Lentinus edodes mycelium polysaccharide inhibits AGEs-induced HUVECs pyroptosis by regulating LncRNA MALAT1/miR-199b/mTOR axis and NLRP3/Caspase-1/GSDMD pathway.

A novel Lentinus edodes mycelia polysaccharide (LMP) prepared in our laboratory has been identified to be effective in inhibiting the damage of islet ß cells induced by glucose toxicity. However, whether it can effectively alleviate the pyroptosis of human umbilical vein endothelial cells (HUVECs) induced by advanced glycation end products (AGEs) remains unclear. Bioinformatics and cell biology techniques were used to explore the mechanism of LMP inhibiting AGEs-induced HUVECs damage. The results indicated that AGEs significantly increased the expression of LncRNA MALAT1, decreased cell viability to 79.67 %, increased intracellular ROS level to 248.19 % compared with the control group, which further led to cell membrane rupture. The release of LDH in cellular supernatant was increased to 149.42 %, and the rate of propidium iodide staining positive cells increased to 277.19 %, indicating the cell pyroptosis occurred. However, the above trend was effectively retrieved after the treatment with LMP. LMP effectively decreased the expression of LncRNA MALAT1 and mTOR, promoted the expression of miR-199b, inhibited AGEs-induced HUVECs pyroptosis by regulating the NLRP3/Caspase-1/GSDMD pathway. LncRNA MALAT1 might be a new target for LMP to inhibit AGEs-induced HUVECs pyroptosis. This study manifested the role of LMP in improving diabetes angiopathy and broadens the application of polysaccharide.

Detoxification of tetracycline and synthetic dyes by a newly characterized Lentinula edodes laccase, and safety assessment using proteomic analysis.

Fungal laccase has strong ability in detoxification of many environmental contaminants. A putative laccase gene, LeLac12, from Lentinula edodes was screened by secretome approach. LeLac12 was heterogeneously expressed and purified to characterize its enzymatic properties to evaluate its potential use in bioremediation. This study showed that the extracellular fungal laccase from L. edodes could effectively degrade tetracycline (TET) and the synthetic dye Acid Green 25 (AG). The growth inhibition of Escherichia coli and Bacillus subtilis by TET revealed that the antimicrobial activity was significantly reduced after treatment with the laccase-HBT system. 16 transformation products of TET were identified by UPLC-MS-TOF during the laccase-HBT oxidation process. Gas chromatography-mass spectrometry (GC-MS) analysis revealed that LeLac12 could completely mineralize ring-cleavage products. LeLac12 completely catalyzed 50 mg/L TET within 4 h by adding AG (200 mg/L), while the degradation of AG was above 96% even in the co-contamination system. Proteomic analysis revealed that central carbon metabolism, energy metabolism, and DNA replication/repair were affected by TET treatment and the latter system could contribute to the formation of multidrug-resistant strains. The results demonstrate that LeLac12 is an efficient and environmentally method for the removal of antibiotics and dyes in the complex polluted wastewater.

Analysis of Volatile Aroma Components in Different Parts of Shiitake Mushroom (Lentinus edodes) Treated with Ultraviolet C Light-Emitting Diodes Based on Gas Chromatography-Ion Mobility Spectroscopy.

Further assessment of ultraviolet C light-emitting diode (UVC-LED) irradiation for influencing shiitake mushrooms' (Lentinus edodes) volatile and sensory properties is needed. In this study, a comparison of UVC-LED irradiation treatment on the flavor profiles in various parts of shiitake mushrooms was conducted using gas chromatography-ion mobility spectrometry (GC-IMS) and sensory analysis. Sixty-three volatile compounds were identified in shiitake mushrooms. The fresh shiitake mushrooms were characterized by the highest values of raw mushroom odors. After UVC-LED treatment, the content of C8 alcohols decreased, especially that of 1-octen-3-ol, while the content of aldehydes increased, especially the content of nonanal and decanal. The score of fatty and green odors was enhanced. For fresh samples, the mushroom odors decreased and the mushroom-like odors weakened more sharply when treated in ethanol suspension than when treated with direct irradiation. The fruit odors were enhanced using direct UVC-LED irradiation for fresh mushroom samples and the onion flavor decreased. As for shiitake mushroom powder in ethanol suspension treated with UVC-LED, the sweaty and almond odor scores decreased and the vitamin D2 content in mushroom caps and stems reached 668.79 µg/g (dw) and 399.45 µg/g (dw), respectively. The results obtained from this study demonstrate that UVC-LED treatment produced rich-flavored, quality mushroom products.

Edodin: A New Type of Toxin from Shiitake Mushroom (Lentinula edodes) That Inactivates Mammalian Ribosomes.

Ribosome-inactivating proteins (RIPs) are a group of proteins with rRNA N-glycosylase activity that irreversibly inhibit protein synthesis and consequently cause cell death. Recently, an RIP called ledodin has been found in shiitake; it is cytotoxic, strongly inhibits protein synthesis, and shows rRNA N-glycosylase activity. In this work, we isolated and characterized a 50 kDa cytotoxic protein from shiitake that we named edodin. Edodin inhibits protein synthesis in a mammalian cell-free system, but not in insect-, yeast-, and bacteria-derived systems. It exhibits rRNA N-glycosylase and DNA-nicking activities, which relate it to plant RIPs. It was also shown to be toxic to HeLa and COLO 320 cells. Its structure is not related to other RIPs found in plants, bacteria, or fungi, but, instead, it presents the characteristic structure of the fold type I of pyridoxal phosphate-dependent enzymes. Homologous sequences have been found in other fungi of the class Agaricomycetes; thus, edodin could be a new type of toxin present in many fungi, some of them edible, which makes them of great interest in health, both for their involvement in food safety and for their potential biomedical and biotechnological applications.

Near-gapless genome and transcriptome analyses provide insights into fruiting body development in Lentinula edodes.

Fruiting body development in macrofungi is an intensive research subject. In this study, high-quality genomes were assembled for two sexually compatible monokaryons from a heterokaryotic Lentinula edodes strain WX1, and variations in L. edodes genomes were analyzed. Specifically, differential gene expression and allele-specific expression (ASE) were analyzed using the two monokaryotic genomes and transcriptome data from four different stages of fruiting body development in WX1. Results revealed that after aeration, mycelia sensed cell wall stress, pheromones, and a decrease in CO2 concentration, leading to up-regulated expression in genes related to cell adhesion, cell wall remodeling, proteolysis, and lipid metabolism, which may promote primordium differentiation. Aquaporin genes and those related to proteolysis, mitosis, lipid, and carbohydrate metabolism may play important roles in primordium development, while genes related to tissue differentiation and sexual reproduction were active in fruiting body. Several essential genes for fruiting body development were allele-specifically expressed and the two nuclear types could synergistically regulate fruiting body development by dominantly expressing genes with different functions. ASE was probably induced by long terminal repeat-retrotransposons. Findings here contribute to the further understanding of the mechanism of fruiting body development in macrofungi.

The improvement effects of Lentinus edodes powder marination on sous vide cooked chicken patties: Physicochemical attributes, oxidative properties and flavor characteristics.

The improvement effects of Lentinus edodes powder (LEP) marination with different concentrations (0, 6-14 %) on physicochemical, oxidative and flavor quality of chicken patties were evaluated. Greater pH, redness, yellowness, water holding capacity and their strong correlations were observed in LEP-marinated samples. Changed water distribution, inhibited lipid oxidation and enhanced protein oxidation occurred through LEP marination. The highest gel strength and resilience and the lowest hardness and chewiness were obtained in 10 % LEP-marinated sample. Meanwhile, taste activity values of amino acids and saltiness peaked and umami rose in this sample. 124 volatiles were detected and 16 compounds were simultaneously detected by gas chromatography-ion mobility spectrometry and gas chromatography-mass spectrometry. Hexanal, 1,2,4-trithiolane and 1-hexanol were considered as the key differential aroma-active compounds according to odor activity values and chemometric analysis. This study confirmed LEP as a prospective ingredient to improve the quality of meat products.

The Tumor Growth Inhibitory Effect of a Standardized Extract of Cultured Lentinula edodes Mycelia Using Patient Derived Xenograft Model.

Patient derived xenograft (PDX) is a powerful tool to confirm pharmacological efficacy in non-clinical studies for the development of various drugs including anti-cancer agents and therapeutic research. A standardized extract of cultured Lentinula edodes mycelia, a product name AHCC® is produced by Amino Up Co., Ltd. (Sapporo, Japan). In this study, we investigated the inhibitory effect of AHCC® on the growth of tumor PDX in Super SCID (severe combined immunodeficiency) mice. Effects of AHCC® and BCG administration on the growth of renal cancer PDX implanted in Super SCID mice were evaluated by PDX growth curve. Tendency for the effects on the growth of renal cancer PDX in Super SCID by administration of AHCC® and BCG before implanting the PDX were demonstrated. The effects of the oral administration of AHCC® on the growth of renal, invasive and non-invasive breast cancer PDX in Super SCID mice were studied. In Super SCID mice transplanted with renal cancer PDX, AHCC® significantly suppressed tumor proliferation from the day 48 to 83 after transplantation. In two types of breast cancer PDX, tendency of the growth inhibitory effects of AHCC® were shown by PDX growth curve. Significant inhibitory effect was found at only one time point for during proliferation in each PDX. Super SCID-PDX model has the potential to be a useful tool to investigate for the effect of functional foods.

In vitro evaluation of methanol extracts of edible fungi Pleurotus ostreatus and Lentinula edodes against Rhyssomatus nigerrimus Fahraeus.

The study aimed to evaluate the insecticidal activity of extracts of edible mushrooms Pleurotus ostreatus and Lentinula edodes against Rhyssomatus nigerrimus. Methanol:water extracts (70:30) of P. ostreatus and L. edodes were made and evaluated in two in vitro tests (exposure and immersion toxic effect) against R. nigerrimus. Subsequently, the toxicity test of the extracts against Artemia salina was evaluated. These extracts were subjected to colorimetric tests and gas chromatography-mass spectrometry. The results showed a mortality effect against R. nigerrimus of 50% for the P. ostreatus 2 extracts at a concentration of 20% in the immersion test. Likewise, in the toxic effect test, 90% mortality was observed after five days of exposure to a concentration of 10%. On the other hand, for the toxicity test, the extract that showed the values with the highest mortality against A. salina was P. ostreatus, starting with 80% mortality at 100µg/mL. The functional groups present in the extracts were saponins, coumarins, and alkaloids. Furthermore, the presence of more than 7 compounds in the mushroom extracts evaluated is reported. This study demonstrates the insecticidal activity of P. ostreatus and L. edodes fungal extracts and indicates the importance of using different in vitro tests to elucidate the mechanism of action for future studies.

Anti-inflammatory and immunomodulatory properties of polysaccharides in mushrooms.

Mushrooms are distinguished as important food-containing polysaccharides possessing potent anti-inflammatory and immunomodulating properties. These compounds belong mostly to polysaccharides that are mostly ß-D-glucans. Among them, ß-1,3-glucan with ß-1,6 side chains of glucose residues, has more important roles in their properties. In this review, we have introduced polysaccharides mainly from Lentinula edodes and Pleurotus citrinopileatus with anti-inflammatory and immunomodulating properties. In addition, the mechanisms of activation of their physiological properties and signal cascade are also reviewed.

Hepatoprotection of a Standardized Extract of Cultured Lentinula edodes Mycelia against Liver Injury Induced by Ischemia-Reperfusion and Partial Hepatectomy.

A standardized extract of cultured Lentinula edodes mycelia (ECLM, AHCC®) has been shown to have beneficial effects on organ metabolism. ECLM has been indicated to have liver protective properties by suppressing inflammatory responses. The pathogenesis of hepatic ischemia-reperfusion injury is thought to involve the induction of inflammatory mediators. However, whether ECLM affects inflammatory mediators caused by warm hepatic ischemia-reperfusion injury and partial hepatectomy (HIRI+PH) has not been clarified. In this study, we evaluated the protective effects of ECLM against liver damage caused by HIRI+PH. Rats were fed a normal diet (HIRI+PH) or a normal diet with 2% ECLM (HIRI+PH and ECLM) for ten days, then the liver and duodenal ligament were clamped and subjected to 15 min of hepatic ischemia. After 70% hepatectomy, the inflow occlusion was released, and liver and blood samples were collected at 3, 6, and 24 h. The effect of ECLM on mortality induced by 30 min of ischemia and hepatectomy was evaluated. The results showed that ECLM attenuated pathological liver damage, including apoptosis, in the rats treated with HIRI+PH, and decreased serum aminotransferase activity; ECLM decreased mRNA levels of the inflammation-related genes inducible nitric oxide synthase and C-X-C motif chemokine ligand 1, and increased mRNA levels of interleukin 10, an anti-inflammatory cytokine; ECLM increased hepatocyte growth factor mRNA levels and Ki-67 labeled nuclei in the liver at 24 h; ECLM significantly reduced HIRI+PH-induced mortality. In conclusion, ECLM may prevent HIRI+PH-induced liver injury in part by suppressing various inflammatory responses and promoting liver regeneration.

Peach palm shells (Bactris gasipaes Kunth) bioconversion by Lentinula edodes: Potential as new bioproducts for beef cattle feeding.

This paper aims to develop and assess the in vitro effects on ruminal fermentation and greenhouse gas parameters of new bioproducts for beef cattle diets, carried out by solid-state fermentation of peach palm shells colonized by Lentinula edodes (SSF) and after Shiitake mushroom cultivation in axenic blocks (SMS). In vitro experiments were performed to assess the in vitro gas production, digestibility, and fiber degradation of formulated total diets. Bioproducts presented high ß-glucans (9.44---11.27 %) and protein (10.04---8.35 %) contents, as well as similar digestibility to conventional diets. SMS diet had the lowest methane and carbon dioxide (19.1 and 84.1 mM/g OM) production, and the SSF diet presented lower carbon dioxide production (98.9 mM/g OM) than other diets, whereas methane was similar. This study highlighted a sustainable use of byproducts for beef cattle diets, promising for digestibility, nutritional value, ß-glucans incorporation, and environmental impact mitigation, favoring the circular bioeconomy.

Photoregulation of the biosynthetic activity of the edible medicinal mushroom Lentinula edodes in vitro.

The findings of the study demonstrate the impact of low-intensity laser and quasi-monochromatic light on the biosynthetic activity of the edible medicinal fungus L. edodes during submerged cultivation. An artificial lighting installation based on matrices of light-emitting diodes (LED) emitting light at 470 nm (blue), 530 nm (green), 650 nm (red), and argon gas laser (488 nm) was used. Irradiation with blue and red LED and laser led to a shortening of the lag phase by 2 days and an increase in the mycelial mass. Irradiation with laser light resulted in the highest mycelial mass yield (14.1 g/L) on the 8th day of cultivation. Irradiation in all used wavelength ranges caused an increase in the synthesis of both extracellular and intracellular polysaccharides. Laser light at 488 nm and LED at 470 nm proved to be the most effective. Irradiation with red, green, and blue laser light caused an increase in the total amount of fatty acids in the mycelial mass compared to the control. A significant distinction in qualitative composition was observed: short-chain acids C6‒C12 compounds were produced under red light irradiation, whereas long-chain C20‒C24 were formed under green light irradiation. The most significant changes in the aromatic profile of the mycelial mass and culture liquid were recorded upon irradiation with green light. The content of aromatic components increased 24.6 times in the mycelial mass and 38.5 times in the culture liquid. The results suggest the possibility of using low-intensity quasi-monochromatic light for targeted regulation of L. edodes biosynthetic activity.

Modulatory effects of supplementation of Lentinula edodes mycelia extract and l-arginine on the therapeutic efficacy of immunogenic chemotherapy in colon cancer-bearing mice.

Some chemotherapeutic drugs can induce cancer cell death and enhance antitumor T-cell immunity in cancer-bearing hosts. Immunomodulatory reagents could augment such chemotherapy-induced effects. We previously reported that oral digestion of Lentinula edodes mycelia (L.E.M.) extract or  l-arginine supplementation can augment antitumor T-cell responses in cancer-bearing mice. In this study, the effects of L.E.M. extract with or without  l-arginine on the therapeutic efficacy of immunogenic chemotherapy by 5-fluorouracil (5-FU)/oxaliplatin (L-OHP) and/or cyclophosphamide (CP) are examined using two mouse colon cancer models. In MC38 and CT26 cancer models, therapy with 5-FU/L-OHP/CP significantly suppressed tumor growth, and supplementation with L.E.M. extract halved the tumor volumes. However, the modulatory effect of L.E.M. extract was not significant. In the CT26 cancer model, supplementation with L.E.M. extract and  l-arginine had no clear effect on tumor growth. In contrast, their addition to chemotherapy halved the tumor volumes, although the effect was not significant. There was no difference in the cytotoxicity of tumor-specific cytotoxic T cells generated from CT26-cured mice treated by chemotherapy alone versus chemotherapy combined with L.E.M. extract/ l-arginine. These results indicate that the antitumor effects of immunogenic chemotherapy were too strong to ascertain the effects of supplementation of L.E.M. extract and  l-arginine, but these reagents nonetheless have immunomodulatory effects on the therapeutic efficacy of immunogenic chemotherapy in colon cancer-bearing mice.

Effects of different cooking methods on proximate composition, digestion characteristics, and antioxidant activity of Lentinus edodes.

Lentinus edodes (SM) are highly appreciated by the food industry together with consumers for nutrition and flavour. It is hypothesized the results of our study can indicate to the suitable cooking method for maximal nutrient retention, this study investigated the effects of six cooking methods on the proximate composition, digestive properties, and antioxidant activities of SM in vitro simulated digestion. The results revealed that the ash and protein contents of cooked samples were reduced excluding steaming and roasting, and likewise minerals during deep-frying and boiling (e.g., Mg from 1080.07 to 629.03 mg/kg, deep-frying). A conspicuous rise in fat and energy was found in deep-frying and stir-frying. Steaming retained more reducing sugar (3.80 mg/mL). Roasting improved bioaccessibility of most amino acids (e.g., 82.61%, Asp). Using oil as a medium was associated with higher antioxidant activities using water. Steaming and roasting were better for preserving the nutrient composition of cooked Lentinus edodes.

Effect of postharvest storage time on quality characteristics of explosion puffing dried whole shiitake mushroom (Lentinula edodes) crisps.

BACKGROUND: Non-fried shiitake mushroom (Lentinula edodes) crisps fabricated by explosion puffing drying (EPD) are receiving worldwide attention because of their crispness, convenience, nutrition and health functions. The quality of mushroom crisps varies with storage time of fresh L. edodes. Therefore, the effect of postharvest storage time (ranging from 0 to 14 days) of fresh L. edodes on quality characteristics of EPD- processed mushroom crisps was evaluated. RESULTS: The weight loss and total color difference of fresh L. edodes were increased to 2.95% and 24.66, but moisture content, firmness and lightness were reduced by 6.14%, 40.70% and 43.57%, respectively, after 14 days storage. The puffing degree of mushroom crisps was initially increased to its highest value (55.95%) on the 4th day storage and thereafter decreased. The highest rehydration ratio (2.36) and crispness (63.67), and lowest hardness (102.95 N) of mushroom crisps were fabricated with L. edodes on the 4th day of storage. Free water was predominant in fresh L. edodes, which was decreased for fresh L. edodes, whereas it increased initially to the maximum value and decreased thereafter for osmotic dehydrated and heat pump pre-dried L. edodes with increasing storage time. Principal component analysis and hierarchical cluster analysis confirmed that fresh L. edodes stored at different times had a remarkable effect on quality characteristics of mushroom crisps. CONCLUSION: Fresh L. edodes stored at 4 ± 1 °C for 4 days is recommended for fabrication of mushroom crisps with superior quality. This study provides a theoretical basis for selection of a suitable storage time for fresh L. edodes before EPD of crisps. © 2023 Society of Chemical Industry.

Production of Extracellular Enzymes by Lentinula edodes Strains in Solid-State Fermentation on Lignocellulosic Biomass Sterilized by Physical and Chemical Methods.

Two methods of sterilization of lignocellulosic biomass were performed in this study. Eucalypt waste (EW) supplemented with rice bran (RB) was added in the proportions 80:20 and 90:10 in dry weight. The compositions were sterilized by physical method (autoclaving) and by chemical method (H2O2). The production of extracellular enzymes by Lentinula edodes strains was compared within the two methods. Inactivation of catalase present in RB was achieved with 250 mM H2O2. The use of H2O2, when compared by physical method, favored high production of hydrolytic enzymes such as endoglucanase (1,600 IU/kg), twofold higher, ß-glucosidase (1,000 IU/kg), fivefold higher, xylanase (55,000 IU/kg), threefold higher and ß-xylosidase (225 IU/kg), similar results. Oxidative enzymes, MnP and laccase, were produced within a different profile between strains, with shorter times for laccase (2,200 IU/kg) by SJC in 45 days and MnP (2,000 IU/kg) by CCB-514 in 30 days. High production of extracellular enzymes is achieved by the use of the chemical method of sterilization of lignocellulosic biomass; in addition to no energy consumption, this process is carried out in a shorter execution time when compared to the physical process. The use of H2O2 in sterilization does not produce toxic compounds from the degradation of hemicellulose and cellulose such as furfural and hydroxy-methyl-furfural that cause inhibition of microorganisms and enzymes.

Water concentration and rate of decrease in shiitake cultivation log during fruiting body development, as measured by MRI.

Large shiitake mushrooms (Lentinula edodes, pileus > 8 cm in diameter) are difficult to cultivate and account for only 3-5% of the total harvest. This study focused on the water absorption process within a log during the growth of fruiting bodies in order to increase the yield of large shiitake mushrooms. Konara oak logs (Quercus serrata, 85-95 mm in diameter, 290 mm in length) were inoculated with shiitake mycelium plugs and nine months later, young fruiting bodies developed, at which point the log was analyzed using magnetic resonance imaging (MRI) over a period of two weeks. The signal intensity and T1 and T2 relaxation time constants were determined from the acquired images, along with the distribution of water concentration within the entire log. The axial distributions of water concentrations in the log were higher in the 80 mm region around the fruiting body. The rate of decrease in water concentration indicated that water was supplied to the fruiting body from 80 mm axially in the upper half of the sapwood in the log. On the other hand, the water concentration in the heartwood did not decrease and the heartwood did not contribute to the water supply to the fruiting bodies.

A non-invasive method for measuring time-series of moisture concentrations in mycelial blocks during shiitake mushroom development using magnetic resonance imaging.

The moisture concentration in mycelial block is an important factor for increasing the yield of high-quality shiitake mushrooms (Lentinula edodes) with a pileus of 4-5cm or more in mycelial block cultivation. Here, we show a novel way to measure moisture concentration in mycelial blocks using magnetic resonance imaging (MRI). The culture medium was inoculated with Hokken No. 607 and mycelial blocks were incubated and their moisture concentration was measured using MRI. A method was developed to calculate the spatial distribution of moisture concentration inside the mycelial blocks by measuring the total water mass in the mycelial block using mass method and creating a calibration line. During the maturation phase of the mycelial block (46-98 days of incubation), the moisture concentration in the top region of the mycelial block decreased once at 66 days of incubation and then gradually increased. The increase in moisture concentration was due to mycelia decomposing the culture medium and producing water. During the growth period of the fruiting body, the moisture concentration in the periphery of the fruiting body increased and, conversely, the moisture concentration in the whole mycelial block decreased because water in the mycelial block moved into the fruiting body.

Lentinula edodes Cultured Extract and Rouxiella badensis subsp. acadiensis (Canan SV-53) Intake Alleviates Immune Deregulation and Inflammation by Modulating Signaling Pathways and Epigenetic Mechanisms.

Puberty is a critical developmental period of life characterized by marked physiological changes, including changes in the immune system and gut microbiota development. Exposure to inflammation induced by immune stressors during puberty has been found to stimulate central inflammation and lead to immune disturbance at distant sites from the gut; however, its enduring effects on gut immunity are not well explored. Therefore, in this study, we used a pubertal lipopolysaccharides (LPS)-induced inflammation mouse model to mimic pubertal exposure to inflammation and dysbiosis. We hypothesized that pubertal LPS-induced inflammation may cause long-term dysfunction in gut immunity by enduring dysregulation of inflammatory signaling and epigenetic changes, while prebiotic/probiotic intake may mitigate the gut immune system deregulation later in life. To this end, four-week-old female Balb/c mice were fed prebiotics/probiotics and exposed to LPS in the pubertal window. To better decipher the acute and enduring immunoprotective effects of biotic intake, we addressed the effect of treatment on interleukin (IL)-17 signaling related-cytokines and pathways. In addition, the effect of treatment on gut microbiota and epigenetic alterations, including changes in microRNA (miRNA) expression and DNA methylation, were studied. Our results revealed a significant dysregulation in selected cytokines, proteins, and miRNAs involved in key signaling pathways related to IL-17 production and function, including IL-17A and F, IL-6, IL-1ß, transforming growth factor-ß (TGF-ß), signal transducer and activator of transcription-3 (STAT3), p-STAT3, forkhead box O1 (FOXO1), and miR-145 in the small intestine of adult mice challenged with LPS during puberty. In contrast, dietary interventions mitigated the lasting adverse effects of LPS on gut immune function, partly through epigenetic mechanisms. A DNA methylation analysis demonstrated that enduring changes in gut immunity in adult mice might be linked to differentially methylated genes, including Lpb, Rorc, Runx1, Il17ra, Rac1, Ccl5, and Il10, involved in Th17 cell differentiation and IL-17 production and signaling. In addition, prebiotic administration prevented LPS-induced changes in the gut microbiota in pubertal mice. Together, these results indicate that following a healthy diet rich in prebiotics and probiotics is an optimal strategy for programming immune system function in the critical developmental windows of life and controlling inflammation later in life.