Metabolites from Resistant and Susceptible <i>Pinus thunbergii</i>after Inoculation with Pine Wood Nematode

Pine wilt disease (PWD), which is caused by pine wood nematodes (PWN), is one of the most serious forest diseases worldwide. To clarify the mechanism of resistance to PWD, we compared metabolites from resistant and susceptible Japanese black pine (Pinus thunbergii) families after inoculation with PWN. After 2 weeks to 1 month post inoculation, the number of PWN dramatically increased in susceptible plants, but not in resistant plants. At this PWN-proliferation phase, ethyl acetate soluble fractions extracted from PWN-inoculated plants were analyzed by gas chromatogramphy-mass spectrometry (GC-MS). Although most compounds were qualitatively and quantitatively similar between resistant and susceptible plants, resistant plants accumulated 2.0-fold more linoleic acid (LA) than susceptible plants. On the other hand, benzoic acid (BA) was barely detected in resistant plants, but it accumulated in susceptible plants as the number of PWN increased. Susceptible plants contained greater levels of the nematicidal compounds pinosylvin and pinosylvin monomethyl ether, compared with resistant plants. These results suggested that LA is involved in the resistance reaction against PWN-proliferation, and that BA could be a good biomarker for PWD.

Evaluation of the Production of Young Green Barley Plants Containing Functional Ingredients

We conducted experiments to establish the growing conditions of hydroponic young green barley plants in which functional ingredients were applied and absorbed during the cultivation period. No studies have conducted experiments on functional ingredients applied during the cultivation of young green barley. In this study, glucosamine (GlcN) and collagen (Cgn) were used, both of which are widely known as functional ingredients and are in high market demand. In the GlcN application experiment, young green barley plants were cultivated with only liquid fertilizer during the early growing period for 9 days, and then the plants were cultivated in 0.25% GlcN-water solution for the following 3 days. The plants used in this experiment absorbed 0.60% of GlcN. Furthermore, an experiment was conducted to test plant absorption of collagen. Collagen absorption was evaluated using hydroxyproline (Hyp) as an indicator ingredient. Under control conditions, the Hyp content was 0.04% after 14 days of cultivation. Meanwhile, in the application experiment of Cgn derived from pigs (average molecular weight is 3000), plants were cultivated in 1% Cgn-water solution for 14 days. As a result, the Hyp content increased to 0.28%. Thus, this study clarifies, for the first time, that barley plants can absorb exogenous functional ingredients applied from the outside. The nutrient component contents in young green barley plants were measured. The amino acid and vitamin C content in hydroponic young green barley significantly increased, as compared to those grown in organic soil. Furthermore, in 0.1% Cgn-water solution cultivation, glutamic acid and aspartic acid, both of which are a functional amino acids, increased approximately three folds as compared to the plant grown in organic soil under natural sunlight. Furthermore, K and Na, which are inorganic ingredients in young green barley, were measured. The K content in the plants grown in 1% Cgn-water solution cultivation for 14 days decreased by 80% as compared with the control hydroponic plant grown with liquid fertilizer. It was inferred that the plant might be used as food for dialysis patients.

Randomized clinical trial of an ethanol extract of Ganoderma lucidum in men with lower urinary tract symptoms

Aim： To evaluate the safety and efficacy of an extract of Ganoderma lucidum that shows the strongest 5α-reductase inhibitory activity among the extracts of 19 edible and medicinal mushrooms by a double-blind, placebo-controlled, randomized and dose-ranging study in men with lower urinary tract symptoms （LUTS）. Methods： In this trial, we randomly assigned 88 men over the age of 49 years who had slight-to-moderate LUTS to 12 weeks of treatment with G. lucidum extract （6 mg once a day） or placebo. The primary outcome measures were changes in the International Prostate Symptom Score （IPSS） and variables of uroflowmetry. Secondary outcome measures included changes in prostate size, residual urinary volume after voiding, laboratory values and the reported adverse effects. Results： G. lucidum was effective and significantly superior to placebo for improving total IPSS with 2.1 points decreasing at the end of treatment （mean difference, -1.18 points; 95% confidence interval, -1.74 to -0.62; P 〈 0.0001）. No changes were observed with respect to quality of life scores, peak urinary flow, mean urinary flow, residual urine, prostate volume, serum prostate-specific antigen or testosterone levels. Overall treatment was well tolerated with no severe adverse effects. Conclusion： The extract of G. lucidum was well tolerated and improved IPSS scores. These results encouraged a further, large-scale evaluation of phytotherapy for a long duration using the extract of G. lucidum on men with LUTS.

Effect of an extract of Ganoderma lucidum in men with lower urinary tract symptoms： a double-blind, placebo-controlled randomized and dose-ranging study

Aim： To conduct a double-blind, placebo-controlled randomized and dose-ranging study to evaluate the safety and efficacy of the extract of Ganoderma lucidum （G. lucidum） in men with lower urinary tract symptoms （LUTS）. Methods： We enrolled male volunteers （〉 50 years） with an International Prostate Symptom Score （IPSS; questions 1-7） 〉 5 and a prostate-specific antigen （PSA） value 〈 4 ng/mL. Volunteers were randomized into groups of placebo （n = 12）, G. lucidum of 0.6 mg （n = 12）, 6 mg （n = 12） or 60 mg （n = 14）, administered once daily. Efficacy was measured as a change from baseline in IPSS and the peak urine flow rate （Qmax）. Prostate volume and residual urine were estimated by ultrasonography, and blood tests, including PSA levels, were measured at baseline and at the end of the treatment. Results： The overall administration was well tolerated, with no major adverse effects. Statistical significances in the magnitude of changes between the experimental groups were observed at weeks 4 and 8. No changes were observed with respect to Qmax, residual urine, prostate volume or PSA levels. Conclusion： The extract of G. lucidum was well tolerated and an improvement in IPSS was observed. The recommended dose of the extract of G. lucidum is 6 mg in men with LUTS.

The effect of photo-irradiation on the growth and ingredient composition of young green barley (<i>Hordeum vulgare</i>)

We clarified that photo-irradiation of young green barley from three different light sources, natural light, 100% red light-emitting diodes (R-LEDs), and a mixture of 90% red-LEDs + 10% blue-LEDs (RB-LEDs), had significantly different results in growth degree (weight and height) and in components of young green barley. Barley that has sprouted for 15 days after germination did not show any apparent difference in height in response to irradiation by the three tested light sources, but by the 20th day of sprouting the height showed a positive effect by R-LEDs irradiation. By 15 days of sprouting the barley had achieved the heaviest weight by natural light irradiation, while the barley irradiated by R-LEDs had made remarkable progress at 20 days of sprouting. On the other hand, the irradiation by RB-LEDs showed a suppressive tendency after 15 days or more. The amino acid content, as indicated by dry weight conversion, was greatest in the barley irradiated by RB-LEDs, followed by R-LEDs, and natural light, which showed that LEDs irradiation is effective. In addition, four cyanogenic glucosides were isolated, identified, and quantified, as they are components frequently assessed in barley research. With regard to vitamin E, R-LEDs irradiation increased γ-tocopherol. Our results indicate that irradiation by LEDs would be effective for the enhancement of the functionality of young green barley.

Production of Citric Acid from Corncobs with Its Biological Evaluation

Corncobs could serve as a substrate for citric acid production using solid state fermentation technique. The culture optimization concerning substrate concentration, culture duration, pH, temperature and substrate hydrolysis was carried out for maximum productivity of citric acid. Under the optimized conditions, 48.4 g of citric acid was produced from 1 kg dry corncobs. Biological evaluation was carried out for citric acid such as melanin synthesis inhibitory, anti-allergy, anti-bacterial, and hyaluronic acid production activities. The results showed that citric acid has potent melanin inhibitory activity, good inhibition for β-hexosaminidase release and potent stimulatory effect for the production of hyaluronic acid. These activities (melanin synthesis inhibitory, anti-allergy and hyaluronic acid productive activities) of citric acid have been reported for the first time.

Model Examination for the Effect of Treading Stress on Young Green Barley (<i>Hordeum vulgare</i>)

Treading barley (“Mugifumi” in Japanese) is a barley culturing process that is unique and indispensable in Japan. Generally, roller machines or human feet are used to tread young barley several times (Figure 1). As a model examination, a stainless steel instrument was used for treading barley in this study (Figure 2(a)). Treading damages plants and decreases growth, such as the height and weight of aerial parts, which then recover gradually. In our study, the total amino acid contents increased with the treading stress. The treading stress induced a 1.7-fold increase in aspartic acid contents and an approximately 1.6-fold increase in glutamic acid, proline, cystine, and methionine contents. Isolation and purification suggested that the main components of the methanol-eluted fraction from the young green barley were six known phenolic compounds. Saponarin and lutonarin were the main components. The saponarin content was about 160 mg/g (methanol-soluble fraction) at 10 days after germination. We found that the higher antioxidant activity was due to the increased lutonarin/saponarin ratio from 10% to 24%. Therefore, our results suggest that treading stress is useful for young green barley.

Enzymatic Degradation of Organochlorine Insecticide Chlordane by Phlebia brevispora

[ Objective ] This study aimed to clarify the mechanism of enzymatic degradation of organochlorine insecticide chlordane by the white rot fungus Pldebia brevispora. [ Method ] The degradation characteristics of chlordane were determined by the crude enzyme extracted from P. brevispora strain by pure culture, ultrasonic fragmentation and centrifuge separation. [ Result] About 33.2% and 10.4% of chlordane were degraded respectively by intracellular enzyme and extracellular enzyme within 20 min. At pH 5.0 and 3℃, the crude enzyme showed the greatest degradation activity （49.9%）, with its Michaelis-Mentn＇s constant （Km ） and maximum degradation rate （ Vmax ） of 3.49 and 8.38 ttmol/min, respectively. [ Conclusion] Extracellular enzyme can degrade chlordane through dehydrogenation or dehydrochlorination. Intracellular enzyme plays a major role in chlordane degradation. It can transform chlordane to some hydroxylated metabolites through hydroxylation or substitution of chlorine atom by hydroxyl group.

Effects of Irradiation with Light of Different Photon Densities on the Growth of Young Green Barley Plants

In recent years, young green barley has been increasingly used as a functional food ingredient. Studies have shown that cultivated young green barley has different levels of amino acids, vitamin C, and polyphenol content depending on the harvest time. Furthermore, some studies have investigated the effect of the growing conditions on cultivated barley in particular the use of light-emitting diodes (LED). Young green barley was exposed to three light sources of different wavelengths (sun light [ASL], light-emitting diode[LED] - Red 9 + Blue 1 [LED-R9:B1], LED - Red 4 + Green 1+ Blue 1 [LED-R4: G1:B1]). Under light with photon densities of 200 μmolmL-2·s-1, the dry weight of young green barley was not affected, while the differences were observed in the morphology of the underground portion of the plant (roots) depending on the light source. The roots of the plant grown in ASL weighed twice as much as those that were grown under LED irradiation. Furthermore, LED irradiation caused an increase in the amino acid content in plants;the amino acid content of plants grown under LED-R9:B1 was twice as much as that grown under ASL irradiation. Four kinds of cyanogenicglucosides (CGs) were isolated, identified, and their levels were measured. The plant grown under LED-RGB irradiation, including LED-G, produced approximately 20% more CGs as compared to the plants grown under the other two light sources. Thus, it was inferred that young green barley exhibited a stress response under LED-G light and accumulated CGs in the stems and leaves with prepare for any damage that may occur on the leaf surface. The nitrogen (N) content in the root was the lowest, while the CG content was the highest in the plant grown under LED-RGB irradiation. It is inferred that the N content decreased because N was eliminated from the roots to allow for accumulation of CGs response to stress. In general, the growth experiments to use an irradiation condition in which the distance between the light source and plant floor is fixed. It is not clear whether the photon density level received by the top leaves is fixed according to plant growth, by changing the height of plant growing shelf. Therefore, against this background, when the photon density was increased from 200 to 300 μmolmL-2·s-1?under ASL irradiation, only a minimal change was observed in the root weight, while the weight of the part of the plant above the ground surface (stems and leaves) increased by approximately 30%, with a 10% increase in the amino acid content.

The Melanin Biosynthesis Stimulating Compounds Isolated from the Fruiting Bodies of <i>Pleurotus citrinopileatus</i>

In the course to find a stimulating compound for melanin biosynthesis, which should be useful for a gray and a white hair-preventive agent or tanning agent, we evaluated the effects of the methanol extract from mushroom of Pleurotus citrinopileatus on melanin production in B16 melanoma cells without theophylline. Activity-guided fractionation led to isolate myo-inositol (3) and D-mannitol (4) as the stimulating compounds on melanin production in B16 melanoma cells. Also, ergosterol (1), uracil (2), and D-glucose (5) have been isolated from the methanol extract of P. citrinopileatus and showed no effect on melanin production in B16 melanoma cells. These results indicated that myo-inositol (3) and D-mannitol (4) are potential candidates that could be useful such as a gray and a white hair-preventive agent or tanning agent.

Melanin Biosynthesis Inhibitory Activity of Compounds Isolated from Unused Parts of <i>Ammi visinaga</i>

Ten compounds have been isolated from the unused parts of Ammi visinaga. The isolated compounds were identified as tetracosanoic acid (1), β-sitosterol (2), visnadine (3), khellin (4), β-sitosterol glucoside (5), norkhellol (6), khellol (7), rhamnazin (8), cimifugin (9), and cis-khellactone-3’-β-glucopyranoside (10). The chemical structures of these compounds were elucidated based on spectroscopic data (NMR, UV, MS and IR spectra). This is the first report on the identification of tetracosanoic acid (1), norkhellol (6) and cimifugin (9) in the Ammi genus. The melanin biosynthesis inhibitory activities of khellin (4), khellol (7), visnadine (3), cimifugin (9), β-sitosterol (2) and β-sitosterol glucoside (5) were evaluated. Khellin (4) exhibited a potent melanin inhibitory effect compared to arbutin with less toxicity.