reveals that carnosine is the substance in the muscle extract that is responsible for the protective activity. The analysis of the expression of zinc- inducible gene indicates that carnosine acts as an intracellular protective substance in GT1-7 cells. We also demonstrated that the eel skin contained unknown substance(s) that prevent zinc-induced cell death.

During the water-soaking process to remove arsenic from dried Hijiki at 30°C, some other beneficial elements are also lost. During this process, the amounts of calcium, iron, magnesium and zinc released into water were 49 %, 32 %, 77 % and 70 % of those originally present, respectively. Thus, water-soaking for longer than 30 min at room temperature is recommended to eliminate arsenic from Hijiki, but it should be cooked together with other materials, such as soybeans, carrot etc, to supply the lost beneficial elements.

The serine racemase from Oryza sativa L. is a pyridoxyal 5’-phosphate containing enzyme that catalyzes a racemization and a dehydratation of serine. In this study, we constructed the high expression system of the serine racemase from Oryza sativa L. in Escherichia coli, and examined the effect of the metal ions on the activity. The serine racemase activity increased by addition of Mg²⁺ or Ca²⁺, while the dehydratase activity decreased by Mg²⁺ or Cu²⁺. The kinetic analysis showed that Mg²⁺ does not affect on the Vmax value of the racemase activity but decreases the Km value for L-serine. These results suggest that the serine racemase and dehydratase activities are regulated by Mg²⁺.

To analyze soil and plant diversification in Yaku-shima, we measured magnesium (Mg), aluminium (Al), calcium (Ca), manganese (Mn), sodium (Na), iron (Fe), nickel (Ni), and zinc (Zn) contents in tea leaves collected from nine different tea plants by ICP-Atomic Emission Spectrometry (ICPS1000IV). Water, nitrogen, positive ion and phosphoric acid contents and electric conductivity, pH and cation exchange capacity of soils picked up from corresponding tea plants were also evaluated. Multiple classification analysis of tea leaves and soils showed significant difference in seasonality of tea leaves, even though there was no difference in nine soils.

Molybdenum status was examined in rats fed experimental diets containing graded level of molybdenum. Male 5-week Wistar rats were divided into three groups. One group was fed a basal AIN93G diet without ammonium molybdate (molybdenum content, 0.08 μg/g) and other groups were fed the basal diet supplemented with 0.1 or 0.5 μg/g of molybdenum as ammonium molybdate for 4 weeks. Molybdenum concentrations in liver and kidney of the rats were not varied with dietary molybdenum intake level but the serum molybdenum concentration was gradually increased with an increase of dietary molybdenum level. Dietary molybdenum level did not effect on copper concentrations in the liver, kidney and serum as well as xanthine oxidase activity in the liver and the serum uric acid level. These results indicate that variation of dietary molybdenum level in the present study (0.08 to 0.58 μg/g) did not effect on molybdenum status other than serum molybdenum concentration.

It has been reported that the cellular Cu²⁺ ion and metallothionein (MT) expression contribute to the acquisition of cisplatin (CDDP)-resistance phenotype in the tumor cells. However, the mechanism of cellular MT induction by CDDP has not been clear. Previously, we reported that the nuclear proteins prepared from HeLa cells with the elevated MT level after treatment of 12-O-tetradecanoyl phorbol-13-acetate (TPA) or the tumor necrosis factor α (TNFα) bind to the metal responsive elements (MREs) as the regulatory DNA sequences in human MT-IIA gene promoter, in the presence of Cu²⁺ ion. In this study, the copper effect on binding of MREs to the nuclear proteins extracted from CDDP-resistant HeLa cells which were maintained for 3 months in Dulbecco’s modified Eagle (DMEM) medium supplemented with 10 % FBS and 1 μM or 5 μM CDDP. The expression level of MT-IIA mRNA in CDDP-resistant cells was compared with that of the normal HeLa cells, the resistant cells expressed more MT-IIA mRNA than the HeLa cells. The plasmid constructed the human MT-IIA promoter gene in puC19 were digested with the restriction enzymes. The bindings of linear DNA fragments produced by the enzyme digestion to the nuclear extracts prepared from the CDDP-resistant cells or the cells treated with 1 μM CDDP for 1 h in the presence of Cu²⁺ ion, was examined by the gel-mobility shift assay. It was suggested that Cu²⁺ ion may play an important role in the binding of the proteins in nuclear extracts prepared these cells to 3’-end DNA fragments of MRE_d.

In the previous study, we found that the exposure of tributyltin chloride (TBTC) made to disturb the balance of trace elements in brain tissues remarkably and to accumulate excessive calcium in olfactory bulb and olfactory epithelium^1-5). Moreover, under the conditions, pathological cell death of neuron, especially granular neuron, was observed in olfactory bulb and olfactory epithelium^1-5). Therefore, we have predicted that organotin-induced cell death of the olfactory neurons may be caused by the accumulation of excessive calcium. However, a mechanism on the cell death by the higher level concentration of calcium and the location of intracellular calcium have not been clear.

In this study, the olfactory neurons were isolated from olfactory bulb and the movement and location of intracellular calcium in the olfactory neurons after the exposure of TBTC were investigated by an in vitro experimental system. The movement and location of calcium in the olfactory neurons after the exposure of TBTC or the higher level concentration of calcium were observed by a confocal laser scanning microscopic system. As the results, transference of cytosolic calcium to nucleus in neurons after organotin-exposure was observed in the fluorescent images. This result suggests the transference of excessive calcium to the nucleus is strongly associated with the activation of endonuclease, DNase I in the nucleus, followed by the induction of apoptosis.

Zinc (Zn) is an essential trace element and a cofactor of over 300 enzymes. Various symptoms such as growth retardation and dermatitis are associated with Zn deficiency. In recent years, it has been suggested that Zn deficiency causes alteration in the biological defense system. To investigate effects of Zn deficiency on lungs, we examined morphological changes and expression of TNF-α mRNA in the lung tissue after high pressure exposure to oxygen. In the present study, two special diets were prepared; a Zn deficient diet and a standard diet (0.01 % Zn). Seven- weeks-old SD strain male rats were fed either on a Zn deficient (Zn-D group, n = 16) or a standard (Control group, n = 16) diets for 5 weeks. Eight of 16 Zn-D rats and 8 of 16 Control rats were exposed to 2 ATA pure oxygen for 5 hours (ZnD-O₂ and Control-O₂ groups). Then, determination of the number of white blood cells (WBC) and serum CRP level, histopathological examination of lung tissue and detection of TNF-α expression in the tissue were carried out.

Severe pathological changes such as congestion, destruction of alveoli and the appearance of inflammatory cells were seen in ZnD-O₂ rats, suggesting that Zn deficiency reduces the threshold level of oxygen toxicity in the pulmonary tissue. On the other hand, increase in TNF-α mRNA was not detected in ZnD-O₂ animals, although severe damage in the lung was observed. Further studies including localization of TNF-α mRNA expression within the tissue are required to clarify the mechanism of the synergistic effects of Zn deficiency on the lung lesions caused by inhalation of high pressure oxygen.

Effect of dietary zinc (Zn) level on tissue Zn distribution in mice was examined over two generations. Male and female eight-week-old ddy mice were fed diets containing 10 or 100 ppm of Zn as zinc sulfate or an oyster extract; there were four dietary groups consisted of both male and female mice. The groups were designated as follows: 10 ppm Zn as zinc sulfate, SL; 100 ppm Zn as zinc sulfate, SH; 10 ppm Zn as oyster extract, OL; 100 ppm Zn as oyster extract, OH. After 2 weeks of feeding, the mice were mated in each dietary group and then fed each diet for 6 weeks further. The second generation of mice born during these 6 weeks were nursed by the mother mouse and then fed each diet for 3 weeks. In the first generation mice, serum Zn levels of the SH and OH groups were higher than those of the SL and OL groups, but there were no difference among the four dietary groups in Zn levels of the liver, kidney, spleen, tibia, testis and hair. In the second generation, Zn levels in tibia, testis, hair and serum of the SH and OH groups were higher than those of the SL and OL groups. Differences in the sex and dietary Zn source (zinc sulfate or oyster extract) did not influence the serum, organ or hair Zn levels. These findings indicate that organ and hair Zn levels remain constant in mature female mice throughout their pregnancy and lactation periods even though they are fed diets with Zn levels varying from 10 to 100 ppm. However, organ and hair Zn levels in immature mice varied with dietary Zn intake.

It is known that biotin is intrinsic for maintaining reproductive function. Biotin deficiency induces both external and skeletal malformations in the embryos of fowl, and maternal biotin deficiency is severely teratogenic in mammals. In mice, maternal biotin deficiency results in cleft palate, micrognathia and limb hypoplasia. However, the relationship between morphogenesis and biotin is not sufficiently clear. This study was conducted to elucidate the mechanism of biotin transport from dams to embryos and the nutritional roles of biotin in ICR mice. It became obvious that biotin was supplied from dams growing embryos during morphogenesis. In particular, a large amount of biotin was transported to palates and mandibles on days 12-15 of gestation. In fetuses, the transportation of biotin to embryos differed among embryonic growth periods and organs, and biotin may play a principal role in the formation of tissues and organs.

These results suggest that biotin is an essential nutrient and may play an important role in reproductive performance.

Biotin has an important role in gluconeogenesis, amino acid metabolism, and energy metabolism. In the Dietary Reference Intakes for Japanese (2005), adequate intake of biotin in adults was set at 45 μg/day although the scientific evidence concerning biotin is inadequate to designate a set value. In the present study, we examined the effect of the protein sources on bioavailability of biotin in healthy women. Eleven healthy women were enrolled in the balance study of biotin for 16 days. The diets were mainly seafood as the protein sources. The dietary intake of biotin ranged from 24 to 56 μg/day. Serum concentration and urinary excretion of biotin did not change for the balance periods. Biotin excretion in feces was 1.3 times higher than the intake. These results indicate that the fecal excretion of biotin should be considered when evaluating biotin balance.

To establish a microbiological assay for biotin using Lactobacillus plantarum (ATCC8014) in a 96-well microtiter plate, we studied the influence of several pretreated biotin standard solutions on the microbial growth. Then, we measured serum biotin levels by our method, and assessed dietary biotin intake by a 3-day weighed food record to evaluate the biotin status in young Japanese women. A series of pretreatments, including an acid hydrolysis, inhibited growth of the organism. An inverse relationship was observed between growth inhibition and the dilution ratio of biotin standard solutions. These results indicated that biotin standard solutions should be pretreated in a way similar to that of the examination samples. Serum biotin levels (48.3 ± 17.4 μg/mL) were above normal (1.0 μg/mL) and did not change after 14-day biotin supplementation (60 μg/day). Daily biotin intake (48.3 ± 17.4 μg/day) calculated using the German food composition table was higher than the adequate intake (AD; 45 μg/day) of Dietary Reference Intakes for Japanese. There was no correlation between biotin intake and serum biotin level. These results suggest that AD of biotin is sufficient to maintain normal biotin levels. However, further investigations considering other factors such as urine output and supply from enteric bacteria, are necessary to evaluate a more accurate biotin status.

We studied the effect of biotin deficiency on insulin secretion from pancreas in rats with varying degrees of biotin-deficiency: moderate biotin deficiency (MBD), biotin deficiency (BD), and severe biotin deficiency (SBD). The plasma insulin levels were reduced with the severity of the deficiency. There was a significant positive correlation between the plasma insulin and plasma biotin levels. (r = 0.723, p < 0.001). To investigate the insulin secretion from pancreas, the isolated pancreas was perfused with 20 mM glucose or 10 mM arginine, which is a non-metabolic stimulus for insulin secretion. Compared to the control, the levels of insulin response to glucose in MBD, BD, and SBD rats were approximately 43 %, 35 %, and 22 %, respectively (p < 0.01). However, in MBD rats, the insulin content of the pancreas did not decrease significantly as compared with the control, and no retrogressive pathological change was observed in pancreas. In contrast, the level of insulin response to 10 mM arginine in MBD rats was approximately 76 % of the control (not significant). These results indicate that biotin directly acts in the glucose metabolic pathway for ATP production, which is a key mediator of nutrient-induced insulin secretion. Our study suggests that biotin is an essential factor and plays a pivotal role in the normal glucose-induced insulin secretory response.

The purpose of this study was to elucidate the dietary factors that influenced serum folate. Data were obtained from 36 vegetarians and 70 omnivorous middle-aged voluntary women. The vegetarian diet consisted mainly of unpolished rice, green vegetables, and tofu (bean curd), under a physician’s guidance, and there were individual variations. Serum folate concentrations were measured by chemiluminescent immunoassay. The BMI, % body fat, waist circumference, and diastolic pressure in the vegetarian group were significantly lower than those in the non vegetarian group. The folate intakes from cereals, nuts and seeds, pulses and green vegetables in the vegetarian group were significantly more than those in the non vegetarian group. The serum folate and the blood rheology in the vegetarian group were significantly higher than in the non vegetarian group. In dietary behavior, the point of preference for excess (4 items), recognition of constitution (3 items), and snack intention (3 items) in the vegetarian group were lower than those in the non vegetarian group. Serum folate levels were significantly correlated with the intakes of folate from nuts and seeds, pulses, fruits and green vegetables. Serum folate levels were significantly negative correlated with the intakes of energy, fat, carbohydrate, vitamin D and B12. These data suggest that the vegetarian diet increase the serum folate, and subsequently decreased the risk of cardiovascular disease and Alzheimer disease.

Hospitalized cancer patients that are treated with chemotherapy often suffer from nutritional problems called hos- pital malnutrition. In order to assess the nutritional condition in patients who were admitted to Ashiya Municipal Hospital for cancer chemotherapy, we collected the laboratory data and questionnaires asking for changes in the taste and preference for foods. These hospitalized cancer patients, irrespective of the organs affected, were shown to have low total protein (TP) and hemoglobin (Hb) values at admission, during hospitalization, and on discharge. Questionnaires told that 67 % of the cancer patients developed changes in sense of taste and preference for foods after chemotherapy. These findings suggest that nutritional assessment during hospitalization and a nutrition control system administered by a nutrition support team designed for each patient are necessary.

Prooxidant and apoptosis-inducing effects of a polyphenol resveratrol that is a principal constituent in grape epicarp on HL60 cells were examined. Resveratrol produced reactive oxygen species (ROS) in HL60 cells in a dose-dependent manner. ROS generation was enhanced by addition of transition metals. Treatment with 10 μM resveratrol for 20 h induced an apoptotic cell death of HL60 cells, in a dose-dependent manner. Resveratrol induced depolarization of mitochondrial membrane in accordance with activation of caspase-3. These results indicate that resveratrol mediated intracellular production of ROS is closely related to the induction of an apoptosis of HL60 cells.

In this study, in order to elucidate the characteristics of “umami” from bouillon extracted from boiled smoke-dried and molded bonito (Katsuodashi), we compared the composition and number of amino acids of different production districts in bonitos and seaweeds, as well as in different types of water (tap water and RO film filtration water). There was a large difference among the production districts of bonito bouillon in terms of amino acid composition and concentration. A specific kind of amino acid was contained in each bonito bouillon. The amino acid composition of seaweed bouillon also differed by producing district. A large amount of glutamic acid and aspartic acid was contained in the products of Rausu (170.3 and 118.9 μmol/mL, respectively) and Rishiri (159.7 and 100.2 μmol/mL, respectively). On the other hand, the products of Rausu (10.4 and 10.4 μmol/mL) and Hidaka (13.0 and 10.2 μmol/mL) have sweetness-producing alanine and proline in high concentrations. Thus, seaweed bouillon has umami-yielding glutamic and aspartic acid not contained in bonito bouillon. Also, there were differences in the amino acid concentrations of bonito and seaweed bouillons. From these findings, we confirmed that combining bonite and seaweed bouillons can compensate for each of their insufficient tastes, producing a bouillon high in umami as a result.