Analysis of effective components of Agaricus blazei Murill extract by LC-MS and network pharmacology analysis in treatment of chronic myeloid leukemia / 中国药理学通报

Aim To explore the potential targets and related signaling pathways of Agaricus blazei Murill (AbM ) extract in the treatment of chronic myeloid leukemia (CML) based on liquid chromatography mass spectrometry ( LC-MS ), network pharmacology, molecular docking, and were further verified by experiments in vitro. Methods The active components of AbM extract were retrieved from LC-MS, Swiss Target Prediction database was used to predict related targets, and CML disease target genes were obtained from Gen- eCards and DisGeNET databases. After screening the common targets of drug and CML, the protein-protein interaction network of the common targets was performed by STRING, and GO and KEGG enrichment a- nalysis were done by DAVID database. Cytoscape software was used to construct the network of target protein. Molecular docking was carried out by DockThor, and the Pymol software was used to make a visual picture. The inhibitory effect of AbM extract on leukemia cells K562 was determined by CCK-8 experiment, and the effect of AbM extract on the expression and phosphorylation level of related proteins was verified by Western blot. Results The prediction results showed that 126 active components of AbM extract, and 172 common targets were collected. KEGG pathway analysis results showed that PI3K/Akt/mTOR signaling pathway might play an important role in the treatment of CML disease. The IC

Effect of Agaricus blazei Murill on exploratory behavior of mice-model

Abstract Increased anxiety and depressive symptoms have reported to be its association with long term illness. Because of having unwanted effects of newly available drugs, patients administering anxiolytic drugs usually discontinue the treatment before they are completely recovered. Therefore, there is a serious need to develop new anxiolytic drugs. The anxiolytic effect of hydro-alcoholic extract of Agaricus blazei in animal models was assessed. 24 male mice (Mus musculus genus) were included in the study. Four groups were prepared and each group contained six animals. The groups were vehicle control, positive control (diazepam 1.0 mg/kg, i.p.) as well as two treatment groups receiving Agaricus blazei hydro-alcoholic extract at a dose of 136.50 mg/kg and 273.0 mg/kg orally. The Marble burying test, Nestlet shredding test and Light and Dark box test used to assess anxiolytic activity. Mice administered with diazepam 1.0 mg/kg, i.p. while hydro-alcoholic extract of AbM (136.50 and 273.0 mg/kg, respectively) was administered via oral route which exhibited marked reduction in number of marbles-burying as compared to vehicle control group. Mice administered with diazepam 1.0 mg/kg, i.p. and Oral administration of hydro-alcoholic extract of AbM (136.50 and 273.0 mg/kg, respectively) exhibited significant decrease in nestlet shredding in comparison to vehicle control group. The oral administration of hydro-alcoholic extract at a dose of 136.5mg/kg and 273mg/kg showed elevation in time spent in light box and was comparable to standard treated group while time spent by mice following oral administration of hydro-alcoholic extract of Agaricus blazei at a dose of 273.0 mg/kg also showed elevation and was found to be more near to standard treated group (diazepam 1 mg/kg, i.p.).

Resumo O aumento da ansiedade e dos sintomas depressivos têm relatado sua associação com doenças de longa duração. Por causa dos efeitos indesejáveis dos novos medicamentos disponíveis, os pacientes que administram medicamentos ansiolíticos geralmente interrompem o tratamento antes de estarem completamente recuperados. Portanto, há uma necessidade séria de desenvolver novos medicamentos ansiolíticos. Foi avaliado o efeito ansiolítico do extrato hidroalcoólico de Agaricus blazei em modelos animais. Vinte e quatro camundongos machos (gênero Mus musculus) foram incluídos no estudo. Quatro grupos foram preparados, e cada grupo continha seis animais. Os grupos foram controle de veículo, controle positivo (diazepam 1,0 mg/kg, i.p.), bem como dois grupos de tratamento recebendo extrato hidroalcoólico de Agaricus blazei na dose de 136,50 mg/kg e 273,0 mg/kg por via oral. O teste de enterrar Marble, o teste de retalhamento Nestlet e o teste de caixa clara e escura são usados para avaliar a atividade ansiolítica. Camundongos foram administrados com diazepam 1,0 mg/kg, i.p., enquanto o extrato hidroalcoólico de AbM (136,50 e 273,0 mg/kg, respectivamente) foi administrado por via oral, que exibiu redução acentuada no número de mármores enterrados em comparação com o grupo de controle de veículo. Camundongos administrados com diazepam 1,0 mg/kg, i.p. e a administração oral de extrato hidroalcoólico de AbM (136,50 e 273,0 mg/kg, respectivamente) exibiu diminuição significativa na trituração de ninhos em comparação ao grupo de controle de veículo. A administração oral de extrato hidroalcoólico na dose de 136,5mg/kg e 273mg/kg mostrou elevação no tempo gasto na caixa de luz e foi comparável ao grupo tratado padrão, enquanto o tempo gasto por camundongos após a administração oral de extrato hidroalcoólico de Agaricus blazei na dose de 273,0 mg/kg também mostrou elevação e foi mais próximo do grupo tratado padrão (diazepam 1 mg/kg, ip).

Effect of Agaricus blazei Murill on exploratory behavior of mice-model / Efeito de Agaricus blazei Murill no comportamento exploratório de modelo de camundongo

Abstract Increased anxiety and depressive symptoms have reported to be its association with long term illness. Because of having unwanted effects of newly available drugs, patients administering anxiolytic drugs usually discontinue the treatment before they are completely recovered. Therefore, there is a serious need to develop new anxiolytic drugs. The anxiolytic effect of hydro-alcoholic extract of Agaricus blazei in animal models was assessed. 24 male mice (Mus musculus genus) were included in the study. Four groups were prepared and each group contained six animals. The groups were vehicle control, positive control (diazepam 1.0 mg/kg, i.p.) as well as two treatment groups receiving Agaricus blazei hydro-alcoholic extract at a dose of 136.50 mg/kg and 273.0 mg/kg orally. The Marble burying test, Nestlet shredding test and Light and Dark box test used to assess anxiolytic activity. Mice administered with diazepam 1.0 mg/kg, i.p. while hydro-alcoholic extract of AbM (136.50 and 273.0 mg/kg, respectively) was administered via oral route which exhibited marked reduction in number of marbles-burying as compared to vehicle control group. Mice administered with diazepam 1.0 mg/kg, i.p. and Oral administration of hydro-alcoholic extract of AbM (136.50 and 273.0 mg/kg, respectively) exhibited significant decrease in nestlet shredding in comparison to vehicle control group. The oral administration of hydro-alcoholic extract at a dose of 136.5mg/kg and 273mg/kg showed elevation in time spent in light box and was comparable to standard treated group while time spent by mice following oral administration of hydro-alcoholic extract of Agaricus blazei at a dose of 273.0 mg/kg also showed elevation and was found to be more near to standard treated group (diazepam 1 mg/kg, i.p.).

Resumo O aumento da ansiedade e dos sintomas depressivos têm relatado sua associação com doenças de longa duração. Por causa dos efeitos indesejáveis dos novos medicamentos disponíveis, os pacientes que administram medicamentos ansiolíticos geralmente interrompem o tratamento antes de estarem completamente recuperados. Portanto, há uma necessidade séria de desenvolver novos medicamentos ansiolíticos. Foi avaliado o efeito ansiolítico do extrato hidroalcoólico de Agaricus blazei em modelos animais. Vinte e quatro camundongos machos (gênero Mus musculus) foram incluídos no estudo. Quatro grupos foram preparados, e cada grupo continha seis animais. Os grupos foram controle de veículo, controle positivo (diazepam 1,0 mg/kg, i.p.), bem como dois grupos de tratamento recebendo extrato hidroalcoólico de Agaricus blazei na dose de 136,50 mg/kg e 273,0 mg/kg por via oral. O teste de enterrar Marble, o teste de retalhamento Nestlet e o teste de caixa clara e escura são usados ​​para avaliar a atividade ansiolítica. Camundongos foram administrados com diazepam 1,0 mg/kg, i.p., enquanto o extrato hidroalcoólico de AbM (136,50 e 273,0 mg/kg, respectivamente) foi administrado por via oral, que exibiu redução acentuada no número de mármores enterrados em comparação com o grupo de controle de veículo. Camundongos administrados com diazepam 1,0 mg/kg, i.p. e a administração oral de extrato hidroalcoólico de AbM (136,50 e 273,0 mg/kg, respectivamente) exibiu diminuição significativa na trituração de ninhos em comparação ao grupo de controle de veículo. A administração oral de extrato hidroalcoólico na dose de 136,5mg/kg e 273mg/kg mostrou elevação no tempo gasto na caixa de luz e foi comparável ao grupo tratado padrão, enquanto o tempo gasto por camundongos após a administração oral de extrato hidroalcoólico de Agaricus blazei na dose de 273,0 mg/kg também mostrou elevação e foi mais próximo do grupo tratado padrão (diazepam 1 mg/kg, ip).

Study on the Mechanism of Multi-Drug Resistance of Agaricus Blazei Extract FA-2-b-β Mediated Wnt Signaling Pathway to Reverse Acute T Lymphoblastic Leukemia / 中国实验血液学杂志

OBJECTIVE@#To investigate the mechanism of drug reversing resistance of Agaricus blazei extract FA-2-b-β on T cell acute lymphoblastic leukemia (T-ALL) cell lines.@*METHODS@#Cell proliferation was detected by CCK-8 assay; the apoptosis, cell cycle mitochondrial membrane potential, and intracellular rhodamine accumulation were detected by flow cytometry, and apoptosis-related gene and protein expression were detected by qPCR and Western blot; the membrane surface protein MDR1 was observed by immunofluorescence microscopy.@*RESULTS@#Different concentrations of FA-2-b-β significantly inhibited proliferation and induced apoptosis of CCRF-CEM and CEM/C1 (P<0.05), and CCRF-CEM cell cycle were arrested at S phase, and CEM/C1 cells were arrested at G0/G1 phase. Western blot and qPCR results show that FA-2-b-β inhibited ABCB1、ABCG2、CTNNB、MYC and BCL-2 expression, but upregulated Bax expression. In addition, FA-2-b-β reversed the resistance characteristics of CEM/C1 drug-resistance cells, which decreased mitochondrial membrane potential, and significantly increased the intracellular rhodamine accumulation, and weakening of the expression of the membrane surface protein MDR1. With the Wnt/β-catenin inhibitor (ICG001), the process was further intensified.@*CONCLUSION@#Agaricus Blazei Extract FA-2-b-β inhibits cell proliferation, promotes apoptosis, regulates the cell cycle, reduces mitochondrial energy supply, and down-regulate MDR1 expression to reverse the resistance of CEM/C1, which all suggest it is through regulating the Wnt signaling pathway in T-ALL.

In-Vivo Study on Anxiolytic Effects of Hydroalcoholic Extract of Agaricus blazei Murill

Background: Anxiety involved panic attacks either having or not having social fear, social anxiety disorder, generalized anxiety disorder as well as separation anxiety disorder is known to be marked mental diseases. It is related to high medical cost and a significant load of disease. Agaricus blazei Murill (AbM) is a mushroom and possesses immunemodulating and antimicrobial effects both in-vivo and in-vitro and as well as it has been used to treat cancer, hepatitis, dermatitis, and hyperlipidemia traditionally. Method: In this experiment evaluation of anxiolytic effect of AbM on mice has been done by using Elevated Plus Maze test, open field test and motor co-ordination test by rotarod. Mice (Mus musculus) weighing 22-25 grams, were divided into 4 groups (n=6). Oral administration of hydro-alcoholic extract of AbM was utilized in 2 doses i.e. 136.5 mg/kg and 273 mg/kg. Group, I received vehicle (distilled water 10 ml/kg), p.o. Group II received standard (diazepam 1 mg/kg), i.p. Group III and IV orally received hydro-alcoholic extract of AbM (136.5 mg/kg and 273 mg/kg, respectively). Result: In Elevated Plus Maze test, oral administration of hydro-alcoholic extract of AbM (136.5 mg/kg and 273 mg/kg, respectively) exhibited significant (p<0.01) elevation in the percentage of number of open arm entries (48.0 ± 1.1% and 48.93 ± 2.1% respectively) and time spent in open arm (14.92 ± 1.9% and 84.17 ± 2.4%). Conclusion: Hence it is concluded that hydro-alcoholic extract of AbM can be a new therapeutic agent to treat anxiety.

Cryopreservation at -75 °C of Agaricus subrufescens on wheat grains with sucrose

Abstract Agaricus subrufescens is a basidiomycete which is studied because of its medicinal and gastronomic importance; however, less attention has been paid to its preservation. This study aimed to evaluate the effect of sucrose addition to substrate and cryotube on the viability of Agaricus subrufescens cryopreserved at -20 °C and at -75 °C for one and two years. Zero, 10% or 20% sucrose was added to potato dextrose agar or wheat grain. The mycelia were cryopreserved in the absence of cryoprotectant or with sucrose solutions at 15%, 30% or 45%. After one or two years at -75 °C or at -20 °C, mycelia were thawed and evaluated about viability, initial time of growth, colony diameter and genomic stability. Cryopreservation at -20 °C is not effective to keep mycelial viability of this fungus. Cryopreservation at -75 °C is effective when sucrose is used in substrates and/or cryotubes. Without sucrose, cryopreservation at -75 °C is effective only when wheat grains are used. Physiological characteristic as mycelial colony diameter is negatively affected when potato dextrose agar is used and unaffected when wheat grain is used after two-year cryopreservation at -75 °C. The fungus genome does not show alteration after two-year cryopreservation at -75 °C.

Immunity-enhancing effect of polysaccharide ABP-AW1 isolated from Agaricus blazei Murill on immunosuppressive mice / 中国药理学与毒理学杂志

OBJECTIVE To explore the immunity-enhancing effect of ABP-AW1, a low-molecular-mass polysaccharides isolated from Agaricus blazei Murill, on immunosuppressive mice. METHODS ICR mice were ip injected cyclophosphamide 80 mg · kg-1, once daily for 3 d, to establish an immuno?suppressive mouse model. Then, ABP-AW1125, 250 and 500 mg · kg-1 were ig given to the immuno?suppressive mice,respectively, once daily for 7 d. The mouse thymus index and spleen index were calcu?lated, and the phagocytic function of phagocytes was determined using carbon clearance test. Splenic lym?phocyte proliferation was measured by MTT method. The interleukin 2 (IL-2) and interferon γ (IFN-γ) production from splenic lymphocytes was examined by ELISA. The splenic lymphocyte CD4+/CD8+ratio was determined by flow cytometric analysis. RESULTS Compared with normal control group, the thymus index, spleen index and phagocytic index of phagocytes in the immunosuppressive model mice declined (P<0.05). ABP-AW1250 and 500 mg·kg-1 treatment significantly increased the thymus index, spleen index and phagocytic index in immunosuppressive mice (P<0.05). Compared with normal control group, concanavalin A (Con A) and lipopolysaccharide (LPS) induced T and B lymphocyte proliferation, respectively, and IL-2 and IFN-γproduction from splenic lymphocytes in the immunosuppressive model mice was lower (P<0.05). Compared with model group, ABP-AW1250 and 500 mg·kg-1 promoted Con A and LPS induced T and B lymphocyte proliferation (P<0.05) , and elevated IL-2 and IFN-γ production from splenic lymphocytes (P<0.05). In addition, ABP-AW1250 and 500 mg·kg-1 reversed the decreased splenocyte CD4+/CD8+ratio in immunosuppressive model mice (P<0.05). CONCLUSION ABP-AW1 has immuneity-enhancing effect on cyclophosphamide-induced immunosuppressive mice.

Protective Effect of Different Extraction of Agaricus blazei Murrill on HaCaT Cells Damaged by UVB / 浙江中医药大学学报

[Objective] To investigate the protective effects of different extraction of Agaricus blazei Murrill on HaCaT cells damaged by UVB. [Methods] The cell viability which was detected by MTT method was used to determine the impact of different dose of UVB and different extraction of Agaricus blazei Murrill on normal HaCaT cells and HaCaT cells damaged by UVB. [Results] ①The HaCaT cells viability declined after irradiation with UVB, and it was a dose-dependent manner. The cell viability was 53.23%when the dose of UVB was 20mj/cm2. ②The water extraction at the concentration of 0.2~4mg·mL-1, aqueous extraction at the concentration of 0.1~2mg·mL-1 and crude polysaccharide at the concentration of 0.2~1mg·mL-1 could promote the proliferation of HaCaT cells, while aqueous extraction and crude polysaccharide inhibited the proliferation of HaCaT cells at higher concentrations.③The water extraction at the concentration of 1~4mg·mL-1, aqueous extraction at the concentration of 1~2mg·mL-1 and crude polysaccharide at the concentration of 0.5~1mg·mL-1 could enhance the cells viability which was irradiated by 20mj/cm2 UVB (P<0.01). [Conclusion] The different extraction of Agaricus blazei Murrill all could relieve the photo-damage caused by UVB, and it has the potential of anti-photoaging.

Effect of Low Molecular Polysaccharide from Agaricus Blazei on Oxidative Injury in Hippocampal Neuronal Cells of Rats / 医药导报

Objective To investigate the effect and the potential mechanisms of low molecular polysaccharide from agaricus blazei (LMPAB) on H2O2-induced oxidative injury in hippocampal neuronal cells of rats.Methods Hippocampal neuronal cells were isolated from SD rats (24 h) and grew in culture.Cultured cells were divided into normal control group (added the same amount of nutrient solution), model control group (added 500 μmol·L-1H2O2 solution) and LMPAB high, medium, low dose groups (added 20,10,5 mg·L-1 LMPAB solution, respectively, then added 500 μmol·L-1 H2O2 solution each).The hippocampal neuron cell activity was detected with MTT method.The hippocampus neuron mitochondrial membrane potential (MMP) was detected by flow cytometry.According to the reagent instruction methods, malondialdehyde (MDA), catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) activities were detected.Results The activities of cell, CAT, SOD, GSH-PX and MMP in normal control group and the LMPAB high dose group were significantly higher than those of model control group (P<0.01);The content of MDA in normal control group and LMPAB high dose group was significantly lower than that of model control group (P<0.01).Conclusion The protective effect of LMPAB on hippocampal neurons with H2O2-induced injury may be related with the mechanism of enhancing the neuronal antioxidative capacity.

Agaricus brasiliensis polysaccharides stimulate human monocytes to capture Candida albicans, express toll-like receptors 2 and 4, and produce pro-inflammatory cytokines

Agaricus brasiliensis é um cogumelo medicinal com atividades imunomoduladoras e antitumorais atribuídas aos ß-glucanos presentes na fração polissacarídica de seu corpo de frutificação. Uma vez que os ß-glucanos aumentam a imunorresponsividade celular, neste estudo objetivamos avaliar o efeito de uma fração rica em polissacarídeos tratados com ácido (ATF) de A. brasiliensis sobre a capacidade de monócitos humanos de aderir / fagocitar células de levedura C. albicans . expressão de receptores de reconhecimento de padrões e sua capacidade de produzir citocinas. Métodos: A adesão / fagocitose de C. albicans marcada com FITC foi avaliada por citometria de fluxo. As células foram incubadas com anticorpos marcados com fluorocromo específicos para TLR2 e 4, ßGR e MR e também avaliadas por citometria de fluxo. Os monócitos foram cultivados com ATF, e os sobrenadantes da cultura foram coletados para análise da produção de citocinas in vitro por ELISA (TNF-α, IL-1ß, IL-12 e IL-10). Resultados: ATF aumentou significativamente a aderência / fagocitose de C. albicans por monócitos e isso foi associado com expressão aumentada de TLR2 e TLR4, enquanto nenhum efeito foi observado em ßGR ou MR. Além disso, a expressão de TLR4 e TLR2 foi associada a níveis mais elevados de produção in vitro de TNF-α e IL-1, respectivamente. A produção de IL-10 também foi aumentada pelo tratamento com ATF, mas não encontramos associação entre sua produção e a expressão de receptores Toll-like. Conclusão: Nossos resultados nos forneceram evidências de que polissacarídeos de A. brasiliensis afetam monócitos humanos provavelmente através da modulação de receptores Toll-like.(AU)

Agaricus brasiliensis polysaccharides stimulate human monocytes to capture Candida albicans, express toll-like receptors 2 and 4, and produce pro-inflammatory cytokines

Abstract Background Agaricus brasiliensis is a medicinal mushroom with immunomodulatory and antitumor activities attributed to the -glucans presented in the polysaccharide fraction of its fruiting body. Since -glucans enhance cellular immunoresponsiveness, in this study we aimed to evaluate the effect of an acid-treated polysaccharide-rich fraction (ATF) of A. brasiliensis on the ability of human monocytes to adhere/phagocyte C. albicans yeast cells, their expression of pattern recognition receptors and their ability to produce cytokines. Methods Adhesion/phagocytosis of FITC-labeled C. albicans was evaluated by flow cytometry. Cells were incubated with specific fluorochrome-labeled antibodies for TLR2 and 4, GR and MR and also evaluated by flow cytometry. Monocytes were cultured with ATF, and culture supernatants were collected for analysis of in vitro cytokine production by ELISA (TNF-, IL-1, IL-12 and IL-10). Results ATF significantly increased the adherence/phagocytosis of C. albicans by monocytes and this was associated with enhanced expression of TLR2 and TLR4, while no effect was observed on GR or MR. Moreover, expression of TLR4 and TLR2 was associated with higher levels of in vitro production of TNF- and IL-1, respectively. Production of IL-10 was also increased by ATF treatment, but we found no association between its production and the expression of Toll-like receptors. Conclusion Our results provided us with evidence that A. brasiliensis polysaccharides affect human monocytes probably through the modulation of Toll-like receptors.

Antimicrobial properties of the mushroom Agaricus blazei – integrative review

ABSTRACT Infectious diseases associated with antimicrobial resistance are considered to represent an important public health problem. In this regard, the mushroom Agaricus blazei Murrill contains several bioactive substances that promote significant functional properties, among them, antimicrobial activity, which has attracted the interest of the scientific community. Thus, the aim of this study was to determine whether evidence of the antimicrobial action of A. blazei has been reported in the literature. In this integrative review, manuscripts held in research databases available online were examined with a view to answering the question “Does the mushroom A. blazei exert antimicrobial activity against Gram-negative and/or Gram-positive bacteria?” Only eight scientific articles that have addressed the antimicrobial properties of A. blazei, in vitro and in vivo, were found, all characterized as pre-clinical, i.e., with level VII evidence. Most authors have found that the A. blazei extract promotes an antimicrobial effect against peritonitis, as well as deadly oral infections, especially those caused by Gram-positive bacteria. However, the scientific data currently available are not sufficient to verify the antimicrobial aspect of the mushroom A. blazei and thus further investigation is required.

Bioacumulação de zinco em micélio de Agaricus subrufescens / Zinc bioaccumulation in Agaricus subrufescens mycelium / Bioacumulación de zinc en micelio de Agaricus subrufescens

Fungos basidiomicetos têm a capacidade de bioacumular metais pesados, no entanto existem poucos trabalhos sobre bioacumulação de zinco em micélio de Agaricussubrufescens. O objetivo deste trabalho foi avaliar a bioacumulação de zinco em micélio vegetativo de A. subrufescens cultivado em meio sólido e líquido. O fungo foi crescido em meio sólido ou em meio líquido a base de extrato de malte adicionado de ZnSO4 a fim de obter zero; 2,5; 5; 7,5; 10; 15 ou 20 ppm de zinco. Os meios foram inoculados e após 14 dias foi determinada a biomassa e a bioacumulação de zinco. A adição de zinco no meio de cultivo inibiu o crescimento micelial e induziu a bioacumulação na biomassa tanto no cultivo sólido como no líquido. Adições acima de 7,5 ppm de zinco inibiram totalmente o crescimento micelial. O fungo crescido em meio de cultivo líquido sofre maior inibição do crescimento com a adição de zinco e maior bioacumulação que no meio sólido...

Fungi basidiomycetes have the ability to bioaccumulate heavy metals, but there are few studies on zinc bioaccumulation in the mycelium of Agaricus subrufescens. The objective of this study is to evaluate the zinc bioaccumulation in the mycelium of A. subrufescens cultivated in solid and liquid culture media. Mycelium was grown on solid or liquid medium in malt extract base added with ZnSO4 to obtain zero, 2.5, 5, 7.5, 10, 15 or 20-ppm zinc. Mycelial biomass and zinc bioaccumulation were determined 14 days after inoculation in the culture media. Addition of zinc in culture medium inhibited mycelial growth and induced biomass bioaccumulation both in solid and in liquid culture. Additions higher than 7.5-ppm zinc completely inhibited mycelial growth in culture medium. Mycelial growth in liquid culture presented greater increase of growth inhibition with the addition of zinc and greater bioaccumulation than in solid medium...

Hongos basidiomicetos tienen la capacidad de bioacumular metales pesados, sin embargo hay pocos estudios sobre la bioacumulación de zinc en el micelio de Agaricus subrufescens. El objetivo de este estudio ha sido evaluar la bioacumulación de zinc en el micelio de A. subrufescens cultivado en medio sólido y líquido. El hongo ha crecido en medio sólido o líquido a base de extracto de malta agregado de ZnSO4 para obtener cero; 2.5, 5, 7.5, 10, 15 o 20 ppm de zinc. Los medios fueron inoculados y después de 14 días se determinó la biomasa y la bioacumulación de zinc. La adición de zinc en el medio del cultivo inhibió el crecimiento micelial y indujo la bioacumulación de la biomasa tanto en el cultivo sólido como en el líquido. Adiciones superiores a 7.5 ppm de zinc inhibieron completamente el crecimiento del micelio. El hongo crecido en medio de cultivo líquido sufre mayor inhibición del crecimiento con la adición de zinc y mayor bioacumulación que en el medio sólido...

Anti-carcinogenic actions of glycoprotein conjugated with isoflavones from submerged-liquid culture of Agaricus blazei mycelia through reciprocal expression of Bcl-2 and Bax proteins / 동물의과학연구지

Glycoproteins isolated from fruit bodies and mycelial cultures of mushrooms exhibit anti-carcinogenic actions in human cancer cells and animal tumor cells by induction of apoptosis. Here, we report that isoflavone-conjugated glycoproteins (designate Gluvone), exhibit strong anti-carcinogenic effects on human breast cancer MCF-7 cells by induction of apoptosis. Gluvone with 9.4 kDa of molecular weight was isolated from submerged-liquid culture of Agaricus blazei mycelia (ABM) in soy flake-containing liquid medium. MCF-7 cells were incubated with various amounts of Gluvone (0~250 microM) for a period of 6 days. Gluvone exhibited anti-proliferative actions in a dose-dependent manner and 62% growth inhibition at 200 microM for 4 days relative to control. Hoechst 33258 staining analysis revealed that Gluvone induced formation of apoptotic bodies. Gluvone was associated with down-regulation of anti-apoptotic Bcl-2 protein expression as well as up-regulation of pro-apoptotic Bax protein expression. Gluvone treatment induced proteolytic activation of caspase-9 and caspase-3 through cytochrome c release from mitochondria to cytosol as well as concomitant degradation of poly (ADP-ribose) polymerase (PARP). In addition, Gluvone induced activation of caspase-8. Taken all together, these results indicate that the anti-proliferative effect of Gluvone is associated with induction of apoptotic cell death through the mitochondrial dysfunction pathway mediated by enhancement of Bax protein expression and suppression of Bcl-2 protein expression.

Antioxidant activity of Agaricus brasiliensis basidiocarps on different maturation phases

Different maturation phases of basidiocarp could affect the bioactivity and concentration of some active substances. A. brasiliensis Wasser et al.(A. blazei Murrill) has shown antitumor activity that could be related to the antioxidant activity. However there is no information of the best basidiocarp maturation phase for extracting antioxidant substances in order to determine the moment of harvesting in mushroom cultivation. The objective of this work was to evaluate the antioxidant activity of A. brasiliensis strains on different basidiocarp maturation phases. The best condition for extraction of A. brasiliensis antioxidants is with methanol as solvent at 60 ºC for 60 min. Strains with closed basidiocarp have higher antioxidant activity than with opened basidiocarp. Antioxidant activity varies in each strain. It was concluded that A. brasiliensis is a natural source of antioxidant compounds. Also there is higher antioxidant activity in closed than opened caps and consequently higher functional activity. It reinforces the synergic action among different A. brasiliensis compounds as a functional food and the importance of further investigation for isolation and characterization of antioxidant substances of A. brasiliensis. It also determines the best harvest period in order to obtain the highest antioxidant activity from basidiocarp.

Tratamentos térmicos do calxisto para uso como camada de cobertura no cultivo de Agaricus brasiliensis / Thermal treatments on lime schist casing layer for Agaricus brasiliensis cultivation

A escolha da camada de cobertura é uma das mais importantes etapas do cultivo de Agaricus brasiliensis. Apesar dessa importância, poucos estudos relatam o uso de diferentes tratamentos térmicos para o controle da microbiota em camadas de cobertura alternativas. Assim, o objetivo deste trabalho foi avaliar o efeito da pasteurização e da autoclavagem do material alternativo calxisto para utilização como camada de cobertura no cultivo de A. brasiliensis. O fungo foi inicialmente crescido em grãos de trigo e transferido para meio de cultivo previamente compostado. Após a completa colonização, a camada de cobertura (calxisto) pasteurizada ou autoclavada foi adicionada. Avaliaram-se a eficiência biológica, o número e a biomassa de cogumelos produzidos e o fluxo de produção. Concluiu-se que a camada de cobertura com calxisto autoclavado reduzem o tempo de produção, a eficiência biológica e o número e a biomassa de cogumelos cultivados. Entretanto, a camada de cobertura com o calxisto pasteurizado é a mais eficiente para o cultivo de A. brasiliensis.

Casing layer choice is one of the most important phases on Agaricus brasiliensis cultivation. Besides the importance of it few studies report the use of different heat treatments to control the microbiota in alternative casing layers. Thus, the objective of this work was to evaluate the effect of pasteurized or autoclaved lime schist as an alternative casing layer on A. brasiliensis cultivation. The fungus was previously grown on wheat grains and transferred to a substratum previously composted. After substratum mycelium colonization a pasteurized or autoclaved lime schist casing layer was added on. It was evaluated the biological efficiency, the number and mass of produced mushroom and the production flush along cultivation. It was concluded that autoclaved lime schist casing layer decreases period of production, biological efficiency, number and mass of cultivated mushrooms. However pasteurized lime schist casing layer is the most efficient on A. brasiliensis cultivation.

Antineoplasic activity of Agaricus brasiliensis basidiocarps on different maturation phases

The fungus Agaricus brasiliensis is a Basidiomycete studied because of its immunomodulation and/or antitumor substances. The objective of this study was to verify the Agaricus brasiliensis antineoplasic activity in vivo on different basidiocarp maturation phases on Sarcoma 180 cells implanted in mice. Sarcoma cells were implanted in mice and after seven days mice were divided in three groups. The first group was treated with saline solution, the second group was treated with closed basidiocarp extract solution and the third group was treated with opened basidiocarp extract solution. After 30 days of being daily orally treated with these three solutions all animals suffered euthanasia, and the splenic index, tumor mass and volume were determined. No significant differences of the tumor growth inhibition in function of the different basidiocarp maturation phases for the Agaricus brasiliensis strain were observed. The in vivo basidiocarp antineoplasic average activity was 89.22 percent.

Produção de agaricus blazei ss. Heinemann (a. Brasiliensis) em função de diferentes camadas de cobertura e substratos de cultivo / Production of agaricus blazei ss. Heinemann (a. Brasiliensis) in function of different casing layers and composts / Producción de agaricus blazei ss. Heinemann (a. Brasiliensis) en funcción de diferentes camadas de cobertura y sustratos de cultivo

Avaliou-se o efeito de duas formulações de composto, com palha de tifton (Cynodon sp.) e com palha de aveia (Avena sativa) e três camadas de cobertura (testemunha, testemunha + gel de acrilamida e testemunha + farelo de soja) na produção, eficiência biológica, massa e número de basidiomas de A. blazei. O delineamento experimental foi em esquema fatorial 3´2 inteiramente casualizado, com quatro 4 repetições. A unidade experimental constou de 12-12,5kg de composto úmido. A presença de gel de acrilamida como camada de cobertura tendeu a apresentar maior produtividade e eficiência biológica. O composto à base de tifton apresentou uma tendência de menor produção e eficiência biológica em relação ao composto à base de aveia.

The aim of this study was to evaluate yield, biological efficiency, fresh weight and number of mushrooms of Agaricus blazei grown in two kinds of compost, prepared with Tifton (Cynodon sp.) grass and with oat (Avena sativa) straw, and three casing layers (control, control + acrilamyde gel and soil and control + soybean meal). The experiment consisted of four replicates (each with 12-12,5kg of fresh weight compost) arranged in a factorial scheme 3´2. The results showed a tendency for higher yield and biological efficiency when adding acrilamyde gel to the casing soil. The composts made with oat straw presented a tendency for higher yield and biological efficiency compared to the Tyfton.

Se evaluó el efecto de dos formulaciones de composta, con paja de césped Tifton (Cynodon sp.) y con paja de avena (Avena sativa) y tres camadas de cobertura (control, control + gel de acrilamida y control + salvado de soya) en la producción, eficiencia biológica, masa y número de basidiomas de A. blazei. El diseño experimental fue un esquema factorial 3´2 enteramente aleatorio con cuatro repeticiones. La unidad experimental consistió de 12-12,5kg de composta húmeda. La presencia de gel de acrilamida como camada de cobertura resultó en una tendencia a presentar mayor productividad y eficiencia biológica. La composta a base de Tifton presentó una tendencia a menor producción y eficiencia biológica en relación a la composta en base a avena.

Multi-Potential Cancer Preventive Efficacy and the Current Safety Status of Agaricus blazei Murill Products / 日本補完代替医療学会誌

<i>Agaricus blazei</i> Murill (ABM), widely known as “Himematsutake” in Japan, has been one of the most popular dietary supplements, especially among the aged-population. ABM has been traditionally used to treat age-related disease such as cardiovascular disease, diabetes, and cancers. Previously, ABM has been shown to have immunostimulatory, antioxidant, antimutagenic, and antitumorigenic properties. In contrast to antitumor effects in rodent tumor cell lines, ABM has only minimal to moderate antitumor effects in a few human cancer cell lines, and antitumor effects were thought to be attributed to a high molecular weight fraction containing highly branched β-glucans, while a low molecular weight fraction of ABM (ABM F500) is a very effective cancer preventive agent. The question of genetic toxicity with agaritine has been scrutinized by using the transgenic F344 rat LacI, a state-of-the-art mutagenicity testing system. Neither K-ABM nor agaritine proved mutagenic. Furthermore, expected DNA adducts (8-HMPdGuo and 8-HMPdAdo) with the postulated mutagenic metabolite 4-(methylhydroxy)phenylhydrazine (HMPD) of agaritine were not found. These new mutagenicity test results provide irrefutable evidence that the earlier positive Ames mutagenicity test results with K-ABM were in fact false positive. Thus, the most recent mutagenicity test results together with the 2-year chronic carcinogenic bioassay strongly suggest neither agaritine nor quality controlled ABM are mutagenic or carcinogenic. Clinical improvement of QOL among cancer patients undergoing chemotherapy complimented by ABM consumption appears to be due to immunostimulation of immunosuppressed patients. Retrospective QOL analysis of 782 cancer patients further supports the thesis that ABM consumption improves QOL in cancer patients. Thus, it is reasonable to expect ABM treatment in combination with cancer chemotherapy may synergistically enhance the outcome of such chemotherapy. Complimentary clinical contribution of ABM to conventional therapy or type II diabetes is also expected. Further clinical studies are necessary to evaluate and validate the chemopreventive potential of ABM F500 as well as clinical merits of ABM as a complimentary medicine in cancer chemotherapy.<br>

Cytotoxicity and genotoxicity of Agaricus blazei methanolic extract fractions assessed using gene and chromosomal mutation assays

Functional food investigations have demonstrated the presence of substances that could be beneficial to human health when consumed. However, the toxic effects of some substances contained in foods have been determined. Reported medicinal and nutritive properties have led to the extensive commercialization of the basidiomycete fungi Agaricus blazei Murrill (sensu Heinemann), also known as Agaricus brasiliensis Wasser et al., Agaricus subrufescens Peck or the Brazilian medical mushroom (BMM). Different methanolic extract fractions (ME) of this mushroom were submitted to the cytokinesis-block micronucleus (CBMN) clastogenic assay and the hypoxanthine-guanine phosphoribosyl transferase locus (HGPRT) assay for gene mutation, both using Chinese hamster ovary cells clone K1 (CHO-K1). The results suggest that all the fractions tested possess cytotoxic and mutagenic potential but no clastogenic effects. Further information is needed on the biochemical components of the A. blazei methanol fractions to identify any substances with cytotoxic and/or mutagenicity potential. These findings indicate that A. blazei methanolic extract should not be used due to their genotoxicity and care should be taken in the use of A. blazei by the general population until further biochemical characterization of this fungi is completed.