Ultrasound Intensify the Flavonoid Production of the Willow Bracket Mushroom, Phellinus igniarius (Agaricomycetes), Fermentation Mycelia.

This research aimed to use a novel and effective ultrasound (US) approach for obtaining high bio-compound production, hence proposing strategies for boosting active ingredient biosynthesis. Furthermore, the US promotes several physiological effects on the relevant organelles in the cell, morphological effects on the structure of Phellinus igniarius mycelium, and increases the transfer of nutrients and metabolites. One suitable US condition for flavonoid fermentation was determined as once per day for 7-9 days at a frequency 22 + 40 kHz, power density 120 W/L, treated 10 min, treatment off time 7 s. The flavonoid content and production increased about 47.51% and 101.81%, respectively, compared with the untreated fermentation (P < 0.05). SEM showed that sonication changes the morphology and structure of Ph. igniarius mycelium; TEM reveals the ultrasonic treatment causes organelle aggregation. The ultrasound could affect the metabolism of the biosynthesis of the active ingredients.

Optimization of the Solid-State Culture Conditions and Chemical Component Analysis of Poria cocos (Agaricomycetes).

The optimal cultivation conditions and chemical components of Poria cocos fruiting bodies were examined by employing the single factor and response surface methods to screen for optimal conditions for artificial cultivation. The differences in chemical composition among the fruiting bodies, fermented mycelium, and sclerotia of P. cocos were compared using UV spectrophotometry and high-performance liquid chromatography (HPLC). The optimal growth conditions for P. cocos fruiting bodies were 28.5°C temperature, 60% light intensity, and 2.5 g pine sawdust, which resulted in the production of numerous basidiocarps and basidiospores under microscopic examination. Polysaccharides, triterpenoids, and other main active components of P. cocos were found in the fruiting bodies, sclerotia, and fermented mycelium. The triterpenoid components of the fruiting bodies were consistent with those of the sclerotia. The content of pachymic acid in the fruiting bodies was significantly higher than that in the sclerotia, with a value of 33.37 ± 0.1902 mg/g. These findings provide novel insights into the sexual breeding and comprehensive development and utilization of P. cocos.

Hydrophilic Metabolite Composition of Fruiting Bodies and Mycelia of Edible Mushroom Species (Agaricomycetes).

Mushrooms have two components, the fruiting body, which encompasses the stalk and the cap, and the mycelium, which supports the fruiting body underground. The part of the mushroom most commonly consumed is the fruiting body. Given that it is more time consuming to harvest the fruiting body versus simply the mycelia, we were interested in understanding the difference in metabolite content between the fruiting bodies and mycelia of four widely consumed mushrooms in Taiwan: Agrocybe cylindracea (AC), Coprinus comatus (CC), Hericium erinaceus (HE), and Hypsizygus marmoreus (HM). In total, we identified 54 polar metabolites using 1H NMR spectroscopy that included sugar alcohols, amino acids, organic acids, nucleosides and purine/pyrimidine derivatives, sugars, and others. Generally, the fruiting bodies of AC, CC, and HM contained higher amounts of essential amino acids than their corresponding mycelia. Among fruiting bodies, HE had the lowest essential amino acid content. Trehalose was the predominant carbohydrate in most samples except for the mycelia of AC, in which the major sugar was glucose. The amount of adenosine, uridine, and xanthine in the samples was similar, and was higher in fruiting bodies compared with mycelia, except for HM. The organic acid and sugar alcohol content between fruiting bodies and mycelia did not tend to be different. Although each mushroom had a unique metabolic profile, the metabolic profile of fruiting bodies and mycelia were most similar for CC and HE, suggesting that the mycelia of CC and HE may be good replacements for their corresponding fruiting bodies. Additionally, each mushroom species had a unique polar metabolite fingerprint, which could be utilized to identify adulteration.

Extraction and characterization of polysaccharide from fermented mycelia of Coriolus versicolor and its efficacy for treating nonalcoholic fatty liver disease.

Coriolus versicolor, a popular traditional Chinese medicinal herb, is widely used in China to treat spleen and liver diseases; however, the beneficial effects of C. versicolor polysaccharides (CVPs) on nonalcoholic fatty liver disease (NAFLD) remain elusive. Herein we isolated and purified a novel CVP (molecular weight, 17,478 Da) from fermented mycelium powder. This CVP was composed of mannose, galacturonic acid, glucose, galactose, xylose, and fucose at a molar ratio of 22:1:8:15:10:3. Methylation, gas chromatography-mass spectrometry, and nuclear magnetic resonance analyses indicated that the CVP backbone consisted of →1)-ß-D-Man-(6,4→1)-α-D-Gal-(3→1)-α-D-Man-(4→1)-α-D-Gal-(6→, with branches of →1)-α-D-Glc-(6→1)-α-D-Man-(4,3→1)-ß-D-Xyl-(2→1)-ß-D-Glc on the O-6 position of →1)-ß-D-Man-(6,4→ of the main chain. The secondary branches linked to the O-4 position of →1)-α-D-Man-(4,3→ with the chain of →1)-α-D-Fuc-(4→1)-α-D-Man. Further, CVP treatment alleviated the symptoms of NAFLD in an HFD-induced mice model. CVP altered gut microbiota, predominantly suppressing microbes associated with bile acids both in the serum and cecal contents. In vitro data showed that CVP reduced HFD-induced hyperlipidemia via farnesoid X receptor. Our results improve our understanding of the mechanisms underlying the cholesterol- and lipid-lowering effects of CVP and indicate that CVP is a promising candidate for NAFLD therapy.

Potential bioactivities via anticancer, antioxidant, and immunomodulatory properties of cultured mycelial enriched ß-D-glucan polysaccharides from a novel fungus Ophiocordyceps sinensis OS8.

Background: Ophiocordyceps sinensis is well-known worldwide as a traditional medicine. An alternative natural source of O. sinensis is provided by mycelial cultivation. However, the bioactivities of cultured mycelial-enriched ß-D-glucan polysaccharides from a novel fungus O. sinensis OS8 are still unknown. Methods: We investigated the potential bioactivities via anticancer, antioxidant, and immunomodulatory polysaccharides (OS8P) produced from cultured mycelia of O. sinensis OS8. This strain is a novel fungus isolated from natural O. sinensis, which is further cultured by submerged mycelial cultivation for polysaccharide production. Results: The yield of mycelial biomass was 23.61 g/l, and it contained 306.1 mg/100 g of adenosine and 3.22 g/100 g of polysaccharides. This OS8P was enriched with ß-D-glucan at 56.92% and another form of α-D-glucan at 35.32%. The main components of OS8P were dodecamethyl pentasiloxane, 2,6-bis (methylthiomethyl) pyridine, 2-(4-pyrimidinyl)-1H-Benzimidazole, and 2-Chloro-4-(4-nitroanilino)-6-(O-toluidino)-1,3,5-triazine at the rates of 32.5, 20.0, 17.5, and 16.25%, respectively. The growth of colon cancer cells (HT-29) was significantly inhibited by OS8P, with IC50 value of 202.98 µg/ml, and encouraged apoptosis in HT-29 cells as confirmed by morphological change analysis via AO/PI and DAPI staining, DNA fragmentation, and scanning electron microscopic observations. In addition, significant antioxidant activity was demonstrated by OS8P through DPPH and ABTS assays, with IC50 values of 0.52 and 2.07 mg/ml, respectively. The OS8P also exhibited suitable immunomodulatory activities that significantly enhanced (P< 0.05) the induction of splenocyte proliferation. Conclusion: The OS8P enriched with ß-D-glucan polysaccharides and produced by submerged mycelial culture of a new fungal strain of O. sinensis OS8 strongly inhibited the proliferation of colon cancer cells without any cytotoxicity against normal cells. The potential effect of the OS8P on the cancer cells was due to the stimulation of apoptosis. Also, the OS8P exhibited good antioxidant and immunomodulatory activities. The results indicate that OS8P has promising applications in the functional food industry and/or therapeutic agents for colon cancer.

Antioxidant and Antimicrobial Properties of Fruiting Body and Submerged Mycelium of Medicinal Mushroom Phellinus robiniae (Agaricomycetes).

This study was conducted to evaluate extraction yield, antioxidant content, antioxidant capacity and antibacterial activity of extracts obtained from submerged mycelium (ME) and fruiting body (FBE) of Phellinus robiniae NTH-PR1. The results showed that yields of ME and FBE reached 14.84 ± 0.63 and 18.89 ± 0.86%, respectively. TPSC, TPC, and TFC were present in both mycelium and fruiting body, and the more contents of them were found in fruiting body. The concentrations of TPSC, TPC and TFC in ME and FBE were 17.61 ± 0.67 and 21.56 ± 0.89 mg GE g-1, 9.31 ± 0.45 and 12.14 ± 0.56 mg QAE g-1, and 8.91 ± 0.53 and 9.04 ± 0.74 mg QE g-1, respectively. EC50 values for DPPH radical scavenging revealed FBE (260.62 ± 3.33 µg mL-1) was more effective than ME (298.21 ± 3.61 µg mL-1). EC50 values for ferrous ion chelating in ME and FBE were 411.87 ± 7.27 and 432.39 ± 2.23 µg mL-1, respectively. Thus, both extracts were able to inhibit Gram-positive and Gram-negative pathogenic bacterial strains, at concentrations ranging in 25-100 mg mL-1 of ME and 18.75-75 mg mL-1 of FBE for Gram-positive bacteria; ranging in 75-100 mg mL-1 of ME and 50-75 of FBE for Gram-negative bacteria. Overall submerged mycelial biomass and fruiting bodies of Ph. robiniae NTH-PR1 can be considered as useful natural sources for development of functional food, pharmaceuticals and cosmetic products or cosmeceuticals.

Antiprotozoal Activity of Mycelial Extracts of Several Medicinal Agaricomycetes Mushrooms against Giardia duodenalis.

Despite knowledge on the therapeutic properties of fungal bio-compounds, few studies have been reported on their anti-parasitic activities. The anti-parasitic activity (APA) of mycelial extracts from seven medicinal agaricomycetous mushrooms (Polyporus lipsiensis, Ganoderma applanatum, Pleurotus ostreatus, P. flabellatus, Oudemansiella canarii, Lentinula edodes, and Pycnoporus sanguineus) against Giardia duodenalis, and identification of chemical compounds produced by mycelium P. lipsiensis mycelium, have been reported. The extracts of mycelia and fermented culture broths of tested mushroom species were evaluated against G. duodenalis by biological assays. P. lipsiensis showed the highest APA. The chemical analysis of mycelial extract of P. lipsiensis by gas chromatography-mass spectrometry (GC-MS) identified 73 molecules, including steroids, terpenes, and lipids. According to literature data, among these molecules, 11 possess APA. The present study revealed the diversity of compounds with anti-protozoal potential produced by mycelia of Agaricomycetes mushrooms, particularly P. lipsiensis against G. duodenalis.

Development of a mass spectrometry-based metabolomics workflow for traceability of wild and cultivated Cordyceps sinensis.

Cordyceps sinensis, as an expensive traditional Chinese medicine and edible fungus mycelium, lacks an effective quality evaluation method, especially and cultivated Cordyceps sinensis. In this study, a feasible workflow method was developed for traceability evaluation of wild and cultivated Cordyceps sinensis, based on mass spectrometry-based metabolomics. Mass spectrometry data were firstly acquired from Cordyceps sinensis, samples by liquid chromatography-quadrupole and time of flight mass spectrometry. Characteristic mass spectrometry peaks were extracted by applying the MZmine. Then significant markers were obtained from Cordyceps sinensis samples by orthogonal partial least square discriminant analysis. Then, identification of significant markers were identified by MS-FINDER data analytics. The results showed that Changdu, the other four wild origins (Naqu, Xinghai, Yushu and Guoluo) and cultivated samples could be significantly distinguished. This identified significant markers of Cordyceps sinensis, including 174 special significant markers for the wild samples, 204 special significant markers for the cultivated samples and 87 share significant markers. Number of 87 shared significant markers were identified in the wild and cultivated Cordyceps sinensis, especially 28 confident significant compounds, such as adenosine, riboflavin, tyrosine, arginine and glutamine. These shared significant markers might support the quality control of multi-targets of Cordyceps sinensis, compared with a single target in the Chinese Pharmacopoeia. The special significant markers indicated that cultivated Cordyceps sinensis was different from the wild based on mass spectrometry-based metabolomics. In the comparison of chromatographic fingerprint technology, it was found that the established feasible workflow method was easy to acquire significant markers and traceability of Cordyceps sinensis. This feasible workflow method has great potential to be successful for comprehensive and traceability evaluation of the wild and cultivated Cordyceps sinensis.

Biological Characterization and Antimicrobial Bioactives of Mycelium Extracts from Medicinal Mushrooms Phellinus linteus and Pleurotus albidus (Agaricomycetes).

Bioactivity is defined as the intrinsic property of compounds that enables their participation in specific biological reactions. This study aimed to evaluate the antimicrobial capacity and to separate and characterize bioactives from aqueous and hydroalcoholic extracts obtained from the mycelium of medicinal mushrooms Pleurotus albidus and Phellinus linteus. Antimicrobial activity, through the disc diffusion method, was found against strains of Bacillus cereus, B. subtilis, Pseudomonas aeruginosa, and Staphylococcus aureus. P. albidus extracts showed better activity against Bacillus strains, whereas Ph. linteus extracts had greater effectiveness against S. aureus and P. aeruginosa. Aqueous extraction was best for obtaining bioactive compounds of P. albidus, whereas 30% hydralcoholic extraction performed best for obtaining Ph. linteus. Mass spectrometry analyses allowed the identification of the main chemical compounds extracted from the fungal biomasses, including glutathione oxidase, leucovorin, and riboflavin. Taking these findings into consideration, P. albidus and Ph. linteus might be used as sources of bioactive molecules for the development of novel drugs or nutraceuticals, contributing to the improvement of public health.

Fruit body formation and intra-species DNA polymorphism in Japanese Wolfiporia cocos strains.

Poria, the dried sclerotium of Wolfiporia cocos, is a medicinal mushroom that is widely used in traditional Japanese medicine. The fruit body of W. cocos is rarely found in the natural environment in Japan, therefore an optimized technique for fruit body formation is essential for producing new strains through crossbreeding and for biological research. Here, we developed a cultivation technique for fruit body formation of W. cocos using three strains collected from different areas of Japan. When mycelia were cultured on sawdust-based medium after liquid medium culture, all strains successfully formed fruit bodies as a brown honeycomb-like structure. Furthermore, we analyzed single nucleotide polymorphisms of the three strains using the STE3-like pheromone receptor protein gene, STE3.2, and found a genetic marker for discriminating one strain from the others. The results are expected to promote extensive studies on crossbreeding and domestic production of W. cocos.

Bioactivity and Mycochemical Profile of Extracts from Mycelial Cultures of Ganoderma spp.

Fungal mycelium cultures are an alternative to natural sources in order to obtain valuable research materials. They also enable constant control and adaptation of the process, thereby leading to increased biomass growth and accumulation of bioactive metabolites. The present study aims to assess the biosynthetic potential of mycelial cultures of six Ganoderma species: G. adspersum, G. applanatum, G. carnosum, G. lucidum, G. pfeifferi, and G. resinaceum. The presence of phenolic acids, amino acids, indole compounds, sterols, and kojic acid in biomass extracts was determined by HPLC. The antioxidant and cytotoxic activities of the extracts and their effects on the inhibition of selected enzymes (tyrosinase and acetylcholinesterase) were also evaluated. The total content of phenolic acids in the extracts ranged from 5.8 (G. carnosum) to 114.07 mg/100 g dry weight (d.w.) (G. pfeifferi). The total content of indole compounds in the extracts ranged from 3.03 (G. carnosum) to 11.56 mg/100 g d.w. (G. lucidum) and that of ergosterol ranged from 28.15 (G. applanatum) to 74.78 mg/100 g d.w. (G. adspersum). Kojic acid was found in the extracts of G. applanatum and G. lucidum. The tested extracts showed significant antioxidant activity. The results suggest that the analyzed mycelial cultures are promising candidates for the development of new dietary supplements or pharmaceutical preparations.

Biochemical characterization and biological properties of mycelium extracts from Lepista sordida GMA-05 and Trametes hirsuta GMA-01: new mushroom strains isolated in Brazil.

The objective of this study was to evaluate the antioxidant activity, determine and quantify the phenolic compounds and other compounds, and evaluate the cellular cytotoxicity of mycelium extracts of two new Basidiomycete mushrooms strains isolated in Brazil and identified as Lepista sordida GMA-05 and Trametes hirsuta GMA-01. Higher amounts of proteins, free amino acids, total and reducing carbohydrates, and phenolic compounds as chlorogenic, ferulic, caffeic, and gallic acids were found in extracts of T. hirsuta and L. sordida. Protocatechuic acid was found only in aqueous extracts of L. sordida. The TLC of the extracts showed the predominance of glucose and smaller amounts of xylose. It was observed through UPLC-MS higher amounts of phenolic compounds. The aqueous extract from T. hirsuta had the most noteworthy results in the antioxidant assays, especially the ABTS test. The cytotoxic activity was evaluated using two different cell lineages and showed higher toxicity for L. sordida in macrophages J774-A1. However, in Vero cells, it was 12.6-fold less toxic when compared to T. hirsuta. Thus, both mushrooms show potential as functional foods or additives, presenting phenolic content, antioxidant activity, and low cytotoxic activity in the tested cells.

Selective Biological Effects of Selenium-Enriched Polysaccharide (Se-Le-30) Isolated from Lentinula edodes Mycelium on Human Immune Cells.

A common edible mushroom Lentinula edodes, is an important source of numerous biologically active substances, including polysaccharides, with immunomodulatory and antitumor properties. In the present work, the biological activity of the crude, homogenous (Se)-enriched fraction (named Se-Le-30), which has been isolated from L. edodes mycelium by a modified Chihara method towards human peripheral blood mononuclear cells (PBMCs) and peripheral granulocytes, was investigated. The Se-Le-30 fraction, an analog of lentinan, significantly inhibited the proliferation of human PBMCs stimulated with anti-CD3 antibodies or allostimulated, and down-regulated the production of tumor necrosis factor (TNF)-α by CD3+ T cells. Moreover, it was found that Se-Le-30 significantly reduced the cytotoxic activity of human natural killer (NK) cells. The results suggested the selective immunosuppressive activity of this fraction, which is non-typical for mushroom derived polysaccharides.

Purification and comparative study of bioactivities of a natural selenized polysaccharide from Ganoderma Lucidum mycelia.

The development of selenized polysaccharides is a promising strategy for the dietary selenium supplementation. The purpose of this research is to determine the influence of selenium on the structure and bioactivity of a polysaccharide fraction (MPN) isolated from Ganoderma lucidum mycelia. After biological selenium enrichment, the selenium content in the selenized polysaccharide (SeMPN) was 18.91 ± 1.8 µg/g. SeMPN had a slightly lower molecular weight than MPN, but the carbohydrate content and monosaccharide composition remained identical. Additionally, the band at 606 cm-1 in MPN changed to 615 cm-1 in SeMPN as revealed by FT-IR spectra. No significant changes were observed in the types and ratios of glycosidic linkages, as determined by NMR spectroscopy. Extracellular and intracellular antioxidant assays demonstrated that SeMPN was more effective than MPN in scavenging free radicals, inhibiting AAPH-induced erythrocyte hemolysis, and protecting catalase (CAT) and glutathione peroxidase (GSH-Px) activity in H2O2-injured PC12 cells. Additionally, SeMPN had a higher increase effect on RAW 264.7 cells's pinocytic and phagocytic capacity, as well as their production of NO, TNF-α, and IL-6. SeMPN could be as potential functional selenium supplementation.

Immunostimulatory effects of the intracellular polysaccharides isolated from liquid culture of Ophiocordyceps sinensis (Ascomycetes) on RAW264.7 cells via the MAPK and PI3K/Akt signaling pathways.

ETHNOPHARMACOLOGICAL RELEVANCE: Chinese Cordyceps (DongChong XiaCao), a parasitic complex of a fungus Ophiocordyceps sinensis and a caterpillar, is a traditional Chinese medicine. Polysaccharides extracted from O. sinensis have immunomodulatory effects on macrophages. However, the mechanism of polysaccharides on macrophage and the composition of polysaccharides are not known. AIM OF STUDY: We aimed to investigate composition and structure of the intracellular polysaccharides from O. sinensis mycelia (designed as OSP), and evaluate its the immunomodulatory effect on macrophages and its underlying mechanism. MATERIALS AND METHODS: We performed a liquid-state fermentation of O. sinensis to produce mycelia. The DEAE-Sephadex-A25 cellulose column and Sephadex-G100 gel column chromatography were employed to purify and character the intracellular OSP. Macrophages RAW264.7 cells were employed to evaluate OSP's immunomodulatory activity and the possible mechanism responsible for the activation of macrophages in vitro. RESULTS: The average molecular weight of OSP was distributed at 27,972 Da, OSP was composed of xylose, mannose, glucose, and galactose with the ratio of 2.9 : 6.6 : 166 : 2.6, with a trace amount of fucose, arabinose and rhamnose. The phagocytosis of RAW264.7 cells was improved significantly and remarkable changes were observed in the morphology with OSP-treated cells. Real-time quantitative polymerase chain reaction (RT-qPCR) analysis demonstrated that OSP had an ability to regulate the mRNA expression of pro-inflammatory and anti-inflammatory cytokines, and to induce the mRNA expression level of iNOS in a concentration dependent manner in RAW264.7 cells. Western blotting analysis showed that the regulation of NO and cytokines was mediated through mitogen-activated protein kinase (MAPK) and PI3K/Akt signaling pathways. CONCLUSION: This study demonstrated that OSP was with a capacity to activate macrophage cells RAW264.7 for an improvement of immunomodulation activities, which was through regulation of inflammatory mediators via MAPK and PI3K/Akt signaling pathways.

Effect of Cytokinin-Containing Extracts from Some Medicinal Mushroom Mycelia on HepG2 Cells In Vitro.

Endogenous cytokinins in mycelia of medicinal mushrooms Hericium coralloides and Fomitopsis officinalis grown in vitro were identified using high-performance liquid chromatography coupled with mass spectrometry. High amounts of zeatin-type cytokinins and isopentenyladenine were found. The qualitative composition and quantitative content of cytokinins were species-specific traits of mushrooms. Optical microscopy was used to perform a comparison analysis of the influence of crude extracts and purified cytokinin fractions from both species' mycelial biomass on HepG2 tumor cell growth in vitro and morphology. The results showed that purified cytokinin fractions from H. coralloides and F. officinalis mycelia demonstrated a cytotoxic effect on HepG2 cells, unlike crude extracts. Under the influence of all mushroom extracts, similar patterns of changes in HepG2 cell morphology were observed, but they were more pronounced for H. coralloides compared with F. officinalis. Purified fractions of both mushroom species caused an increased level of apoptosis compared to crude extracts. Some increase in glucose uptake by cultured cells was found in all investigated samples wherein the influence of H. coralloides extracts was approximately twice the effect of the corresponding F. officinalis extracts. The data obtained confirm the assumption that cytokinins are involved in the expression of therapeutic effects of medicinal mushrooms and indicate the need to take into consideration the methods of cytokinin extraction when preparing pharmacologically active drugs based on fungal raw materials. Thus, extracts from H. coralloides and F. officinalis mycelial biomass are promising in the search for anticancer agents.

Enhanced Mycelium Production of Phellinus igniarius (Agaricomycetes) Using a He-Ne Laser with Pulsed Light.

This study used a He-Ne laser with pulsed light irradiation to produce mutant strains of Phellinus igniarius strain JQ9 with enhanced characteristics for fermentation (17.685 ± 3.092 g/L) compared with the parent strain (12.062 ± 1.119 g/L). The combined treatment conditions were as follows: He-Ne laser irradiation for 30 min using a spot diameter of 10 mm, pulsed light treatment power set at 100 J, a treatment distance of 14.5 cm, and a flash frequency of 0.5 s. The production of bioactive polysaccharides and small biocompounds such as polyphenols, flavonoids, and triterpenes increased together with mycelium production. The results showed that polyphenol content was significantly correlated with L*, a*, and b* values (R = -0.594, P < 0.01; R = 0.571, P < 0.01; and R = 0.500, P < 0.05; respectively). Antagonistic and random amplified polymorphic DNA analyses indicated that the genetic material of the screened mutants was altered. The mutant screening using a He-Ne laser with pulsed light irradiation could be an effective method for the development of Phellinus strains and could thus improve mycelium production.

Extraction and characterization of polysaccharide-enriched fractions from Phoma dimorpha mycelial biomass.

Ultrasound-assisted extraction (UAE) and pressurized hot water extraction (PHWE) were tested as advanced clean methods to obtain polysaccharides from Phoma dimorpha mycelial biomass. These methods were compared to conventional extraction (hot water extraction, HWE) in terms of polysaccharides-enriched fractions (PEF) yield. A central composite rotational design was performed for each extraction method to investigate the influence of independent variables on the yield and to help the selection of the condition with the highest yield using water as an extraction solvent. The best extraction condition of PEF yielded 12.02 wt% and was achieved when using UAE with direct sonication for 30 min under the intensity of 75.11 W/cm2 and pulse factor of 0.57. In the kinetic profiles, the highest yield (15.28 wt%) was obtained at 50 °C under an ultrasound intensity of 75.11 W/cm2 and a pulse factor of 0.93. Structural analysis of extracted polysaccharide was performed using Fourier-transform infrared spectroscopy, X-ray diffraction, scanning electron microscopy, and thermal property. The water solubility index, water holding capacity, and emulsification index of PEF were 31.3 ± 1.5%, 138.1 ± 3.2%, and 62.9 ± 2.3%, respectively. The submerged fermentation demonstrates the huge potential of Phoma dimorpha to produce polysaccharides with bioemulsifying properties as a biotechnologically cleaner alternative if compared to commercial petroleum-derived compounds. Furthermore, UAE and PHWE are green technologies, which can be operated at an industrial scale for PEF extraction.

Intraocular pressure-lowering effect of Cordyceps cicadae mycelia extract in a glaucoma rat model.

Glaucoma is a leading cause of irreversible blindness worldwide. This study evaluates the reduction of intraocular pressure (IOP) induced by C. cicadae mycelia extract in a steroid-induced rat model of glaucoma. Cordyceps cicadae mycelia is a well-known and valued traditional Chinese herbal medicine. C. cicadae mycelia were cultured using a liquid fermentation technique. The harvested C. cicadae mycelia were then lyophilized and extracted with two solvents, water and ethanol. The aqueous extract (CCM-DW) and ethanolic extract (CCM-EtOH) of the mycelia were obtained through lyophilization. Sprague Dawley rats were randomly divided into four groups (n = 6 in each group): a normal group, a control group, and experimental groups treated with CCM-DW, or CCM-EtOH (both at 50 mg/kg/body weight). Except for those in the normal group, all rats received a subconjunctival injection of betamethasone to induce high IOP. The rats in the experimental groups received a daily administration of CCM by oral gavage for four consecutive weeks. IOP reduction is the known treatment for glaucoma. The results revealed that steroid treatment caused a significant increase in the animals' IOP (control group). Elevated IOP decreased significantly after treatment with CCM-DW and CCM-EtOH (p < 0.01), and CCM-DW was more effective than CCM-EtOH. CCM-DW and CCM-EtOH were capable of causing significant decreases in high IOP-induced lesions in pathological studies in which it was shown that the efficacy of CCM-DW surpassed that of CCM-EtOH. After CCM-DW administration for 28 days, there were significant decreases in malondialdehyde and lactate dehydrogenase levels and significant increases in catalase, superoxide dismutase, and glutathione peroxidase levels. In summary, C. cicadae mycelia may be beneficial for preventing or treating glaucoma due to its significant IOP-lowering and antioxidant activities.

Antioxidation, anti-inflammation and anti-fibrosis effect of phosphorylated polysaccharides from Pleurotus djamor mycelia on adenine-induced chronic renal failure mice.

The mycelia polysaccharides (MPS) from Pleurotus djamor were prepared and purified by anion exchange column chromatography, and the phosphate content of phosphorylated MPS (PMPS) was 15.22 ± 0.37%. FT-IR spectra, HPLC and 1H and 13C-NMR results showed the PMPS contained α-pyranose structure and the peak area percentage composition of galacturonic acid and glucose were 13.01% and 85.82%, respectively. Animal experiment investigated the antioxidant, anti-inflammation, anti-fibrosis effects of PMPS on kidney in adenine-induced chronic renal failure (CRF) mice. All results including serum biochemical indices, histopathological observation, qRT-PCR, western blotting, immunohistochemical staining manifested the kidney injury could be remitted by PMPS interventions. This experiment suggested that PMPS could remit CRF and other kidney injury related diseases as one kind of dietary supplements and functional foods without toxic side effects.