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Inhibition by the mushroom toxins alpha-amanitin and phalloidin of hepatopoietin-induced 3H-thymidine incorporation into rat liver DNA and of plasma protein production in hepatocyte cultures.

In primary cultures of rat hepatocytes the induction of 3H-thymidine uptake into DNA of liver cells by the liver cell proliferation factor hepatopoietin demonstrates that this factor is active not only in vivo but also in vitro. Addition of the mushroom toxins alpha-amanitin or phalloidin to liver cell culture decreased the uptake of 3H-thymidine into hepatocytes (in the absence or presence of hepatopoietin) as well as the attachment of the hepatocyte cultures. Mushroom toxins also inhibited the production of plasma proteins in hepatocyte cultures. The inhibition, observed at toxin concentrations from 10(-5) to 10(-7) M, was dose-dependent. At low concentrations of phalloidin the inhibition appears to be selective for certain proteins.

Amanitins↗

Some properties of flammutoxin from the edible mushroom Flammulina velutipes.

A cytolytic toxin from the basidiocarps of the edible mushroom Flammulina velutipes was purified to homogeneity. The lysin, flammutoxin, is a single polypeptide chain of Mr 32,000 and pK about 5.4. It contains unusually large amounts of tryptophan, serine and glycine, and few or none of the sulfur-containing amino acids. Erythrocytes of rat, rabbit, guinea pig, man, mouse, cat and dog were sensitive to lysis, in that order, whereas erythrocytes of sheep, ox, goat, swine and horse were largely or completely resistant to lysis. The toxin appears not to be a phospholipase and it was not inhibitable by any of a variety of lipids. Hemolysis probably involves alteration of the erythrocyte membrane, with formation of submicroscopic ion channels, and it appears to be of the osmotic type. In some respects flammutoxin resembles phallolysin, a cytolytic toxin obtained from the mushroom Amanita phalloides.

Amino Acids↗

Data on arsenic and cadmium contents of some common mushrooms.

The arsenic and cadmium contents of 88 samples of mushrooms were determined. The majority of samples have a very low (practically zero) arsenic level; however, significant accumulations were found in the Agaricus species and in Macrolepiota rhacodes (which is related to the Agaricaceae family) and in three Tricholomataceae species (Flammulina velutipes, Lepista nebularis and Clitocybe, inversa). The average cadmium content of all samples was 4.91 ppm (0.28-86 ppm) on a dry weight basis. The highest concentration (34.9 ppm) was found to be characteristic of genus Agaricus. The accumulation potential of genus Russula is lower, and it appears that this content is more characteristic in three sections (Ingratea, Heterophyllae and Xerampelinae), whereas the others have a low (normal) cadmium level. These data confirm that the accumulation ability is genetically coded, thus, only certain taxonomical groups of fungi play a toxicological role. Our data offer new information about the concentration of two toxic elements of particular mushroom species as well as in other taxonomic groups. These data are of great importance in view of toxicology, food chemistry and, partly, environmental protection.

Agaricus↗

Concentrations of lead, cadmium, mercury and copper in mushrooms in the vicinity of a lead smelter.

The concentrations of four heavy metals in 149 samples of mushroom fruiting bodies, representing 11 species, mainly all edible, were determined by atomic absorption spectroscopy. The mushrooms were collected up to a distance of 6 km from a lead smelter in central Bohemia (Czechoslovakia) in operation since 1786. Lead was accumulated extensively by Lepiota rhacodes and Lepista nuda. Among other species, significant accumulation was found up to a distance of 1 km from the source. Concentrations of greater than 100 mg kg-1 dry matter were often determined. The safe limit of 5 mg kg-1 dry matter was exceeded in most samples collected at distances of up to 6 km from the source. Concentrations of cadmium in the polluted area were generally significantly higher than in other parts of Bohemia. Cadmium was extensively accumulated by the toxic Amanita muscaria, but also by the edible Boletus edulis and Amanita rubescens, with mean values 28.6, 15.2 and 12.3 mg Cd kg-1 dry matter, respectively. The Czechoslovakian statutory limit is 0.5 mg Cd kg-1 dry matter. Statistically significant linear correlations between lead and cadmium concentrations were found only for Boletus edulis and Paxillus involutus. Mercury was accumulated by Lepista nuda and Lepiota rhacodes; mean values of 11.9 and 6.5 mg Hg kg-1 dry matter, respectively, were found. Concentrations of mercury in most species from the study area were higher than in those from other parts of Bohemia. Lepiota rhacodes and Lepista nuda also accumulated copper extensively with mean values of 280 and 193 mg Cu kg-1 dry matter, respectively.

Basidiomycota↗

Stability of necatorin, a highly mutagenic compound from Lactarius necator mushroom.

The mutagenic compound necatorin [7-hydroxycoumaro(5,6-c)cinnoline] was present in fresh Lactarius necator mushrooms at concentrations ranging from 3 to 20 mg/kg. Blanching decreased the concentration of necatorin in the mushrooms to about 25% of the original value. Pure necatorin was shown to be susceptible to decomposition by light, especially at high pH. The destruction of pure necatorin by boiling was most effective at pH 5.0, whereas at the other pH values studied (0.5 and 13.5) necatorin was relatively stable during boiling.

Agaricales↗

A study of the mutagenicity of some commercially canned Spanish mushrooms.

The mutagenicity of two wild (Lactarius deliciosus and Boletus luteus) and two cultivated (Agaricus bisporus and Pleurotus ostreatus) mushrooms, preserved by canning and widely consumed in Spain, was studied in the Ames Salmonella/microsome test and in the CHO/HPRT assay system using mammalian cells. The mushroom extracts did not show mutagenicity in the microbial Ames test nor in the mammalian CHO-K1 cells, and this response was not modified by the presence of S-9 mix in the assay mixtures. Only the extracts from P. ostreatus showed a weak mutagenic activity in the CHO/HPRT assay in the presence of a metabolic activation system (S-9).

Animals↗

Psychoactive mushroom use in Koh Samui and Koh Pha-Ngan, Thailand.

This paper presents the results of recent ethnomycological exploration in southern Thailand. Field observations, interviews and collection of fungi specimens were carried out primarily on two islands, Koh Samui and Koh Pha-ngan, situated in the western region of the Gulf of Siam. Some fieldwork was also conducted in the northern Thai province of Chiang Dao and in the southern Thai province of Surat Thani. During five separate excursions (1989-90), observations were made of occurrence, harvesting, use, and marketing of psychoactive fungi by local Thai natives (males and females, adults and children), foreign tourists, and German immigrants. The first records of psychoactive Psilocybe subcubensis and Copelandia dung fungi in Thailand are presented in this paper. These fungi exhibited intense bluing reactions when handled, indicating the presence of psilocybin and/or psilocin. Seven collections of Psilocybe cubensis (Earle) Singer and/or Psilocybe subcubensis Guzman and four collections of Copelandia sp. were harvested and sun-dried for herbarium deposit. These fungi are cultivated or occur spontaneously, often appearing in the decomposed manure of domesticated water buffalo (Bubalus bubalis) and at least three different species of cattle (Bos indicus, B. guarus, and B. sundaicus). The psychoactive fungi are cultivated in clandestine plots, both indoors and outdoors, in the uplands and villages on Koh Samui by both Thai natives and some foreigners. The sale of psychoactive fungi directly to tourists and to resort restaurants for use in edible food items such as omelettes and soups is discussed in detail. The preparation and sale of mushroom omelettes adulterated with artificial hallucinogens in some restaurants is also discussed. In addition, the marketing of items such as hand painted T-shirts, post cards, and posters bearing mushroom related motifs in Thailand is described.

Animals↗

Soma siddhas and alchemical enlightenment: psychedelic mushrooms in Buddhist tradition.

In the legendary biographies of some Buddhist adepts from the 2nd- and 9th-centuries there are some clues which can be interpreted to reveal that the adepts were consuming psychedelic Amanita muscaria, 'fly agaric', mushrooms to achieve enlightenment. This secret ingredient in the alchemical elixir they used to attain 'realization' was, of course, unnamed, in keeping with their vows to maintain the secrecy of their practices. Its identity was concealed behind a set of symbols, some of which appeared in the Soma symbol system of the Rg Veda, some other symbols possibly passed down from a time of earlier shamanic use of the mushroom in the forests of Northern Eurasia, and some symbols that may be unique to these Buddhist legends. The congruity of these sets of symbols from Northern and Southern Asian traditions will be shown to be reflected in the Germanic tradition in some characteristics of the Oldest God, Odin.

Amanita↗

First electron spin resonance evidence for the production of semiquinone and oxygen free radicals from orellanine, a mushroom nephrotoxin.

Orellanine is the tetrahydroxylated and di-N-oxidized bipyridine toxin from several Cortinarius mushrooms. The mechanism responsible for its lethal nephrotoxicity was unknown until now. Our present ESR spectroscopic study of the redox properties of the toxin is an original contribution to the knowledge of its toxicity. It was achieved in particular by comparison of the properties of orellanine to that of other bipyridine compounds. After a one-electron oxidation (e.g., photochemical oxidation upon visible light), a radical form of orellanine is observed at physiological pH under aerobic or anaerobic conditions. This radical, identified as ortho-semiquinone anion radical, can also be generated by oxidation with biological oxidizing agents or enzymatic systems. Production of superoxide and hydroxyl radicals is shown by the spin-trapping method using DMPO as a spin trap. Bioreducing agents like GSH and cysteine involve in vitro the semiquinone radical and orellanine in a redox cycling process resulting in the production of glutathionyl and oxygen free radicals. This process leads in vitro to a large oxygen consumption and to a dramatic depletion of glutathione level. The formation of an apparently stable ortho-semiquinone anion radical and of reactive oxygen radical species is observed for the first time with a mushroom toxin. It is due to the catechol-like functions borne by the di-N-oxidized bipyridine structure of the toxin and may at least partly explain the toxicity of orellanine.

2,2'-Dipyridyl↗

Polysaccharide-peptide complexes from the cultured mycelia of the mushroom Coriolus versicolor and their culture medium activate mouse lymphocytes and macrophages.

The aim of this study was to investigate the effects of the mushroom Coriolus versicolor on cells of the immune system. The cultured mycelia of the mushroom Coriolus versicolor and their culture medium were separately extracted with boiling water. The resulting polysaccharopeptide preparations were designated intramycelial (IM) and extramycelial materials (EM), and were separated by gel filtration before determining their effects on lymphocytes and macrophages in vitro and in vivo. After gel filtration on Sepharose 6B, only a single peak with a molecular weight of 13-19 KDa was obtained. Gel filtration of IM and EM on Sephadex G-50 revealed the presence of a larger peak of 28 KDa (from IM) and 15 KDa (from EM) and a smaller peak of 3.5 KDa. IM, EM and their large molecular peaks enhanced the mitogenic response of T-cells from BALB/c mice in vitro. Splenocytes from C57BL/6 mice pre-treated by force-feeding with IM and EM demonstrated an augmented mitogenic response to Con A. The macrophages of C57BL/6 mice that had been pre-treated with IM or EM showed an enhanced production of nitrite ions. The results indicate that both mouse lymphocytes and macrophages were activated by preparations of polysaccharopeptide from cultured mycelia and culture medium of C. versicolor. However, no direct cytotoxic activity against fibroblasts, hepatoma cells and choriocarcinoma cells could be demonstrated.

Animals↗

Microkeratome-assisted mushroom keratoplasty with minimal endothelial replacement.

PURPOSE: To report the outcome of a new keratoplasty technique aimed at treating full-thickness opacities with minimal removal of recipient endothelium. DESIGN: Interventional case report. METHODS: A deep central scar was removed using microkeratome-assisted mushroom-shaped keratoplasty, consisting of a large anterior stromal lamella (9.0-mm in diameter) and a small posterior button (5.0-mm in diameter) including deep stroma and endothelium. Complete suture removal was performed 3 months after surgery. Visual acuity, refraction, and computerized corneal topography were evaluated pre- and postoperatively. RESULTS: Best spectacle-corrected visual acuity improved from 20/60 to 20/20 at 6 months postoperatively. Postoperative refraction was -2.50-1.00 x 20 degrees. CONCLUSIONS: Microkeratome-assisted mushroom keratoplasty may offer visual and refractive advantages over conventional keratoplasty surgery. Because most of the recipient endothelium is preserved and may spread onto the posterior surface of the small donor button, graft decompensation secondary to immunologic rejection may be avoided with this technique.

Adult↗

New covered mushroom-shaped metallic stent for managing anastomotic leak after esophagogastrostomy with a wide gastric tube.

PURPOSE: To evaluate the clinical results of a new covered mushroom-shaped metallic stent for managing gastroesophageal anastomotic leak after esophagogastrostomy with a wide gastric tube and gastric pull-up. DESCRIPTION: The stent is a self-expanding prosthesis especially designed for occlusion of the esophago-thoracic anastomotic leaks after esophagogastrostomy. From January 2002 to September 2005, 8 patients with gastroesophageal anastomotic leaks were treated with stents. Information about the technical success of stent placement, definitive closure of leak, stent removal, and complications were obtained. EVALUATION: Stent placement was technically successful in all patients, without immediate procedural complications. The stent completely sealed off the fistula in all patients, and 30-day mortality was nil. Follow-ups at 7 to approximately 30 months showed that all leaks were healed without stent-related complications, and the stents were removed after approximately 18 to 48 days. Two patients died, and the remaining 6 patients were alive with no evidence of disease at the time of this report. CONCLUSIONS: The use of a new covered mushroom-shaped metallic stent proved expedient, safe, and effective in the treatment of gastroesophageal anastomotic leaks.

Adult↗

Purification of a novel ribonuclease from dried fruiting bodies of the edible wild mushroom Thelephora ganbajun.

A ribonuclease, with a molecular mass of 30 kDa and a potent inhibitory activity toward HIV-1 reverse transcriptase (IC50=300 nM), was isolated from dried fruiting bodies of the edible wild mushroom Thelephora ganbajun. The ribonuclease exhibited a unique polyhomoribonucleotide specificity, with the highest activity toward poly(U), about 50% and 25% as much activity toward poly(A) and poly(C), respectively, and minimal activity toward poly(G). Unlike other mushroom RNases, the ribonuclease was adsorbed on DEAE-cellulose and Q-Sepharose, and unadsorbed on CM-cellulose. A temperature of 40 degrees C and a pH of 6-7 were required for maximal activity of the enzyme. The enzyme was characterized by an N-terminal sequence without any homology to known proteins.

Agaricales↗

Inhibitory effects of cis- and trans-isomers of 3,5-dihydroxystilbene on the activity of mushroom tyrosinase.

The effects of cis- and trans-isomers of 3,5-dihydroxystilbene on the activity of mushroom tyrosinase have been studied. The results show that both cis- and trans-isomers of 3,5-dihydroxystilbene can inhibit the diphenolase activity of the enzyme and the inhibition type was reversible. The IC(50) values were estimated as 0.405+/-0.013 and 0.705+/-0.017 mM, respectively. Kinetic analysis showed that the inhibition of cis-3,5-dihydroxystilbene and trans-3,5-dihydroxystilbene on the diphenolase activity of the enzyme belonged to competitive type, and the inhibition constants (K(I)) were determined to be 0.232+/-0.015 and 0.395+/-0.020 mM, respectively. In this investigation, the inhibitory effects of cis-3,5-dihydroxystilbene and trans-3,5-dihydroxystilbene on the diphenolase activity of mushroom tyrosinase were compared. The inhibitory capacity of cis-isomer was stronger than that of corresponding trans-isomer. Nevertheless, the trans-3,5-dihydroxystilbene was used more frequently than its corresponding cis-form compound. This research may offer some references for designing and synthesizing some novel and effective tyrosinase inhibitors. Furthermore, it may improve the use of stilbenes on the field of food preservation and depigmentation.

Agaricales↗

Irreversibly inhibitory kinetics of 3,5-dihydroxyphenyl decanoate on mushroom (Agaricus bisporus) tyrosinase.

3,5-Dihydroxyphenyl decanoate (DPD) is found to inhibit the diphenolase activity of tyrosinase from mushroom (Agaricus bisporus). The effects of DPD on the diphenolase activity of mushroom tyrosinase have been studied. The results show that the enzyme activity decreases very slowly with an increase in DPD concentrations at lower concentrations of DPD (between 5 and 60 microM). But at higher concentrations of DPD, DPD can strongly inhibit the diphenolase activity of the enzyme and the inhibition is irreversible. The IC50 value was estimated to be 96.5 microM. The inhibition mechanism of DPD has been investigated and the results show that DPD can bind to the free enzyme molecule and enzyme-substrate complex and lose the enzyme activity completely. The inhibition kinetics has been studied in detail by using the kinetic method of the substrate reaction described by Tsou. The microscopic rate constants of the enzyme inhibited by DPD at higher concentrations have been determined.

Agaricus↗

Protective effect of an extract of the oyster mushroom, Pleurotus ostreatus, on antioxidants of major organs of aged rats.

This study was undertaken to investigate the putative antioxidant activity of the oyster mushroom, Pleurotus ostreatus, on lipid peroxidation and antioxidant status of major organs of aged (24 month old) rats when compared to young (4 month old) rats. Elevated levels of malondialdehyde (MDA) and significantly lowered levels of reduced glutathione (GSH) and of vitamins C and E were observed in the liver, kidneys, heart and brain of aged rats, when compared to values in young rats. Quantitative analysis of the activities of the antioxidant enzymes catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (Gpx) revealed significantly lower values in the liver, kidneys, heart and brain of aged rats. An analysis of isozyme pattern of these enzymes in aged rats also revealed variations in relative concentration, presumably due to oxidative stress. Administration of the extract of P. ostreatus to aged rats resulted in elevated levels of reduced glutathione and vitamins C and E and in increased activities of CAT, SOD and Gpx so that the values in most of these parameters did not differ significantly from those in young rats. In addition, the level of MDA was lowered on administration of mushroom extract to aged rats. These results suggest that treatment with an extract of P. ostreatus can improve the antioxidant status during ageing, therein minimizing the occurrence of age-associated disorders associated with involvement of free radicals.

Aging↗

Effects of extracts from Brazilian sun-mushroom (Agaricus blazei) on the NK activity and lymphoproliferative responsiveness of Ehrlich tumor-bearing mice.

Agaricus blazei Murrill, is an edible and medicinal mushroom which is popularly consumed due to its antitumoral properties. The immunomodulatory effects of methanol (METH), dichloromethane (DM) and n-hexane (HEX) extracts of this mushroom were evaluated in Ehrlich tumor-bearing mice. Subcutaneous inoculation of Ehrlich tumor cells inhibited the natural killer (NK) activity of spleen cells (specific lysis=6.18+/-2.56%) compared with normal mice (17.59+/-7.77%). Treatment of tumor-bearing mice with the extracts for 10 days restored the natural killer activity against Yac-1 target cells and the best results were observed by treatment with the HEX extract (21.48+/-5.26%). Treatment of the animals with the HEX extract for 10 days was also able to stimulate the mitogen-induced lymphoproliferative activity of spleen cells. Thirty days after the treatment, all groups presented low proliferative activity. Specific antibody production was observed to be higher in the groups treated with the DM or METH extract 30 days after the treatment. Analysis of the 3 extracts by gas chromatography mass spectrum (GCMS) and magnetic nuclear resonance (MNR) showed that the HEX extract contains mainly sugar and fatty acids and that the METH extract also contains sugar and possibly amino acids.

Agaricus↗

Cloning and developmental expression of a metzincin family metalloprotease cDNA from oyster mushroom Pleurotus ostreatus.

A cDNA clone, PoMTP, encoding a putative metzincin family metalloprotease was isolated from the expressed sequence tags of a basidiomycete Pleurotus ostreatus. The 5'-end sequence of PoMTP was determined by the 5'-RACE method. Full-length cDNA sequence (1140 bp) of PoMTP contained a 870 bp open reading frame encoding a protein product of 290 amino acids in addition to a 99 bp of 5'-untranslated sequence and a 171 bp of 3'-untranslated sequence with a poly(A) tail. The deduced amino-acid sequences of PoMTP contained an extensive zinc-binding consensus sequence and a so-called Met-turn sequence which are typical for the metzincin family of metalloproteases, indicating that the PoMTP protein belongs to the metzincin metalloproteases. Four cysteine residues were also observed in the zinc-binding region of PoMTP amino-acid sequence, which are known to be important for the structure and the function of some subfamilies of the metzincins. Comparison of the PoMTP in sequence database showed no significant homology with functionally known metalloproteases of Armillaria mellea, Grifola frondosa, Lentinula edodes, Pleurotus ostreatus, Schizophyllum commune and Tricholoma saponaceum in mushroom. Northern blot and qunatitative RT-PCR analyses indicated the PoMTP mRNA to be abundant at primordial and fruit body stages, but scarce at the mycelial stage, suggesting that the PoMTP metalloprotease plays an important role in mushroom fruiting.

Amino Acid Sequence↗