Antioxidant activity of Phellinus igniarius fermentation mycelia contributions of different solvent extractions and their inhibitory effect on α-amylase.

Phellinus spp. have historically been used as traditional medicines to treat various diseases owing to their antioxidant, antitumor, and antidiabetic activities. Polysaccharides exhibit antidiabetic activity. In the present study, the polysaccharide contents of four Phellinus strains were compared. Phellinus igniarius QB72 possessed higher polysaccharide production, stronger 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, and α-amylase inhibitory activity. The three polysaccharides were sequentially extracted and partially purified from the fermentation mycelia using hot water, 1 % (NH4)2C2O4, and 1.25 M NaOH. Hot water extract polysaccharides exhibited higher DPPH radical scavenging and strong inhibitory activity against α-amylase with an IC50 value of 6.84 ± 0.37 mg/mL. The carbohydrate content of A1 (approximately 17457 Da) was approximately 88.28 %. The α-amylase inhibitory activity IC50 was decreased (3.178 ± 0.187 mg/mL) after DEAE water elution. P. igniarius QB72 hot-water extracts of partially purified polysaccharides have great potential as α-amylase inhibitors in food and medication-assisted additives.

Species Diversity of Lycoperdaceae (Agaricales) in Israel, with Some Insights into the Phylogenetic Structure of the Family.

The diversity of Lycoperdaceae in Israel was studied. Molecular phylogenetic relationships within the family, and genus Lycoperdon in particular, were inferred using original ITS rDNA sequences of 58 samples belonging to 25 species from Israel and six other countries, together with 66 sequences stored in the GenBank database. The current molecular phylogenetic study recovered the family Lycoperdaceae as a monophyletic group, which was supported in both maximum likelihood and Bayesian analyses. The clades corresponding to the genera Apioperdon, Bovista, Calvatia, Disciseda, and Lycoperdon were revealed. The taxonomic structure of the named genera was partially resolved. Within the genus Lycoperdon, some species received significant statistical support; however, their relationships, as well as the problem of the genus monophyly, mostly remained questionable. As a result of a thorough literature survey, extensive sample collection, and studies of the material stored in the Herbarium of the Institute of Evolution, University of Haifa (HAI, Israel), fifteen species representing five genera were found in the territory of Israel. Six species, namely Apioperdon pyriforme, Bovista aestivalis, Calvatia candida, Lycoperdon decipiens, L. niveum, and L. perlatum, are new additions to the diversity of Lycoperdaceae in Israel. Detailed macro- and micromorphological descriptions, ecology, geography, and critical notes, together with light microscopy photos and SEM micrographs, are provided. In-depth discussion on some taxonomically challenging species is presented.

Effect of the Lingzi or Reishi Medicinal Mushroom Ganoderma lucidum (Agaricomycetes) on Hyperglycemia and Dyslipidemia with Experimental Metabolic Syndrome.

The effect of Ganoderma lucidum hot water extract of submerged cultivated mycelium suspensia on carbohydrate metabolism and lipid profile during fructose-induced metabolic syndrome in rats was studied. The outbred white male Wistar rats, in which metabolic syndrome was induced by consuming a 10% fructose solution instead of drinking water for 42 days, were used. After the induction of metabolic syndrome, the mycelium of G. lucidum in the form of water suspension (a dose of 1 g/kg of the animal's body weight) was administered to animals per os for 7 and 14 days. Glucose concentration was determined using the glucose oxidase method. The content of glycosylated hemoglobin in erythrocytes was determined by the colorimetric method. The concentration of triglycerides, cholesterol, high-density lipoproteins, and low-density lipoproteins in blood plasma was determined by enzymatic methods. A significant decrease in the content of glycosylated hemoglobin was established in animals with metabolic syndrome against the background of administration of the studied suspension. Under the conditions of experimental metabolic syndrome, the administration of mycelium for 7 and 14 days led to a decrease in the concentration of triglycerides by 17.8 and 44.8%, cholesterol by 10.7 and 21.3%, low-density lipoproteins by 14.8 and 28.4%, and to an increase in high-density lipoproteins concentration by 11.9 and 21.5%, compared with metabolic syndrome. The obtained results demonstrate the corrective effect of the suspension of the G. lucidum powdered mycelium on carbohydrate and lipid metabolism, which was directly proportional to the duration of administration.

Antioxidant Activities of Hot Water Extracts from Mycelial Biomass of Different Combinations of Medicinal Agaricomycetes Mushrooms.

This study aimed to determine the free radical scavenging and antioxidant potential of hot water extracts prepared from different combinations and ratios of submerged cultivated mycelial biomass of medicinal mushrooms. Total phenolic compounds, flavonoid content, and antioxidant activity were evaluated for combined crude hot water extracts from medicinal higher Basidiomycetes mushrooms belonging to ten genera. The results demonstrate that almost all tested combinations were good sources of phenolic compounds and flavonoids, ranging between 16.42 and 18.83 gallic acid equivalents/g and 1.5 and 4.34 mg rutin equivalents/g, respectively. Moreover, free radical scavenging properties were evaluated with the DPPH and ABTS assays and metal chelating effects were investigated. All tested samples and/or extracts demonstrated significant free radical scavenging properties and antioxidant potential.

Striatal Isolated from Cyathus striatus Extracts Induces Apoptosis in Human Pancreatic Cancer Cells.

The aim of the present study was to identify the structure of active compounds in Cyathus stratus that previously demonstrated anti-pancreatic cancer activity. The active compounds were purified from a crude extract by a series of RP-18 preparative chromatography using homemade octadecyl silica gel column. HPLC injection of the crude extract revealed a chromatogram with three main peaks with retention times (RT) 15.6, 18.2, and 22.5 min. Each fraction that exhibited promising activity in vitro was further separated using various available chromatographic techniques. The purified compound with the ultimate anti-cancer activity appeared at RT of 15.8 in the HPLC chromatogram with more than 90% purity. The main peak at the mass spectra appeared at m/z = 446.2304 with the calculated molecular formula of C25H34O7. One- and two-dimensional NMR analyses indicated that the structure of the active molecule (peak 15.8 min in HPLC) was identified as striatal C. Exposure of human pancreatic cancer cells to purified striatal C resulted in induction of apoptosis. Further studies are needed in order to develop a method for the synthesis of striatal in order to use it in clinical studies for treatment of cancer.

Reappraisal of the Genus Exsudoporus (Boletaceae) Worldwide Based on Multi-Gene Phylogeny, Morphology and Biogeography, and Insights on Amoenoboletus.

The boletoid genera Butyriboletus and Exsudoporus have recently been suggested by some researchers to constitute a single genus, and Exsudoporus was merged into Butyriboletus as a later synonym. However, no convincing arguments have yet provided significant evidence for this congeneric placement. In this study, we analyze material from Exsudoporus species and closely related taxa to assess taxonomic and phylogenetic boundaries between these genera and to clarify species delimitation within Exsudoporus. Outcomes from a multilocus phylogenetic analysis (ITS, nrLSU, tef1-α and rpb2) clearly resolve Exsudoporus as a monophyletic, homogenous and independent genus that is sister to Butyriboletus. An accurate morphological description, comprehensive sampling, type studies, line drawings and a historical overview on the nomenclatural issues of the type species E. permagnificus are provided. Furthermore, this species is documented for the first time from Israel in association with Quercus calliprinos. The previously described North American species Exsudoporus frostii and E. floridanus are molecularly confirmed as representatives of Exsudoporus, and E. floridanus is epitypified. The eastern Asian species Leccinum rubrum is assigned here to Exsudoporus based on molecular evidence, and a new combination is proposed. Sequence data from the original material of the Japanese Boletus kermesinus were generated, and its conspecificity with L. rubrum is inferred as formerly presumed based on morphology. Four additional cryptic species from North and Central America previously misdetermined as either B. frostii or B. floridanus are phylogenetically placed but remain undescribed due to the paucity of available material. Boletus weberi (syn. B. pseudofrostii) and Xerocomus cf. mcrobbii cluster outside of Exsudoporus and are herein assigned to the recently described genus Amoenoboletus. Biogeographic distribution patterns are elucidated, and a dichotomous key to all known species of Exsudoporus worldwide is presented.

Chemical Composition and Antioxidant Properties of Different Combinations of Submerged Cultured Mycelia of Medicinal Mushrooms.

This research describes the investigation of submerged cultivated mycelial biomass and hot water extracts prepared from different combinations and ratios of medicinal mushroom (MM) dry powders, comprising various biologically active compounds/secondary metabolites. In particular, it was evaluated the proximate composition (moisture, ash, crude protein, fat, total carbohydrates, and total energy), γ-aminobutyric acid (GABA) and ergothioneine (ERG), amino acid content of mycelia of 16 higher Basidiomycetes MM species. The results obtained demonstrate that almost all tested combinations were found to be good sources of polysaccharides, with content varying in the ranges of 4.73 ± 1.33% and 58.46 ± 4.15%. Total protein contents varied in 1.97 ± 0.40% - 5.37 ± 0.40% range. ERG was detected in all tested samples, while GABA existed only in eight samples out of 15 and varied from 0.03 ± < 0.01 to 0.61 ± 0.03 mg/g, and from 0.16 ± 0.03 to 5.69 ± 0.41 mg/g respectively. Analyses of total phenolic and flavonoid contents demonstrate considerable content in all samples (15.53 ± 0.23 - 18.88 ± 0.34 mg gallic acid equivalents/g and 1.23 ± 0.04 - 4.34 ± 0.73 mg rutin equivalents/g respectively). In present research the complexity of samples/extracts were evaluated by multiple antioxidant assays to verify their antioxidant capacity. Determination of in vitro antioxidant activity was successfully carried out by several different methods such as 2,2-diphenyl-1-picrylhydrazyl scavenging activity, reducing power, chelating ability, hydroxyl radical scavenging activity, and 2,2'-azino-bis(3-ethylbenzothi-azoline-6-sulfonic acid scavenging activity. Therefore, all tested samples confirm the capable antioxidant activities of bioactive compounds extracted from MMs.

Cyathus striatus Extract Induces Apoptosis in Human Pancreatic Cancer Cells and Inhibits Xenograft Tumor Growth In Vivo.

Pancreatic cancer is a highly lethal disease with limited options for effective therapy and the lowest survival rate of all cancer forms. Therefore, a new, effective strategy for cancer treatment is in need. Previously, we found that a culture liquid extract of Cyathus striatus (CS) has a potent antitumor activity. In the present study, we aimed to investigate the effects of Cyathus striatus extract (CSE) on the growth of pancreatic cancer cells, both in vitro and in vivo. The proliferation assay (XTT), cell cycle analysis, Annexin/PI staining and TUNEL assay confirmed the inhibition of cell growth and induction of apoptosis by CSE. A Western blot analysis demonstrated the involvement of both the extrinsic and intrinsic apoptosis pathways. In addition, a RNAseq analysis revealed the involvement of the MAPK and P53 signaling pathways and pointed toward endoplasmic reticulum stress induced apoptosis. The anticancer activity of the CSE was also demonstrated in mice harboring pancreatic cancer cell line-derived tumor xenografts when CSE was given for 5 weeks by weekly IV injections. Our findings suggest that CSE could potentially be useful as a new strategy for treating pancreatic cancer.

Medicinal Mushrooms against Influenza Viruses.

This review provides results obtained by scientists from different countries on the antiviral activity of medicinal mushrooms against influenza viruses that can cause pandemics. Currently, the search for antiviral compounds is relevant in connection with the coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Medicinal mushrooms contain biologically active compounds (polysaccharides, proteins, terpenes, melanins, etc.) that exhibit an antiviral effect. The authors present the work carried out at the State Research Center of Virology and Biotechnology Vector in Russia, whose mission is to protect the population from biological threats. The research center possesses a collection of numerous pathogenic viruses, which allowed screening of water extracts, polysaccharides, and melanins from fruit bodies and fungal cultures. The results of investigations on different subtypes of influenza virus are presented, and special attention is paid to Inonotus obliquus (chaga mushroom). Compounds produced from this mushroom are characterized by the widest range of antiviral activity. Comparative data are presented on the antiviral activity of melanin from natural I. obliquus and submerged biomass of an effective strain isolated in culture against the pandemic strain of influenza virus A/California/07/09 (H1N1 pdm09).

Botanical drugs and supplements affecting the immune response in the time of COVID-19: Implications for research and clinical practice.

In times of health crisis, including the current COVID-19 pandemic, the potential benefit of botanical drugs and supplements emerges as a focus of attention, although controversial efficacy claims are rightly a concern. Phytotherapy has an established role in everyday self-care and health care, but, since botanical preparations contain many chemical constituents rather than single compounds, challenges arise in demonstrating efficacy and safety. However, there is ample traditional, empirical, and clinical evidence that botanicals can offer some protection and alleviation of disease symptoms as well as promoting general well-being. Newly emerging viral infections, specifically COVID-19, represent a unique challenge in their novelty and absence of established antiviral treatment or immunization. We discuss here the roles and limitations of phytotherapy in helping to prevent and address viral infections, especially regarding their effects on immune response. Botanicals with a documented immunomodulatory, immunostimulatory, and antiinflammatory effects include adaptogens, Boswellia spp., Curcuma longa, Echinacea spp., Glycyrrhiza spp., medicinal fungi, Pelargonium sidoides, salicylate-yielding herbs, and Sambucus spp. We further provide a clinical perspective on applications and safety of these herbs in prevention, onset, progression, and convalescence from respiratory viral infections.

Caterpillar Medicinal Mushroom, Cordyceps militaris (Ascomycetes), Mycelia Attenuates Doxorubicin-Induced Oxidative Stress and Upregulates Krebs Cycle Dehydrogenases Activity and ATP Level in Rat Brain.

Post-chemotherapy-induced cognitive dysfunction remains one of the challenges in cancer survivors. Cytokine-induced neurotoxicity manifests in subjects at any time after doxorubicin (DOX) chemotherapy. We examined the effect of bioactive Cordyceps militaris mycelia extract (CM) on the energy status, oxidative stress, and acetylcholinesterase activity in the brain of DOX treated rats. The CM (150 and 300 mg/kg b.w.) and DL-α lipoic acid (LA, 100 mg/kg b.w) were administered orally once daily for 5 days to male Wistar rats prior to the DOX administration (18 mg/kg as 3 doses of 6 mg/kg, i.p. b.w.) and continued for 6 more days. Cellular antioxidant status, Krebs cycle dehydrogenases, electron transport chain complexes (ETC) (I, III, and IV), adenosine triphosphate (ATP) level, advanced oxidation of protein products (AOPP), and acetylcholinesterase (AchE) activities were determined in the brain homogenate. The DOX alone treated group of animals showed significant decrease (p < 0.05) of brain antioxidant levels, Krebs cycle dehydrogenases activities, ETC complex activities, and decreased ATP level, while lipid peroxidation and AOPP levels were elevated. CM at 300 mg/kg b.w. or LA at 100 mg/kg b.w. elevated antioxidant status, Krebs cycle dehydrogenases, and complex activities and thus alleviated the toxicity. CM also inhibited the AchE activity in brain. The experimental results thus reveal that CM possessed excellent capacity to attenuate oxidative stress, upregulate respiratory chain complex activity and ATP levels, as well as inhibition of AchE activity.

Antioxidant Activities of Selected Medicinal Mushroom Submerged Cultivated Mycelia.

Medicinal mushrooms fruiting bodies have been used as food or medicine for years but cultured mycelium is faster to grow and costs less. This research studied the antioxidant activities of three species (five strains) of medicinal mushroom mycelia (Cordyceps militaris, Ganoderma tsugae I and II, Trametes versicolor I and II). Two-stage extractions were performed: first the sample was extracted with 70% ethanol, and then the residue was extracted with 95°C hot water. Both ethanolic and hot water extracts showed effective concentration (EC50) values of 0.29-4.22 mg/mL, indicating that these extracts were remarkably effective in antioxidant activities. The ethanolic extracts displayed more effective reducing power, scavenging, and chelating ability (EC50 0.33-2.37 mg/mL) than hot water extracts (EC50 0.58-4.22 mg/g). Besides, ethanolic extracts contained higher total phenol content (75.49-144.99 GAE mg/g) than the hot water extracts (22.77-58.68 GAE mg/g). Furthermore, the ethanolic extracts contained flavonoids but not the hot water extract. Overall, these mycelia were highly effective in the antioxidant activities and might be potent antioxidants.

Professor Shu-Ting Chang on His 90th Birthday.

This issue is dedicated to the 90th birthday of Professor Shu-Ting Chang, a prominent scientist in the field of mushroom biology, including cultivation and nutritional values of mushrooms, medicinal mushroom science, and environmental impact of mushrooms. Professor Shu-Ting Chang is also one of the initiators and has been an editor of our journal from its inception (in 1999) until now.

Professor Shu-Ting Chang, Cancer Mycotherapy and Le Chatelier Principle.

In the jubilee paper devoted to Professor Shu-Ting Chang, we aimed to outline the applicability of the Le Chatelier principle to various fields, especially to cancer biology and immunotherapy of malignant neoplasms. Cancer progression has been shown to be associated with a response to cell stress and survival agents. It was proclaimed that cancer immunotherapy, using fungal metabolites, should be carried out as a mode of compensation for imbalances that arise during the basic treatment. The challenges of immunotherapy in recent years have also received mention.

Profiles of Little-Known Medicinal Polypores: Haploporus odorus (Agaricomycetes).

The purpose of this study was to compile a comprehensive characterization of little-known polypores, which have recently been found to possess anticancer activity and thus can also be used in cancer target therapy. Haploporus odorus is a polypore of Holarctic distribution and has been found by harvesters working in taiga floodlands and broadleaf forests of the Northern Hemisphere. A substance known as haploporic acid A was determined in methylene chloride extract from the dried basidiomata of H. odorus. This substance can be used in cancer therapy; more details of its health benefits could be used in mycotherapy.

Cancer without Pharmacological Illusions and a Niche for Mycotherapy (Review).

In this review we outline a framework in which mycotherapy is effective in the field of oncology. We suppose that irreversible epigenomic changes in cancer cells and achieving their replicative immortality when cancer-specific targets are absent should take away any illusions about a fundamental possibility of pharmacological blockage of the cancer process once ontogenesis begins. At the same time, however, we believe that effects of both traditional and alternative medicines on cancer clonogenic units within a particular range can lead to prolonged remission; with this in mind, we carefully consider the various possibilities of mycotherapy in controlling cancer activity. The aforementioned range is limited to nondisseminated cancer processes and depends on the absence of large secondary tumor nodes and the inexhaustibility of immune depots after chemotherapeutic treatment. The main therapeutic effect of fungal bioactive complexes is dectin-1-mediated immunity, including the reprogramming of dendritic cells, which significantly increases the period during which tumors generate immune tolerance. An inhibitory effect of fungal bioactive complexes on some molecular mediators of proliferative signaling and components of proinflammatory (synergistic with cancer) immunity can be considered less significant. The effect of fungal bioactive complexes on vital (including overexpressed) targets of cancer cells is even more limited. The results of this study stress that mycotherapy is only one of the tools that can be used to balance remission. Palliative mycotherapy is associated with polyphenols composites, which contribute to detoxification and to the suppression of inflammation and pain sensation.

The Brown Roll-Rim Mushroom, Paxillus involutus (Agaricomycetes), as a Promising Biomedical Research Resource.

The brown roll-rim mushroom (Paxillus involutus) quickly produces biomass in nature, although, being a mycorrhizal fungus, it is rather poorly maintained in culture. Information about its toxic properties is controversial. Until the mid-20th century, the species was considered as an edible fungus; however, data later accumulated regarding its poisonous properties, leading to the term "Paxillus syndromeP. involutus. Since mushrooms can have quite a few unidentified antigens complementary to B-lymphocyte receptors, this is a hidden danger of using unfractionated mushroom raw materials for preventive and oncotherapy purposes, and we hope that this article stimulates immunological groups worldwide to identify the "X" antigen related to the Paxillus syndrome. Oncotherapy effects of the known bioactive complexes of P. involutus are associated with a specific inhibition of some growth receptors of the cancer cell, whereas experimentation with purified substances of P. involutus and various families of growth receptors of cancer cell has good prospects. A clear speciation is fixing within the P. involutus complex. The key for identification of species of P. involutus complex is given and cultural characteristics of P. involutus strains kept at Komarov Botanical Institute Basidiomycetes Culture Collection are presented.

Profiles of Little-Known Medicinal Polypores: Funalia trogii (Agaricomycetes).

The purpose of this study was to comprehensively characterize little-known polypores that have recently been found to possess anticancer activity and thus can be used in targeted cancer therapy. Funalia trogii is a polypore with bipolar distribution and has been found by harvesters working in taiga forests, broadleaf forests, and forest-steppes of the Holarctic, and in semiarid temperate forests of the Southern Hemisphere. Substances such as gibberellic acid, abscisic acid, indole-3-acetic acid, and natural cytokinin were determined in culture media of F. trogii. Also, laccases and peroxidases of spare action have been reported in F. trogii culture media. All of the aforementioned substances can be used in targeted cancer therapy, but further investigation of F. trogii is recommended; more details of its health benefits could expand its use in mycotherapy.

Current and Future Research Trends in Agricultural and Biomedical Applications of Medicinal Mushrooms and Mushroom Products (Review).

Here, we analyze the status and future trends in the study of medicinal mushrooms (MMs) in agriculture and biomedicine. Fruiting bodies of mushrooms are heterotrophic organisms that obtain all their nutritive requirements from the substrate. Mushroom substrates are agro-industrial plant residues and forest byproducts, which are usually called lignocellulosic materials. A good substrate for mushroom growth must be suitable both chemically and physically, as well as have the proper conditions for microbial activities. Under suitable conditions, mushrooms can excrete key enzymes for unlocking indigestible lignocellulosic biomasses to help provide sources of nourishment. The production of enzymes by mushroom mycelia plays a crucial part in the colonization process and is an important determinant of mushroom yields. The sense of purpose and vision for mushroom industries is also briefly discussed. Special attention is given to the use of mushroom extracts with antiphytopathologenic and insecticidal properties in modern agriculture. In the second part of this article, we summarize biomedical applications of medicinal mushrooms, which are currently used as 1) dietary food, 2) dietary supplement products, 3) a new class of drugs called "mushroom pharmaceuticals or mushroom drugs", 4) natural biocontrol agents in plant protection demonstrating insecticidal, fungicidal, bactericidal, herbicidal, nematocidal, and antiphytoviral activities, and 5) cosmeceuticals. We also aimed to draw attention to many critically important unsolved problems in the future development of medicinal mushroom science in the 21st century, including problems of production, standardization, and safety of mushroom dietary supplement products, as well as to discuss the problems of developing new medicinal mushroom drugs based not only on beta-glucans polysaccharides but also on low-molecular-weight secondary metabolites.

Enzymatic System of Antioxidant Protection of Erythrocytes in Diabetic Rats Treated with Medicinal Mushrooms Agaricus brasiliensis and Ganoderma lucidum (Agaricomycetes).

Excessive glucose concentrations in blood and cells promote the intensification of auto-oxidation. This is one of the mechanisms through which free radicals form in hyperglycemia. As a result of hyperglycemia, oxidative stress develops and lipid peroxidation (LPO) is enhanced. Erythrocytes are particularly susceptible to reactive oxygen species and LPO, which can violate cell functions. This article describes the analysis of the influence of mycelia from the medicinal mushrooms Agaricus brasiliensis and Ganoderma lucidum on the enzymatic link of the antioxidant system in rat erythrocytes under streptozotocin-induced diabetes mellitus. Oxidative stress was strengthened in red blood cells of diabetic rats, as evidenced by decreased activity of antioxidant enzymes such as superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase, and by increased amounts of thiobarbituric acid-positive products, which are markers of LPO. Administration of A. brasiliensis and G. lucidum submerged cultivated mycelial powder to animals with streptozotocin-induced diabetes restored superoxide dismutase, catalase, and glutathione peroxidase activity and reduced the amounts of thiobarbituric acid-positive products to control values, but did not affect the activity of glutathione reductase.