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).

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

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.

H2S is synthesized via CBS/CSE pathway and triggered by cold conditions during Agaricus bisporus storage

BACKGROUND: H2S is proved to be functioning as a signaling molecule in an array of physiological processes in the plant and animal kingdom. However, the H2S synthesis pathway and the responses to cold conditions remain unclear in postharvest mushroom. RESULTS: The biosynthesis of H2S in the Agaricus bisporus mushroom tissues exhibited an increasing tendency during postharvest storage and was significantly triggered by cold treatment. The cystathionine clyase (AbCSE) and cystathionine b-synthase (AbCBS) genes were cloned and proved responsible for H2S biosynthesis. Furthermore, transcriptional and posttranscriptional regulation of AbCSE and AbCBS were crucial for the enzyme activities and subsequent H2S levels. However, the AbMST was not involved in this process. Moreover, the AbCSE and AbCBS genes displayed low identity to the characterized genes, but typical catalytic domains, activity sites, subunit interface sites, and cofactor binding sites were conserved in the respective protein sequences, as revealed by molecular modeling and docking study. The potential transcription factors responsible for the H2S biosynthesis in cold conditions were also provided. CONCLUSIONS: The H2S biosynthetic pathway in postharvest mushroom was unique and distinct to that of other horticultural products.

Improving the yield of Anoectochilus roxburghii by Bacillus licheniformis cultured in Agaricus bisporus industrial wastewater

BACKGROUND: There is a large amount of industrial wastewater produced by the mushroom industry during the canning processing each year, which could provide abundant carbon, nitrogen and inorganic salts for microbial growth. The aim of this study was to optimize the culture conditions for Bacillus licheniformis cultured in the Agaricus bisporus industrial wastewater to produce the agricultural microbial fertilizer. RESULTS: In this work, the maximal biomass of B. licheniformis could be obtained under the following culture conditions: 33.7°C, pH 7.0, 221 rpm shaking speed, 0.5% wastewater, 2 (v:v, %) inoculum dose, loading liquid of 60 mL/250 mL and a culture time of 24 h, and the average experimental value obtained was 1.35 ± 0.04 × 109 Obj/mL, which was within the 95% confidence interval of the predicted model (1.29­1.38 × 109 Obj/mL), and met the national microbial fertilizers' standard in China. Furthermore, the field experiment results showed that the fermentation broth of B. licheniformis could significantly improve the yield of Anoectochilus roxburghii. CONCLUSIONS: Agaricus bisporus industrial wastewater can be used to produce agricultural microbial fertilizer.

Effect of protease inhibitor from Agaricus bisporus on glucose uptake and oxidative stress in 3T3-L1 adipocytes

Objective: To explore the effect of the protease inhibitor from Agaricus bisporus (J.E. Lange) Imbach (AbPI) on glucose uptake and oxidative stress in 3T3-L1 adipocytes. Methods: Adipocytes were differentiated and stained with Oil-Red-O staining to confirm adipogenesis. The toxic/protective effect of AbPI on the adipocytes was determined by MTT assay, intracellular reactive oxygen species generation through flow cytometry, and morphologically through confocal microscopy using propidium iodide, 4,6-diamino-2-phenylindol dihydrochloride, and 2',7'-dichlorofluorescein diacetate dyes. The uptake of fluorescent glucose analog, 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino]-2-deoxy-d-glucose by adipocytes was also studied through confocal microscopy. Results: MTT assay showed that the cell survival rate was (28.00±3.00)%, (92.33±2.60)%, and (71.34±2.10)% in the presence of 2 mM H

Effect of protease inhibitor from Agaricus bisporus on glucose uptake and oxidative stress in 3T3-L1 adipocytes

Objective: To explore the effect of the protease inhibitor from Agaricus bisporus (J.E. Lange) Imbach (AbPI) on glucose uptake and oxidative stress in 3T3-L1 adipocytes. Methods: Adipocytes were differentiated and stained with Oil-Red-O staining to confirm adipogenesis. The toxic/protective effect of AbPI on the adipocytes was determined by MTT assay, intracellular reactive oxygen species generation through flow cytometry, and morphologically through confocal microscopy using propidium iodide, 4,6-diamino-2-phenylindol dihydrochloride, and 2',7'-dichlorofluorescein diacetate dyes. The uptake of fluorescent glucose analog, 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino]-2-deoxy-d-glucose by adipocytes was also studied through confocal microscopy. Results: MTT assay showed that the cell survival rate was (28.00±3.00)％, (92.33±2.60)％, and (71.34±2.10)％ in the presence of 2 mM H2O2, AbPI alone, and AbPI and H2O2 both, respectively, in comparison to the control. Oil-Red-O staining indicated that AbPI enhanced adipogenesis. AbPI stimulated the glucose uptake by adipocytes similar to the drug rosiglitazone, and showed insulin-sensitizing effect in the presence of insulin, but failed to stimulate the uptake in the absence of insulin. Intracellular reactive oxygen species generation was reduced in differentiating adipocytes upon AbPI treatment. Confocal microscopy showed that the damaged cell population rose to 3.50％, 117.84％, and 261.50％ in the presence of AbPI alone, AbPI with H2O2, and H2O2 alone, respectively. Conclusions: The protease inhibitor enhances glucose uptake by adipocytes and exhibits a cytoprotective effect on them.

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.

Micetismos: Parte 3: Síndromes tempranos gastrointestinales / Mycetisms Part 3: Early Gastrointestinal Syndromes

En esta Parte 3 de la serie de cuatro artículos sobre micetismos se analizan los síndromes tempranos con síntomas gastrointestinales que se caracterizan por presentar un período de latencia muy corto, de menos de 6 horas después de la ingestión de los macromicetos. Los restantes síndromes tempranos con sintomatología compleja serán tratados en la Parte 4 de la serie. Actualmente se conocen más de 200 especies responsables de síndromes gastrointestinales, pero en este trabajo se abordarán solamente diez ejemplos que involucran los géneros Boletus [Boletus satanas (o Rubroboletus satanas) y Boletus venenatus (o Neoboletus venenata)], Hypholoma, Agaricus (Agaricus xanthodermus), Omphalotus, Lactarius, Russula, Entoloma, Chlorophyllum (Chlorophyllum molybdetes) y Leucoprinus (Leucoprinus birnbaumii). Las toxinas involucradas en estos casos presentan gran variedad estructural, desde proteínas hasta terpenoides, en particular sesquiterpenoides y triterpenoides, vinilglicina, fenol y azocompuestos, pero todas generan la misma sintomatología. Estas sustancias y otros componentes químicos de los hongos suelen ser indigestos, con una susceptibilidad variable entre los consumidores. El tratamiento es de apoyo y es estrictamente para esos casos con cuadros más graves de deshidratación. Normalmente, los casos evolucionan favorablemente después de 12 a 48 horas. Se analizan los síntomas, las toxinas involucradas, los mecanismos de acción, cuando se conocen y las especies causantes de los micetismos.

This part 3 of the series of four articles on mushroom poisoning refers to early-onset gastrointestinal syndromes, which are characterized by a very short latency period of less than 6 hours after mushroom ingestion. The remaining early-onset syndromes with complex symptoms will be treated in Part 4 of the series. Currently, more than 200 species responsible for gastrointestinal syndromes are known, but in this paper only ten examples will be addressed involving the genera Boletus [e.g., Boletus satanas (or Rubroboletus satanas), and Boletus venenatus (or Neoboletus venenata)], Hypholoma, Agaricus (e.g., Agaricus xanthodermus), Omphalotus, Lactarius, Russula, Entoloma, Chlorophyllum (e.g., Chlorophyllum molybdetes), and Leucoprinus (e.g., Leucoprinus birnbaumii). The toxins involved in these cases have a great structural variety, from proteins to terpenoids, in particular sesquiterpenoids and triterpenoids, vinylglycine, phenol, and azocompounds, but all show the same symptoms. These substances and other mushroom chemical constituents are usually indigestible, with varying consumer susceptibility. The treatment is supportive and is strictly for those cases with more severe dehydration. Usually, the cases progress favourably after 12 to 48 hours.The symptoms, toxins involved, mechanisms of action when known, and the species of mushrooms responsible for the mycetisms are analysed.

Nesta parte 3 da série de quatro artigos sobre intoxicação por cogumelos são analisadas as síndromes precoces com sintomas gastrointestinais que se caracterizam por apresentar um período de latência muito curto, de menos de 6 horas, após a ingestão de cogumelos. As síndromes precoces restantes com sintomatologia complexa serão tratadas na Parte 4 da série. Atualmente, são conhecidas mais de 200 espécies responsáveis por síndromes gastrointestinais, mas neste trabalho serão abordados apenas dez exemplos que envolvem os gêneros Boletus [Boletus satanas (ou Rubroboletus satanas) e Boletus venenatus (ou Neoboletus venenata)], Hypholoma, Agaricus (Agaricus xanthodermus), Omphalotus, Lactarius, Russula, Entoloma, Chlorophyllum (Chlorophyllum molybdetes) e Leucoprinus (Leucoprinus birnbaumii). As toxinas envolvidas nestes casos têm uma grande variedade estrutural, desde proteínas até terpenóides, em particular sesquiterpenóides e triterpenóides, vinilglicina, fenol e azo compostos, mas todas apresentam a mesma sintomatologia. Essas substâncias e outros constituintes químicos dos cogumelos costumam ser indigestos, com uma suscetibilidade variável entre aqueles que os consomem. O tratamento é de suporte e é rigorosamente para esses casos com quadros mais graves de desidratação. Normalmente, os casos evoluem favoravelmente após 12 a 48 horas. São analisados os sintomas, as toxinas envolvidas, os mecanismos de ação, quando conhecidos, e as espécies de cogumelos responsáveis pelas intoxicações.

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.

Green Synthesis, Characterization and Antioxidant Activity of Silver Nanoparticles in Extracts of Acorus calamus and Agaricus bisporus

Nanoparticles are in increasing commercial demand due to their wide applicability in various areas such as catalysis, chemistry, energy and medicine. Green chemistry is the best option to opt for the synthesisof nanoparticles.Agaricus bisporusand Acorus calamusare used medicinally in Ayurvedic medicines. The present work conducted endeavored to determine antioxidant activity of the extracts made. Silver nanoparticles were synthesized by using an aqueous extract of Acorus calamus & Agaricus bisporus.They were characterized by using UV spectrophotometer, Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM).Plant extracts were incubated with Silver Nitrate Solution (AgNO3) and the color change was observed as dark reddish brown. The presence of Ag nanoparticles was analyzed between the wavelengths 400-550 nm. SEM & TEM determined the size of the nanoparticlesdetermined by SEM &TEM. The antioxidant activity was performed by DPPH (1, 1diphenyl-2-picryl hydrazyl) free radical scavenging method for different solvents of Acorus calamus andAgaricus bisporus which showed that methanol extract on higher concentration possesses better antioxidant potential when compared to standard. They exhibited strong DPPH free radical scavenging activity with 66.3% and 59.2% respectively for ascorbic acid equivalence. The absorbance for reducing power was found to be in increasing order with an increase in concentration.Methanolic extract displayed an increased antioxidant activity extract, could be due to the presence of flavonoids and phenols. Also, the study reveals that the silver nanoparticles from methanol extract show better activity than thenormal aqueous extract.

Development of Polymorphic Simple Sequence Repeat Markers using High-Throughput Sequencing in Button Mushroom (Agaricus bisporus)

The white button mushroom (Agaricus bisporus) is one of the most widely cultivated species of edible mushroom. Despite its economic importance, relatively little is known about the genetic diversity of this species. Illumina paired-end sequencing produced 43,871,558 clean reads and 69,174 contigs were generated from five offspring. These contigs were subsequently assembled into 57,594 unigenes. The unigenes were annotated with reference genome in which 6,559 unigenes were associated with clusters, indicating orthologous genes. Gene ontology classification assigned many unigenes. Based on genome data of the five offspring, 44 polymorphic simple sequence repeat (SSR) markers were developed. The major allele frequency ranged from 0.42 to 0.92. The number of genotypes and the number of alleles ranged from 1 to 4, and from 2 to 4, respectively. The observed heterozygosity and the expected heterozygosity ranged from 0.00 to 1.00, and from 0.15 to 0.64, respectively. The polymorphic information content value ranged from 0.14 to 0.57. The genetic distances and UPGMA clustering discriminated offspring strains. The SSR markers developed in this study can be applied in polymorphism analyses of button mushroom and for cultivar discrimination.

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)

Protective effect of Agaricus bisporus intracellular polysaccharides and exopolysaccharides against concanavalin A-induced immunological liver injury in mice / 中国药理学与毒理学杂志

OBJECTIVE To evaluate the protective effect of Agaricus bisporus intracellular polysaccharides(IPS) and exopolysaccharides (EPS) on immunological liver injury induced by concanavalin A (Con A). METHODS Mice were pretreated with IPS and EPS (100, 200 and 400 mg kg- 1, ig) daily for 12 d. Immunological liver injury was induced by Con A 25 mg·kg-1 byinjection via the tail vein of mice.Eight hours after injection of Con A, the indexes of the liver, spleen and thymus, serum level of glutamicpyruvic transaminase (GPT), glutamic-oxalacetic transaminase (GOT), tumor necrosis factor-α (TNF-α) and interferon- γ (IFN- γ), splenic lymphocyte percentages of CD4 + and CD8 + , and liver homogenate content of superoxide dismutase (SOD) and malondialdehyde (MDA) were measured. Liver pathological changes were observed by HE staining. RESULTS Compared with normal group, the autoimmune liver injury in mice induced by Con A resulted in an increase in the liver index (P<0.01) , spleen index (P<0.01), the activity of GPT (P<0.01) and GOT (P<0.01), the content of TNF- α (P<0.01) and IFN- γ (P< 0.01), and the level of MDA (P<0.01), but a decrease in the thymus index (P<0.01), the percentage of CD4+ (P<0.01) , the ratio of CD4+/CD8+ (P<0.01), and SOD activity (P<0.01). Compared with model group, treatment with IPS (200 and 400 mg·kg-1) and EPS (200 and 400 mg·kg-1 ) respectively resulted in an increase in the thymus index (P<0.01) but in a decrease in the liver index and spleen index (P<0.01). Similarly, the activity of GOT and GPT was decreased obviously (P<0.01), and the content of TNF-α and IFN-γ in IPS and EPS 200 and 400 mg·kg-1 groups was decreased. Compared with model group, the activity of SOD in IPS and EPS (200 and 400 mg·kg- 1) group was increased (P<0.01) while MDA was decreased (P<0.01). Moreover, the percentage of CD4 + Iymphocytes decreased (P<0.01), whereas no significant difference was found in the ratio of CD4 +/CD8 + .Pathological changes of the liver were observed under a microscope. Pretreatment with IPS and EPS could effectively reduce the liver injury induced by Con A. CONCLUSION IPS and EPS have certain protective effect on immunological liver injury, which may be related to their ability to clean up free radicals, control lipid peroxidation and regulate the balance of the immune system.

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

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.

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.

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.