Evaluation of Polysaccharide Content in Shiitake Culinary-Medicinal Mushroom, Lentinula edodes (Agaricomycetes), via Near-Infrared Spectroscopy Integrated with Deep Learning.

Polysaccharide is one of the bioactive ingredients extracted from the fruiting body of Lentinula edodes (=L. edodes), which has many medicinal functions. While the content of polysaccharide can be measured by near-infrared (NIR) spectroscopy, the NIR analytical models established previously only covered L. edodes from very limited sources, and thus could not achieve high accuracy for large samples from more varied sources. Strictly, there is a nonlinear relationship between NIR spectral data and chemical label values, and traditional modeling methods for NIR data analysis have problems such as insufficient feature learning ability and difficulty in training. The deep learning model has excellent nonlinear modeling ability and generalization capacity, which is very suitable for analyzing larger samples. In this study, we constructed a novel framework with deep learning techniques on the NIR analysis of the content of polysaccharide in L. edodes. The siPLS model was established based on the combination of the bands 4797-3995 cm-1 and 6401-5600 cm-1, while the one-dimensional convolutional neural network (1D-CNN) model was established with improved feature in the treatment of the spectral data. The comparative experimental results showed that the 1D-CNN model (R2pre = 95.50%; RMSEP =0.1875) outperformed the siPLS model (R2pre = 87.89%, RMSEP = 0.6221). As such, this work has demonstrated that NIR spectroscopy with the integration of deep learning can provide more accurate quantification of polysaccharide in L. edodes. Such method can be very useful for nutritional grading and quality control of diverse L. edodes in the market.

Salt reducing and saltiness perception enhancing strategy for shiitake (Lentinus edodes) bud using novel combined treatment of yeast extract and radio frequency.

This study investigated the potential of yeast extract and radio frequency (RF) treatment as a strategy of reducing salt and enhancing saltiness perception for Lentinus edodes bud. The results of E-nose demonstrated yeast extract and RF treatment improved the saltiness of Lentinus edodes bud. Meanwhile, yeast extract and RF treatment significantly decreased the addition of salt (P < 0.05), and led to the formation of special flavor substances, whereas amino acid nitrogen content decreased. On the other hand, sensory attribute, hardness, total flavonoid and phenolic content, antioxidant capacity of L. edodes buds significantly (P < 0.05) increased after the yeast extract combined with RF treatment. In addition, the modification of water distribution, the formation of dense structure, uniform microstructure and Na+ distribution were observed in treated sample, causing the enhancement of saltiness perception. Accordingly, the alteration of properties contributed to higher sensory properties of texture, taste, flavor, and overall acceptability.

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.

In Vivo Evaluation and Nutritional Quality of By-Products Subjected to Solid-State Fermentation Using Shiitake Culinary-Medicinal Mushroom, Lentinula edodes (Agaricomycetes).

This study evaluates the nutritional quality and in vivo biological activity of a peach palm by-product food ingredient processed via solid-state fermentation by shiitake culinary-medicinal mushroom, Lentinula edodes. The group of Wistar rats that consumed this diet had higher total dietary fiber content, digestibility, rate of protein quality, and protein efficiency. They also presented a late and softer insulinemic peak with an increase in the glycemic index, demonstrating amino acid limitation but with a feasible matrix as a complement protein. Discrete variation on total cholesterol and triglycerides was observed with a reduction in lipid profile, attributed to its high dietary fiber content. Lipids from within the liver and stool revealed that the fermented diet contained the lowest rates of fat in the liver and, consequently, the highest elimination compared to the other control diets. The serum lipid profile suggests a positive modulation of this diet, and that it has good nutritional quality with the potential to positively influence glycemic and lipid profiles.

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.

Combining the Anticancer and Immunomodulatory Effects of Astragalus and Shiitake as an Integrated Therapeutic Approach.

Astragalus root (Huang Qi) and Shiitake mushrooms (Lentinus edodes) are both considered medicinal foods and are frequently used in traditional Chinese medicine due to their anticancer and immunomodulating properties. Here, the scientific literatures describing evidence for the anticancer and immunogenic properties of Shiitake and Astragalus were reviewed. Based on our experimental data, the potential to develop medicinal food with combined bioactivities was assessed using Shiitake mushrooms grown over Astragalus beds in a proprietary manufacturing process, as a novel cancer prevention approach. Notably, our data suggest that this new manufacturing process can result in transfer and increased bioavailability of Astragalus polysaccharides with therapeutic potential into edible Shiitake. Further research efforts are required to validate the therapeutic potential of this new Hengshan Astragalus Shiitake medicinal food.

Nutritional impact of active hexose-correlated compound for patients with resectable or borderline-resectable pancreatic cancer treated with neoadjuvant therapy.

Active hexose-correlated compound (AHCC) is a standardized extract from cultured Lentinula edodes mycelia, used as a potent biological response modifier in cancer treatment. We evaluated the nutritional effect of AHCC, given during neoadjuvant therapy, to patients with pancreatic ductal adenocarcinoma (PDAC). Thirty patients with resectable or borderline-resectable PDAC received neoadjuvant therapy with gemcitabine plus S-1. We compared, retrospectively, the outcomes of 15 patients who received AHCC combined with neoadjuvant therapy with those of 15 patients who did not receive AHCC combined with neoadjuvant therapy. The median changes of the neutrophil-to-lymphocyte ratio (NLR) and prognostic nutrition index (PNI) were significantly better in the AHCC group. The relative dose intensity of neoadjuvant therapy was also significantly higher in the AHCC group. Thus, AHCC may improve the nutritional status during neoadjuvant therapy of patients with pancreatic ductal adenocarcinoma. To validate these results and examine the long-term impact of AHCC, a prospective phase II study for PDAC is ongoing.

Evaluation of the chemical qualities and microstructural changes of Lentinula edodes caused by airborne ultrasonic treatment combined with microwave vacuum drying.

This study analyzed a new drying method using airborne ultrasound combined with microwave vacuum to study its effect on the quality characteristics and microstructure of dehydrated L. edodes. Ultrasonic treatment resulted in many micropores in the product, forming the sponge effect caused by ultrasonic waves, which can promote the rapid evaporation of water in the product. Samples of Lentinula edodes individuals were dried using four methods: hot air drying (HAD), microwave vacuum drying (MVD), microwave vacuum drying after ultrasonic pretreatment (US+MVD) and airborne ultrasonic treatment combined with microwave vacuum drying (USMVD). The results showed that USMVD can reduced the loss of total sugar, total phenol, and total antioxidants in L. edodes, and increased the relative content of ergosterol, sulfur compounds, and free amino acids (p < 0.05). Scanning electron microscope observation revealed that USMVD resulted in a uniform reticular porous structure, which could better maintain desirable levels of nutrients. Therefore, USMVD can produce high quality products. PRACTICAL APPLICATION: Airborne ultrasonic waves combined with MVD provides an innovative drying method for mushrooms, which has not been studied at present. The mixed drying method has great potential in maintaining product quality. It provides a theoretical basis for studying drying technology in the future.

Antioxidant and Anti-Inflammatory Potential of Shiitake Culinary-Medicinal Mushroom, Lentinus edodes (Agaricomycetes), Sporophores from Various Culture Conditions.

Lentinus edodes (= Lentinula edodes) is an edible mushroom grown and marketed for centuries due to its nutritional and medicinal properties. L. edodes has multiple pharmacological activities as an antioxidant and anti-inflammatory. Few studies were performed taking into account the influence of culture conditions to optimize the biological properties of L. edodes on human health. Our work focused on the comparison of antioxidant capacity and anti-inflammatory activity of L. edodes fruit bodies cultivated by three mushroom producers in the French Occitanie region using the same strain in various growing conditions (organic and nonorganic). Sequential extraction was performed on freeze-dried fungal materials. All extracts have a quantifiable but moderate antioxidant activity measured via DPPH and ORAC tests. The anti-inflammatory activity of the ethanol and aqueous extracts was evaluated on a model of inflammatory macrophages. The ethanol extracts inhibit NO production in a dose-dependent manner when the cells are pretreated for 4 h with a 24 h stimulation time.

Purified ß-glucans from the Shiitake mushroom ameliorates antibiotic-resistant Klebsiella pneumoniae-induced pulmonary sepsis.

Bacterial infection remains the main cause of acute respiratory distress syndrome and is a leading cause of death and disability in critically ill patients. Here we report on the use of purified ß-glucan (lentinan) extracts from Lentinus edodes (Shiitake) mushroom that can reduce infection by a multidrug-resistant clinical isolate of Klebsiella pneumoniae in a rodent pneumonia model, likely through immunomodulation. Adult male Sprague-Dawley rats were subjected to intra-tracheal administration of K. pneumoniae to induce pulmonary sepsis and randomized to three groups; vehicle control (Vehicle, n = 12), commercial lentinan (CL, n = 8) or in-house extracted lentinan (IHL, n = 8) were administered intravenously 1 h postinfection. Physiological parameters and blood gas analysis were measured, bacterial counts from bronchoalveolar-lavage (BAL) were determined, along with differential staining of white cells and measurement of protein concentration in BAL 48 h after pneumonia induction. Use of IHL extract significantly decreased BAL CFU counts. Both CL and IHL extractions reduced protein concentration in BAL. Use of IHL resulted in an improvement in physiological parameters compared to controls and CL. In conclusion, administration of lentinan to treat sepsis-induced lung injury appears safe and effective and may exert its effects in an immunomodulatory manner.

Preparation of lentinan-calcium carbonate microspheres and their application as vaccine adjuvants.

Adjuvants improve vaccine potency by enhancing immunogenicity and sustaining long-term immune responses. Lentinan (LNT), a ß-1,3-glucohexaose with ß-1,6-branches, is extracted from the mushroom Lentinus edodes and functions as an effective immunostimulatory drug. Previous studies have demonstrated the adjuvant activity of calcium carbonate (CaCO3) microspheres as well as their use as antigen delivery systems. In this study, we successfully loaded CaCO3 microspheres with LNT and evaluated their physicochemical characteristics prior to the adsorption of ovalbumin. Our experimental results demonstrated that LNT-CaCO3 significantly enhanced lymphocyte proliferation, and boosted the frequency of CD69 + B cells and the ratio of CD4+ to CD8 + T cells in spleen lymphocytes. Moreover, LNT-CaCO3 unexpectedly induced the secretion of IgG and Th-associated cytokines (IL-2, IL-4, IFN-γ, and TNF-α) in immunized mice. Therefore, LNT-CaCO3 microspheres induce robust cellular and humoral immune responses and have potential utility as vaccine delivery systems.

Protective Role of Shiitake Mushroom-Derived Exosome-Like Nanoparticles in D-Galactosamine and Lipopolysaccharide-Induced Acute Liver Injury in Mice.

Fulminant hepatic failure (FHF) is a rare, life-threatening liver disease with a poor prognosis. Administration of D-galactosamine (GalN) and lipopolysaccharide (LPS) triggers acute liver injury in mice, simulating many clinical features of FHF in humans; therefore, this disease model is often used to investigate potential therapeutic interventions to treat FHF. Recently, suppression of the nucleotide-binding domain and leucine-rich repeat related (NLR) family, pyrin domain containing 3 (NLRP3) inflammasome, was shown to alleviate the severity of GalN/LPS-induced liver damage in mice. Therefore, the goal of this study was to find dietary exosome-like nanoparticles (ELNs) with therapeutic potential in curbing FHF by suppressing the NLRP3 inflammasome. Seven commonly consumed mushrooms were used to extract ELNs. These mushrooms were found to contain ELNs composed of RNAs, proteins, and lipids. Among these mushroom-derived ELNs, only shiitake mushroom-derived ELNs (S-ELNs) substantially inhibited NLRP3 inflammasome activation by preventing inflammasome formation in primary macrophages. S-ELNs also suppressed the secretion of interleukin (IL)-6, as well as both protein and mRNA levels of the Il1b gene. Remarkably, pre-treatment with S-ELNs protected mice from GalN/LPS-induced acute liver injury. Therefore, S-ELNs, identified as potent new inhibitors of the NLRP3 inflammasome, represent a promising class of agents with the potential to combat FHF.

Enhancing the Extraction Yield from Shiitake Culinary-Medicinal Mushroom, Lentinus edodes (Agaricomycetes), by Pulsed Electric Fields Treatment.

Medicinal mushrooms contain highly valuable substances with proven positive effects on human health. To extract these components, different methods are available. Most of them suffer from individual disadvantages, therefore making them economically unviable. Pulsed electric fields (PEFs) could provide an opportunity to improve these processes. PEFs cause pore formation of cell membranes, facilitating substance transport out of cells. Thus, the influence of this technique on the extraction yield of medicinal mushrooms was studied for the first time. Lentinus edodes was used as model case and PEF treatment was compared with standard Soxhlet extraction alone. A square pulse generator (Electro Square Porator™ ECM 830) with a voltage of 3 kV and pulse length of 100 µs was used for PEF treatment. Extraction was studied for fresh and dried fruiting bodies, and dichloromethane and hot water extracts were analyzed. Extracts were quantified gravimetrically, and carbohydrate yields were also determined qualitatively with GC-MS and quantitatively with anthrone method. PEFs could increase in particular the yield of water-soluble compounds of fresh mushroom material. However, the lipid fraction was not affected by PEF in neither fresh nor dried material.

Biovalorization of soybean residue (okara) via fermentation with Ganoderma lucidum and Lentinus edodes to attain products with high anti-osteoporotic effects.

Okara, despite being a soybean processing by-product, still holds many nutrients. Thus, considerable attention has been recently paid to its reuse. In this study, solid-state fermentation was performed using Ganoderma lucidum and Lentinus edodes. Antioxidant activity and bioactive compound levels in G. lucidum-fermented okara (GLFO) and L. edodes-fermented okara (LEFO) were assayed. Antiosteoporosis bioactivity was evaluated using an animal model. The results demonstrated that solid-state fermentation significantly improved the antioxidant activity and bioactive compound levels. Furthermore, GLFO and LEFO increased trabecular bone volume, although only the GLFO-treated group exhibited significantly improved trabecular separation compared with the bilateral ovariectomy-treated control group. GLFO-related outcomes were superior to those of LEFO. The results demonstrate that okara products are effective for treating postmenopausal osteoporosis in humans.

Effects of UV-C treatment and ultrafine-grinding on the biotransformation of ergosterol to vitamin D2, physiochemical properties, and antioxidant properties of shiitake and Jew's ear.

In the present study, the impact of ultraviolet (UV)-C treatment and ultrafine grinding on the conversion of ergosterol to vitamin D2, physiochemical properties, and antioxidant properties of shiitake and Jew's ear was assessed. After exposure to UV-C, vitamin D2 contents of both the mushroom samples has increased significantly (p < 0.05). Whereas, ultrafine grinding along with UV-C treatment has a synergistic effect on bioconversion of ergosterol to vitamin D2 and this effect is more prominent in low dose UV-C irradiation groups (2 kJ/m2). Ultrafine grinding significantly (p < 0.05) improved the water holding capacity (WHC), water solubility index (WSI) and polysaccharide dissolution rate (PDR). However, UV-C treatment led to insignificant changes in the physiochemical properties of mushroom samples. A significant improvement was also observed in the antioxidant profiles especially tannin contents of mushrooms followed by the ultrafine grinding and UV-C treatment.

The Inhibitory Properties of Ethanol Extracts of Some Culinary-Medicinal Mushrooms on the Secretion of Interleukin-8 and Vascular Endothelial Growth Factor by PC3 Cancer Cells.

Interleukin (IL)-8, a cytokine produced by immune and non-immune cells, induces angiogenesis via increased vascular endothelial growth factor (VEGF) secretion; both cytokines promote tumor growth. IL-8 and VEGF plasma levels correlate with prostate cancer severity, suggesting that therapeutic options aimed at their downregulation may modulate tumor growth. Available data suggest that Agaricus bisporus (white button mushroom [WBM]) extracts inhibit cancer cell proliferation through aromatase inhibition. However, the extent to which they affect IL-8 and VEGF remains to be elucidated. The aims of this study were to (1) investigate the antiproliferative properties of WBM, brown A. bisporus (portabella), and Lentinus edodes (shiitake mushroom) on PC3 cancer cells; (2) demonstrate that these properties are exerted through the regulation of both IL-8 and VEGF; and (3) determine the role of NFκB activation in the antiproliferative process of mushroom extracts. Cytokine secretion in the supernatant, NFκB activity, and cell proliferation were measured in PC3 cells incubated with 0-100 µg/mL of ethanol extracts of mushrooms. Mushroom extracts decreased IL-8 secretion and cell proliferation (P < .05), and also tended to decrease VEGF (P < .09). Decreased cell proliferation did not appear to result from cell death because trypan blue exclusion tests showed comparable cell viability among cultures. Mushroom extracts also decreased nuclear and total NFκB activity, and the ratio of nuclear to cytoplasmic activity (P < .05) suggesting altered translocation from the cytoplasm to the nucleus. Our data suggest that the three types of studied mushrooms may modulate tumor growth through inhibition of IL-8, VEGF, and NFκB pathways.

HS-SPME/GC-MS Assisted Analysis of Volatile Constituents in Different Strains of Shiitake Culinary- Medicinal Mushroom, Lentinus edodes (Agaricomycetes).

The shiitake culinary-medicinal mushroom Lentinus edodes is one of the most consumed species worldwide because it has nutritional, medicinal, and palatable properties. Its organoleptic characteristics contribute substantially to its high popularity. The pleasant aromas result from the presence of volatile compounds. The objective of the present work was to study the profiles of volatile constituents of fresh fruiting bodies of five strains of L. edodes. The volatile compounds were extracted by solid phase microextraction method and analyzed by gas chromatography and mass spectrometry. The aromatic profiles of the strains revealed variability. Both alcohols and sulfides were the most abundant volatile compounds. LE6 strain presented the highest number of volatile compounds with predominance of sulfides (dimethyl pentasulfide, s-tetrathiane) and for LE2, the aldehydes were the most representative chemical class with the main volatile being (E)-2-octen-1-al.

Selenium-containing polysaccharides from Lentinula edodes-Biological activity.

We hypothesized that selenium(Se)-enriched polysaccharides would possess superior biological activity when compared to those non-enriched. To verify this hypothesis, we obtained by biotechnological methods a Se-enriched analog of Japanese anticancer drug lentinan and, as a reference, the non-Se-enriched fraction. We tested the effects of the obtained fractions on the proliferation of human peripheral blood mononuclear cells. The results suggested a selective immunosuppressive activity, non-typical for mushroom derived polysaccharides. Both fractions caused significant inhibition of human T lymphocyte proliferation induced by mitogens, without significant effects on B lymphocytes. The inhibitory effect was not due to the toxicity of the examined polysaccharides. In normal (HUVEC) or malignant (HeLa) cells tested fractions significantly enhanced cell viability and protected the cells from oxidative stress conditions. However, we observed no effect of the polysaccharide fractions on the production of reactive oxygen species by granulocytes in vitro. The selenium content increased the biological activity of the tested polysaccharide fractions.

Enhanced Z-isomerization of tomato lycopene through the optimal combination of food ingredients.

In tomatoes, most lycopene is present in the all-E-configuration and shows very low bioavailability, whereas the Z-isomers show higher bioavailability. Hence, for health reasons, it is expected that the ingestion of lycopene Z-isomers is preferable. Very recently, it was reported that onion and possibly garlic promoted thermal Z-isomerization of (all-E)-lycopene but there are no reports for other food ingredients. Here we show new food ingredients that enhance thermal Z-isomerization of lycopene in tomatoes and from the results, we guessed some causative components having the Z-isomerization promoting effect. A comprehensive investigation of food ingredients revealed that some vegetables (Allium sp., Brassica sp., and Raphanus sp.), shiitake mushroom (Lentinus edodes), and some edible seaweeds (Saccharina sp. and Ecklonia sp.) markedly promoted Z-isomerization of (all-E)-lycopene in tomato puree with heating at 80 °C for 1 h. Moreover, it was revealed that polysulfides, isothiocyanates, carbon disulfide, and iodine, which were commonly contained in the above food ingredients in considerable quantity, enhanced thermal Z-isomerization of (all-E)-lycopene. Our findings on the food ingredients and the food-derived catalysts having a carotenoid Z-isomerization promoting effect are important, not only for the food, drink, and dietary supplement manufacturing industries, but also for daily home cooking.

Lentinan administration relieves gut barrier dysfunction induced by rotavirus in a weaned piglet model.

Rotavirus (RV) is a pathogen that induces severe diarrhea in infants and young animals. Shiitake mushroom is a traditional food, which can improve physiological function, including gut health. Lentinan (LNT) is the main functional component of Shiitake mushroom. This study aimed to verify whether LNT administration could improve intestinal barrier function, thereby decreasing RV-induced diarrhea in a porcine model. According to initial weight and origin, a total of 28 weaned piglets were randomly fed 2 diets containing 0 or 84 mg kg-1 LNT for 19 d (n = 14). On day 15, RV was orally infused to half of the pigs in each group. RV-induced diarrhea (P < 0.05), the positive rate of RV non-structural protein 4 (NSP4), impaired intestinal morphology, antioxidant capacity and microbiota (P < 0.05), and increased apoptosis of jejunal epithelial cells (P < 0.05) were assessed in the piglets. Dietary LNT supplementation was found to improve intestinal morphology, permeability, antioxidant capacity and microbiota (P < 0.05). Supplementation also further alleviated the effects of RV infection on diarrhea, intestinal morphology, permeability, antioxidant capacity, microbiota and apoptosis of jejunal epithelial cells in piglets (P < 0.05). Thus, these results suggest that LNT administration relieved RV-induced diarrhea in piglets, which could be due to the increase in antioxidant capacity, reduction in apoptosis and improvement of the microbiota-increased gut barrier.