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The separation of triterpene acids was achieved from Poria cocos (Schw.) Wolf by TLC after treatment with Ph2CNN or RX under mild condition followed by selective hydrolysis of ester groupings. Three triterpene acids, 3 beta-acetoxyl-16 alpha-hydroxy-lanosta-8,24(31)-diene-21-oic acid (I), 3-oxo-16 alpha-hydroxy-lanosta-7,9(11), 24(31)-trien-21-oic acid (II) and 3 beta, 16 alpha-dihydroxylanosta-7,9(11), 24(31)-trien-21-oic acid (III) were isolated.

Three new compounds, 5 alpha-lanosta-7,9(11),24-triene-3beta-hydroxy-26-al (1), 5 alpha-lanosta-7,9(11),24-triene-15 alpha-26-dihydroxy-3-one (2), and 8 alpha,9 alpha-epoxy-4,4,14 alpha-trimethyl-3,7,11,15,20-pentaoxo-5 alpha-pregnane (3), were isolated from Ganoderma concinna along with 12 known compounds. The structures of compounds 1 and 2 were determined on the basis of MS and NMR studies. The structure of 3 was determined by MS, NMR, and single-crystal X-ray diffraction. Compounds 1, 2, and 3 induce apoptosis in human promyelocytic leukemia HL-60 cells, as indicated by examining the morphological features of cells and detection of DNA fragmentation by gel electrophoresis.

We recently discovered that the triterpene acid compound dehydrotrametenolic acid promotes adipocyte differentiation in vitro and acts as an insulin sensitizer in vivo. This natural product has been isolated from dried sclerotia of Poria cocos WOLF (Polyporaceae), a well-known traditional Chinese medicinal plant. We examined the effects of dehydrotrametenolic acid on plasma glucose concentration in obese hyperglycemic db/db mice. Dehydrotrametenolic acid can reduce hyperglycemia in mouse models of noninsulin-dependent diabetes mellitus (NIDDM) and act as an insulin sensitizer as indicated by the results of the glucose tolerance test. These terpenoids and thiazolidine type of antidiabetic agents such as Ciglitazone, although structurally unrelated, share many biological activities: both induce adipose conversion, activate peroxisome proliferator-activated receptor gamma (PPAR gamma) in vitro, and reduce hyperglycemia in animal models of NIDDM. Dehydrotrametenolic acid is a promising candidate for a new type of insulin-sensitizing drug. This finding is very important for the development of insulin sensitizers that are not of the thiazolidine type.

We have reported that D-Fraction extracted from maitake (Grifola frondosa), activates immune competent cells, and indicates anti-tumor activities. The D-Fraction was observed to induce angiogenesis in vivo and to enhance the proliferation capability and migration capability of human vascular endothelial cell in vitro. The D-Fraction also increased plasma vascular endothelial growth factor (VEGF) concentration significantly. Also VEGF and TNF-alpha production by the activated peritoneal macrophages were enhanced. These results suggest that the anti-tumor activity of the D-Fraction is not only associated with the activation of the immuno-competent cells but also possibly related to the carcinoma angiogenesis induction.

Aim: To study the structure of polysaccharides from Coriolus versicolor., Methods: By chromatography a homogeneous polysaccharide B-1-3 was obtained. The structure of B-1-3 was elucidated using the method of total acid hydrolysis, methylation, and 1D and 2D NMR., Results: B-1-3 was found to be mainly composed of Glc by total acid hydrolysis. Its molecular weight was estimated to be 3.16 x 10(5) by HPLC method. Methylation analysis and 1H and 13CNMR indicated the main chain of B-1-3 consisted of beta-D-1,4-Glc and beta-D-1,3-Glc, and branch chains were situated, beta-D-1,3,6-Glc and beta-D-1,4,6-Glc. 1H and 13CNMR also showed that beta-D-Glc-1-->3-beta-D-Glc-beta-D-Glc-1-->4-beta-D-Glc and beta-D-Glc-1-->4-beta-D-Glc-beta-D-Glc-1-->4-beta-D-Glc fragments existed in B-1-3., Conclusion: B-1-3 is a glucan which main chain was composed of beta-D-1,4-Glc and beta-D-1,3-Glc beta-D-1,6-Glc.

Two new triterpenoids, lucidenic acid N (1) and methyl lucidenate F (2), together with four known compounds, lucidenic acid A, lucidenolactone, lucidenic acid C, and ganoderic acid E, were isolated from the dried fruiting bodies of Ganoderma lucidum. Their structures were elucidated by spectral and chemical transformation studies. Among them, lucidenic acid N (1), lucidenic acid A, and ganoderic acid E showed significant cytotoxic activity against Hep G2, Hep G2,2,15, and P-388 tumor cells.

A new 9-methyl-sphinga-4,8-dienine-containing glucocerebroside (1), together with two additional known analogs, cerebrosides B and D, was isolated from the chloroform-soluble lipid fraction of the ethanol and chloroform/methanol extract of the fruiting bodies of the basidiomycete Polyporus ellisii Berk. and characterized. The structure and relative stereochemistry of the new compound were identified as (2S,3R,4E,8E-1-(beta-D-glucopyranosyl)-3-hydroxy-2-[(R)-2'-hydroxyheptadecanoyl]amino-9-methyl-4,8-octadecadiene by means of spectroscopic (1H,13C, and two-dimensional nuclear magnetic resonance; mass spectrometry) and chemical methods.

A new benzofuran glycoside, masutakeside I (1) and a new C10 acetylenic acid, masutakic acid A (2) were isolated from the fruiting bodies of the fungus Laetiporus sulphureus var. miniatus. Their structures were established by spectroscopic and chemical methods. The known compounds 3-6 were also obtained and identified as egonol, demethoxyegonol, egonol glucoside and egonol gentiobioside. Some of these compounds exhibited cytotoxicity against Kato III cells.

We have previously reported that rats with diabetes induced by injecting streptozotocin into neonates showed remarkably lower blood glucose, urine volume, and glucosuria after administration of Maitake (Grifola frondosa). In the present study, we investigated the effects of Maitake on insulin concentration, organ weight, serum composition, and islets of Langerhans in streptozotocin-induced diabetic rats using the same method. The diabetic rats were produced by injecting 80 mg/kg B.W. streptozotocin into 2-d-old neonates. From the age of 9 wk, the rats were given experimental diets for 100 d. The diabetes and control groups were given either diets containing 20% Maitake (DM and CM groups) or control diets (D and C groups). During administration of the experimental diets, we measured body weight, food intake, amount of feces, and serum insulin concentration at glucose loading. The glucose tolerance test was performed at the 10th week after the start of the experimental diets. The D group had an initial fasting blood glucose of 225+/-49 mg/dL, and a maximum blood glucose of 419+/-55 mg/dL at 60 min. In the DM group, however, the initial fasting blood glucose was 170+/-23 mg/dL, and the maximum blood glucose was 250+/-41 mg/dL at 15 min. Both values were markedly lower than those in the D group (p<0.05). The insulin concentration at 15 min. after glucose loading in the DM group was 41+/-16 microU/mL, which was significantly higher than that in the D group (15+/-7 microU/mL) (p<0.05). After the 100-d experimental period, blood samples were collected. The fructosamine level was significantly lower in the DM group (152+/-21 mmol/L) than in the D group (185+/-13 mmol/L). The concentration of 1.5-A.G. (1.5-anhydro glucitol) was significantly higher in the DM group (9.33+/-2.42 microg/mL) than in the D group (1.33+/-0.52 microg/mL). Observation of insulin antibody stain in the Langerhans cells of the pancreas using ABC method showed a decrease insulin antibody stain in the D group. The cells of the DM group were stained more darkly than those of the D group. From these results, we postulated that the bioactive substances present in Maitake can ameliorate the symptoms of diabetes.

A preparation of water soluble components (EA) was made from carpophores of Elfvingia applanata (Pers.) Karst and its in vitro antiviral activity on vesicular stomatitis virus [(Indiana serotype, VSV(IND)] was investigated by plaque reduction assay. EA exhibited potent antiviral activity on VSV(IND) growth and negligible cytotoxicity on Vero cells, 50% effective concentration (EC50) of 104 microg/ml and 50% cytotoxic concentration (CC50) of 3,793 microg/ml, respectively. Selectivity index (SI, CC50/EC50) of EA on Vero cell and VSV(IND) was about 36.5. EA did not display either a direct virucidal effect on VSV(IND) or induction of antiviral substance by Vero cells upon its treatment. Thus, the mode of antiviral activity of EA was studied at steps of viral adsorption onto cell. When both EA and virus were added to cell monolayers, titer of cell-free virus in culture supernatant increased in ca. 30-40% compared with that of control group and titer of cell-associated virus was 60-100% higher than that of control group. These results suggested that antiviral activity of EA on VSV(IND) might be due to the hindrance of viral entry to cells at either endocytosis or loss of envelope.

The ability of Pycnoporus sanguineus to adsorb heavy metals from aqueous solution was investigated in fixed-bed column studies. The experiments were conducted to study the effect of important design parameters such as column bed height, flow rate and initial concentration of solution. The breakthrough profiles were obtained in these studies. A mathematical model based on external mass transfer and pore diffusion was used for the prediction of mass transfer coefficient and effective diffusivity of metals in macro-fungi bed. Experimental breakthrough profiles were compared with the simulated breakthrough profiles obtained from the mathematical model. Bed Depth Service Time (BDST) model was used to analyse the experimental data and evaluated the performance of biosorption column. The BDST model parameters needed for the design of biosorption columns were evaluated for lead, copper and cadmium removal in the column. The columns were regenerated by eluting the metal ions using 0.1 M hydrochloric acid solution after the adsorption studies. The columns were subjected to repeated cycles of adsorption of same metal ions and desorption to evaluate the removal efficiency after adsorption-desorption.

Four new phytosphingosine-type ceramides, (2S,3S,4R)-2-[(2'R)-2'-hydroxydocosanoylamino]-1,3,4-octadecane triol (1), (2S,3S,4R)-2-[(2'R)-2'-hydroxytricosanoylamino]-1,3,4-octadecan etriol (2), (2S,3S,4R)-2-[(2'R)-2'-hydroxypentacosanoylamino]-1,3,4-octadec anetriol (3) and (2S,3S,4R)-2-[(2'R)-2'-hydroxyhexacosanoylamino]-1,3,4-octadeca netriol (4), have been isolated from the fruit bodies of Grifola frondosa. The structures of the new compounds were elucidated on the basis of their spectral data.

Distillery wastewaters generated by ethanol production from fermentation of sugar-cane molasses, named vinasses, lead to important ecological impact due to their high content of soluble organic matter and their intense dark-brown color. Taking advantage of the well-known ability of white-rot fungi to degrade an extensive variety of organic pollutants, the capacity of Trametes sp. I-62 (CECT 20197) to detoxify this type of effluents was evaluated. In this work, pyrolysis/gas chromatography/mass spectrometry was applied to the chemical characterization of several fractions of Cuban distillery wastewater as well as to monitoring the changes which occurred after fungal treatment with this white-rot basidiomycete. Maximum effluent decolorization values and chemical oxygen demand reduction attained after seven days of fungal treatment were 73.3 and 61.7%, respectively, when 20% (v/v) of distillery vinasses was added to the culture medium. Under these conditions a 35-fold increase in laccase production by Trametes sp. I-62 was measured, but no manganese peroxidase activity could be detected. The pyrolysis/gas chromatography/mass spectrometry results showed a decrease in a number of pyrolysis products after seven days of fungal treatment, mainly furan derivatives. The decrease in the relative areas of these compounds could be related to the vinasse color-removal associated with melanoidin degradation. All these results indicated the potential use ofTrametes sp. I-62 in the detoxification of recalcitrant distillery vinasses., (Copyright 2000 John Wiley & Sons, Ltd.)

Objective: To study the chemical composition and structure of polysaccharides from the mycelium of Hericium erinaceum., Method: DEAE-cellulose and DEAE-Sephadex column chromatography were employed for isolation and purification. Sephadex column chromatography and agarose gel electrophoresis were used to measure the purity. Gel filtration was employed to measure MW. Acid hydrolysis, enzymolysis, GC and spectral analysis etc. were employed for determination of the composition and construction of the unique homogeneous polysaccharides., Result: The five homogeneous polysaccharides were obtained from the mycelium. The main compounds(HEPA1, HEPA4 and HEPB2) were identified., Conclusion: HEPA1 (MW 62,000) is a heteropolysaccharide consisting mainly of glucose. HEPA4 (MW 26,000) is a heteropolysaccharide peptide. HEPB2 (MW 12,000) is a heteropolysaccharide consisting mainly of glucose. The main linkage of all the afore-said homogeneous monosaccharides is alpha-linkage.

Fractionation of the 50% ethanol extract of Polyporus umbellatus Fries by column chromatography on Amberlite XAD-2, silica gel, Sephadex LH-20 and octadecyl silica gel (ODS) (C18)) monitored by a hair-regrowth activity assay, afforded three active principles, 1, 2 and 3. The structures of 1, 2 and 3 were determined as acetosyringone, polyporusterone A, and polyporusterone B by comparison of their spectral data with that of authentic samples, respectively. The effects of several compounds related to acetosyringone, 3,4-dihydroxybenzaldehyde or polyporusterone A on hair regrowth were also investigated.

To prove the relationship between the fluctuation in serum beta-glucuronidase level and hepatotoxicity, an inhibitor of beta-glucuronidase from G. lucidum was isolated and its hepatoprotective activity was investigated. The ether fraction of G. lucidum, which had potent beta-glucuronidase-inhibitory activity, protected against CCl4-induced liver injury. From this ether fraction, ganoderenic acid A, was isolated as the potent inhibitor of beta-glucuronidase. It had a potent hepatoprotective effect against CCl4-induced liver injury. These results suggest that the beta-glucuronidase seems to be closely related to liver injury, which could be prevented by beta-glucuronidase inhibitors.

Among antifungal products of natural origin, Pycnoporellus fulgens (Fr) Donk, is active against pathogenic yeasts. Our study was carried out to observe the ultrastructural modifications produced by P. fulgens crude extract on Candida glabrata. Yeasts were submitted to different concentrations (50, 100, 200, 400 micrograms ml-1) for 72 h under constant stirring at 35 degrees C. Transmission electron microscopy revealed that the extract acted on the cell envelope (cell wall and plasmalemma). Cell divisions were also affected by thickening of the septum (50 micrograms ml-1) and a deficiency in the daughter cell wall texture. The extent of the antifungal effect clearly depended on the extract concentration.

Sequence analysis of the downstream region of the basidiomycete Lentinus edodes priB gene encoding a protein with a 'Zn(II)2Cys6 zinc cluster' DNA-binding motif (Endo, H., Kajiwara, S., Tunoka, O., Shishido, K., 1994. A novel cDNA, priBc, encoding a protein with a Zn(II)2Cys6 zinc cluster DNA-binding motif, derived from the basidiomycete Lentinus edodes. Gene 139, 117-121) suggested the presence of a Saccharomyces cerevisiae URA6 gene homologue encoding UMP kinase. We isolated a corresponding cDNA from a mature fruiting-body cDNA library of L. edodes. The nucleotide sequence of this was determined and compared with that of the genomic DNA, revealing that the URA6 gene homologue encodes 227 amino acids (aa) and is interrupted by four small introns. The deduced aa sequence showed an overall identity of 51.1% to that of the S. cerevisiae URA6 gene product. The URA6 homologue protein produced in Escherichia coli using the glutathione S-transferase gene fusion system was found to catalyze the phosphoryl transfer from ATP to UMP and CMP efficiently and also to AMP and dCMP with lower efficiencies. Thus, the URA6 gene homologue was designated uck1 and its product UMP-CMP kinase. Northern-blot analysis showed that the uck1 is actively transcribed in the gill tissue of mature fruiting bodies of L. edodes, implying that uck1 may play a role during the formation of basidiospores occurs in the gill tissue.

This paper reports that polysaccharose was prepared by ethanol precipitation of water extracted and by deproteinization with sevage method, purification obtain Fomes marginatus. The polysaccharose which was determined by using sephadex-G, PAGE, ACTME, TLC and PC is single composition. [alpha]D20 = 28.89, it content of polysaccharose is 22.62 percent. It was hydrolied of periodate oxidation, smith degradation and IR analysis, the results show that the main chain is composed of B-(1-->4), (1-->6) linkage.

Aim: To study the role of interleukin-2 (IL-2) and mediobasal hypothalamus (MBH) in analgesia produced by Coriolus versicolor polysaccharide peptide (PSP)., Methods: The IL-2 antiserum was injected i.c.v. or i.p. and the MBH was destroyed electrolytically., Results: PSP i.g. 1 g.kg-1.d-1 for 6 d increased the pain threshold in tail stimulation-vocalization test in rats. This PSP-produced analgesia was blocked by i.c.v., but not i.p., IL-2 antiserum and disappeared after electrolytic lesion of MBH., Conclusion: The analgesia produced by PSP is mediated by IL-2 which is activated by PSP and interacts with IL-2 receptors in the MBH.

The water-soluble lignin in LEM (the extract of the solid culture medium of Lentinus edodes mycelia) has been known to have antiviral and immunopotentiating activities in vivo and in vitro. The water-soluble lignin rich fraction (JLS-18) was prepared from LEM using ultrafiltration and hydrophobic column chromatography. JLS-18 showed about 70 times higher antiviral activity than LEM in vitro. JLS-18 activated the cytotoxicity of NK cells and macrophages, and activated T cells in vitro. JLS-18 also induced interleukin 6 (IL-6) secretion from human leukocytes infected with Sendai virus in vitro. These data showed that JLS-18, the water-soluble rich fraction of LEM, had antiviral and immunopotentiating activities.

To study the effects of three kinds of treatment: Zhuling polysaccharides combined with hepatitis B vaccine (group I), LAK cells, induced by interleukin-2 (IL-2) in vitro, transfusion therapy, (group II), bacille Calmette-Guerin (BCG) combined with persatin (group III) in chronic hepatitis B and the mechanism of their effects, we observed 286 patients with chronic hepatitis B, diagnosed according to the criteria made by The 6th National Meeting for Viral Hepatitis and Substantiated by the results of liver biopsy. The patients were divided into 3 treatment groups randomly, with interferon 3 MIU, thrice a week and 10% glucose 500 ml (i.v.) dripping everyday as two different control groups. The ALT recovery rates in group I, II, III were 64%, 35% and 57%, respectively. At the end of the treatment, the HBeAg and HBV DNA clearence rates in group I, II, III were 43% and 44%, 34% and 30%, 57% and 61%, respectively. After one year of follow up, the HBeAg and HBV DNA clearence rates in group I, II, III were 59%, 58%, 61% and 55%, 70%, 60%, respectively. The effects of these kinds of treatment were similar to these of interferon. We also injected Zhuling polysaccharides combined with hepatitis B vaccine, Zhuling polysaccharides alone and normal saline as control to DHBV vertical transmission duck. It was shown that serum DHBV DNA was cleared and the results were also proved by liver histologic change in the two treatment groups. According to these data, it is suggested that the three kinds of treatment for hepatitis B could restore the ALT level, suppress the replication of HBV and improve the liver histologic change. Their effects were similar to those of interferon.

Feeding of Pleurotus to hypercholesterolemic rabbits resulted in lowering of total lipids, total cholesterol and glyceride levels of plasma and liver whereas heart lipids were not affected. HDL cholesterol/total cholesterol and HDL cholesterol/LDL cholesterol ratios increased in experimental animals proving antiatherogenic potential of this mushroom. The effect was further supported by the severity of tissue damage as evidenced by histopathological studies. An increase in bile acid excretion could be taken as one of the possible attributes to hypocholesterolemic action of P. florida.

A fruiting body-specific cDNA mfbAc, derived from the basidiomycete Lentinus edodes, has been shown to encode a high-molecular-mass (2157 amino acids) cell-adhesion protein MFBA containing the Arg-Gly-Asp (RGD) motif. A 425-amino-acid fragment containing the RGD motif of MFBA (designated MFBA(582-1006) peptide) produced in Escherichia coli exhibited cell-adhesion and spreading activity toward mammalian cells and cell-aggregation activity toward basidiomycetous hyphal cells via the RGD sequence. Here we investigated the biological activity of MFBA(582-1006) peptide in Saccharomyces cerevisiae. The DNA sequence encoding MFBA(582-1006) peptide, introduced into the yeast using an expression vector, resulted in a marked aggregation of the yeast cells. The aggregation was almost completely abolished by replacement of the RGD motif by an RGE motif in the peptide sequence.

One compounds were isolated from the fruiting body of Ganoderma lucidum. Basis on chemical evidences and spectral analysis (MS, UV, IR, 1HNMR, 13CNMR), its structures were deduced as ergosta-7,22 E-dien-3-one.

Seven homogeneous peptidepolysaccharides have been obtained from the fruit-body of Ganoderma lucidum. The main constituents are TGLP-2, TGLP-3, TGLP-6 and TGLP-7, whose molecular weights are 20.9 x 10(4), 4.5 x 10(4), 3.2 x 10(4) and 10 x 10(4) respectively. TGLP-2 is a heteroglycan peptide, TGLP-3 and TGLP-6 are glycan peptides and TGLP-7 is a galactan peptide.

Three compounds have been isolated from the dichloromethane soluble fraction of the fruiting body of Ganoderma lucidum (Fr.) Karst. On basis of spectral analyses (UV, IR, MS, 1HNMR, 13CNMR and 2D-NMR), they were identified as 3, 7-dioxo-lanosta-8, 24(E)-dien-26-oic acid (I), 7 beta-15 alpha-dihydroxy-3, 11, 23-trioxo-5 alpha-lanost-8-en-26-oic acid (II) and 3 beta, 7 beta, 15 alpha-trihydroxy-11, 23-dioxo-5 alpha-lanosta-8-en-26-oic acid (III). Compound I is a new compound named ganoderic acid DM.

ICR mice were treated with a carcinogen, N-butyl-N'-butanolnitrosoamine BBN), every day for 8 consecutive weeks and the effects of oral administration of edible mushrooms on the induction of urinary bladder carcinoma and on the activities of macrophages and lymphocytes were studied. Bladder carcinoma were found in all 10 mice (100%) treated with BBN alone, while we observed carcinoma only in 9 of 17 mice (52.9%), in 7 of 15 mice (46.7%) and 13 of 20 mice (65.0%) treated with Lentinus edodes, Grifola frondosa and Pleurotus ostreatus, respectively. Chemotactic activity of macrophages was suppressed in mice treated with BBN alone but maintained almost the normal level in mice treated with BBN plus Lentinus, Grifola or Pleurotus. Lymphocytes collected from mice treated with BBN plus each mushroom showed almost normal blastogenic response against concanavalin A, although those from mice treated with BBN alone completely retarded their response. Cytotoxic activity of lymphocytes against Yac-1 cells was also maintained at a normal level in mice treated with BBN plus each mushroom. Whereas in mice treated with BBN alone significant depression of NK cell activity occurred. Significantly higher cytotoxic activity against P-815 cells was observed in lymphocytes from mice treated with BBN plus each mushroom than that in lymphocytes from normal mice or mice treated with BBN alone.

For the past few years, we have been investigating polysaccharides from Ganoderma lucidum as antifibrotic agents. In a previous study, we discovered that polysaccharides extracted from G. Iucidum lowered the collagen content in liver but had no effect on serum biochemical parameters in rats subjected to bile duct ligation and scission-induced fibrosis. In this study, we changed the extraction method and obtained polysaccharides extracted from G. Iucidum. The polysaccharide from G. Iucidum reduced the serum aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP) and total bilirubin and also reduced the collagen content in liver and improved the morphology. Pentoxifylline, which is reported to exhibit an antifibrotic effect in pigs with fibrosis induced by yellow phosphorus, did not have any antifibrotic effects in fibrosis induced by biliary obstruction. Glycyrrhizin, which is used in the treatment of hepatitis, reduced serum ALT and AST values but there was no significance. It had no effect on liver hydroxyproline content which implies that glycyrrhizin has no antifibrotic effect in the rats with fibrosis induced by bile duct ligation and scission. These data suggest that the polysaccharide from Ganoderma lucidum could be a promising antifibrotic agent. However, further study is needed to understand the inhibition mechanism of collagen deposition of polysaccharides from Ganoderma Iucidum and its clinical applicability remains to be established.

Objective: To investigate the effect of treatment of cachexia with Umbellatus Polyporus Polysaccharide (UPP)., Methods: UPP was used to treat cachexia induced by toxohormone-L., Results: Toxohormone-L from ascites fluid with primary hepatoma could induce lipolysis in vitro significantly, inhibit the intake behavior, decrease the level of blood zinc, increase that of blood copper (P < 0.01), upp could inhibit these effects., Conclusion: Toxohormone-L was closely related with neoplastic cachexia and UPP could inhibit this effect induced by toxohormone-L significantly.

Two lipid peroxidation inhibitors, designated as betulinans A (1) and B (2), were isolated from the MeOH extract of Lenzites betulina. The structures of these compounds have been determined to be 2,5-diphenyl-3,6-dimethoxy-p-benzoquinone and 2-phenyl-3-methoxy-[1H-2-benzopyran][4,3-e][p]benzoquinone, respectively, on the basis of various spectral data. Betulinans A and B inhibited lipid peroxidation with IC50 values of 0.46 and 2.88 micrograms/mL, respectively.

The hydroalcoholic extract of Poria cocos and two lanostane derivatives isolated from it, pachymic acid (1) and dehydrotumulosic acid (2), were active as inhibitors of phospholipase A2 from snake venom when a polarographic method was used. Dehydrotumulosic acid exhibited an IC50 of 0.845 mM. These two compounds are structurally related to certain triterpenoids from Ganoderma and Schinus that have previously been described as competitive inhibitors of phospholipase A2. These comprise a new group of natural potential antiinflammatory agents due to their interaction with that enzyme.

A novel lectin was isolated from mycelia of the basidiomycete Pleurotus cornucopiae grown on solid medium. The lectin was purified to homogeneity by mucin-Sepharose affinity chromatography. The molecular mass of the lectin was 40 kDa under reducing conditions, but the subunits were polymerized through disulfide bridges under physiological conditions. Hemagglutinating activity of this lectin was completely inhibited by 2-mercaptoethanol, indicating that the multimer is active. The activity was also inhibited by EDTA, and restored by CaCl2. N-Acetyl-D-galactosamine was the most potent hapten inhibitor. N-terminal amino acid sequence analysis revealed that the mycelial lectin was different from the fruit body lectin of this organism. The mycelial lectin appeared prior to fruit body formation and disappeared during the formation of fruit bodies. The lectin was localized on the surface of solid-medium-grown mycelia, and only dikaryotic, and not monokaryotic, mycelia produced the lectin. These results suggest that the appearance of this lectin is associated with fruit body formation.

The effect of the diet containing 5% of powdered oyster mushroom (Pleurotus ostreatus) or an equivalent amount of mushroom ethanolic extract on cholesterol content in serum and liver, on its distribution in lipoproteins, absorption and turnover was studied in male Wistar rats (initial body weight about 70 g) fed a diet with 0.3% cholesterol. 12 weeks of feeding with whole oyster mushroom or mushroom extract reduced cholesterol level in serum by 52 and 33%, respectively. However, cholesterol content in liver was reduced only by whole oyster mushroom (by 20%). Diminished serum cholesterol level was mediated in 60% by reduction of cholesterol in very-low-density lipoproteins. Both whole oyster mushroom and mushroom extract increased the concentration of cholesterol in high-density lipoproteins. Consuming whole oyster mushroom decreased cholesterol absorption (estimated by dual-isotope plasma ratio method) by nearly 16% while no significant effect of mushroom extract could be demonstrated. Feeding the diet containing whole oyster mushroom or its extract reduced the half-times of decay curve of cholesterol-4-14C by 29 and 35%, respectively and reciprocally increased the fractional catabolic rate of plasma cholesterol.

Many immune modulating compounds have been isolated from fungal extracts, but the molecular mechanisms of their action have rarely been elucidated. In this study we isolated a proteoglycan from cultured mycelia of Lentinus lepideus and tested its effects on murine spleen cells. The acidic-polysaccharide fraction was obtained by extraction with hot water followed by purification using DEAE-cellulose anion exchange. The molecular mass of the compound was determined by Sepharose CL-4B gel filtration to be approximately 47 kDa. When cultured in the presence of the compound, spleen cells from C3H mice underwent rapid cell proliferation and cell aggregation. Treatment with the compound also caused a 10-fold increase in [3H]-thymidine incorporation compared to a control, confirming cell proliferation. Flow cytometry analysis indicated that the affected cell population was mainly B cells. As one approach to understanding the molecular mechanism of this action, we investigated the effects of the compound on cellular transcription factors which are known to control the proliferation of immune cells. Using gel retardation assays, we found that the compound significantly activated NF-kappa B but not AP-1 in spleen cells. Taken together, the data suggest that the proteoglycan compound is a biological response modifier that stimulates B cell proliferation, probably by regulating cellular transcription factors such as NF-kappa B.

Aim: The nervous mechanism of the immune potentiating effect of Coriolus versicolor polysaccharides peptides (PSP) was studied in Wistar rats., Methods: The unit discharge of the mediobasal hypothalamus (MBH) neurons was recorded extracellularly and the lymphocyte proliferation was measured., Results: PSP 1 g.kg-1 ig for 5 d increased the T-lymphocytes and promoted T-lymphocyte proliferation in spleen and peripheral blood. This promoting effect of PSP was blocked by MBH lesion. PSP increased the discharge frequency of MBH neurons, but no increase in discharge frequency was observed after treatment of PSP plus immune inhibitor, cyclosporin A., Conclusion: MBH is involved in the immune-potentiating effect of PSP.

The cytotoxic effect of two aqueous extracts of Inonotus obliquus on human cervical uteri cancer cells (Hela S3) in vitro was evaluated. It was concluded that Inonotus extracts at a concentration of 10 micrograms/ml to 2000 micrograms/ml inhibited cancer cells growth. In cultures with extracts of the fungus a decrease of the cell proteins and mitotic index was observed. Moreover, the extracts disturbed mitoses by elevating the number of mitotic cells in metaphase. Aqueous extracts of Inonotus effected not only mitoses but also the 8/G phase of the cell cycle.

The actions of Polyporus polysaccharide (PPS), mycobacterium polysaccharide (MPS) and lentinan (LEN) on lymphokine-activated killer (LAK) cell activity in vitro were investigated in this study. Human peripheral blood mononuclear cells (PBMC) were cultured for 96 hours with medium containing different concentrations of the above-mentioned drugs in combination with recombinant interleukin 2 (rIL-2). Then cell-mediated lysis was determined by 1H-TdR release assay including NK sensitive and NK resistant target cells. The results demonstrated that, when combined with rIL-2 in a certain concentration, all three kinds of polysaccharides could enhance the LAK activity by 42%-56.9%, and reduce the dose of rIL-2 by 50% (P < 0.05-0.01). It suggested that the PPS, MPS and LEN could be used as bioactivity regulators in LAK cell therapy in tumor treatment.

The extracellular polysaccharides produced in liquid cultures of the ligninolytic fungi Pleurotus cornucopiae, P. eryngii, P. floridanus, P. ostreatus, P. pulmonarius and P. sajor-caju showed similar composition and structure. They included a minor water-soluble fraction, containing D-glucose, D-mannose, D-galactose and protein (rich in amide- and hydroxyl-containing residues which can be involved in protein-polysaccharide linkages). The structural analysis of this fraction mainly showed alpha-(1-->4)-linked glucose units. The structure of the major fraction (amounting to more than 80% of the total extracellular polysaccharide) was investigated in detail. Its low solubility was attributed to the high M(r). For its characterization a procedure of progressive dissolution and dialysis was set up. The methylation analysis showed that it was a (1-->3)-beta-D-glucan and that approximately 25% of the total units were C-6 branched. This result was confirmed by the intensities of the signals assigned to C-3 and C-6, free and substituted, in 13C-NMR spectra. Additional studies showed linear polysaccharides after one-cycle Smith degradation or after glucan acetolysis. Both results, as well as glucose release after the latter treatment, indicated that the C-6 linked side chains are constituted by single glucose residues, on every second or third unit of the main chain of the (1-->3)-beta-D-glucan.

Three new steroids, zhankuic acids A [1], B [2], and C [3], were isolated from the fruiting bodies of Antrodia cinnamomea by bioassay-guided fractionation. The structures of these compounds were elucidated by chemical reactions and detailed analysis of their 1H- and 13C-nmr spectra. Biological studies revealed that 1 exhibited cytotoxic activity against P-388 murine leukemia cells and 2 showed weak anticholinergic and antiserotonergic activities.

Mice sarcoma S180 cell membranes, [r-32 P]ATP and plant polysaccharides (PPS, ASPS, TF and lentinan) were incubated together for 5 min at 30 degrees C. Phosphatidylinositides were isolated and subjected to scintillation fluid counting. The results showed that PPS and ASPS, especially the latter, could inhibit the phosphatidylinositol turnover obviously, while TF and lentinan had no such effect.

A number of common edible fungi were analysed for their contents of Pg, Cd, Cr, Ni, Co, Zn, Cu, Mn and Se. The results indicate that the Cd level in the species Rozites caperata can be as high as several mg/kg fresh weight. In Lycoperdon perlatum and Macrolepiota procera the Pb level was found to be up to 2.5 mg/kg. The Ni level was at or below 0.1 mg/kg in all species, with the exception of Albatrellus ovinus, which had a mean concentration of 0.72 mg/kg. The level of Cr was highest in M. procera with a mean of 0.091 mg/kg. In a single sample of the species Agaricus augustus, the concentration of Co was found to be exceptionally high at 0.28 mg/kg. Zn and Cu levels were highest in L. perlatum and M. procera. Mn levels were fairly similar in the different species. The results of several surveys show that there can be large differences in the concentrations of the same metal (e.g. Pb) in the same species. The Se level was found to be very high in the species Boletus edulis, as has been shown by others. In the other species analysed the level of Se was low.

World wide about 200,000 tons of shii-take mushrooms (Lentinus edodes) are produced per year. Different positive biological effects are known (anticarcinogenic, anticholesterol, immunostimulating effects), but the mineral contents and amino acid composition of caps and stipes are still little investigated. The concentrations of minerals are in general lower than those in the cultivated white mushroom (Agaricus bisporus) and in the oyster mushroom (Pleurotus ostreatus). The greatest differences are found in the concentrations of potassium, phosphorus, calcium, copper, strontium, manganese and zinc. The concentrations are higher in caps than in stipes. The total amino acid content is 15.24% in caps and 11.35% in stipes (dry matter) and thus in general half of the concentration of cultivated champignon. The amounts of Phe, Gly, His, Arg, Ile and Met are relative higher than in Agaricus fruit bodies.

In the fruiting bodies of the fungus Pleurotus ostreatus, also called the oyster mushroom, we found a competitive inhibitor of 3-hydroxy-3-methylglutaryl-coenzyme A reductase-lovastatin. The appearance of the inhibitor during the development of fruiting bodies was followed and lovastatin determined in the vegetative mycelium, in the primordia, as well as in different parts of sporocarps of different sizes. Less lovastatin was found in stripes as compared to pili or in mature stages in the lamellae and basidiospores.

The complete amino acid sequences of two isomeric endogenous inhibitors, IA-1 and IA-2, both of which specifically inhibit an intracellular serine proteinase (proteinase A) purified from the fruiting bodies of a higher basidiomycete, Pleurotus ostreatus, were determined. Both inhibitors are acidic polypeptides with respective molecular masses of 8307 and 8244 Da, as determined by plasma desorption mass spectral analyses, and their N-terminal serine residue is blocked by acetylation. The fragments generated from the inhibitors by proteolytic and chemical cleavages were subjected to amino acid composition, sequence, and mass spectral analyses. The sequence and molecular mass information for the peptides established that the inhibitors both consisted of 76 amino acid residues and differed from each other in that aspartic acid and glutamic acid residues at residues 12 and 15 of IA-1 were replaced by glycine and aspartic acid in IA-2, respectively. The molecular masses of IA-1 and IA-2 were calculated to be 8309 and 8237, based on the sequence data. The action of carboxypeptidase Y on IA-1 resulted in a complete loss of the inhibitory activity along with successive release of glutamine and threonine from the C-terminus. Cyanogen bromide cleavage of Met38-Pro39 and Met41-Lys42 in IA-1 and hydroxylamine degradation of IA-2 completely abolished their inhibitory activity. These results suggest that the whole molecules of both inhibitors are essential to their inhibitory activities. Their structural resemblance to propeptides of subtilisin family proteinases revealed their mechanism of action.

The technical feasibility of using agricultural wastes (mango and date industry wastes) as a substrate for the cultivation of Pleurotus ostreatus NRRL-0366 is evaluated. When comparing the biological efficiency of mushroom production, the highest yield of fruiting bodies was obtained using a mixture of date waste and rice straw at a ratio (1:1) (11.96%), followed by a mixture 3:1 (11.16%). The lowest one was the mixture 2:1 (9.19%). Fungus Pleurotus ostreatus NRRL-0366 can also be cultivated on mango waste supplemented with rice straw at a different ratio. The best one was the 1:1 mixture (10.18%), whereas the lowest was a mixture 3:1 (6.4%). Comparing the results obtained favored the use of date waste as a substrate for growing Pleurotus ostreatus NRRL-0366. Spawn was cultured on three different substrates as follows: Date waste alone (I); 1:1 (by wt) date waste and rice straw (II); 1:1:1 date waste, rice straw, and corncobs (III). Final dry weight and composition of the fruiting bodies are tabulated for the three sets of conditions. Date waste and rice straw mixture (II) is a good source of nonstarchy carbohydrate (67%) and protein (27.44%) containing amounts of essential amino acids, especially lysine and low RNA (3.81%). Elemental analysis were studied in the fruit bodies of the three media.

The fruit body of Grifola frondosa (maitake), Basidiomycetes was confirmed to contain substances with anti-diabetic activity. When 1 g/d of powdered fruit body of maitake was given orally to a genetically diabetic mouse (KK-Ay), blood glucose reduction was observed, in contrast to the control group in which the blood glucose increased with ageing. Moreover, levels of insulin and triglyceride in plasma demonstrated a change similar to blood glucose with feeding of maitake. Ether-ethanol-soluble (ES) and hot water-soluble (WS) fractions were prepared from the fruit body and their hypoglycemic activity was examined. Blood glucose-lowering activity was found when ES-fraction or WS-50% ethanol float (X) fraction was administered orally, but other WS-fractions were inactive. These results suggest that the anti-diabetic activity was present not only in the ES-fraction consisting of lipid but also in the X-fraction of peptidoglycan (sugar:protein = 65:35).

The brown-rot fungus Gloeophyllum odoratum was collected from spruce stumps in southern Finland. The volatiles in the fruiting body and fungal cultures grown in malt extract and liquid medium were investigated. Chitin, chitosan and D-(+)-glucosamine at a concentration of 450 mgl-1 medium were used as elicitors. Chitosan completely inhibited growth in the solid medium. The main volatile(s) according to GC and GC-MS analysis were either linalool, citronellol, geraniol and methyl p-methoxyphenylacetate or drimenol depending on the culture type and elicitor. The composition of volatiles in the natural fungus differed slightly from that of the cultivated fungus since the major compound was methyl p-methoxyphenylacetate. The volatile oils were toxic to larvae of the brine shrimp, Artemia salina, indicating that they may possess insecticidal and cytotoxic activity.