Edodes Mycelia Research Articles

Untargeted Metabolite Profiling of Antimicrobial Compounds in the Brown Film of Lentinula edodes Mycelium via LC-MS/MS Analysis.

The brown film (BF) of Lentinula edodes mycelium has been reported to exert biological activities during mushroom cultivation; however, to date, there is limited information on its chemical composition. In this study, untargeted metabolomics analysis was performed via liquid chromatography–mass spectrometry (LC–MS), and the results were used to screen the antimicrobial compounds. A total of 236 differential metabolites were found among the BF stages compared with the white hyphal stage. Among them, five important antimicrobial metabolites related to antimicrobial activities, namely, 6-deoxyerythronolide B, tanikolide, hydroxyanthraquinone, benzylideneacetone, and 9-OxooTrE, were present at high levels in the BF samples. The score plots of the principal component analysis indicated that the samples from four time points could be classified into two groups. This study provided a comprehensive profile of the antimicrobial compounds produced during BF formation and partly clarified the antibacterial and antifungal mechanism of the BF of L. edodes mycelium.

Fruiting bodies of selected edible mushrooms as a potential source of lovastatin

Agaricus bisporus, Cantharellus cibarius, Imleria badia, and Lentinula edodes are among the most popular species of edible mushrooms in Poland. These edible mushrooms are an important source of biologically active substances exhibiting beneficial (e.g., antioxidant, antitumor, antimicrobial, anti-inflammatory) effects on the human body. The fruiting bodies of edible mushrooms are also a valuable source of lovastatin, which belongs to a group of compounds, called statins, commonly used as cholesterol-lowering drugs. Due to the presence of lovastatin, edible mushrooms can be useful in the prevention of hypercholesterolemia. Therefore, the aim of this study was to determine the content of lovastatin in the selected species of edible mushrooms and to evaluate its release into artificial digestive juices. This study was the first to determine the release of lovastatin into digestive juices after the extraction of materials obtained from edible mushrooms. The largest amount of lovastatin was found in the fruiting bodies of C. cibarius (67.89 mg/100 g d.w.), and the smallest in those of L. edodes (0.95 mg/100 g d.w.). The amount of lovastatin released from the extracts of the examined species into digestive juices was found to be relatively low. The highest content after incubation in artificial gastric juice was detected for the fruiting bodies of L. edodes (0.02 mg/100 g d.w.) and after incubation in the intestinal juice for the mycelium from the in vitro cultures of L. edodes (0.51 mg/100 g d.w.). Thus, the results of the present study showed that due to its ability to accumulate lovastatin from culture medium, L. edodes mycelium can be used to obtain a product with increased hypolipidemic activity.

A Quality-of-Life Study in Healthy Adults Supplemented with Lentinex® Beta-Glucan of Shiitake Culinary-Medicinal Mushroom, Lentinus edodes (Agaricomycetes).

Lentinus edodes (=Lentinula edodes) is a culinary-medicinal mushroom with a long tradition of use in Asia. The major active substance in L. edodes is a beta-(1-3,1-6)-glucan (lentinan). GlycaNova produces Lentinex®, which contains this beta-glucan from L. edodes mycelia, in a proprietary process that maintains the triple helix molecular structure and high molecular weights. This study was carried out to investigate the effect of Lentinex supplementation on the well-being of adults. We evaluated the effect of Lentinex in healthy adult subjects in a randomized, placebo-controlled, double-blind trial. Sixty-three subjects, randomly allocated to two groups, took orally either 1-2 mL/day Lentinex (1-2 mg beta-glucan) or placebo for four weeks. The participants completed a well-being questionnaire prior to commencing supplementation and again at the end of the in-home study. The results showed an important and statistically significant improvement in well-being over the period in the treated group compared with the placebo group. The degree of improvement in the treated group, including relative to placebo, was higher for subjects who had lower initial well-being than for subjects with higher initial well-being. In conclusion, the lower an individual's initial well-being, the more Lentinex helped.

Variations in Nuclear Number and Size in Vegetative Hyphae of the Edible Mushroom Lentinula edodes.

In basidiomycete fungi, the number of nuclei and their ploidy level per nucleus can vary tremendously among species; however, within species, nuclear number and ploidy levels are traditionally considered fixed in their vegetative hyphae. In the edible mushroom Lentinula edodes, the hyphae are classified as either monokaryotic or dikaryotic, with each monokaryotic hyphal cell containing one haploid nucleus, and each dikaryotic hyphal cell containing two haploid nuclei. The dikaryotic hyphae are the results of mating between two genetically distinct monokaryons with different mating types. In this study, we examined the nuclear number and size (a potential correlate to ploidy) of L. edodes mycelia throughout its vegetative growth. We found that the number of nuclei within individual hyphal cells varied widely from non-nucleated to uninucleated, dinucleated, and multinucleated. Additionally, different nuclei within the same cell appeared very different in size, with a maximum nucleus cross-sectional area of 4.94 μm2 and the minimum nucleus cross-sectional area at only 0.37 μm2. Moreover, as culture time increased, more cells appeared to be devoid of any nuclei, with transmission electron microscopy and terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assays of late-stage cultures showing autophagosomes fusing and dissolving the nuclei and resulting in a large number of TUNEL-positive DNA fragments in non-nucleated cells. These results indicated that non-nucleated cells were likely caused by autophagy and apoptosis-like activities within aging L. edodes hyphae.

The Effect of Hemicellulose and Lignin on Properties of Polysaccharides in Lentinus edodes and Their Antioxidant Evaluation.

Lentinus edodes, whose polysaccharides possess diverse bioactivities, commonly grows on hardwood sawdust composed of hemicellulose, lignin and cellulose. In this study the effect of hemicellulose and lignin on the growth of mycelia, as well as the physicochemical properties of polysaccharides from L. edodes mycelia (LEPs) were investigated. The antioxidant properties of LEPs were evaluated through radical scavenging assays in vitro and through the Caenorhabditis elegans model in vivo. The results showed that hemicellulose at a concentration of 4% increased the yield of the mycelia biomass to twice that of the control group. Meanwhile, when cultured with 4.0% hemicellulose, the polysaccharide content of the mycelia was raised by 112.2%. In addition, the appropriate concentration of lignin could stimulate mycelia growth and polysaccharide biosynthesis in L. edodes. Monosaccharide composition analysis showed that a higher content of xylose was found when mycelia were cultured with higher concentrations of hemicellulose. The molecular structure, including the molecular weight distribution and configuration type, was affected by hemicellulose and lignin. Antioxidant assays indicated that LEPs supplemented with hemicellulose and/or lignin possessed higher radical scavenging abilities in vitro and exhibited a thermal resistance effect on C. elegans, implying that the antioxidant effect is potent in vivo. In summary, the addition of hemicellulose and lignin improved the biosynthesis and bioactivity of LEPs.

GC–MS-Based Nontargeted and Targeted Metabolic Profiling Identifies Changes in the Lentinula edodes Mycelial Metabolome under High-Temperature Stress

To clarify the physiological mechanism of the Lentinula edodes (L. edodes) response to high-temperature stress, two strains of L. edodes with different tolerances were tested at different durations of high temperature, and the results showed that there were significant changes in their phenotypes and physiology. To further explore the response mechanism, we established a targeted GC–MS-based metabolomics workflow comprising a standardized experimental setup for growth, treatment and sampling of L. edodes mycelia, and subsequent GC–MS analysis followed by data processing and evaluation of quality control (QC) measures using tailored statistical and bioinformatic tools. This study identified changes in the L. edodes mycelial metabolome following different time treatments at high temperature based on nontargeted metabolites with GC-MS and further adopted targeted metabolomics to verify the results of the analysis. After multiple statistical analyses were carried out using SIMCA software, 74 and 108 differential metabolites were obtained, respectively. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that the metabolic pathways with significant changes included those related to the following: amino acid metabolism, the glycolysis pathway, the tricarboxylic acid (TCA) cycle, and sugar metabolism. Most amino acids and carbohydrates enriched in these metabolic pathways were upregulated in strain 18, downregulated in strain 18N44, or the synthesis in strain 18 was higher than that in strain 18N44. This result was consistent with the physiological phenotypic characteristics of the two strains under high-temperature stress and revealed the reason why strain 18N44 was more heat-sensitive. At the same time, under high temperature, the decrease of intermediate products in glycolysis and the TCA cycle resulted in carbon starvation and insufficient energy metabolism, thus inhibiting the growth of L. edodes. In addition, the results also showed that the metabolites produced by different L. edodes strains under high-temperature stress were basically the same. However, different strains had species specificity, so the changes in the content of metabolites involved in the response to high-temperature stress were different. This provides a theoretical basis for further understanding the mechanism of the L. edodes response to high temperature and can be used to establish an evaluation system of high-temperature-resistant strains and lay the foundation for molecular breeding of new L. edodes strains resistant to high temperature.

Lentinula edodes Mycelium as Effective Agent for Piroxicam Mycoremediation.

Pollution of the environment with inorganic and organic substances is one of the main problems in the world. For this reason, it is necessary to conduct researches for effective methods of biodegradation of xenobiotics, including drugs whose unmetabolized forms are introduced into the environment, especially into water. One possible solution to this problem may be the use of white rot fungi, such as Lentinula edodes. This is an edible species used in medicine because of its beneficial anti-cancer, hypocholesterolemic, hypotensive, hypoglycemic and antioxidant effects. Due to the fact that the mycelium of L. edodes produces enzymes with oxidizing properties that can degrade xenobiotics. The aim of the work was verification if in vitro cultures of L. edodes can be used for bioremediation of non-steroidal, anti-inflammatory drug: piroxicam. For this purpose, the in vitro culture of L. edodes was derived and the mycelial cultures of this species enriched with piroxicam were analyzed. The biodegradation pathway of piroxicam by L. edodes mycelium was carried out by the UPLC/MS/MS method. The degradation process of piroxicam was found to affect primarily the linker between the thiazine and the piperidine ring, leading to its oxidation and cleavage. Additionally, oxidation of the benzothiazine moiety was observed, leading to hydroxylation and oxidation of the phenyl ring as well as oxidation of the thiazine ring leading to partial or complete removal of the sulfonamide moiety. It seems that the degradation process led finally to 2-hydroxybenozquinone, which may be further oxidized to inorganic compounds. What’s more, concentration of piroxicam in in vitro cultures of L. edodes was not correlated with effectiveness of biodegradation of this compound – on each experimental series, the level of degradation was the same. The results confirm the possibility of using the investigated L. edodes mycelium for remediation of piroxicam.

Potential Reduction of Salt Consumption by Preparing Noodles with Entrapped NaCl in Mycelial Cell Wall Cavities of Lentinus edodes

For the reduction of sodium consumption from foods with soups or sauces for noodles, NaCl mixture (LMC-salt) was prepared with the Lentinus edodes mycelium cell wall components from mycelia grown in submerged culture. NaCl was entrapped in their cavities and also deposited on the surface of mycelium cell wall. Noodles prepared with LMC-salt (Nm) enhanced the saltiness perception up to 40%, potentially by reducing “severance of flavor” as a consequence of slower release of entrapped NaCl in L. edodes cell wall, when compared to noodles prepared with NaCl (Nc) only. Compared to Nc, lightness was lower and redness was slightly higher resulting in higher color intensity in Nm. Overall textural properties did not show significant differences. Weight and volume of the cooked Nm were decreased, while turbidity of the soup was increased compared to Nc after cooking. Overall preference was slightly lower, but did not have significant specific negative attributes. These results could support a strategic approach to reduce total sodium consumption by enhancing saltiness of noodle in noodle dishes and consequently reducing uptake of soups or sauces that are leading sources of sodium intake.

English

In this study, the effect of three carbon sources (mannitol, minced potato and sucrose), two immobilization substrates (alginate and wood cylinders), and three surfactants (Tween 20, Tween 40 and Tween 80), were evaluated on eritadenine production using shiitake (Lentinula edodes) mycelium under submerged cultivation, in shake flasks within 20 days. Eritadenine and biomass were measured by HPLC and gravimetrically, respectively. Alginate immobilization of mycelium promoted significant enhancement of eritadenine yields of 88 mg/L, compared to the control (8.7 mg/L) and wood immobilization (14.8 mg/L). Likewise, eritadenine yields (72.4 mg/L) were enhanced by adding surfactant tween 20 to the broths in 0.5%, than control (8.7 mg/L) without surfactant. Tween 40 and 80 did not improve eritadenine yields, but both produced better biomass values (superior to 5 g/L) than the control (3.9 g/L). All carbon sources (sucrose, mannitol, mince potato, and glucose as control) produced similar low eritadenine yields, with best results (10.2 mg/L) by sucrose, although glucose produced the best biomass yields of 3.9 g/L. Also, carbon sources and the best biomass values did not show significant effect on eritadenine production. pH values in the best eritadenine yielding fermentations went down from 6 to 3-4, but pH had a low correlation with eritadenine yields. Finally, all data obtained in the present study are useful for optimizing culture conditions, towards industrialization of this important health improver metabolite (eritadenine). Key words: Eritadenine, shiitake mushroom, submerged culture, immobilization, surfactants.

Reference Gene Selection for Quantitative Real-Time PCR of Mycelia from Lentinula edodes under High-Temperature Stress.

Housekeeping genes are important for measuring the transcription expression of functional genes; 10 traditional reference genes, TUB, TUA, GADPH, EF1, 18S, GTP, ACT, UBI, UBC, and H2A, were tested for their adequacy in Lentinula edodes (L. edodes). Using specific primers, mRNA levels of these candidate housekeeping genes were evaluated in mycelia of L. edodes, which were treated with high-temperature stress at 37°C for 0, 4, 8, 12, 18, and 24 hours. After treatment, expression stability of candidate genes was evaluated using three statistical software programs: geNorm, NormFinder, and BestKeeper. According to geNorm, TUB had the lowest M values in L. edodes strains 18 and 18N44. Using NormFinder, the best candidate reference gene in strain 18 was TUB (0.030), and the best candidate reference gene in strain 18N44 was UBI (0.047). In BestKeeper analysis, the standard deviation (SD) values of UBC, TUA, H2A, EF1, ACT, 18S, and GTP in strain 18 and those of GADPH and GTP in strain 18N44 were greater than 1; thus, these genes were disqualified as reference genes. Taken together, only UBI and TUB were found to be desirable reference genes by BestKeeper software. Based on the results of three software analyses, TUB was the most stable gene under all conditions and was verified as an appropriate reference gene for quantitative real-time polymerase chain reaction in L. edodes mycelia under high-temperature stress.

Bioprocessed polysaccharide from Lentinus edodes mycelia cultures with turmeric represses the proinflammatory cytokine expression of the porcine macrophage infected by Salmonella Choleraesuis

Turmeric is known for its ability to enhance immunity via anti-inflammatory and anti-oxidant effects. Salmonella enterica species contain a large number of pathogenic serotypes that are adapted to a broad range of vertebrate hosts. Our previous study revealed that bioprocessed polysaccharides from the liquid culture Lentinus edodes fungal mycelia containing turmeric (BPP-turmeric) is able to alter chicken macrophage responses and increases chick survival against Salmonella enterica infection. In this study, we examined the immunomodulatory effects of BPP-turmeric on the porcine macrophage 3D4/31 cell line infected with Salmonella enterica subsp. enterica serovar Choleraesuis (S. Choleraesuis) or S. Enteritidis. Our experimental analyses demonstrated that BPP-turmeric (i) does not alter phagocytic and killing activity of 3D4/31 against either Salmonella serotypes, but that it (ii) represses mRNA transcription of interleukin (IL)-6, IL-8, and tumor necrosis factor α in response to Salmonella infection. Collectively, these results imply that BPP-turmeric has an immunomodulatory effect that represses pro-inflammatory cytokine expression in porcine macrophages, suggesting that it may protect swine from salmonellosis via controlling Salmonella-induced hyperinflammation.

Effect of Lignin-Containing Media on Growth of Medicinal Mushroom Lentinula Edodes

Abstract The effect of lignocellulose and lignin on growth of mycelium of mushroom Lentinula edodes and laccase activity in cultivation medium was studied. It was shown that cultivation of L. edodes mycelium in liquid nutrient medium with addition of 0.25-0.5% of kraft lignin increased mycelium biomass yield approximately two times compared with reference conditions without addition of lignin. Similar results were obtained in experiments in which 0.5% lignocellulose that remained after obtaining furfural, and 0.5% lignin that remained after obtaining furfural and glucose from wheat straw, were added to the nutrient medium. This effect was greater in the conditions of cultivation with good aeration, compared with static culture. Laccase activity in medium increased after addition of wheat straw lignocellulose or lignin only in the case of mycelium cultivation with aeration. In the case of mushroom cultivation on solid nutrient medium, addition of wheat straw lignocellulose and lignin promoted growth of mycelium only during the first 7 days of cultivation.

Observation of the curative effect of compound brucea Javanica preparation combined with lentinus edodes mycelia polysacharide tablets in the treatment of condyloma acuminatum

目的 观察复方鸦胆子联合香菇菌多糖片治疗男性尖锐湿疣的疗效。 方法 选择黄州区人民医院皮肤性病科门诊2013年3月至2014年5月就诊的尖锐湿疣患者121例为研究对象，将入选患者按随机数字表法分为两组，观察组(61例)给予复方鸦胆子外搽，2次/d，同时内服香菇菌多糖片10 mg，3次/d；对照组60例，单用复方鸦胆子外搽，2次/d。治疗4周后比较两组的治愈率、复发率及不良反应。 结果 随访12周，观察组治愈率为90.5%，复发率0.95%；对照组治愈率为58.3%，复发率41.7%。两组治愈率、复发率比较差异均有统计学意义(χ2＝0.95、7.12，均P<0.05)。 结论 采用复方鸦胆子联合香菇菌多糖片治疗男性尖锐湿疣治愈率高，复发率低，不良反应轻，值得基层医院推广应用。

がん患者におけるシイタケ菌糸体抽出物配合顆粒のQOL改善作用：多施設共同研究

シイタケ菌糸体抽出物は癌化学療法との併用時に，患者のQOLを維持・改善することが報告されている．本研究では，様々な治療背景および様々なステージのがん患者のQOLに及ぼすシイタケ菌糸体抽出物の影響を検討した．16施設で73症例を対象にシイタケ菌糸体抽出物を4週間（1,200 mg/day）連日経口摂取させ，摂取前後のQOLをEORTC-QLQ-C30調査票でスコア化した．シイタケ菌糸体抽出物摂取後のQOLスコアは被験者全体では，摂取前スコアに比べ，心理的，疲労スコアで有意に改善した．特にステージ3，4の被験者では，総体的，身体的スコアの改善も観察された．シイタケ菌糸体抽出物の経口摂取の併用は，進行性がん患者のQOLを改善することが示唆された．

Mycelial Extracts as Sensitive Element for Acoustoelectronic Gas Sensor

The main problem in development of the acoustoelectronic gas sensors is the search of the elements which are sensitive to the presence of different gases. In this paper we suggest to use for this purpose the piezoelectric quartz resonators with the mycelium films of mushroom Lentinula edodes strain F-249, which was cultivated in synthetic medium. The extract of mycelium was deposited on the surface of the resonator. The measurements of the frequency dependencies of the real and imaginary parts of electrical impedance of the resonator allowed finding the density, elastic constants and viscosity of the films under study. The influence of various gases (ammonia, formaldehyde, ethylacetate) and volatile liquids (acetone, acetic acid, chloroform) on physical properties of the extracts of mushroom Lentinula edodes mycelium has been investigated. It has been found that it is possible to use the mycelium films of mushroom Lentinula edodes prepared by different ways as sensitive film to ammonia, formaldehyde and ethyl acetate.

In vitro 및 In vivo에서 표고버섯, 표고버섯균사체배양쌀 추출물의 항당뇨와 항비만 작용

This study investigated the ability of extracts from Lentinus edodes (LE) and rice with Lentinus edodes mycelium (LEM) to inhibit diabetes and obesity. Lipid accumulation significantly decreased by 78% and 74% upon treatment with 300 μg/mL of LE and LEM, respectively (p<0.01). Cholesteryl ester transfer protein (CETP) inhibition activity increased by 94% and 99% upon treatment with 300 μg/mL of LE and LEM, respectively. In order to investigate the effect of LE and LEM on diabetes, the inhibition of protein tyrosine phosphate 1B (PTP1B) activity from the LE and LEM extracts at various concentrations (1, 3, 10, 30, 100, 300 μg/mL) was assessed. PTP1B activity by treatment with 10, 30, and 100 μg/mL of LE, was inhibited at a rate of 7, 9, and 7% respectively. Also, PTP1B activity from treatment with increasing concentration of LEM led to a significant concentration-dependent inhibition of PTP1B activity (p<0.01). LE and LEM were orally administered for 28 days after a high fat diet (HFD). LE and LEM significantly reduced triglyceride, cholesterol, HDL-cholesterol and LDL-cholesterol levels. GOT and GPT were not significantly effected. These results indicate that extracts of LE and rice with LEM have potent activities useful in the treatment of obesity and diabetes mellitus.

Biological Availability and Preliminary Selenium Speciation in Selenium-Enriched Mycelium of Lentinula edodes (Berk.)

Our goal is to obtain a new food supplement — a chemopreventive preparation derived from selenium (Se)-enriched Lentinula edodes mycelium. In the present study the bioavailability of selenium from Se-enriched mycelium preparations was tested in vitro and in vivo. Three different preparations of selenated mycelium were compared: dried mycelium, lyophilized mycelium, and lyophilized-autolyzed mycelium. In vitro, estimated Se bioavailabilities were 60%, 82%, and 98%, respectively. In vivo bioavailability was determined in rats. As reference, sodium selenite and Se yeast formulations were used at an Se-equivalent dose. The pharmacokinetic data for tested mycelial preparations suggest a rapid but incomplete Se absorption, and rapid elimination, without risk of accumulation. The speciation of selenium in Se-enriched mycelial cultures was carried out by specific oxido-reduction reactions. For tested mycelium the main part of Se was in the 0 and IV oxidation states in inorganic form or combined in a lipid or carbohydrate structure, about 47% was in the –II state in Se-amino acids or in other undefined water- or alcohol soluble organic compounds. Differences in pharmacokinetic data for Se yeast and L. edodes mycelial formulations probably arise from differences in Se speciation. Our data suggest that formulations of selenized Lentinula edodes mycelium could be used in chemoprevention as food supplements.

Supplementation with a Soluble Beta-Glucan Exported from Shiitake Medicinal Mushroom, Lentinus edodes (Berk.) Singer Mycelium: a Crossover, Placebo-Controlled Study in Healthy Elderly

Lentinus edodes (Shiitake) is a medicinal mushroom with a long tradition of use in Asia. The major active substance in L. edodes is a (1-6,1-3)-beta-glucan (lentinan). No clinical controlled studies have yet investigated the effect of orally administered lentinan on the immune response in healthy, elderly Caucasian subjects. We evaluated the effect and the safety of a beta-glucan from L. edodes mycelium, Lentinex, in healthy, elderly subjects in a double blind, crossover, placebo-controlled trial. Forty-two subjects were randomly allocated to two groups given orally either 2.5 mg/day Lentinex or placebo for 6 weeks; then after a washout period of 4 weeks, the alternate supplementation was given for 6 weeks. The changes in the number of B-cells were significantly different between the groups. The number ofNK cells increased significantly in both groups, but there was no significant difference between the groups. Other factors of the immune response (immunoglobulins, complement proteins, cytokines) were not altered. The safety blood variables (differential cell count, liver function, kidney function, and other blood chemistry) were not influenced by Lentinex, and the number, nature, and severity of adverse events were similar to placebo. Lentinex given orally to elderly subjects was safe and induced an increase in the number of circulating B-cells.

Relationship Between Selenium Accumulation and Mycelial Cell Composition in Lentinula edodes (Berk.) Cultures

It was postulated that fractions enriched in selenium (Se) isolated from Lentinula edodes mycelium polysaccharide might possess higher biological activity than the non-enriched fractions currently used to treat cancer. In order to obtain Se-enriched mycelial preparations, L. edodes cultures were cultivated in media enriched with sodium selenite. In order to determine whether the concentration of Se in the culture medium affected the biosynthesis and composition of cell wall and cell membrane, concentrations of the exopolysaccharide (EPS), chitin, and sterol (ergosterol) were measured in harvested mycelia. In addition, the relationship between Se accumulation and content of polyphenols and vitamin D2 in L. edodes mycelium was examined. The effects of Se levels on the mycelium cell composition were determined in culture media enriched with Se at concentrations ranging from 0 to 30 μg/ml. In each culture mycelial growth, total Se and Se distribution were determined between mycelial fractions of different polarity. The EPS, polyphenolics, and ergosterol content in harvested mycelia rose in proportion to Se concentration in the culture medium. The chitin content in mycelia increased with Se concentrations in the range 0–5 μg/ml, but at higher concentrations chitin levels decreased. Data showed that Se in culture medium exerted potent effects on the composition of the mushroom cell wall and semipermeable membrane, and on the content of polyphenolics that are involved in detoxification processes. Our findings indicate the optimal concentration of Se required in the culture medium for maximal yield of immunostimulatory-active selenated exopolysaccharides.

Hepatoprotective Effect of Syringic Acid and Vanillic Acid on CCl4-Induced Liver Injury

The mycelia of the edible mushroom Lentinula edodes can be cultured in solid medium containing lignin, and the hot-water extracts (L.E.M.) is commercially available as a nutritional supplement. During the cultivation, phenolic compounds, such as syringic acid and vanillic acid, were produced by lignin-degrading peroxidase secreted from L. edodes mycelia. Since these compounds have radical scavenging activity, we examined their protective effect on oxidative stress in mice with CCl(4)-induced liver injury. We examined the hepatoprotective effect of syringic acid and vanillic acid on CCl(4)-induced chronic liver injury in mice. The injection of CCl(4) into the peritoneal cavity caused an increase in the serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels. The intravenous administration of syringic acid and vanillic acid significantly decreased the levels of the transaminases. Four weeks of CCl(4) treatment caused a sufficiently excessive deposition of collagen fibrils. An examination of Azan-stained liver sections revealed that syringic acid and vanillic acid obviously suppressed collagen accumulation and significantly decreased the hepatic hydroxyproline content, which is the quantitative marker of fibrosis. Both of these compounds inhibited the activation of cultured hepatic stellate cells, which play a central role in liver fibrogenesis, and maintained hepatocyte viability. These data suggest that the administration of syringic acid and vanillic acid could suppress hepatic fibrosis in chronic liver injury.