Glucan polysaccharides isolated from Lactarius hatsudake Tanaka mushroom: Structural characterization and in vitro bioactivities.

Commercially available mushroom polysaccharides have found widespread use as adjuvant tumor treatments. However, the bioactivity of polysaccharides in Lactarius hatsudake Tanaka (L. hatsudake), a mushroom with both edible and medicinal uses, remains relatively unexplored. To address this gap, five L. hatsudake polysaccharides with varying molecular weights were isolated, named LHP-1 (898 kDa), LHP-2 (677 kDa), LHP-3 (385 kDa), LHP-4 (20 kDa), and LHP-5 (4.9 kDa). Gas chromatography-mass spectrometry, nuclear magnetic resonance, and atomic force microscopy, etc., were employed to determine their structural characteristics. The results confirmed that spherical aggregates with amorphous flexible fiber chains dominated the conformation of the LHP. LHP-1 and LHP-2 were identified as glucans with α-(1,4)-Glcp as the main chain; LHP-3 and LHP-4 were classified as galactans with varying molecular weights but with α-(1,6)-Galp as the main chain; LHP-5 was a glucan with ß-(1,3)-Glcp as the main chain and ß-(1,6)-Glcp connecting to the side chains. Significant differences were observed in inhibiting tumor cell cytotoxicity and the antioxidant activity of the LHPs, with LHP-5 and LHP-4 identified as the principal bioactive components. These findings provide a theoretical foundation for the valuable use of L. hatsudake and emphasize the potential application of LHPs in therapeutic tumor treatments.

Development of innovative active packaging films using gelatin/pullulan-based composites incorporated with cinnamon essential oil-loaded metal-organic frameworks for meat preservation.

This study aimed to investigate the effect of cinnamon essential oil (CEO)-loaded metal-organic frameworks (CEO@MOF) on the properties of gelatin/pullulan (Gel/Pull)-based composite films (Gel/Pull-based films). The incorporation of CEO@MOF into Gel/Pull-based films demonstrated significant antimicrobial activity against S. aureus, S. enterica, E. coli, and L. monocytogenes. Additionally, CEO@MOF integrated film exhibited a 98.16 % ABTS radical scavenging, with no significant change in the mechanical properties of the neat Gel/Pull film. The UV blocking efficiency of the composite films increased significantly from 81.38 to 99.56 % at 280 nm with the addition of 3 wt% CEO@MOF. Additionally, Gel/Pull/CEO@MOF films effectively extended the shelf life of meat preserved at 4 °C by reducing moisture loss by 3.35 %, maintaining the pH within the threshold limit (6.2), and inhibiting bacterial growth by 99.9 %. These results propose that CEO@MOF has significant potential as an effective additive in active packaging to improve shelf life and food safety.

Fabrication and characterization of pullulan/tapioca starch-based antibacterial films incorporated with Litsea cubeba essential oil for meat preservation.

Active packaging is a novel technology that utilizes active materials to interact with products and the environment, improving food shelf life. The purpose of this work was to fabricate a multifunctional film using Litsea cubeba essential oil (LC-EO) (1 %, 3 %, 5 %, and 7 %) as the active ingredient and pullulan(P)/tapioca starch (TS) as the carrier material. Adding essential oil improves the films properties, such as barrier ability, anti-oxidant, and antibacterial activity. However, tensile strength (TS) and elongation at break (EAB) were slightly reduced from 28.94 MPa to 11.29 MPa and 15.36 % to 12.19 %. The developed PTS3% films showed the best performance in mechanical properties, especially EAB (14.26 %), WVP (3.26 %) and OP (3.13 %), respectively. The inhibitory zone diameters in the agar-well diffusion test were 18.59 mm for Staphylococcus aureus and 17.32 mm for Escherichia coli. Further study was conducted to compare the preservation effects of film with low-density polyethylene bag (LDPE) on chilled beef. Remarkably, PTS3% film decreased the bacterial population in beef meat while maintaining the pH, color, texture, and TBARS levels within an acceptable range for ten days of storage at 4 °C rather than in a low-density polyethylene bag. The outcomes indicated the potential of PTS3% films in food packaging applications.

An Overview on Mushroom Polysaccharides: Health-promoting Properties, Prebiotic and Gut Microbiota Modulation Effects and Structure-function Correlation.

Mushroom polysaccharides are recognized as "biological response modifiers". Besides several bioactivities, a growing interest in their prebiotic potential has been raised due to the gut microbiota modulation potential. This review comprehensively summarizes mushroom polysaccharides' biological properties, structure-function relationship, and underlying mechanisms. It provides a recent overview of the key findings in the field (2018-2024). Key findings and limitations on structure-function correlation are discussed. Although most studies focus on ß-glucans or extracts, α-glucans and chitin have gained interest. Prebiotic capacity has been associated with α-glucans and chitin, while antimicrobial and wound healing potential is attributed to chitin. However, further research is of utmost importance. Human fecal fermentation is the most reported approach to assess prebiotic potential, indicating impacts on intestinal biological, mechanical, chemical and immunological barriers. Gut microbiota dysbiosis has been directly connected with intestinal, cardiovascular, metabolic, and neurological diseases. Concerning gut microbiota modulation, animal experiments have suggested proinflammatory cytokines reduction and redox balance re-establishment. Most literature focused on the anticancer and immunomodulatory potential. However, anti-inflammatory, antimicrobial, antiviral, antidiabetic, hypocholesterolemic, antilipidemic, antioxidant, and neuroprotective properties are discussed. A significant overview of the gaps and research directions in synergistic effects, underlying mechanisms, structure-function correlation, clinical trials and scientific data is also given.

Role of beta-(1→3)(1→6)-D-glucan derived from yeast on natural killer (NK) cells and breast cancer cell lines in 2D and 3D cultures.

BACKGROUND: Beta-(1,3)(1,6)-D-glucan is a complex polysaccharide, which is found in the cell wall of various fungi, yeasts, bacteria, algae, barley, and oats and has immunomodulatory, anticancer and antiviral effects. In the present study, we investigated the effect of beta-(1,3)(1,6)-D-glucan derived from yeast on the proliferation of primary NK cells and breast cancer cell lines in 2D and 3D models, and on the cytotoxicity of primary NK cells against breast cancer cell lines in 2D and 3D models. METHODS: In this study, we investigated the effects of different concentrations of yeast-derived beta-(1→3)(1→6)-D-glucan on the proliferation and cytotoxicity of human NK cells and breast cancer cell lines in 2D and 3D models using the XTT cell proliferation assay and the CellTiter-Glo® 2.0 assay to determine the cytotoxicity of human NK cells on breast cancer cell lines in 2D and 3D models. RESULTS: We found that the co-incubation of NK cells with beta-glucan in the absence of IL2 at 48 h significantly increased the proliferation of NK cells, whereas the co-incubation of NK cells with beta-glucan in the presence of IL2 (70 U/ml) increased the proliferation of NK cells but not significantly. Moreover, beta-glucan significantly inhibited the proliferation of breast cancer cell lines in 2D model and induced a weak, non-significant growth inhibitory effect on breast cancer multicellular tumor spheroids (3D). In addition, the cytotoxicity of NK cells against breast cancer cell lines was examined in 2D and 3D models, and beta-glucan significantly increased the cytotoxicity of NK cells against MCF-7 (in 2D). CONCLUSIONS: Yeast derived beta-(1,3)(1,6)-D-glucan could contribute to the treatment of cancer by enhancing NK cell immune response as well as contributing to inhibition of breast cancer cell growth.

Natural Polymer-Polyphenol Bioadhesive Coacervate with Stable Wet Adhesion, Antibacterial Activity, and On-Demand Detachment.

Medical adhesives are emerging as an important clinical tool as adjuvants for sutures and staples in wound closure and healing and in the achievement of hemostasis. However, clinical adhesives combining cytocompatibility, as well as strong and stable adhesion in physiological conditions, are still in demand. Herein, a mussel-inspired strategy is explored to produce adhesive coacervates using tannic acid (TA) and methacrylate pullulan (PUL-MA). TA|PUL-MA coacervates mainly comprise van der Waals forces and hydrophobic interactions. The methacrylic groups in the PUL backbone increase the number of interactions in the adhesives matrix, resulting in enhanced cohesion and adhesion strength (72.7 Jm-2), compared to the non-methacrylated coacervate. The adhesive properties are kept in physiologic-mimetic solutions (72.8 Jm-2) for 72 h. The photopolymerization of TA|PUL-MA enables the on-demand detachment of the adhesive. The poor cytocompatibility associated with the use of phenolic groups is here circumvented by mixing reactive oxygen species-degrading enzyme in the adhesive coacervate. This addition does not hamper the adhesive character of the materials, nor their anti-microbial or hemostatic properties. This affordable and straightforward methodology, together with the tailorable adhesivity even in wet environments, high cytocompatibility, and anti-bacterial activity, enables foresee TA|PUL-MA as a promising ready-to-use bioadhesive for biomedical applications.

PURIFICATION, CHARACTERIZATION, AND IN VITRO ANTITUMOR ACTIVITY OF A NOVEL GLUCAN FROM PHOENIX DACTYLIFERA L. FRUITS.

In this study, ß- glucan was extracted by the hot water extraction method followed by ethanol precipitation and purified using ion and gel filtration chromatography, then evaluate the anticancer effects of ß- glucan that purified from Phoenix dactylifera on cancer cell line. Ahmed Nahi Glioblastoma Multiform (ANGM) cancer cell line was used in the in vitro study. Cell line exposure times were calculated after 24, 48, and 72 hours in a micro titration plate under absolutely sterile conditions. High molecular weight ß-glucans can be obtained using the hot water extraction method without having to use strong agents to change their structure, like alkalis or acids. Anti-cancer property of ß-glucan derived from Phoenix dactylifera fruits on cancer cell lines has been reported. In this work, the ANGM cell line was treated with different concentrations of ß-glucan (31.25, 62.5, 125, 250, 500 and 1000 µg/mL). and the inhibition of the cells was investigated using the MTT assay after 24, 48 and 72 hours. The result obtained showed time and concentration dependent cytotoxic effect, and the higher concentrations at 48 hrs of exposure gave significantly (p<0.05) higher cytotoxic effect.

Effect of Polycan, a ß-Glucan from Aureobasidium pullulans SM-2001, on Inflammatory Response and Intestinal Barrier Function in DSS-Induced Ulcerative Colitis.

Ulcerative colitis (UC), a subtype of inflammatory bowel disease, is a chronic gastrointestinal inflammatory disease with unclear etiology and pathophysiology. Herein, we determined the effects of extracellular polysaccharides purified from Aureobasidium pullulans SM-2001 (Polycan) on tight junction protein expression, inflammation, and apoptosis in a dextran sodium sulfate (DSS)-induced acute colitis model. Fifty mice were divided into normal, DSS, DSS + Polycan 250 mg/kg (Polycan 250), DSS + Polycan 500 mg/kg (Polycan 500), and DSS + 5-aminosalicylic acid 100 mg/kg (5-ASA) groups. Their body weights, colon lengths, histological changes in colon tissue, and tight junction function were observed. Results showed that Polycan 250, Polycan 500, and 5-ASA significantly inhibited body weight loss compared with DSS. Similar to 5-ASA, Polycan 500 exhibited preventive effects on colon length shortening and histological changes in colon tissues. Polycan inhibited the DSS-induced decrease in fluorescein isothiocyanate-dextran permeability and myeloperoxidase activity. Moreover, Polycan significantly recovered serum cytokine (e.g., tumor necrosis factor-α, interleukin (IL)-6, and IL-1ß) or mRNA expression in colon tissue compared with DSS. Polycan also inhibited apoptosis by reducing caspase-3 activity and the Bcl-2 associated X/B-cell lymphoma 2 (Bcl-2) ratio. Additionally, DSS treatment significantly reduced microbial abundance and diversity, but the administration of Polycan reversed this effect. Collectively, Polycan protected intestinal barrier function and inhibited inflammation and apoptosis in DSS-induced colitis.

Aminated yeast ß-D-glucan for macrophage-targeted delivery of CpG oligodeoxynucleotides and synergistically enhanced cancer immunotherapy.

CpG oligodeoxynucleotides (CpG ODNs) activate immune system and show strong potential in cancer immunotherapy. However, therapeutic efficacy of CpG ODNs is hampered due to rapid nuclease degradation and insufficient cellular uptake. Delivery of CpG ODNs into antigen presenting cells (APCs) is vital to enhance their therapeutic efficacy. Herein, we developed a super-convenient yet efficient strategy for macrophage-targeted delivery of CpG ODNs and synergistically enhanced cancer immunotherapy. Aminated yeast ß-D-glucan (NH2-Glu) was simply synthesized through functionalization of ß-D-glucan with DETA, which exhibited a dendrimer-like shape with size of about 80 nm. NH2-Glu complexed negatively-charged CpG ODNs. The as-prepared NH2-Glu/CpG complexes were positively charged, uniformly dispersed and exhibited good stability against nuclease degradation. Due to the specific recognition with dectin-1 expressed on macrophages, NH2-Glu/CpG complexes targeted macrophage and exhibited significantly enhanced cellular uptake due to dectin-1-mediated endocytosis. NH2-Glu/CpG complexes showed potent immunostimulatory activity. Contributed by the inherent immunostimulatory and antitumor activity, yeast ß-D-glucan functioned synergistically with CpG ODNs in inducing antitumor immunity. NH2-Glu/CpG complexes remarkably inhibited tumor growth without causing toxic effect. In summary, this work provides a facile yet efficient macrophage-targeted CpG ODNs delivery system for cancer immunotherapy.

Synthesis, characterization and anti-inflammatory activity of selenium nanoparticles stabilized by aminated yeast glucan.

Improving the dispersed stability of selenium nanoparticles (SeNPs) is the key to its application. In this study, yeast glucan with different degrees of amination (BNs) were used as stabilizers and capping agent to prepare dispersed SeNPs. The size, storage stability, and morphology of BNs/SeNPs were characterized. Results show that BNs/SeNPs presented positive potential and spherical morphologies with average particle size about 100-300 nm and kept stable at room temperature for a long time. The CCK-8 assay showed that BNs/SeNPs had significantly lower toxicity to RAW264.7 cells than SeNPs. Moreover, BNs/SeNPs could inhibit the generation of NO, IL-1ß and IL-6 effectively in RAW 264.7 macrophages induced by LPS, and down-regulate the mRNA transcription of iNOS, IL-1ß, IL-6 and chemokines (CCL2 and CCL5), indicating that BNs/SeNPs had good anti-inflammatory activity. Therefore, aminated yeast glucan could improve the stability and bioactivity of SeNPs simultaneously, which is a promising stabilizer for SeNPs.

Effects of dietary soluble ß-1,3-glucan on the growth performance, antioxidant status, and immune response of the river prawn (Macrobrachium nipponense).

The effects of dietary ß-1,3-glucan on the growth performance, body composition, hepatopancreas tissue structure, antioxidant activities, and immune response of the river prawn (Macrobrachium nipponense) were investigated. In total, 900 juvenile prawns were fed one of five diets with different contents of ß-1,3-glucan (0%, 0.1%, 0.2%, and 1.0%) or 0.2% curdlan for 6 weeks. The growth rate, weight gain rate, specific growth rate, specific weight gain rate, condition factor, and hepatosomatic index of juvenile prawns fed 0.2% ß-1,3-glucan were significantly higher than those fed 0% ß-1,3-glucan and 0.2% curdlan (p < 0.05). The whole-body crude lipid content of prawns supplemented with curdlan and ß-1,3-glucan was significantly higher than that of the control group (p < 0.05). The antioxidant and immune enzyme activities of superoxide dismutase (SOD), total antioxidant capacity (T-AOC), catalase (CAT), lysozyme (LZM), phenoloxidase (PO), acid phosphatase (ACP), and alkaline phosphatase (AKP) in the hepatopancreas of juvenile prawns fed 0.2% ß-1,3-glucan were significantly higher than those of the control and 0.2% curdlan groups (p < 0.05), and tended to increase and then decrease with increasing dietary ß-1,3-glucan. The highest malondialdehyde (MDA) content was observed in juvenile prawns without ß-1,3-glucan supplementation. The results of real-time quantitative PCR indicated that dietary ß-1,3-glucan promoted expression of antioxidant and immune-related genes. Binomial fit analysis of weight gain rate and specific weight gain rate showed that the optimum ß-1,3-glucan requirement of juvenile prawns was 0.550%-0.553%. We found that suitable dietary ß-1,3-glucan improved juvenile prawns growth performance, antioxidant capacity, and non-specific immunity, which provide reference for shrimp healthy culture.

Effects of grape seed extract on the properties of pullulan polysaccharide/xanthan gum active films for apple preservation.

Grape seed extract (GSE) was added to pullulan polysaccharide (PP)/xanthan gum (XG) as composite film (PP/XG/GSE or PXG). The observed composite morphology indicated their biocompatibility. Sample PXG100 (contain 100 mg/L GSE) demonstrated the best mechanical properties, with tensile strength of 16.62 ± 1.27 MPa, and the elongation at break of (22.60 ± 0.48)%. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical scavenging activity of PXG150 were the highest at (81.52 ± 1.57)% and (90.85 ± 1.54)%, respectively. PXG films also demonstrated inhibitory effects on Staphylococcus aureus, Escherichia coli, and Bacillus subtilis. The PXG films could also prolong the shelf life of fresh-cut apples because it could decrease the rate of weight loss and retain more vitamin C and total polyphenol even on the 5th day. The weight loss rate of PXG150 was decreased from (8.58 ± 0.6)% (control) to (4.15 ± 0.19)%. It was able to achieve vitamin C and total polyphenol retention rate of 91 % and 72 %, respectively, which was significantly higher that the control sample. Therefore, GSE had contributed in enhancing the antibacterial, antioxidant properties, mechanical strength, UV protection and water resistance in PXG composite films. This effectively extend the shelf life of fresh-cut apples, which it will be an excellent food packaging material.

A Natural Glucan from Black Bean Inhibits Cancer Cell Proliferation via PI3K-Akt and MAPK Pathway.

A natural α-1,6-glucan named BBWPW was identified from black beans. Cell viability assay showed that BBWPW inhibited the proliferation of different cancer cells, especially HeLa cells. Flow cytometry analysis indicated that BBWPW suppressed the HeLa cell cycle in the G2/M phase. Consistently, RT-PCR experiments displayed that BBWPW significantly impacts the expression of four marker genes related to the G2/M phase, including p21, CDK1, Cyclin B1, and Survivin. To explore the molecular mechanism of BBWPW to induce cell cycle arrest, a transcriptome-based target inference approach was utilized to predict the potential upstream pathways of BBWPW and it was found that the PI3K-Akt and MAPK signal pathways had the potential to mediate the effects of BBWPW on the cell cycle. Further experimental tests confirmed that BBWPW increased the expression of BAD and AKT and decreased the expression of mTOR and MKK3. These results suggested that BBWPW could regulate the PI3K-Akt and MAPK pathways to induce cell cycle arrest and ultimately inhibit the proliferation of HeLa cells, providing the potential of the black bean glucan to be a natural anticancer drug.

Synergistic effect of p53 gene/DOX intracellular delivery and P-gp inhibition by pullulan thiomers on cancer cells: in vitro and in vivo evaluations.

Numerous reports emphasize the inverse relationship between the mutant p53 protein and P-glycoprotein overexpression, which adversely affects the chemosensitivity of cancer cells. In this study, the cationised pullulan polysaccharide was conjugated with dithiobutyric acid (PPDBA) for the intracellular delivery of doxorubicin and the p53 gene. The transfection efficiency of PPDBA using the apoptotic gene p53 and its ability to modulate efflux pumps in the presence and absence of glutathione and the subsequent drug retention were studied in different cell lines. The percentage cell death mediated by the PPDBA/p53 nanoplex (4 : 1 ratio) was 59%, and by DOX alone a 50% cell death was attained at 3.13 µM in C6 cells, but the percentage cell death mediated by PPDBA/p53 (4 : 1) in combination with 1 µM DOX was as high as 98%. The effect of PPDBA II/p53/DOX nanoplexes on the mouse tumor model was evaluated in BALB/c mice which demonstrated good efficacy when compared with the drug or gene alone.

Extraction, characterization, and anti-nonalcoholic steatohepatitis activity of a (1,3) (1,6)-ß-D-glucan from the Polyporus umbellatus (Pers.) Fries.

Nonalcoholic steatohepatitis (NASH) is a chronic liver disease characterized by inflammation and hepatic steatosis that may coincide with fibrotic activity. To date, no pharmacological agents have been approved for NASH treatment. Here, a homogeneous (1,3),(1,6)-ß-D-glucan (PUP-W-1, Mw: 41.07 kDa) was successfully purified from Polyporus umbellatus (Pers.) Fries sclerotia and characterized. The analysis showed that the PUP-W-1 backbone consisted of a repeating chain of eight →3)-ß-D-Glcp-(1 â†’ units, with branched chains of four ß-D-Glcp residues, joined by repeating 1,6-linkage units at the O-6 position of the backbone. The pharmacological effects of PUP-W-1 treatment in the context of NASH pathogenesis were explored using a methionine choline-deficient (MCD) diet-induced murine steatohepatitis model. The MCD model mice exhibited pronounced steatohepatitis, inflammatory activity, steatosis, stellate cell activation, and mild fibrotic activity. Treatment of the mice for three weeks with PUP-W-1 prevented the development of NASH due to the suppression of inflammation, lipid accumulation, and fibrosis. As suggested by these findings, PUP-W-1 may hold promise as a natural drug candidate or precursor for the treatment of NASH.

Maitake α-glucan promotes differentiation of monocytic myeloid-derived suppressor cells into M1 macrophages.

AIMS: Myeloid-derived suppressor cells (MDSCs) are major components of the tumor microenvironment and systemically accumulate in tumor-bearing hosts and patients with cancer, facilitating cancer progression. Maitake macromolecular α-glucan YM-2A, isolated from Grifola frondosa, inhibits tumor growth by enhancing immune responses. The present study investigated the effects of YM-2A on the immunosuppressive potential of MDSCs. MAIN METHODS: YM-2A was orally administered to CT26 tumor-bearing mice, and the number of immune cells in the spleen and tumor was measured. Splenic MDSCs isolated from the CT26 tumor-bearing mice were treated with YM-2A and co-cultured with T cells to measure their inhibitory effect on T cell proliferation. For adoptive transfer of monocytic MDSCs (M-MDSCs), YM-2A-treated M-MDSCs mixed with CT26 cells were implanted subcutaneously in the mice to measure the tumor growth rate. KEY FINDINGS: YM-2A selectively reduced the accumulation of M-MDSCs but not that of polymorphonuclear MDSCs (PMN-MDSCs) in CT26 tumor-bearing mice. In tumor tissues, YM-2A treatment induced the polarity of immunostimulatory M1-phenotype; furthermore, it increased the infiltration of dendritic, natural killer, and CD4+ and CD8+ T cells. YM-2A treatment of purified M-MDSCs from CT-26 tumor-bearing mice induced dectin-1-dependent differentiation into M1 macrophages. YM-2A-treated M-MDSCs lost their inhibitory activity against proliferation and activation of CD8+ T cells. Furthermore, adoptive transfer of M-MDSCs treated with YM-2A inhibited CT26 tumor growth. SIGNIFICANCE: YM-2A promotes the differentiation of M-MDSCs into immunostimulatory M1 macrophages, thereby enhancing the efficacy of cancer immunotherapy.

Dietary yeast beta 1,3/1,6 glucan supplemented to adult Labrador Retrievers alters peripheral blood immune cell responses to vaccination challenge without affecting protective immunity.

Yeast-derived 1,3/1,6 ß-glucans may alter host immunity to produce robust and quickly resolved responses that align with companion animal health goals. In adult dogs, immunomodulation by yeast 1,3/1,6 ß-glucans in extruded kibble diet have not been well documented. The study objective was to evaluate systemic immune responses in dogs fed kibble diets with two yeast 1,3/1,6 ß-glucans doses before and after vaccine challenge. Twenty-four adult Labrador Retrievers were assigned to three dietary treatments consisting of a basal diet (control) supplemented with 0.012% or 0.023% (0.5 or 1×, respectively) yeast 1,3/1,6 ß-glucan with equal sex representation within each treatment (8 dogs/diet). Animals were fed experimental diets for a 29-d acclimation period, after which baseline blood samples were collected before administration of a combination canine distemper virus, parvovirus, and adenovirus-2 vaccine. Blood samples were collected weekly for 21 d following vaccination with whole blood for CBC analysis, serum for titer and cytokine assays, and peripheral blood mononuclear cells (PBMC) isolated for flow cytometric immune cell profiling. Data were analyzed using the MIXED procedure with diet and timepoint fixed effects. Serum titer was analyzed by Kruskal-Wallis test (SAS 9.4; P ≤ 0.05). Prior to vaccination, ß-glucan diets did not affect serum cytokines, antibody titer, or immune cell populations. In the first 7 d post-vaccination (dpv), PBMC CD21low B cells increased 36.5% to 58.1% in all groups but the magnitude of change was lesser in the 0.5× ß-glucan diet resulting in 25.6% lower CD21low populations compared to control-fed dogs (P = 0.007). By 21 dpv, B-cell populations recovered to baseline levels in dogs fed 1× ß-glucan, but CD21high cells remained elevated 50.5% in dogs fed 0.5× ß-glucan diets compared with baseline (P < 0.0001). While no differences in serum titer or cytokines were observed, feeding both ß-glucan diets maintained stable blood monocytes, whereas a 53.0% decrease between baseline and 14 dpv was observed in control-fed dogs (P = 0.01). Collectively, these outcomes suggest that a 1× dose of 1,3/1,6 yeast ß-glucan in extruded kibble diets altered monocytes associated with trained immunity, did not reduce PBMC CD21low B-cell responsiveness, and simultaneously contributed to B-cell population resolution by 21 dpv in adult dogs. Additional research to assess the functionality of these changes is needed.

Companion animal food trends reflect the growing demand for healthy, functional foods. Yeast-derived ß-glucans have been shown to train the human immune system to respond and resolve inflammation quickly, which aligns with companion animal health goals of providing disease protection without excessive inflammation. In this work, 24 healthy adult Labrador Retrievers were fed diets without or with yeast ß-glucans at two doses (0.5 and 1×). After feeding experimental diets for 4 wk, blood samples were collected to establish baseline immunity before dogs were challenged with a commercially available vaccine. Blood samples were collected weekly for 3 wk post-vaccination to determine changes, if any, to serum antibody, cytokine production, and blood counts. All dogs achieved protective antibody titers within 1 wk post-vaccination. Dogs fed the 1× ß-glucans inclusion showed potentially reduced reliance on cells associated with early immune responses without prolonging responses by antibody-producing cells. These outcomes suggest beneficial responses to dietary yeast ß-glucans in vaccinated adult Labrador Retrievers, but further research and refinement of immunological assessment in companion animals are needed.

Yeast ß-D-glucan functionalized graphene oxide for macrophage-targeted delivery of CpG oligodeoxynucleotides and synergistically enhanced antitumor immunity.

Immunostimulatory CpG oligodeoxynucleotides (CpG ODNs) show strong potential in cancer immunotherapy. However, therapeutic efficacy of CpG ODNs is hindered due to rapid nuclease degradation and insufficient cellular uptake. Transfecting CpG ODNs into antigen presenting cells (APCs) is vital to enhance their therapeutic efficacy while reduce the potential side effects. Herein, a multifunctional CpG ODNs vector was fabricated through functionalization of graphene oxide (GO) with yeast ß-D-glucan, and its potential in cancer immunotherapy was further investigated. GO-ß-D-glucan protected CpG ODNs from nuclease digestion. ß-D-glucan endowed the delivery system with targeting ability for macrophage due to its recognition with dectin-1. Thus, GO-ß-D-glucan enhanced the delivery of CpG ODNs into RAW264.7 cells due to dectin-1-mediated endocytosis. More importantly, ß-D-glucan functioned synergistically with CpG ODNs in inducing antitumor immunity. GO-ß-D-glucan/CpG ODNs inhibited the tumor cells growth more effectively. This work provides a macrophage-targeted CpG ODNs delivery system for cancer immunotherapy. Graphic abstract.

The carboxymethylated derivative of laminaran from brown alga Saccharina cichorioides: Structure, anticancer and anti-invasive activities in 3D cell culture.

Polysaccharides' derivatives are promising biologically active compounds for biotechnology, nutrition, industries, and are becoming increasingly important in medicine and pharmacy. Laminaran from brown alga Saccharina cichorioides (ScL) was chemically modified to obtain the carboxymethylated derivative (ScLCM) with improved structure and bioactivity. ScLCM was identified as (1 â†’ 3)-ß-D-glucan with -CH2-COOH groups at some positions 2, 4, and 6 of glucose residues. The anticancer activity of ScLCM was studied on the models of viability and invasion of 3D human melanoma SK-MEL-28, breast cancer T-47D, and colorectal carcinoma DLD-1 cells in comparison with native laminaran or its sulfated or aminated derivatives. ScLCM had the highest anticancer and anti-invasive effects among investigated polysaccharides. ScLCM significantly suppressed the viability and invasion of 3D SK-MEL-28 cells via the regulation of the activity of matrix metalloproteinase 9 (MMP 9) and protein kinases of ERK/MAPK signaling pathway. These findings may contribute to the reported anticancer effects of algal polysaccharides' derivatives.

Ultrasound-assisted development and characterization of novel polyphenol-loaded pullulan/trehalose composite films for fruit preservation.

A novel food packaging film was developed by incorporating a tea polyphenols-loaded pullulan/trehalose (TP@Pul/Tre) into a composite film with ultrasound-assisted treatment of dual-frequency (20/35 kHz, 40 W/L) for 15 min to assess the physicochemical and mechanical properties of a composite film. The optimized ultrasound-assisted significantly increases elongation at break, tensile strength, and improves the composite film's UV/water/oxygen barrier properties. Structure analysis using attenuated total reflectance-Fourier transform infrared, X-ray diffraction and thermal stability revealed that these improvements were achieved through ultrasound-enhanced H-bonds, more ordered molecular arrangements, and good intermolecular compatibility. Besides, the ultrasound-assisted TP@Pul/Tre film has proven to have good antibacterial performance against Escherichia coli and Staphylococcus aureus, with approximately 100 % lethality at 4 h and 8 h, respectively. Moreover, the ultrasound-assisted TP@Pul/Tre film effectively delayed moisture loss, oxidative browning, decay, and deterioration in fresh-cut apples and pears, thereby extending their shelf life. Thus, ultrasound has proved to be an effective tool for improving the quality of food packaging films, with a wide range of applications.