Evaluation of the Effect of Lentinula Edodes Mycelia Extract on Immune Function in Healthy Subjects Placebo-controlled, Parallel-group Study / 日本補完代替医療学会誌

In this study, we examined the effects of Lentinula edodes mycelia (LEM) on fatigue and immune function in healthy subjects. 22 people were randomly assigned in a 1:1 ratio to the LEM ingestion group and the placebo group. Daily oral ingestion of LEM for 4 weeks (600 mg/day) significantly improved the VAS fatigue score compared with before treatment (p<0.01). Additionally, NK cell activity was increased by ingestion of LEM (p<0.01) and was higher level compared to the placebo group (p<0.05). These results indicate that oral ingestion of LEM may improve the feeling of fatigue and immune function.

Antimicrobial activity of crude extracts prepared from fungal mycelia

Objective To evaluate the in vitro antimicrobial property of three different partitioned extracts (petroleum ether, ethanol and water) prepared from some fungal mycelia. Methods Seven fungal mycelia were prepared, initially extracted with acidified ethanol (0.2 mol/L HCl in 80% ethanol), yielding the raw crude extracts. The obtained extracts were then further partitioned with petroleum ether (F1), ethanol (F2) and water (F3). All the fractions were tested for antimicrobial activity using the disc diffusion assay. Results Our data showed that all the fractions could inhibit the testing bacteria. However, the inhibitory activity was found to be dependent on (i) the fungal strains used; (ii) the solvent extracted; and (iii) the testing bacteria assayed. In general, the ethanolic extracts (F2) derived from all fungi displayed highest inhibitory activity against the testing bacteria except for Chaetomium sp. Conclusions The findings of the present study concluded that the extracts prepared from the fungal mycelia had the bioactive compounds with antibacterial property. This study is a pioneering work and further study should be carried out for development of the new drug leads.

HPLC-ELSD fingerprints of oligosaccharide sites from mycelia of Hericium erinaceum solid cultures and Weilening Tablets / 中成药

AIM To establish the HPLC-ELSD fingerprints of oligosaccharide sites from mycelia of Hericium erinaceum solid cultures and Weilening Tablets.METHODS The analysis of aqueous extract from samples was performed on a 80 ℃ thermostatic Waters XBridge TM Amide column (4.6 mm × 150 mm,3.5 μm),with the mobile phase comprising of acetonitrile-0.2％ ammonium acetate flowing at 1 mL/min in a gradient elution manner.RESULTS There were eight and nine common peaks in two HPLC-ELSD fingerprints with the similarties of 0.994-0.966 and 0.990-0.997,respectively.Three of them were mannitol,lactose and trehalose,which showed good linear relationships within their own ranges (r ≥ 0.999 0),the average recoveries were 95.08％-104.82％ with the RSDs of 1.12％-2.90％.CONCLUSION This simple and accurate method can be used for the rapid quality control of mycelia of Hericium erinaceum solid cultures and Weilening Tablets.

Antimicrobial activity of crude extracts prepared from fungal mycelia

Objective: To evaluate the in vitro antimicrobial property of three different partitioned extracts (petroleum ether, ethanol and water) prepared from some fungal mycelia. Methods: Seven fungal mycelia were prepared, initially extracted with acidified ethanol (0.2 mol/L HCl in 80%ethanol), yielding the raw crude extracts. The obtained extracts were then further partitioned with petroleum ether (F1), ethanol (F2) and water (F3). All the fractions were tested for antimicrobial activity using the disc diffusion assay. Results: Our data showed that all the fractions could inhibit the testing bacteria. However, the inhibitory activity was found to be dependent on (i) the fungal strains used;(ii) the solvent extracted; and (iii) the testing bacteria assayed. In general, the ethanolic extracts (F2) derived from all fungi displayed highest inhibitory activity against the testing bacteria except for Chaetomium sp. Conclusions: The findings of the present study concluded that the extracts prepared from the fungal mycelia had the bioactive compounds with antibacterial property. This study is a pioneering work and further study should be carried out for development of the new drug leads.

Evaluation of Quality of Life among Patients with Various Types of Cancer Treated with Oral Lentinula Edodes Mycelia Extract / 日本補完代替医療学会誌

Extract of Lentinula edodes mycelia (LEM) reportedly maintains and improves the quality of life (QOL) of patients with cancer who undergo chemotherapy. We investigated the influence of LEM extract on the QOL of 73 Japanese patients with different backgrounds and stages of cancer who were undergoing various types of therapy at 16 institutions. All patients were orally administered with LEM (1200 mg/day) for four weeks. We compared changes in their QOL before and after taking LEM using the EORTC-QLQ-C30. The QOL scores for emotional function and fatigue were significantly improved after LEM intake. Global QOL and physical function scores improved only in patients with cancer at stages III and IV. These findings suggest that oral LEM intake improves the QOL of patients with advanced cancer.

Antioxidant activity of polysaccharides extracted from Lentinus edodes mycelia and their protective effects on INS-1 cells / Atividade antioxidante de polissacarídeos extraídos de Lentinus edodes micélio e seus efeitos protectores em células INS-1

The disruption of the delicate balance between the generation of reactive oxygen species (ROS) and antioxidant scavenging systems can lead to many health problems. Polysaccharides extracted from fungi fruit body and myceliums have several potential health benefits. In this study, the antioxidant capacity and cell protective effects of a polysaccharide isolated from Lentinus edodes mycelia (LMP) were investigated. The antioxidant properties of LMP were screened using radical scavenging (1, 1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl, and superoxide anion), reducing power and inhibition of lipid peroxidation assays. Intracellular reactive oxygen species (ROS), in response to H2O2, was determined by the dichloro-dihydro-fluorescein diacetate (DCFH-DA) assay. Rat insulinoma (INS-1) cells apoptosis was detected using flow cytometry analysis. The results showed that LMP exhibited significantly strong radical scavenging activity. At the concentration of 2.0 mg mL-1, DPPH radical, hydroxyl radical, and superoxide anion radical scavenging rates are 80.32 ± 2.58%, 92.56 ± 3.11% and 93.73 ± 2.82%, respectively. The reducing power and inhibition of lipid peroxidation activity of LMP were as strong as the positive control. LMP alleviated the intracellular reactive oxygen species level and inhibited islet cell apoptosis significantly. The results open perspectives for studies of LMP in the treatment of diabetes.

O estresse oxidativo tem uma função biológica importante em muitos organismos. A ruptura do equilíbrio delicado entre a geração de espécies reativas de oxigênio (ROS) e sistemas de eliminação de antioxidantes pode levar a muitos problemas de saúde. Polissacarídeos extraídos do corpo fungos frutas e micélio têm vários benefícios potenciais para a saúde. Neste estudo, foram investigados os efeitos celulares e capacidade de proteção antioxidante de um polissacarídeo isolado a partir de Lentinus edodes micélios (LMP). As propriedades antioxidantes da LMP foram rastreadas utilizando eliminação de radicais (1, 1-difenil-2-picrilhidrazil (DPPH), hidroxilo e anião superóxido), e a redução da potência de inibição de ensaios de peroxidação lipídica. Espécies reativas de oxigénio intracelulares (ROS), em resposta ao H2O2, foi determinada pelo ensaio de diacetato de dicloro-di-hidro-f luoresceina (DCFH-DA). INS-1 células a apoptose foi detectada utilizando análise de citometria de fluxo. Os resultados mostraram que LMP exibiu significativamente forte atividade de eliminação de radicais. Na concentração de 2 mg mL-1, DPPH radical, radical hidroxilo e anião superóxido velocidades de eliminação de radicais livres são 80,32 ± 2,58%, 92,56 ± 3,11% 93,73 ± 2,82%, respectivamente. O poder redutor e inibição da peroxidação lipídica de LMP foram tão forte quanto o controlo positivo. LMP aliviou a nível de espécies reativas de oxigênio intracelular e apoptose de células da ilhota inibiu significativamente. Sugere-se que LMP pode ser utilizado como um reagente complementar para a diabetes a prevenção e terapia.

Induction of Translocation of Glucose Transporter 4 in Rat Skeletal Muscle Cells by a Water-Soluble Extract from Culture Medium of <i>Ganoderma lucidum</i> Mycelia / 日本補完代替医療学会誌

<b>Objective:</b> The water-soluble extract of <i>Ganoderma lucidum</i> mycelia (WER), which is used as a health food, reduced hyperglycemia and enhanced glucose transporter 4 (GLUT4) translocation to the plasma membrane in skeletal muscle and adipose tissue in KK-<i>A</i>^y<i> </i>mice, a type 2 diabetic animal model with obesity.In order to elucidate the reduction of hyperglycemia by WER, we investigated the translocation of glucose transporter 4, glucose uptake and associating signal transduction in rat skeletal muscle (L6) cells. <b>Method:</b> The glucose uptake was analyzed with radioactive 2-deoxy-D-glucose.The localization of GLUT4 in L6 cells treated with various concentrations of WER was analyzed with immunohistochemical staining and Western blot technique.As a positive control, insulin or troglitazone was used in these experiments.Furthermore, the activation of intracellular signaling pathways by Western blot analysis and the influence of glucose uptake using four kinds of inhibitors (LY294002 as potent PI3K inhibitor, rapamycin as mTOR inhibitor, Gö6983 as broad PKC inhibitor, compound C as AMPK inhibitor) was evaluated. <b></b><b>Results: </b>GLUT4 protein content in the plasma membrane was induced in a dose-dependent manner of WER without increasing the gene expression and amount of total protein in the L6 cells and the glucose uptake was augmented with increasing the amount of GLUT4 translocated on the plasma membrane.The phosphorylation of phosphatidylinositol-3 kinase (PI3K), Akt and acetyl CoA carboxylase (ACC) were induced in a concentration dependent manner and inhibited by the above inhibitors except rapamycin. <b>Conclusion: </b>These results indicate that the hypoglycemic effect of some material(s) in WER may be due to the enhancement of glucose uptake through GLUT4 translocation on the plasma membrane by activating the PI3K/Akt pathway through improving insulin resistance.

Influence of a Water-Soluble Extract from Culture Medium of<i> Ganoderma lucidum</i> Mycelia (WER) on Carbohydrate Metabolism in the Liver of Type 2 Diabetic Mice / 日本補完代替医療学会誌

<b>Objective:</b> Recently, we reported that long-term intake of a water-soluble extract from culture medium of <i>Ganoderma lucidum</i> mycelia (WER) reduced hyperglycemia and enhanced glucose transporter-4 (GLUT4) translocation to the plasma membrane in skeletal muscles and adipose tissue in KK-<i>A^y</i> mice, a type 2 diabetic animal model with obesity. In the present study, we investigated the effect of WER on hepatic carbohydrate metabolism.<br> <b>Methods: </b>Female KK-<i>A^y</i> mice were given free access to water and high-fat food containing 0.5% WER for 8 weeks, and blood glucose levels were assessed every week. At the end of the experimental period, the expression and activities of sugar metabolic enzymes in the liver were determined by Real Time RT-PCR and each activity measurement method. Also, the amount of glycogen was measured by anthrone-sulfuric acid method. Furthermore, the expression level of GLUT2 and activation of AMP kinase (AMPK) and glycogen synthase kinase 3β (GSk3β) was also determined by western blot analysis.<br> <b>Results: </b>The mice with the high-fat ingestion showed a gradual increase in the levels of blood glucose and body weight. In the WER-treated mice, the blood glucose level was suppressed after 2 weeks of intake. The gene expression and enzyme activities of both glucose-6-phosphatase and phosphoenolpyruvate carboxykinase were suppressed, whereas those of glucokinase were increased in the mice with WER intake and pioglitazone administration. The accumulation of glycogen was increased. Moreover the expression of GLUT2 and phosphorylation levels of AMPK and GSk3β were also increased in the mice with WER intake.<br> <b>Conclusion: </b>These results indicate that WER affects hepatic carbohydrate metabolism, which may derive from the suppression of gluconeogenesis through the modulation of related enzymes and enhancement of glucose uptake, glycolysis and glycogen synthesis.<br>

Medium optimization for palmarumycin C13 production in liquid culture of endophytic fungus Berkleasmium sp. Dzf12 using response surface methodology

Background: Berkleasmium sp. Dzf12, an endophytic fungus from Dioscorea zingiberensis, was a high producer of palmarumycin C13 with various bioactivities. In the present study, the experimental designs based on statistics were employed to evaluate and optimize the medium for palmarumycin C13 production in mycelia liquid culture of Berkleasmium sp. Dzf12. Results: Among various carbon and nitrogen sources, glucose, peptone and yeast extract were found to be the most favourable for palmarumycin C13 production based on the one-factor-at-a-time experiments. After Plackett-Burman test on the medium, glucose, peptone and yeast extract were further verified to be the most significant factors to stimulate palmarumycin C13 accumulation. These three factors (i.e., glucose, peptone and yeast extract) were then optimized through the experiments of central composite design (CCD) and analysis of response surface methodology (RSM). The optimized medium compositions for palmarumycin C13 production were determined as 42.5 g/l of glucose, 6.5 g/l of peptone, 11.0 g/l of yeast extract, 1.0 g/l of KH2PO4, 0.5 g/l of MgSO4 x 7H2O, 0.05 g/l of FeSO4 x 7H2O, and pH 6.5. Under the optimal culture conditions, the maximum palmarumycin C13 yield of Berkleasmium sp. Dzf12 was increased to 318.63 mg/l, which was about 2.5-fold in comparison with that (130.44 mg/l) in the basal medium. Conclusions: The results indicate that the optimum production of palmarumycin C13 in Berkleasmium sp. Dzf12 liquid culture can be achieved by addition of glucose, peptone and yeast extract with their appropriate concentrations in the modified Sabouraud medium.

Isolation and identification for strains from wild Phellinus and analysis on its secondary metabolites / 中草药

Objective: To isolate and identify the strains from wild Phellinus linteus and analyze its secondary metabolites. Methods: Strains isolated and purified were investigated by optical microscope to observe the characteristics of mycelia, ITS sequence was used for molecular identification and its secondary metabolite content was analyzed by chemical colorimetry. Results: The morphological characteristics of mycelia from pinus, Morus alba, Syringa reticulata, and populus were the same. The growth rate of four flamentous colonies and their colors were different, wherein the growth rate of the colony from M. alba was the fastest, up to 0.47 cm/d. Four strains all belonged to genus Phellinus sp, where strain from pinus was identified as P. pini, M. alba was P. linteus, S. reticulata and populus were P. baumii. The species of the secondary metabolites in the sporocarp and mycelia of Phellinus were the same, but the content was different. For the sporocarp, the highest polysaccharide content was in the sporocarp from S. reticulata with the content of 98.20 mg/g, the highest flavonoids, terpenoids, and polyphenols contents were all in the sporocarp from poplar with the contents of 548.49, 1.48, and 33.70 mg/g, respectively. For mycelia, the highest polysaccharide, flavonoids, terpenoids, and polyphenols contents were all in the mycelia of M. alba, the contents were 259.64, 223.11, 43.78, and 24.80 mg/L, respectively. Additionally, the content of terpenoids in the mycelia was higher than that of the sporocarp. Among them, the content of terpenoids in the mycelia from M. alba was about 7 times as that in fruit body. Conclusion: There are no differerences amony form of mycelia from four strains of P. linteus. rDNA ITS sequence analysis could be used for the identification of strains. The species four Phellinus strains and its secondary metabolite content are clarified.

Quality Characteristics of Yanggaeng Supplemented with Sanghwang Mushroom (Phellinus linteus) Mycelia

The principal objective of this study was to examine the quality characteristics of yanggaeng supplemented with powder derived from Sanghwang mushroom (Phellinus linteus) mycelia. We analyzed the potential of utilizing Phellinus linteus mycelia as a functional food material by estimating total polyphenol and flavonoid contents, electron-donating abilities, as well as antioxidative activities of the water and ethanol extracts of Sanghwang mushroom mycelia. The total phenol and flavonoid contents of ethanol extracts from Phellinus linteus mycelia were 0.69 mg/ml and 0.16 mg/ml, respectively, while the contents from the water extract of Phellinus linteus mycelia were 0.66 mg/ml and 0.22 mg/ml, respectively. The electron-donating abilities of ethanol and water extracts from Phellinus linteus mycelia were 88.64 and 90.29%, respectively. The ABTS radical scavenging activities of ethanol and water extracts from Phellinus linteus mycelia were 89.74 and 71.35%, respectively. In terms of color values, as the level of powder increased, the value of L (lightness) decreased, whereas those of a (redness) and b (yellowness) increased. In regard to the mechanical properties of the samples, we noted significant differences in hardness, springiness, chewiness, and gumminess (P<0.05). The results of the sensory evaluation showed that the score from SHM (Sanghwang mushroom mycelia) with 4% powder was significantly higher than other samples in terms of sweetness, color, taste, texture and overall quality (P<0.05). Taken together, the recommended level of Phellinus linteus mycelia powder in yanggaeng is 4% for optimal sensory characteristics.

The proximate, mineral and phytochemical analysis of the leaves of Ocimum gratissimum L., Melanthera scandens A. and Leea guineensis L. and their medicinal value.

The chemical composition of Ocimum gratissimum, Melanthera scanden and Leea guineensis were investigated. The proximate analysis in % showed that Melanthera scanden had the highest amount of ash content of 7.73 and moisture content 6.54 while Ocimum gratissimum had the lowest amount of ash content 5.11 and moisture content 5.04. Ocimum gratissimum had the highest amount of fat 7.75 and carbohydrate 56.16 while Melanthera scanden had the least amount of fat 6.87 and carbohydrate 50.0. Leea guineensis had the highest amount of crude protein 19.3 while Melanthera scanden had the highest amount of crude fibre 12.66. The mineral analysis in mg/100g indicated that the leaves contained calcium, sodium, potassium, magnesium, zinc, iron, manganese, and phosphorus. The Phytochemical analysis of the plants showed that the three medicinal plants contained alkaloid, tannins, saponins, steroid, phlobatannin/ terpenoid, flavonoid cardiac glycoside, while phlobatanin was not found in Melanthera scanden. The medicinal plants also contained antinutrient phytin phosphorus, oxalate, phytic acid and polyphenol.

Protective Effects of a Water-Soluble Extract from Culture Medium of Lentinus Edodes Mycelia against Neuronal Damage after Hypoxia-Ischemia in Mice / 日本補完代替医療学会誌

<b>Objective</b>: <i>Lentinus edodes</i> (Shiitake) is a very popular mushroom in Asian cuisine. The water-soluble extract from culture medium of <i>Lentinus edodes</i> mycelia (LEM), which is commercially available as a nutritional supplement, is prepared by hot-water treatment from a solid medium composed of bagasse and defatted-rice bran overgrown for about 4 months with its mycelia. LEM was previously reported to have antioxidant activity and to suppress various oxidative damages. In this study, the neuroprotective effects of 2-week intake of LEM on cerebral ischemic damage induced by hypoxia/ischemia (H/I) followed by reoxygenation in mice were examined.<br> <b>Method</b>: Male C57BL/6J mice were divided into three groups, fed for two weeks with the control laboratory powder chow, 0.5% LEM-contained chow, or 1% LEM-contained chow, respectively. Cerebral ischemic damage was induced in the mice by H/I (i.e., unilateral ligation of the carotid artery and exposure of 8%O₂ for 30 min). Twenty-four hours after H/I, total plasma oxidative stress, neurological deficits, cerebral infarction volume were evaluated in each group. Furthermore, the number of apoptotic cells in ischemic penumbra, the hippocampal CA1 and CA2, and the somatosensory area of the cortex, were analyzed by TUNEL staining and cleaved caspase-3 immunostaining.<br> <b>Results</b>: The infarct area assessed 24-h after H/I was extended to the corpus striatum and cortex in the control mice. Treatment of LEM dose dependently improved plasma oxidative stress, neurological deficits, and cerebral infarction volume. Moreover, LEM decreased the levels of dihydroethidium activity as an index of super oxide production and the number of apoptotic cells in ischemic penumbra.<br> <b>Conclusion</b>: These results show that chronic intake of LEM relieves the hypoxia-induced cerebral ischemic injury, which may be attributed to the antioxidant effects of LEM.<br>

Inhibitory Effects of a Water-Soluble Extract from Culture Medium of Ganoderma lucidum (Rei-shi) Mycelia on Postprandial Blood Glucose Elevation in Type 2 Diabetic Mice and Additional Effect with .ALPHA.-Glucosidase Inhibitors / 日本補完代替医療学会誌

<b>Objective:</b> The water-soluble extract of <i>Ganoderma lucidum</i> mycelia (WER) is prepared from a solid medium composed of bagasse and rice bran overgrown with<i> Ganoderma lucidum</i> mycelia. Recently, we reported that WER shows a blood glucose-lowering effect in maltose-loaded non-diabetic mice. Here, we investigated the efficacy of WER in type 2 diabetic state using KK-A^y mice. Moreover, the food-drug interactions of WER with α-glucosidase inhibitors, voglibose or acarbose were examined using both <i>in vitro</i> and <i>in vivo </i>experiments.<br> <b>Methods:</b> The glucose-lowering effects of oral administration <i>in vivo</i> of WER alone, or concomitant administration of WER with voglibose/acarbose on the elevation of blood glucose levels by sugar-tolerance tests were examined in KK-A^y mice. The inhibitory effects on α-glucosidase <i>in vitro</i> were also evaluated.<br> <b>Results:</b> Oral administration of WER (1 g/kg), which did not affect fasting blood glucose, significantly suppressed the hyperglycemia after loading of maltose (18% of decrease in AUC) compared to the water-administrated control mice. <i>In vitro</i> study showed that WER inhibited maltase in concentration-dependent manner. The inhibitory effects of lower concentrations of voglibose or acarbose on α-glucosidase activity were additively enhanced by the presence of WER, but those of higher concentrations were not affected. The glucose-lowering effect of voglibose (0.1 mg/kg) disappeared in maltose-loaded KK-A^y mice when the drug was concomitantly administrated with WER (1 g/kg), whereas acarbose (16 mg/kg) with WER showed no significant change in its effect.<br> <b>Conclusion:</b> These results demonstrated that WER shows the glucose-lowering effect in maltose-loaded KK-A^y, which may be based on inhibition of the α-glucosidase activity. The present study suggests that concomitant intake of WER with voglibose may override the therapeutic effect of voglibose on postprandial hyperglycemia by food-drug interaction in diabetic state.<br>

Effect of different metal ions for submerged culture of Shiraia bambusicola / 中草药

Objective: To study the effect of different levels of Cu2+, Fe3+, Cr3+, and Ca2+ on mycelia biomass and the hypocrellin yield by submerged culture of Shiraia bambusicola. Methods: The fungis were cultured under 27°C and 130 r/min for 96 h with each level of metal ions. Results: The biomass and the hypocrellin yield were the highest with Cr3+ of 0.005%; Fe3+ of 0.003% and 0.005% contents were favorable to the biomass and the hypocrellin yield, respectively; Ca 2+ content of 0.03% was suitable for the mycelia ' growth and the hypocrellin yield benefited from the concentration of 0.005%; Cu2+ content of 0.03% was added, both the biomass and the hypocrellin yield reached the best. Conclusion: The highest mycelia dry weight and the hypocrellin yield could reach 15.28 g/L and 8.12 mg/g, respectively in addition of metal ion.

Safety Evaluation of Extract from Cultured Lentinula edodes Mycelia; Study of Acute Toxicity, Genotoxicity and Inhibiting Effect of Drug-Metabolizing Enzyme, Cytochrome P-450 3A4 / 日本補完代替医療学会誌

<i>Objective</i>: Extract from cultured <i>Lentinula edodes</i> mycelia (L.E.M.) is a food ingredient possessing various pharmacologic actions such as immunomodulatory properties, antitumor and hepatoprotective effects. In Japan, it has been used as a health food for 30 years or more.<br> In the present study to evaluate the safety of L.E.M., a genotoxicity study and acute toxicity study were conducted. In addition, the inhibitory effect of drug-metabolizing enzyme by L.E.M. was tested<i> in vitro</i>, to gain insight on the interaction with medicines.<br> <i>Methods</i>: The genotoxicity study was performed using a bacterial reverse mutation assay and a <i>in vivo</i> mammalian bone marrow cell chromosomal mutation assay. The acute toxicity study was performed using a single-dose oral toxicity test in rats. Inhibitory activity of cytochrome P-450 3A4 (CYP3A4), one of the most important drug-metabolizing enzymes, by L.E.M. was tested using a baculovirus-expressed system.<br> <i>Results</i>: In the genotoxicity study, mutagenicity was negative for both bacterial reverse mutation assay and <i>in vivo</i> mammalian bone marrow cell chromosomal mutation assay. In the acute toxicity study, no toxic symptoms were observed by single dose oral administration of L.E.M. at a dose of 10,000 mg/kg BW in rats. This implies LD₅₀>10,000 mg/kg BW. No inhibitory activity of CYP3A4 by L.E.M. was observed at in the <i>in vitro</i> screening system to investigate drug-L.E.M. interaction.<br> <i>Conclusion</i>: It is believed L.E.M. is a safety ingredient for foods used in complementary and alternative medicine, since it was toxicologically safe and showed no inhibitory activity of CYP3A4 in the studies conducted.<br>

Hypoglycemic Effects of a Water-Soluble Extract from Culture Medium of Ganoderma lucidum (Rei-shi) Mycelia in Type 2 Diabetic Mice / 日本補完代替医療学会誌

<i>Objective</i>: The water-soluble extract of <i>Ganoderma lucidum</i> mycelia (WER) is prepared from a solid medium composed of bagasse and rice bran overgrown with <i>Ganoderma lucidum</i> mycelia. Recently, we have reported that WER had glucose-lowering effect in streptozotosin-induced diabetic mice, an animal model of type 1 diabetes. Here, we investigated whether long-term treatment with WER affects hyperglycemia and insulin resistance in KK-A^y mice, a type 2 diabetic animal model with obesity.<br> <i>Methods</i>: Female KK-A^y mice were given free access to water and high-fat food containing 0.5% WER for 8 weeks, with blood glucose and plasma insulin levels assessed every week. At the end of the experimental period, insulin tolerance test (ITT) was performed, and plasma levels of triglyceride, total cholesterol, HDL-cholesterol, AST, ALT and adiponectin were measured. Furthermore, expression of GLUT4 in skeletal muscle cell membrane and adipocytes was also determined by immunostaining and Western blot analysis.<br> <i>Results</i>: The mice with high-fat ingestion showed a gradual increase in levels of blood glucose and body weight. In the WER-treated mice, the blood glucose level was significantly suppressed after 2 weeks of treatment. WER also reduced plasma levels of ALT and insulin, but did not affect the other parameters. Additionally, ITT revealed that WER improved insulin sensitivity. Moreover, expression of GLUT4 in the plasma membrane of skeletal muscle cells and adipocytes of the WER-treated mice was increased.<br> <i>Conclusion</i>: These results indicate that WER has a glucose-lowering effect in type 2 diabetic mice. WER also improved hyperinsulinemia and insulin sensitivity, which may derive from enhancement of glucose uptake through GLUT4 of skeletal muscle cells and adipocytes.<br>

Optimal Culture Conditions for Mycelial Growth and Exo-polymer Production of Ganoderma applanatum

The effect of fermentation parameters and medium composition on the simultaneous mycelial growth and exo-polymer production from submerged cultures of Ganoderma applanatum was investigated in shake-flask cultures. The optimum initial pH for mycelial growth and exo-polymer production was 5.0 and 6.0, respectively. The optimum temperature was 25degrees C and the optimum inoculum content was 3.0% (v/v). The optimal carbon and nitrogen sources were glucose and corn steep powder, respectively. After 12 days fermentation under these conditions, the highest mycelial growth was 18.0 g/l and the highest exo-polymer production was 3.9 g/l.

The Safety of Excessive Intake of the Food Containing Extract of Cultured Lentinula edodes Mycelia (L.E.M.) in Healthy Adult Volunteers / 日本補完代替医療学会誌

<i>Objective</i>: In this study, we evaluated the safety of excessive intake of granular foods containing extract of cultured <i>Lentinula edodes </i>mycelia (L.E.M.) in healthy adult volunteers.<br> <i>Methods</i>: Eleven subjects (8 males and 3 females, ages 33.4 ± 9.4) consumed the test foods containing 5,400 mg L.E.M. a day, three times the recommended daily intake, for 4 weeks.<br> <i>Results</i>: No adverse effect by excessive intake of test foods was observed in physical and clinical exam. No severe symptom was observed, except for mild gastrointestinal symptom such as soft stool in a subject who has hypersensitive intestine.<br> <i>Conclusion</i>: These results suggest that the granular food containing L.E.M. is safe in healthy adults, even if excessive amount up to 5,400 mg a day is consumed.<br>

Antioxidant Properties of a Water-soluble Extract from Culture Medium of Ganoderma lucidum (Rei-shi) Mycelia and Antidiabetic Effects in Streptozotocin-treated Mice / 日本補完代替医療学会誌

<i>Objective</i>: <i>Ganoderma lucidum</i> (Rei-shi) is widely used as an alternative medicine agent to promote health and longevity. The water-soluble extract from culture medium of <i>Ganoderma lucidum</i> mycelia (WER) is prepared from a solid medium composed of bagasse and rice bran overgrown for 3–4 months with <i>Ganoderma lucidum</i> mycelia.<br> <i>Design</i>: In this study, we evaluate antioxidant activity of WER <i>in vitro</i>, and examined the effects of oral treatment of WER on oxidative stress in streptozotocin (STZ)-induced diabetic mice.<br> <i>Method</i>: Male ICR mice were treated with STZ (150 mg/kg, i.p.) and were housed for a week for induction of experimental diabetic state. WER (1 g/kg daily) was orally administered for an additional 9 weeks, and the levels of blood glucose, oxidative stress in plasma, liver and kidney of the mice were assessed.<br> <i>Result</i>: WER inhibited generation of superoxide anion and lipid peroxidation in concentration dependent manner<i> in vitro</i>. Oral administration of WER to diabetic mice significantly reduced the levels of blood glucose, hydroperoxides, triglyceride, ALT, and AST. Moreover, WER prevented the increase in lipid peroxide levels and the decrease in activity of antioxidant enzymes (superoxide dismutase, catalase, glutathione peroxidase) in liver and kidney of diabetic mice. In addition, histochemical studies revealed that treatment of WER precluded the sinusoidal dilation in liver and expansion of mesangial matrix in kidney of diabetic mice. Normal mice treated with WER showed no change in any parameters studied.<br> <i>Conclusion</i>: These data suggested that WER can act as an antioxidant <i>in vivo</i>, and show the antidiabetic effects by relieving diabetic hyperglycemia-induced oxidative stress.<br>