Melatonin is secreted during the hours of darkness and is thought to influence the circadian and seasonal timing of a variety of physiological processes. Serotonin N-acetyltransferase (AA-NAT) which is found to be expressed in pineal gland, retina, and various tissues, catalyses the conversion of serotonin to N-acetylserotonin and is known as the rate-limiting enzyme in the biosynthetic pathway of melatonin. The compounds that modulate the activity of AA-NAT can be used to treat serotonin-and melatonin-related diseases such as insomnia, depression and seasonal affective disorders (SAD). Several assay methods have been developed by which to measure AA-NAT activity. We have also developed a simple, rapid and sensitive AA-NAT assay method that takes advantage of differences in the organic solubilities between acetyl CoA and N-acetyltryptamine. We screened modulators of AA-NAT activity from the water extracts of the medicinal plants. We found MNP1005 which strongly inhibited the activity of AA-NAT ($IC_{50}$=2.2$\mu$M). Enzyme inhibitory kinetic studies revealed that MNP1005 exhibited a noncompetitive inhibition toward tryptamine. The antidepressant effect of MNP1005 was investigated on behavioral despair test so called forced swimming test (FST). MNP1005 significantly increased swimming behavior by reducing immobility with treatment of 10 mg/kg when compared to the vehicle-treated control group (P < 0.05). This suggests that MNP1005 possesses antidepressant activity. The influence of chronic MNP1005 treatment on the expression of brain-derived neurotrophic factor (BDNF) was examined by in situ hybridization and Northern blot. Chronic treatment of MNP1005 blocked the downregulation of BDNF mRNA in the frontal cortex and other cortex regions in response to restraint stress.

L-theanine is a unique amino acid, found almost solely in tea plants. It is the main component responsible for the exotic taste of green tea. In our studies of L-theanine, we have found a variety of biological activities including relaxation and the alleviation of PMS. In general, animals generate very weak electric pulses on the surface of the brain, called brain waves. Brain waves are classified into four types, namely $\alpha$, $\beta$, $\delta$ and $\theta$-waves, based on their frequency. Brain waves correlate with individual mental conditions. For example, generation of $\alpha$-waves is considered an index of relaxation. In human volunteers, $\alpha$-waves were generated on the occipital and parietal regions of the brain surface within 40 minutes after the oral administration of 50 or 200 mg Suntheanine$^{TM}$ without causing drowsiness. Premenstrual Syndrome (PMS) is a symptom unique to women which appears in the luteal phase from the ovulation period through the first day of menstruation. It possesses characteristics of having a peak just prior to menstruation and disappearing 1 - 2 days following the start of menstruation. Symptoms of PMS are generally categorized as mental, physical and social symptoms. When comparing the reported Symptoms of PMS by the methods of MDQ score, the Suntheanine$^{TM}$ group was found to have a lower incidence of PMS symptoms, including physical, mental and social symptoms. Overall, a significant alleviation of PMS symptoms by the administration of 200 mg Suntheanine$^{TM}$ was observed. With the successful industrial production of L-theanine, we are now able to supply Suntheanine$^{TM}$, offering a tremendous opportunity for designing functional foods targeting relaxation and the alleviation of PMS.

Purpose: We performed a randomized, double-blind, placebo-controlled trial in mild to moderate myopia patients to evaluate the benefit of taking a nutrient supplement containing anthocyanoside oligomers for improving nocturnal visiual function and/or clinical symptoms. Methods: The subjects included have refractive error between -lD(Diopters) $\~$-8D in both eyes, symptoms of decreased night vision and asthenopia based on the scoring result of a pre-structured questionnaire, and abnormal results of mesopic contrast sensitivity(MCS) screening test showing abnormal curve of contrast sensitivity in the middle and high frequency level, between 6.0 and 30.0 CPD(Cycles per degree) at mesopic condition(-2$\~$0 log cd/$m^2$). Total 60 people who qualified the criteria above were enrolled and the subjects were instructed to take the investigational product (anthocyanoside or placebo) twice daily for a 4 week period. The enrolled subjects were investigated for nocturnal vision performance by MCS and clinical symptoms at their first visit and re-evaluated at post-intervention (4 weeks later). MCS was measured and improvement of contrast threshold level according to each CPD was calculated by subtracting initial values from final values. Age, refractive error, and MCS were compared between the placebo and anthocyanoside. Results: After 4 weeks of drug administration 22 of the anthocyanoside group showed symptom improvement compared to 1 of the placebo group (p=0.000). Contrast sensitivity levels according to each CPD before and after drug treatment showed significant improvement in the anthocyanoside group but not in the placebo group. Mean MCS change of anthocyanoside group is 2.41$\pm$1.91 which showed significant improvement compared to -0.40$\pm$2.47 of the placebo group(p=0.000). MCS changes of anthocyanoside group showed significant improvement compared to placebo group in all levels of CPD(p<0.05). During our investigation none of the subjects complained of specific side effects related to anthocyanoside use. Conclusion: Our results show that under careful selection of people with significant symptoms and definite MCS abnormalities, anthocyanoside oligomers may improve the subjective symptoms and objective MCS results.

Soybean, rice and nuts are the staple foods in South East Asia. In order to clarify the function of these foods in relation to coronary heart diseases, we have done a series of experiments using animals and humans. Soybean protein preparations (SPI) in comparison with an animal protein, casein (CAS), resulted in reducing atherosclerotic lesion development in apolipoprotein (apo) E deficient mice that develop an advanced lesion similar to that in humans. This action was attributed to the protein, since the ethanol-extracted SPI (EE-SPI), from which isoflavones and saponins removed, lowered the lesion development in apo E-deficient mice. However, The EE-SPI, compared to the SPI, resulted in a decrease of mRNA for key proteins of cholesterol metabolism (low density lipoprotein receptor and cholesterol 7$\alpha$-hydroxylase) in hypercholesterolemic rats, followed by an elevation of the serum cholesterol level, indicating a contribution of isoflavones to the serum cholesterol level. Rice protein (RPI) was also effective to lower the lesion development in apo E deficient mice. Both the SPI and RPI led to an increased level of the serum NOiN03, metabolites of NO. This effect appeared to be attributed to their high content of arginine. Besides, C57BL/6J mice fed a diet containing whole grain rice had an elevation of the serum high-density lipoprotein cholesterol. Furthermore, rice bran oil and the unsaponifiable fraction that is rich in plant sterols were effective to lower serum cholesterol levels in hypercholeserolemic rats. Finally, we tested if walnuts consumption would be hypocholesterolemic for Japanese. A diet containing walnuts effectively lowered the serum total and low density lipoprotein cholesterol in Japanese men and women. These animal and human experiments would be relevant to advocate publics to keep consuming diets including soybean, rice and nuts for their healthful life.

Two opioid peptides, YPLDL and YPLDLF, were isolated from enzymatic digests of spinach ribulose-1, 5-bisphosphate carboxylase/oxygenase (RuBisCO) and named rubiscolin-5 and -6, respectively. These peptides were selective for delta-receptor and the latter was about 3 times more potent than the former. After oral administration in mice at the dose of 100 mg/kg, rubiscolin-6 showed analgesic activity in tail pinch test. It also stimutated learning performance at the same dose in passive avoidance experiment using step-through apparatus. An immunostimulating peptide, MITLAIPVNKPGR, was isolated from a trypsin digest of soybean protein and named soymetide. Immunostimulating activy of soymetide was mediated by fMLP receptor. Interestingly, after oral administration in rats at a dose of 300 mg/kg (po.), soymetide-4 (MITL) protected alopecia (hair-loss) induced by etoposide, a cancer chemotherapy agent. Stimulation of IL-1 release by the peptide was involved in the mechanism. Ovokinin(2-7), RADHPF, is a vasorelaxing peptide released from ovalbumin by the action of chymotrypsin. It lowered blood pressure of spontaneously hypersensive rats (SHR) after oral administration at a dose of 10 mg/kg. RPLKPW, which was designed by replacing 4 amino acid residues in ovokinin(2-7), exhibited hypotensive activity at a dose of 0.1 mg/kg (po.). This peptides was introduced into 3 homologous sites in soybean beta-conglycinin alpha' subunit by site-directed mutagenesis of the cDNA and expressed in E. coli. The minimum effective dose for hypotensive activity of the genetically modified beta-conglycinin alpha' subunit was 10 mg/kg (po.), which is about 1/200 that of ovalbumin.

Evidence shows that the serum level of cholesterol (CH) is decreased with increasing green tea (GT) consumption. This presentation summarizes our recent findings on the effect of GT extract on intestinal absorption of [$^{14}C$-labeled CH and phosphatidylcholine (PC). Ovariectomized (OX) adult rats were infused intraduodenally with lipid emulsions containing radiolabeled lipids [$^{14}C$-CH or $^{14}C$-phosphatidylcholine (PC)] in the presence of GT extract or catechins to determine the rates and amounts of CH absorption and the intestinal hydrolysis and lymphatic output of PC. During lipid infusion, lymph was collected hourly for 8 h. The lymphatic absorption of $^14C$-CH was drastically lowered by infusion of GT extract at two dosage levels (GTl =5.4 mg catechins/h and GT2 = 15.1mg catechins/h). The cumulative lymphatic absorptions of $^{14}C$-CH in rats infused with GT1 and GT2 were 20.7$\pm$4.3 and $4.8{\pm}4.1{\%}$ dose, respectively, whereas the absorption of $^{14}C$-CH in rats infused with no GT extract (GT0) was $36.3{\pm}1.1{\%}$ dose. GT extracts also significantly lowered the absorption of-tocopherol (TP) in a dose dependent manner ($29.6{\pm}4.9{\%}$ dose in GT0, $20.8{\pm}5.8{\%}$ dose in GTl, and $7.9{\pm}5.4{\%}$ dose in GT2 groups). Both (+)-catechin and EGCG significantly lowered the lymphatic outputs of $^{14}C$-radioactivity after intraduodenal $^{14}C$-PC infusion. A significantly higher amount of $^{14}C$-PC remained unhydrolyzed in the intestinal lumen of the EGCG rats ($22.8{\%}$) compared with the (+)-catechin ($15.8\%$) and control groups ($11.9\%$). GT extracts, (+)-catechin, and EGCG significantly reduced the absorption of TP. The inhibitory effect of GT extract and catechins on lipid absorption may be mediated in part through the inhibition of pancreatic PLAz. The findings provide the first direct evidence that green tea and catechins have a profound inhibitory effect on the intestinal absorption of CH in OX rats. Results suggest that green tea and catechins may be used as a dietary or pharmacological means of lowering cholesterol absorption.

The traditional (European) Mediterranean diet is characterized by an abundance of plant foods such as bread, pasta, vegetables, salad, legumes, fruit, nuts; olive oil as the principal source of fat; low to moderate amounts offish, poultry, dairy products and eggs; only small amounts of red meat; low to moderate amounts of wine, normally consumed with meals. This diet is low in saturated fatty acids, rich in carbohydrate and fibre, and has a high content of monounsaturated fatty acids (MUFA). These are primarily derived from olive oil. Despite a wealth of general knowledge concerning the major classes of compounds present in olives and olive oil, detailed knowledge of the phenolic antioxidant content has been lacking. Therefore the aim of the study was to evaluate the phenolic antioxidant content in a range of olive and seed oils. While seed oils were devoid, on average, the olive oils contained $196{\pm}19mg/kg$ total phenolics as judged by HPLC analysis, but the value for extravirgin ($232{\pm}15 mg/kg$) was significantly higher than that of refined virgin olive oil ($62{\pm}12mg/kg$; P<0.0001). Appreciable quantities of simple phenols (hydroxytyrosol and tyrosol) were detected in olive oils, with significant differences between extravirgin ($41.87{\pm}6.17$) and refined virgin olive oils ($4.72{\pm}2.15$; P<0.01). The major linked phenols were secoiridoids and lignans. Although extravirgin contained higher concentrations of secoiridoids ($27.72{\pm}6.84$) than refined olive oils ($9.30{\pm}3.81$) this difference was not significant. On the other hand the concentration of lignans was significantly higher (P<0.001) in extravirgin ($41.53{\pm}3.93$) compared to refined virgin olive oils ($7.29{\pm}2.56$). All classes of phenolics were shown to be potent antioxidants. In future epidemiolgic studies, both the nature and source of olive oil consumed should be differentiated in ascertaining cancer risk.

As dietary intakes levels of Calcium decrease below those recommended by health authorities the incidence of Osteoporosis is set to continue to rise. To address this issue it is becoming necessary to fortify regularly consumed foods such as cereals, biscuits etc. with ingredients such as AquaCal. AquaCal is a natural, organic source of calcium that is produced from calcified seaweed, Phymatolithon and Lithothamnion and can be utilised for Calcium and magnesium supplementation in a broad range of foods and beverages. AquaCal presents a great potential over other calcium sources because of its porous structure and it composition associating key minerals as calcium, magnesium and boron in addition to being neutral in taste in applications. Once consumed the efficacy of AquaCal must then be verified therefore a bioavailability a comparative feeding study to assess effects of AquaCal on bone density was carried out. There was no significant difference in bioavailability based on calcium absorption between AquaCal and Limestone however the weights of the femurs of the animals fed AquaCal did significantly increase over the group fed Limestone. As a natural and organic mineral AquaCal has many different fields of application.

Vitamin deficiency is a result of an inadequate diet. Education on the importance of trace nutrients in diabetic patients with poor blood sugar control is examined. Those who prepare meals must consider the loss of vitamins in the process of cooking. Our study also suggested that marginal vitamin deficiency plays an indirect but important role in the development of diabetic complications. Vitamin C as altering total cholesterol and vitamin E as altering triglyceride could modify diabetic retinopathy. Pharmacologically, niacin might be responsible for the decrease in lipoprotein (a) and vitamin C would inhibit the influence of rapid blood glucose control on diabetic retinopathy.

This study was performed to examine a possible application of the beverage as a bad breath controlling food. To achieve this objective, methods of gas chromatography, electronic nose, sensory analysis and halimeter were used to detect reduction in odor intensities of bad breath caused by the functional beverage as well as its active ingredients. According to results of GC and electronic nose, adding green tea and champignon extracts to bad breath indicators, methylmercaptan and trimethylamine, resulted in significant reduction in headspace concentrations of two indicators. GC results revealed that headspace concentrations of 5 ug/ml of methylmercaptan and 30 ug/ml of trimethylamine added to various concentrations of two extracts were reduced up to $100\%$ after incubating mixtures at $37^{\circ}C$ for 5min. When the functional beverage was properly formulated with green tea extract, champignon extract and $\alpha$-cyclodextrin and evaluated for its deodorizing effect systematically, it also showed distinctive deodorizing activities against bad breath indicators. Conclusively, results obtained from this study might encourage introduction of a new type of bad breath control food in near future.

The ability of microbial strains to confer health benefits to human and animal hosts is not a feature of many organisms. Lactic acid bacteria are the most commonly used bacteria applied as probiotics and there is now strong evidence that certain strains confer tangible benefits to the host. In terms of preventing infection, the ability of probiotic lactobacilli to colonize the tissue site, even temporarily, and inhibit growth and adhesion of pathogens, has been documented. Using molecular tools, such as RAPD and DGGE, probiotic organisms can be tracked through the intestine and in the vagina, and their impact on the flora assessed. Arguably, strains L. rhamnosus GG and GR-l are the most studied probiotic strains in terms of human application to the gut (GG and GR-l) and vagina (GR-1). Combined with L. fermentum RC-14, GR-l provides a two-pronged therapeutic for the intestine and urogenital tract. Care in manufacturing and distribution is essential to ensure that optimal doses of probiotics are accessible to consumers and patients.

Lactic acid producing bacteria were isolated from baby feces and characterized to be used as a probiotic with anti Helicobacter pylori functions. The selected bacteria had inhibition activity on the adherance and growth of H. pylori. These bacteria had additional beneficial characteristics for the probiotic such as antibacterial activity, antitumor activity, immunostimulation activity, resistance to antibiotic and bile salt, ability to bind to the intestinal cells, and safe for the human use.

The objective of this study was to improve the qualities of Saengsik (substitute meal formulated with uncooked cereal powder) such as functionality, safety and sensory preference by using Lactic Acid Bacteria (LAB). The optimum formulation of fermented Saengsik (substitute meal) was verified as $40\%$ of fermented brown rice, $13\%$ of fermented job tears, $5\%$ of yellow soybean, $5\%$ of waxy rice, $5\%$ of sorghum, $3\%$ of Seumok bean, $1\%$ of sesame, $3\%$ of barley, $2\%$ of red bean, $5\%$ of prosomillet, $2\%$ of foxtail millet, $9%$ of black rice, $3\%$ of buckwheat, $1\%$ of chestnut, $2\%$ of pumpkin, $3\%$ of peanut. Fermented Saengsik decreased significantly serum glucose in STZ-induced diabetic rats. However, serum lipid levels were not affected by fermented Saengsik. In rats fed with high-fat diet, serum triglyceride level was slightly decreased and the contents of liver triglyceride were significantly decreased by supplementation of fermented Saengsik.

The importance of probiotics and prebiotics in health-promoting effect on the host is increasing. Recent studies on gastrointestinal system have contributed to further understanding of the mechanisms involved in the interaction between probiotics and prebiotics. In his presentation, the beneficial effects of probiotics and prebiotics, and applications of microencapsulation technique for the application of prebiotics and probiotics are described.

It is a recent observation that about 80 per cent of the body's immune system is localized in the gastrointestinal tract. This explains to a large extent why eating right is important for the modulation the immune response and prevention of disease. I addition it is increasingly recognized that the body has an important digestive system also in the lower gastrointestinal tract where numerous important substances are released by microbial enzymes and absorbed. Among these substances are short chain fatty acids, amino acids, various carbohydrates, polyamines, growth factors, coagulation factors, and many thousands of antioxidants, not only traditional vitamins but numerous flavonoids, carotenoids and similar plant- and vegetable produced antioxidants. Also consumption of health-promoting bacteria (probiotics) and vegetable fibres (prebiotics) from numerous sources are known to have strong health-promoting influence. It has been calculated that the intestine harbours about 300 000 genes, which is much more than the calculated about 60000 for the rest of the human body, indicating a till today totally unexpected metabolic activity in this part of the GI tract. There are seemingly several times more active enzymes in the intestine than in the rest of the body, ready to release hundred thousand or more of substances important for our health and well-being. In addition do the microbial cells produce signal molecules similar to cytokines but called bacteriokines and nitric oxide, with provide modulatory effects both on the mucosal cells, the mucosa-associated lymphoid system (MALT) and the rest of the immune system. Identification of various fermentation products, and often referred to as synbiotics, studies of their role in maintaining health and well-being should be a priority issue during the years to come.