Structural analysis and in vitro fermentation characteristics of an Avicennia marina fruit RG-I pectin as a potential prebiotic.

Avicennia marina (Forssk.) Vierh. is a highly salt-tolerant mangrove, and its fruit has been traditionally used for treating constipation and dysentery. In this study, a pectin (AMFPs-0-1) was extracted and isolated from this fruit for the first time, its structure was analyzed, and the effects on the human gut microbiota were investigated. The results indicated that AMFPs-0-1 with a molecular weight of 798 kDa had a backbone consisting of alternating →2)-α-L-Rhap-(1→ and →4)-α-D-GalpA-(1→ residues and side chains composed of →3-α-L-Araf-(1→-linked arabinan with a terminal ß-L-Araf, →5-α-L-Araf-(1→-linked arabinan, and →4)-ß-D-Galp-(1→-linked galactan that linked to the C-4 positions of all α-L-Rhap residues in the backbone. It belongs to a type I rhamnogalacturonan (RG-I) pectin but has no arabinogalactosyl chains. AMFPs-0-1 could be consumed by human gut microbiota and increase the abundance of some beneficial bacteria, such as Bifidobacterium, Mitsuokella, and Megasphaera, which could help fight digestive disorders. These findings provide a structural basis for the potential application of A. marina fruit RG-I pectic polysaccharides in improving human intestinal health.

Structural characterization of a Chlorella heteropolysaccharide by analyzing its depolymerized product and finding an inducer of human dendritic cell maturation.

Chlorella polysaccharides have been gaining increasing attention because of their high yield from dried Chlorella powder and their remarkable immunomodulatory activity. In this study, the major polysaccharide fraction, CPP-3a, in Chlorella pyrenoidosa, was isolated, and its detailed structure was investigated by analyzing the low-molecular-weight product prepared via free radical depolymerization. The results indicated that CPP-3a with a molecular weight of 195.2 kDa was formed by →2)-α-L-Araf-(1→, →2)-α-D-Rhap-(1→, →5)-α-L-Araf-(1→, →3)-ß-D-Glcp-(1→, →4)-α-D-Glcp-(1→, →4)-α-D-GlcpA-(1→, →2,3)-α-D-Manp-(1→, →3,4)-α-D-Manp-(1→, →3,4)-ß-D-Galp-(1→, →3,6)-ß-D-Galp-(1→, and →2,3,6)-α-D-Galp-(1→ residues, branched at C2, C3, C4, or C6 of α/ß-D-Galp and α-D-Manp, and terminated by α/ß-L-Araf, α-L-Arap, α-D-Galp, and ß-D-Glcp. Biological assays showed that CPP-3a significantly altered the dendritic morphology of immature dendritic cells (DCs). Enhanced CD80, CD86, and MHC I expression on the cell surface and decreased phagocytic ability indicated that CPP-3a could induce the maturation of DCs. Furthermore, CPP-3a-stimulated DCs not only stimulated the proliferation of allogeneic naïve CD4+ T cells and the secretion of IFN-γ, but also directly stimulated the activation and proliferation of CD8+ T cells through cross-antigen presentation. These findings indicate that CPP-3a can promote human DC maturation and T-cell stimulation and may be a novel DC maturation inducer with potential developmental value in DC immunotherapy.

Arginine methylation-dependent cGAS stability promotes non-small cell lung cancer cell proliferation.

Cyclic GMP-AMP synthase (cGAS), promotes non-small cell lung cancer (NSCLC) cell proliferation. However, the specific mechanisms of cGAS-mediated NSCLC cell proliferation are largely unknown. In this study, we found asymmetric dimethylation by protein arginine methyltransferase 1 (PRMT1) at R127 of cGAS. This facilitated the binding of deubiquitinase USP7 and contributed to deubiquitination and stabilization of cGAS. PRMT1-and USP7-dependent cGAS stability, which also played a pivotal role in accelerating NSCLC cell proliferation through activating AKT pathway. We validated that the expression of cGAS and PRMT1 were positive correlated in human non-small cell lung cancer samples. Our study demonstrates a unique mechanism for managing cGAS stability by arginine methylation and indicates that PRMT1-cGAS-USP7 axis is a potential therapeutic target for NSCLC.

Recent progress in marine chondroitin sulfate, dermatan sulfate, and chondroitin sulfate/dermatan sulfate hybrid chains as potential functional foods and therapeutic agents.

Chondroitin sulfate (CS), dermatan sulfate (DS), and CS/DS hybrid chains are natural complex glycosaminoglycans with high structural diversity and widely distributed in marine organisms, such as fish, shrimp, starfish, and sea cucumber. Numerous CS, DS, and CS/DS hybrid chains with various structures and activities have been obtained from marine animals and have received extensive attention. However, only a few of these hybrid chains have been well-characterized and commercially developed. This review presents information on the extraction, purification, structural characterization, biological activities, potential action mechanisms, and structure-activity relationships of marine CS, DS, and CS/DS hybrid chains. We also discuss the challenges and perspectives in the research of CS, DS, and CS/DS hybrid chains. This review may provide a useful reference for the further investigation, development, and application of CS, DS, and CS/DS hybrid chains in the fields of functional foods and therapeutic agents.

Monocarboxylate transporter 4 protects against myocardial ischemia/reperfusion injury by inducing oxidative phosphorylation/glycolysis interconversion and inhibiting oxidative stress.

Myocardial ischemia/reperfusion (I/R) injury is the primary cause of heart damage in the treatment of myocardial infarction, and the imbalance of the energy metabolism in the pathogenesis of myocardial I/R is one of the main triggers of cardiac dysfunction. Monocarboxylate transporter 4 (MCT4) is a key transporter of lactate, which plays a vital role in cellular metabolism. The present study investigated the role and underlying mechanism of MCT4 in myocardial I/R injury. The results of this study showed that MCT4 was upregulated during oxygen-glucose deprivation (OGD) and restored after reoxygenation in cardiomyocytes HL-1. Interestingly, the overexpression of MCT4 increased cell viability and decreased apoptosis of OGD/R-induced HL-1 cells. Furthermore, MCT4 boosted glucose uptake and lactate levels and promoted protein expression of glycolysis regulator LDHA, while also impeding oxidative phosphorylation (OXPHOS) regulators C-MYC and NDUFB8 in OGD/R-induced HL-1 cells. A reduction in reactive oxygen species and oxidative stress markers malonaldehyde and superoxide dismutase was also observed within the OGD/R stimulated HL-1 cells. Additionally, the in vivo exogenous application of MCT4 restored cardiac function, as demonstrated by the reduced infarct size and decreased myocardial apoptosis in I/R rats. OXPHOS and oxidative stress declined, while glycolysis was activated when the I/R mice were injected with AAV-MCT4. Our findings indicate that MCT4 could exert a cardioprotective effect after myocardial I/R injury by inducing OXPHOS/glycolysis interconversion and inhibiting oxidative stress.

Inhibition of EZH2 alleviates SAHA-induced senescence-associated secretion phenotype in small cell lung cancer cells.

Chemotherapy has been widely used in small cell lung cancer (SCLC) treatment in the past decades. However, SCLC is easy to recur after chemotherapy. The senescence of cancer cells during chemotherapy is one of the effective therapeutic strategies to inhibit the progression of cancer. Nevertheless, the senescence-associated secretion phenotype (SASP) promotes chronic inflammation of the cancer microenvironment and further accelerates the progression of tumors. Therefore, inducing the senescence of cancer cells and inhibiting the production of SASP factors during anticancer treatment have become effective therapeutic strategies to improve the anticancer effect of drugs. Here we reported that SCLC cells treated with an FDA-approved HDAC inhibitor SAHA underwent senescence and displayed remarkable SASP. In particular, SAHA promoted the formation of cytoplasmic chromatin fragments (CCFs) in SCLC cells. The increased CCFs in SAHA-treated SCLC cells were related to nuclear porin Tpr, which activated the cGAS-STING pathway, and promoted the secretion of SASP in cancer cells. Inhibition of EZH2 suppressed the increase of CCFs in SAHA-treated SCLC cells, weakened the production of SASP, and increased the antiproliferative effect of SAHA. Overall, our work affords new insight into the secretion of SASP in SCLC and establishes a foundation for constructing a new therapeutic strategy for SCLC patients.

A regular Chlorella mannogalactan and its sulfated derivative as a promising anticoagulant: Structural characterization and anticoagulant activity.

Chlorella is one of the most widely cultivated species of microalgae and has been consumed as a "green healthy food". In this study, a novel polysaccharide (CPP-1) was isolated from Chlorella pyrenoidosa, structurally analyzed, and sulfated as a promising anticoagulant. Structural analyses by chemical and instrumental methods such as monosaccharide composition, methylation-GC-MS and 1D/2D NMR spectroscopy analysis revealed that CPP-1 had a molecular weight of ~13.6 kDa, and mainly consisted of d-mannopyranose (d-Manp), 3-O-methylated d-Manp (3-O-Me-d-Manp), and d-galactopyranose (d-Galp). The molar ratio of d-Manp and d-Galp was 1.0:2.3. CPP-1 consisted of a (1→6)-linked ß-d-Galp backbone substituted at C-3 by the d-Manp and 3-O-Me-d-Manp residues in a molar ratio of 1:1, which was a regular mannogalactan. The sulfated Chlorella mannogalactan (SCM) with sulfated group content of 40.2 % equivalent to that of unfractionated heparin was prepared and analyzed. NMR analysis confirmed its structure, indicating that most free hydroxyl groups in the side chains and partial hydroxyl groups in the backbone were sulfated. Anticoagulant activity assays indicated that SCM exhibited strong anticoagulant activity by inhibiting intrinsic tenase (FXase) with IC50 of 13.65 ng/mL, which may be a safer anticoagulant as an alternative to heparin-like drugs.

Structural Elucidation of a Glucan from Trichaster palmiferus by Its Degraded Products and Preparation of Its Sulfated Derivative as an Anticoagulant.

Echinoderms have been attracting increasing attention for their polysaccharides, with unique chemical structure and enormous potential for preparing drugs to treat diseases. In this study, a glucan (TPG) was obtained from the brittle star Trichaster palmiferus. Its structure was elucidated by physicochemical analysis and by analyzing its low-molecular-weight products as degraded by mild acid hydrolysis. The TPG sulfate (TPGS) was prepared, and its anticoagulant activity was investigated for potential development of anticoagulants. Results showed that TPG consisted of a consecutive α1,4-linked D-glucopyranose (D-Glcp) backbone together with a α1,4-linked D-Glcp disaccharide side chain linked through C-1 to C-6 of the main chain. The TPGS was successfully prepared with a degree of sulfation of 1.57. Anticoagulant activity results showed that TPGS significantly prolonged activated partial thromboplastin time, thrombin time, and prothrombin time. Furthermore, TPGS obviously inhibited intrinsic tenase, with an EC50 value of 77.15 ng/mL, which was comparable with that of low-molecular-weight heparin (LMWH) (69.82 ng/mL). TPGS showed no AT-dependent anti-FIIa and anti-FXa activities. These results suggest that the sulfate group and sulfated disaccharide side chains play a crucial role in the anticoagulant activity of TPGS. These findings may provide some information for the development and utilization of brittle star resources.

Chondroitin Sulfate/Dermatan Sulfate Hybrid Chains from Swim Bladder: Isolation, Structural Analysis, and Anticoagulant Activity.

Glycosaminoglycans (GAGs) with unique structures from marine animals show intriguing pharmacological activities and negligible biological risks, providing more options for us to explore safer agents. The swim bladder is a tonic food and folk medicine, and its GAGs show good anticoagulant activity. In this study, two GAGs, CMG-1.0 and GMG-1.0, were extracted and isolated from the swim bladder of Cynoscion microlepidotus and Gadus morhua. The physicochemical properties, precise structural characteristics, and anticoagulant activities of these GAGs were determined for the first time. The analysis results of the CMG-1.0 and GMG-1.0 showed that they were chondroitin sulfate (CS)/dermatan sulfate (DS) hybrid chains with molecular weights of 109.3 kDa and 123.1 kDa, respectively. They were mainly composed of the repeating disaccharide unit of -{IdoA-α1,3-GalNAc4S-ß1,4-}- (DS-A). The DS-B disaccharide unit of -{IdoA2S-α1,3-GalNAc4S-ß1,4-}- also existed in both CMG-1.0 and GMG-1.0. CMG-1.0 had a higher proportion of CS-O disaccharide unit -{-GlcA-ß1,3-GalNAc-ß1,4-}- but a lower proportion of CS-E disaccharide unit -{-GlcA-ß1,3-GalNAc4S6S-ß1,4-}- than GMG-1.0. The disaccharide compositions of the GAGs varied in a species-specific manner. Anticoagulant activity assay revealed that both CMG-1.0 and GMG-1.0 had potent anticoagulant activity, which can significantly prolong activated partial thromboplastin time. GMG-1.0 also can prolong the thrombin time. CMG-1.0 showed no intrinsic tenase inhibition activity, while GMG-1.0 can obviously inhibit intrinsic tenase with EC50 of 58 nM. Their significantly different anticoagulant activities may be due to their different disaccharide structural units and proportions. These findings suggested that swim bladder by-products of fish processing of these two marine organisms may be used as a source of anticoagulants.

Deaminative-cleaved S. monotuberculatus fucosylated glycosaminoglycan: Structural elucidation and anticoagulant activity.

Stichopus monotuberculatus is a tropical sea cucumber species and used as a folk medicine and tonic food. In this study, a fucosylated glycosaminoglycan (SmFG), the depolymerized SmFG (dSmFG) and its oligosaccharide fractions were prepared. The SmFG and its depolymerized products were comprised of a chondroitin-sulfate-E backbone, and various sulfated fucose side chains, including an unusual disaccharide side chain connected to the C-3 position of D-glucuronic acid (GlcA) or GlcA-ol. A peeling reaction occurred during the deaminative depolymerization process. The dSmFG and its fractions showed strong anticoagulant activity by selectively inhibiting intrinsic tenase complex, and had no anti-factor IIa, Xa and VIIa activity. The anticoagulant activity reduced with the decrease of molecular weight, and the unusual branch and novel reducing end may enhance the anticoagulant activity. These findings can provide significant information for development and utilization of depolymerized products from SmFG in food and pharmaceutical industries.

Simulated digestion, dynamic changes during fecal fermentation and effects on gut microbiota of Avicennia marina (Forssk.) Vierh. fruit non-starch polysaccharides.

Grey mangrove (Avicennia marina (Forssk.) Vierh.) fruit is a traditional folk medicine and health food consumed in many countries. In this study, its polysaccharides (AMFPs) were obtained and analyzed by chemical and instrumental methods, with the results indicating that AMFPs consisted of galactose, galacturonic acid, arabinose, and rhamnose in a molar ratio of 4.99:3.15:5.38:1.15. The dynamic changes in AMFPs during the digestion and fecal fermentation processes were then investigated. The results confirmed that AMFPs were not depolymerized by gastric acid and various digestive enzymes. During fermentation, 56.05 % of the AMFPs were utilized by gut microbiota. Galacturonic acid, galactose, and arabinose from AMFPs, were mostly consumed by gut microbiota. AMFPs obviously decreased harmful bacteria and increased some beneficial microbiota, including Megasphaera, Mistuokella, Prevotella, and Megamonas. Furthermore, AMFPs obviously increased the levels of various short-chain fatty acids. These findings suggest that AMFPs have potential prebiotic applications for improving gut health.

Physicochemical Properties and Anticoagulant Activity of Purified Heteropolysaccharides from Laminaria japonica.

Laminaria japonica is widely consumed as a key food and medicine. Polysaccharides are one of the most plentiful constituents of this marine plant. In this study, several polysaccharide fractions with different charge numbers were obtained. Their physicochemical properties and anticoagulant activities were determined by chemical and instrumental methods. The chemical analysis showed that Laminaria japonica polysaccharides (LJPs) and the purified fractions LJP0, LJP04, LJP06, and LJP08 mainly consisted of mannose, glucuronic acid, galactose, and fucose in different mole ratios. LJP04 and LJP06 also contained minor amounts of xylose. The polysaccharide fractions eluted by higher concentration of NaCl solutions showed higher contents of uronic acid and sulfate group. Biological activity assays showed that LJPs LJP06 and LJP08 could obviously prolong the activated partial thromboplastin time (APTT), indicating that they had strong anticoagulant activity. Furthermore, we found that LJP06 exerted this activity by inhibiting intrinsic factor Xase with higher selectivity than other fractions, which may have negligible bleeding risk. The sulfate group may play an important role in the anticoagulant activity. In addition, the carboxyl group and surface morphology of these fractions may affect their anticoagulant activities. The results provide information for applications of L. japonica polysaccharides, especially LJP06 as anticoagulants in functional foods and therapeutic agents.

Chlorella pyrenoidosa Polysaccharides as a Prebiotic to Modulate Gut Microbiota: Physicochemical Properties and Fermentation Characteristics In Vitro.

This study was conducted to investigate the prebiotic potential of Chlorella pyrenoidosa polysaccharides to provide useful information for developing C. pyrenoidosa as a green healthy food. C. pyrenoidosa polysaccharides were prepared and their physicochemical characteristics were determined. The digestibility and fermentation characteristics of C. pyrenoidosa polysaccharides were evaluated using in vitro models. The results revealed that C. pyrenoidosa polysaccharides were composed of five non-starch polysaccharide fractions with monosaccharide compositions of Man, Rib, Rha, GlcA, Glc, Gal, Xyl and Ara. C. pyrenoidosa polysaccharides could not be degraded under saliva and the gastrointestinal conditions. However, the molecular weight and contents of residual carbohydrates and reducing sugars of C. pyrenoidosa polysaccharides were significantly reduced after fecal fermentation at a moderate speed. Notably, C. pyrenoidosa polysaccharides could remarkably modulate gut microbiota, including the promotion of beneficial bacteria, inhibition of growth of harmful bacteria, and reduction of the ratio of Firmicutes to Bacteroidetes. Intriguingly, C. pyrenoidosa polysaccharides can promote growth of Parabacteroides distasonis and increase short-chain fatty acid contents, thereby probably contributing to the promotion of intestinal health and prevention of diseases. Thus, these results suggested that C. pyrenoidosa polysaccharides had prebiotic functions with different fermentation characteristics compared with conventional prebiotics such as fructooligosaccharide, and they may be a new prebiotic for improving human health.

Improvement of Metabolic Syndrome in High-Fat Diet-Induced Mice by Yeast ß-Glucan Is Linked to Inhibited Proliferation of Lactobacillus and Lactococcus in Gut Microbiota.

There is growing evidence that prevention of metabolic syndrome (MS) by dietary fibers is intricately linked to gut microbiota. In the present work, the mice were fed a high-fat diet (HFD) and orally treated with yeast ß-glucan to further examine the effects of ß-glucan on MS and gut microbiota and the potential relationship between gut microbiota and its activity. After intervention for 10 weeks, it was found that the treatment of yeast ß-glucan could significantly improve the HFD-induced MS. Furthermore, pro-inflammatory cytokines in plasma including IL-6 and IL-1ß were decreased. Yeast ß-glucan could regulate the diversity and composition of HFD-induced gut microbiota. Moreover, the relative abundances of Lactobacillus and Lactococcus, having significant positive correlation with metabolic changes, were decreased by ß-glucan, which might play a critical role in attenuation of MS. Our findings suggest that yeast ß-glucan shows promising application as a prebiotic for preventing MS and regulating gut microbiota.

Low-molecular-weight fucosylated glycosaminoglycan and its oligosaccharides from sea cucumber as novel anticoagulants: A review.

Antithrombotic drugs have some side effects, such as risk of serious bleeding. Development of antithrombotic drugs that inhibit components of the intrinsic coagulation pathway and have a low risk of causing bleeding has recently been a focus of research. Fucosylated glycosaminoglycan (FG), also named as fucosylated chondroitin sulfate (FCS), has potent anticoagulant activity and inhibits intrinsic factor tenase (FXase) complex. Low-molecular-weight FG (LFG) and its oligosaccharides show characteristics of anticoagulant and antithrombotic activities with negligible side effects, such as activation of human FXII, induction of platelet aggregation, and especially, the risk of serious bleeding. They are potential new anticoagulant drugs and have been extensively studied in recent years. This review presents recent findings regarding the preparation, structural analysis, pharmacological activity, and structure-activity relationships of LFG and its derived oligosaccharides, so as to provide a reference for the development of new anticoagulants with low side effects.

Isolation, structures and biological activities of polysaccharides from Chlorella: A review.

Chlorella, a green microalga, has been used as an important ingredient in food and medicine because of its excellent nutritive and functional properties. Polysaccharides, as major active ingredients of Chlorella, have attracted increasing attention due to their various health promotion activities, such as those associated with immunomodulation, antioxidation, anti-hyperlipidemia, antitumor, neuroprotection, and anti-asthmatic effect. The significance of polysaccharides from various species of the Chlorella genus has been extensively reported by the scientific community from the perspectives of extraction, structural features, biological activities, and potential uses, which need to be reviewed to improve the understanding, development and utilization of these species. Therefore, this review aims to comprehensively summarize previous and current references regarding the extraction, purification, structural characterization and biological activities of polysaccharides from Chlorella. Moreover, this review also highlights the challenges of investigation and future considerations for holistic utilization in food and medicine.

Immunoenhancing glucuronoxylomannan from Tremella aurantialba Bandoni et Zang and its low-molecular-weight fractions by radical depolymerization: Properties, structures and effects on macrophages.

In this study, a glucuronoxylomannan named TAP-3 was obtained from high-value Tremella aurantialba Bandoni et Zang. Physicochemical analysis revealed that TAP-3, which had a molecular weight of ∼624 kDa, mainly consisted of d-mannose (Man), d-xylose (Xyl), and d-glucuronic acid (GlcA) in a molar ratio of 3.0:1.0: 1.0. Structural analyses of its depolymerized fragments clarified that TAP-3 contained a (1 → 3) and (1 → 2)-linked α-Manp backbone, side chains formed by ß-Xylp and ß-GlcpA linked to the C-2 position of α-Manp, and acetyl groups connected to the sixth hydroxyl positions of Manp. TAP-3 showed marked immune enhancement activity, promoting NO, IL-1ß and TNF-α secretion from macrophages. The critical membrane receptor of TAP-3 was identified to be TLR4, and the chain length was essential for its immunoregulatory activity. These findings expand knowledge of the structural types of glucuronoxylomannan and illustrate its biological activity as an immunopotentiator.

Emulsifier-Free Acrylate-Based Emulsion Prepared by Reverse Iodine Transfer Polymerization.

The self-emulsifying acrylate-based emulsions with solid content 45 wt.% were prepared in 3.5 h by reverse iodine transfer polymerization (RITP), and the polymer molecular weight (Mn) could be 30,000 g·mol-1. The influences of methacrylic acid (MAA) amount, soft/hard monomer mass ratio, and iodine amount on polymerization and latex were investigated. A moderate amount of ionized MAA was needed to stabilize the emulsion. Glass transition temperature (Tg) was decreased with the increasing mass ratio of soft/hard monomer. A higher iodine amount resulted in lower Mn. The increased Mn after chain extension of the polymer with water-insoluble monomers in iterative one-pot method proved the living of polymer. Compared with conventional emulsion polymerization, molecular weight (Mn) could be controlled, and Mn of polymer synthesized in RITP emulsion polymerization is higher; emulsion of polyacrylate-containing hydroxyl monomer units prepared by RITP emulsifier-free radical polymerization is more stable. Good properties, such as hardness, water resistance, adhesion, and increased value of maximum tensile of films modified by reaction of polyacrylate with melamine-formaldehyde (MF) resin, indicated potential application in baking coating.

Structural characterization of a low-molecular-weight polysaccharide from Angelica pubescens Maxim. f. biserrata Shan et Yuan root and evaluation of its antioxidant activity.

A novel heteropolysaccharide with about twenty sugar residues named DF80-2 was obtained from Angelica pubescens Maxim. f. biserrata Shan et Yuan root, one of the most widely used traditional Chinese medicines for thousands of years in China. The possible structure of DF80-2 was proposed considering the comprehensive results of physicochemical properties, methylation analysis, and 1D/2D NMR spectroscopy, which showed that its main chain was composed of (1→3)-, and (1→4)-linked-α-d-Glcp, (1→4)-linked-ß-d-Galp, (1→6)-linked-α-d-Manp, and (1→3)-linked-α-l-Araf, and the branch was present as the α-d-Glcp-(1→3)-ß-d-GalpA disaccharides stretched from O-6 position of (1→4)-linked-α-d-Glc moiety in the main chain. Congo red analysis, scanning electron microscopy and atomic force microscopy showed that DF80-2 possessed a triple helical conformation, and its branched monomers were interlaced with one another forming a regular network structure. DF80-2 exhibited antioxidant activity by effectively scavenging DPPH, and hydroxyl radicals, and chelating ferrous ions.