Amanitins: The Most Poisonous Molecules of the Fungal World.

Among the toxic metabolites of the fungal world, those that, due to their strong biological effect, can seriously (even fatally) damage the life processes of humans (and certain groups of animals) stand out. Amatoxin-containing mushrooms and the poisonings caused by them stand out from the higher fungi, the mushrooms. There are already historical data and records about such poisonings, but scientific research on the responsible molecules began in the middle of the last century. The goals of this review work are as follows: presentation of the cosmopolitan mushroom species that produce amanitins (which are known from certain genera of four mushroom families), an overview of the chemical structure and specific properties of amanitins, a summary of the analytical methods applicable to them, a presentation of the "medical history" of poisonings, and a summary of the therapeutic methods used so far. The main responsible molecules (the amanitins) are bicyclic octapeptides, whose structure is characterized by an outer loop and an inner loop (bridge). It follows from the unusual properties of amanitins, especially their extreme stability (against heat, the acidic pH of the medium, and their resistance to human, and animal, digestive enzymes), that they are absorbed almost without hindrance and quickly transported to our vital organs. Adding to the problems is that accidental consumption causes no noticeable symptoms for a few hours (or even 24-36 h) after consumption, but the toxins already damage the metabolism of the target organs and the synthesis of nucleic acid and proteins. The biochemical catastrophe of the cells causes irreversible structural changes, which lead to necrotic damage (in the liver and kidneys) and death. The scientific topicality of the review is due to the recent publication of new data on the probable antidote molecule (ICR: indocyanine green) against amanitins. Further research can provide a new foundation for the therapeutic treatment of poisonings, and the toxicological situation, which currently still poses a deadly threat, could even be tamed into a controllable problem. We also draw attention to the review conclusions, as well as the mycological and social tasks related to amanitin poisonings (prevention of poisonings).

The Mushroom Glucans: Molecules of High Biological and Medicinal Importance.

Carbohydrates, including polysaccharide macromolecules, are the main constituents of the fungal cell wall. Among these, the homo- or heteropolymeric glucan molecules are decisive, as they not only protect fungal cells but also have broad, positive biological effects on the animal and human bodies. In addition to the beneficial nutritional properties of mushrooms (mineral elements, favorable proteins, low fat and energy content, pleasant aroma, and flavor), they have a high glucan content. Folk medicine (especially in the Far East) used medicinal mushrooms based on previous experience. At the end of the 19th century, but mainly since the middle of the 20th century, progressively more scientific information has been published. Glucans from mushrooms are polysaccharides that contain sugar chains, sometimes of only one kind (glucose), sometimes having several monosaccharide units, and they have two (α and ß) anomeric forms (isomers). Their molecular weights range from 104 to 105 Da, and rarely 106 Da. X-ray diffraction studies were the first to determine the triple helix configuration of some glucans. It seems that the existence and integrity of the triple helix structure are criteria for their biological effects. Different glucans can be isolated from different mushroom species, and several glucan fractions can be obtained. The biosynthesis of glucans takes place in the cytoplasm, the processes of initiation and then chain extension take place with the help of the glucan synthase enzyme complex (EC 2.4.1.34), and the sugar units are provided by sugar donor UDPG molecules. The two methods used today for glucan determination are the enzymatic and Congo red methods. True comparisons can only be made using the same method. Congo red dye reacts with the tertiary triple helix structure, and the resulting glucan content better reflects the biological value of glucan molecules. The biological effect of ß-glucan molecules is proportional to the integrity of the tertiary structure. The glucan contents of the stipe exceed the values of the caps. The glucan levels of individual fungal taxa (including varieties) differ quantitatively and qualitatively. This review presents in more detail the glucans of lentinan (from Lentinula edodes), pleuran (from Pleurotus ostreatus), grifolan (from Grifola frondose), schizophyllan (from Schizophyllum commune), and krestin (from Trametes versicolor), along with their main biological effects.

The Norsesquiterpene Glycoside Ptaquiloside as a Poisonous, Carcinogenic Component of Certain Ferns.

Previous studies related to the ptaquiloside molecule, a carcinogenic secondary metabolite known from the world of ferns, are summarised. Ptaquiloside (PTA) belongs to the group of norsesquiterpenes of the illudane type. The name illudane refers to the fungal taxa from which the first representatives of the molecular group were identified. Ptaquiloside occurs mainly in Pteridium fern species, although it is also known in other fern taxa. The species of the genus Pteridium are common, frequent invasive species on all continents, and PTA is formed in smaller or larger amounts in all organs of the affected species. The effects of PTA and of their derivatives on animals and humans are of great toxicological significance. Its basic chemical property is that the molecule can be transformed. First, with the loss of sugar moiety, ptaquilosine is formed, and then, under certain conditions, a dienone derivative (pteridienone) may arise. The latter can alkylate (through its cyclopropane groups) certain molecules, including DNA, in animal or human organisms. In this case, DNA adducts are formed, which can later have a carcinogenic effect through point mutations. The scope of the PTA is interdisciplinary in nature since, for example, molecules from plant biomass can enter the body of animals or humans in several ways (directly and indirectly). Due to its physico-chemical properties (excellent water solubility), PTA can get from the plant into the soil and then into different water layers. PTA molecules that enter the soil, but mainly water, undergo degradation (hydrolytic) processes, so it is very important to clarify the toxicological conditions of a given ecosystem and to estimate the possible risks caused by PTA. The toxicoses and diseases of the animal world (mainly for ruminant farm animals) caused by PTA are briefly described. The intake of PTA-containing plants as a feed source causes not only various syndromes but can also enter the milk (and meat) of animals. In connection with the toxicological safety of the food chain, it is important to investigate the transport of carcinogenic PTA metabolites between organisms in a reassuring manner and in detail. This is a global, interdisciplinary task. The present review aims to contribute to this.

Isolation and structural elucidation of a novel brunnein-type antioxidant ß-carboline alkaloid from Cyclocybe cylindracea.

Effect-directed isolation of free radical scavengers from the methanol extract of the freeze-dried fruiting bodies of the cultivated basidiomycetous mushroom, black poplar (Cyclocybe cylindracea), yielded a ß-carboline alkaloid. Its structure was determined based on ESI-TOF-MS/MS, NMR and circular dichroism spectra by comparison with published data. The compound, identified as the C1-S diastereomer of brunnein B, exhibited explicit radical scavenging activity (EC50 = 119.1 ±â€¯1.2 µg/mL). The quantity of the active component was determined with HPLC-MS in the fruiting body (36.2 ±â€¯2.8 ng/g DW, dry weight) and its different tissues such as peel (94.7 ±â€¯1.9 ng/g DW), inner cap (90.5 ±â€¯1.3 ng/g DW), gills (71.5 ±â€¯0.6 ng/g DW), and stipe (162.2 ±â€¯1.7 ng/g DW). It is a ß-carboline alkaloid that was not reported previously.

Nitrate content in a collection of higher mushrooms.

BACKGROUND: Data of mushroom nitrate content from scientific studies is limited. There have been two such recent investigations (mainly regarding certain cultivated species). To obtain comparable analytical data, we analyzed 134 samples of 54 taxa gathered and prepared by our department. RESULTS: The mushroom species were evaluated according to their nutritional types: saprotrophic, mycorrhizal and wood-decaying groups. Low and relatively invariable contents were found in the mycorrhizal (216.5 mg kg(-1) dry matter (DM) and wood-decaying groups (228.6 mg kg(-1) DM), but in the saprotrophic group we observed a wide variability (151.4-12 715 mg kg(-1) DM). CONCLUSION: A considerable nitrate content was found in samples of seven 'accumulator' species (Clitocybe nebularis, C. odora, Lepista nuda, L. personata, L. irina, Macrolepiota rachodes and M. procera). The toxicological relevance of daily uptake of acceptable nitrate content via mushrooms only is not presumable, but the 'accumulator' saprotrophic species can be 'contributors' to our nitrate intake in foods.

Mineral composition of different strains of edible medicinal mushroom Agaricus subrufescens Peck.

Agaricus subrufescens Peck is a well-known Basidiomycota fungus (Royal Sun Agaricus), with rising demand in consumption and production worldwide. This particular mushroom with high medical value has been used successfully in cancer therapy and in the treatment of some bacterial and viral diseases. Four strains of A. subrufescens (Si2.2, 853, 1105, and 2603) were cultivated, and 22 mineral elements of basidiomes (fruit bodies) were analyzed (caps and stipes separately). The data obtained about the mineral compositions were compared to the "reference" Agaricus bisporus (white button mushroom) and to the average of wild growing Agaricus species. The mineral composition of A. subrufescens strains can be characterized by the following: (1) high levels of valuable macroelements, i.e., potassium (28-30,000 mg/kg of dry matter), phosphorus (7-11,000 mg/kg of dry matter), and calcium and magnesium (for both elements, 1,000-1,500 mg/kg of dry matter); (2) significantly higher level of copper (compared to A. bisporus, 70-150 mg/kg of dry matter) and zinc (140-250 mg/kg of dry matter); (3) low quantity of sodium (140-180 mg/kg of dry matter); (4) attention should paid to the detectable amount of cadmium (2-17 mg/kg of dry matter) in strain Si2.2; (5) low or undetectable concentrations of some other poisonous microelements like As, Cr, and V; and (6) the distribution of elements in caps and stipes is characteristic-the majority of beneficial elements have higher contents in caps than in stipes, but some other elements, such as Ca, Fe, and Na, appear in an inverse proportion. In conclusion, it can be said that the mineral composition of A. subrufescens is definitely positive, with the exception of the above-mentioned Cd level.

A biological hazard of our age: bracken fern [Pteridium aquilinum (L.) Kuhn]--a review.

Bracken fern (Pteridium aquilinum) is the fifth most distributed common weed species of the world. Its ecological distribution is very wide, and the plant can grow and spread successfully on many types of soil. The cover of P. aquilinum is--in some cases--remarkable (e.g., in the United Kingdom). Bracken fern contains different poisonous agents: some cyanogen glycosides, factors (agents) of antithiamine character (thermolabile thiaminase and thermostable other compounds) and factors of carcinogenic activity (first of all ptaquiloside). This paper summarises and reviews different toxicological problems and poisonings caused by bracken fern in ruminants (cattle, sheep) and in non-ruminant animals (horses, pigs, rats, mice, etc.). The carcinogenic properties of the norsesquiterpene-type ptaquiloside make bracken fern a potent, living hazard. Recent investigations have shown that ptaquiloside pollution of different soil layers is a distinct possibility. Ptaquiloside may leach from the soil into the drinking water base. This ecotoxicological aspect seems to be the most hazardous phenomenon in relation to P. aquilinum and ptaquiloside. The carcinogenic effect of ptaquiloside is based on its hydrolysis, which leads to the formation of a dienon intermediate. It can produce DNA adducts, which are responsible for inducing carcinoma.

Mineral composition of basidiomes of Amanita species.

Basidiomes of 43 samples of eight Amanita species were gathered from different habitats of Hungary. The mineral composition (22 elements) was analysed by the ICP method in three independent replications, and mineral compositions found as discussed and compared. The Amanita species analysed were very different in As-, Cd-, Cr-, Mo-, Mn-, Se- and mainly in V-content. Other elements (Al, B, Ba, Ca, Co, Cu, Fe, Mg, Na, Ni, Sr, Ti, Zn) occur in the basidiomes in balanced concentrations. The K and P contents have the lowest differences. Summarizing all the data (n = 43), the average mineral status of species of Amanita can be deduced. The lowest variability measured was for K and P, and the highest for chromium, nickel and vanadium. Specific, significant accumulation was found only for vanadium, due to the previously demonstrated occurrence of a binding molecule 'amavadine' in the basidiomes of A. muscaria. Remarkable Cd-levels were estimated in A. pantherina and A. muscaria (11.4 and 12.3 mg kg(-1) D.M., respectively). The higher contents of other elements (e.g. K, practically in all species; Se in A. strobiliformis) are analytical facts but, not accumulations. The mineral compositions of the ectomycorrhizal genus Amanita, of litter decomposing Agaricus and of wood decaying Trametes were compared. Some significant differences were found (AsAmanita < AsAgaricus; CdAmanita < CdAgaricus; CuAmanita < CuAgaricus; PAmanita KTrametes; PAmanita > PTrametes) but it seems that the mineral composition of the basidiomes is practically independent of the ectomycorrhizal habit. The specificities of the fungi-tree symbiotic interactions are known, and well documented (higher uptake and transport of certain elements first of all of P), however, the differences found in the mineral components are due to other factors (e.g. substrates, accumulating ability) and not to the mycorrhizal status.