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A new type of acidic exopolysaccharide (AESP-II) was extracted and separated from the fermentation broth of Cordyceps militaris (C. militaris), which was further purified to elucidate its structural characteristics and immunological activity. AESP-II was confirmed to be an acidic pyranose with a molecular weight of 61.52 kDa, which consisted of mannose, glucuronic acid, rhamnose, galactose acid, N-acetyl-galactosamine, glucose, galactose and arabinose with a molar ratio of 1.07: 5.38: 1: 3.14: 2.23: 15: 6.09: and 4.04. Animal experiment results verified that AESP-II can significantly promote the proliferation of spleen T and B lymphocytes in mice with immune injury caused by cyclophosphamide (CTX). In particular, the promotion of B lymphocytes presented a dose-effect relationship. In addition, the levels of the cytokines IL-2, IL-4, and IFN-γ, which are mainly secreted by T lymphocytes, and immunoglobulin IgG, IgM and IgA, which are mainly secreted by B lymphocytes, were increased after AESP-II treatment. The above results suggest that fluid immunity is involved in the immunomodulatory function of AESP-II. Simultaneously, AESP-II was detected significantly to promote the phosphorylation expression of p38 kinase (p38), extracellular regulated protein kinases (ERK) and c-Jun N-terminal kinase (JNK) by Western blot, further suggesting that the activation of the MAPK signaling pathway mediates the immunoregulatory function of AESP-II.

High mannose has previously associated with insulin resistance and cardiovascular disease (CVD). Our objective is to establish whether mannose is associated with anatomical evidence of coronary artery disease (CAD).Plasma mannose concentrations were measured by liquid chromatography/tandem mass spectrometry in a discovery cohort (n = 513) and a validation cohort (n = 221) of carefully phenotyped individuals. In both cohorts CAD was quantitated using state-of-the-art imaging techniques (coronary computed coronary tomography angiography (CCTA), invasive coronary angiography and optical coherence tomography). Information on subsequent CVD events/death was collected. Associations of mannose with angiographic variables and biomarkers were tested using univariate and multivariate regression models. Survival analysis was performed using the Kaplan-Meier estimator.Mannose was related to indices of CAD and features of plaque vulnerability. In the discovery cohort, mannose was a marker of quantity and quality of CCTA-proven CAD and subjects with a mannose level in the top quartile had a significantly higher risk of CVD events/death (p = 3.6e-5). In the validation cohort, mannose was significantly associated with fibrous cap thickness 65 μm (odds ratio = 1.32 per each 10 μmol/L mannose change [95% confidence interval, 1.05-1.65]) and was an independent predictor of death (hazard ratio for mannose≥vs 84.6 μmol/L: 4.0(95%CI, 1.4-11.3), p = 0.006).The current data add novel evidence that high mannose is a signature of CAD with a vulnerable plaque phenotype, consistently across measures of severity of vessel involvement and independent of the traditional correlates of CVD, and that it is an independent predictor of incident adverse outcomes.

Lectins from plants of the Diocleinae subtribe often exhibit specificity towards mannose/glucose and derived sugars, with some plants also displaying a second lectin specific to lactose/GalNAc. Here, we present a novel lectin from Collaea speciosa, named CsL, that displays specificity for GlcNAc/glucose. The lectin was extracted from Collaea speciosa seeds and purified by a single chromatographic step on a Sephadex G-50 matrix. In solution, the lectin appears as a dimeric protein composed of 25 kDa monomers. The protein is stable at pH 7–8 and dependent on divalent cations. CsL maintained its agglutination activity after heating to 90 °C for 1 h. Glycan array studies revealed that CsL binds to N-glycans with terminal GlcNAc residues, chitobiose and chitotriose moieties. The partial amino acid sequence of the lectin is similar to that of some lactose-specific lectins from the same subtribe. In contrast to other ConA-like lectins, CsL is not toxic to Artemia. Because of its remarkably different properties and specificity, this lectin could be the first member of a new group inside the Diocleinae lectins.

The structural elucidation and investigation of the immunostimulatory effects of crude polysaccharides (ECPs), non-starch polysaccharides (ECPs-I), and deproteinated non-starch polysaccharides (ECPs-II) derived from finger millet (Eleusine coracana) were performed. The extracted ECPs, ECPs-I, and ECPs-II primarily comprised different levels of carbohydrate (82.9, 41.9 and 62.0%), protein (5.3, 38.4 and 22.0%) and uronic acid (0.5, 2.4 and 0.7%). Monosaccharide composition analysis showed that glucose (88.2 and 40.6) was the main sugar unit present in ECPs and ECPs-I, followed by galactose, arabinose, xylose, rhamnose, and mannose; however, ECPs-II contained galactose (42.2%) as a major unit. The average molecular weights of ECPs, ECPs-I, and ECPs-II were found to be 120, 78 and 43 kDa, respectively. The polysaccharides as well as non-starch polysaccharides did not show any toxic effects on RAW 264.7 cells. ECPs-II treatment significantly enhanced the activation of RAW 264.7 cells by inducing nitric oxide (NO) production, upregulating inducible nitric oxide synthase (iNOS) and various pro-inflammatory cytokines including interleukin (IL)-1β, IL-6, IL-10, and tumor necrosis factor alpha (TNF-α). Furthermore, the RAW 264.7 cells were activated after ECPs-II treatment via the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), and mitogen-activated protein kinase signaling (MAPK) pathways induced by a cluster of differentiation 40 (CD40) expression. The structure of ECPs-II mainly consists of (1→3,4)-linked galactopyranosyl and (1→3,5) arabinopyranosyl.

Inflammatory osteolysis is usually linked to the activation of proinflammatory macrophage and the consequent excessive osteoclast formation. Emerging evidence indicates that agents or drugs targeting lipid metabolism in macrophages might be potential in the prevention and treatment of osteolysis. d -mannose, as a natural-existed metabolic regulator, exerts strong effects on attenuating osteopenia and inflammation. However, whether d -mannose is therapeutically effective on osteolysis and whether a metabolic mechanism counts for the effect remain to be addressed. Here, by using an in vivo lipopolysaccharide (LPS)-induced inflammatory osteolysis mouse model as well as an in vitro LPS-induced inflammatory macrophage culture system, we show that d -mannose attenuates inflammatory osteolysis and inhibits excessive osteoclastogenesis by reversing the LPS-induced activation of proinflammatory macrophage. Mechanically, d -mannose recovers LPS-suppressed Cpt1a transcription and promotes lipid metabolism of macrophage. Treatment with etomoxir, an inhibitor of CPT1A, abolishes the effects of d -mannose on LPS-treated macrophage in vitro and eliminates its protection against osteolysis in vivo. Collectively, our results imply that d -mannose attenuates LPS-induced osteolysis by manipulating CPT1A-mediated lipid metabolism in macrophages. Our results disclose the unrecognized utilization of d -mannose as an effective intervention against inflammatory osteolysis and provide evidence to manage inflammatory scenarios by therapeutically targeting lipid metabolism in macrophage.

Carbohydrate-based materials are increasingly investigated for a range of applications spanning from healthcare to advanced functional materials. Synthetic glycopolymers are particularly attractive as they possess low toxicity and immunogenicity and can be used as multivalent ligands to target sugar-binding proteins (lectins). Here, we utilised RAFT polymerisation to synthesize two families of novel diblock copolymers consisting of a glycopolymers block containing either mannopyranose or galactopyranose pendant units, which was elongated with sodium 2-acrylamido-2-methyl-1-propanesulfonate (AMPS) to generate a polyanionic block. The latter enabled complexation of cationic aminoglycoside antibiotic tobramycin through electrostatic interactions (loading efficiency in the 0.5-6.3 wt% range, depending on the copolymer). The resulting drug vectors were characterized by dynamic light scattering, zeta-potential, and transmission electron microscopy. Tobramycin-loaded complexes were tested for their ability to prevent clustering or disrupt biofilm of the Pseudomonas aeruginosa Gram-negative bacterium responsible for a large proportion of nosocomial infection, especially in immunocompromised patients. P. aeruginosa possesses two specific tetrameric carbohydrate-binding adhesins, LecA (PA-IL, galactose/N-acetyl-D-galactosamine-binding) and LecB (PA-IIL, fucose/mannose-binding), and the cell-associated and extracellular adhesin CdrA (Psl/mannose-binding) thus ideally suited for targeted drug delivery using sugar-decorated tobramycin-loaded complexes here developed. Both aliphatic and aromatic linkers were utilised to link the sugar pendant units to the polyacrylamide polymer backbone to assess the effect of the nature of such linkers on bactericidal/bacteriostatic properties of the complexes. Results showed that tobramycin-loaded complexes efficiently suppressed (40 to 60% of inhibition) in vitro biofilm formation in PAO1-L P. aeruginosa and that preferential targeting of PAO1-L biofilm can be achieved using mannosylated glycopolymer-b-AMPSm.

This study aims to determine ovarian cancer (OC) patients with platinum resistance for alternative treatment protocols by using metabolomic methodologies. Urine and serum samples of platinum-resistant and platinum-sensitive OC were analyzed using GC-MS. After data processing of GC-MS raw data, multivariate analyses were performed to interpret complex data for biologically meaningful information and to identify the biomarkers that cause differences between two groups. The biomarkers were verified after univariate, multivariate, and ROC analysis. Finally, metabolomic pathways related to group separations were specified. The results of biomarker analysis showed that 3,4-dihydroxyphenylacetic acid, 4-hydroxybutyric acid, L-threonine, D- mannose, and sorbitol metabolites were potential biomarkers in urine samples. In serum samples, L-arginine, linoleic acid, L-glutamine, and hypoxanthine were identified as important biomarkers. R2Y, Q2, AUC, sensitivity and specificity values of platinum-resistant and sensitive OC patients’ urine and serum samples were 0.85, 0.545, 0.844, 91.30%, 81.08 and 0.570, 0.206, 0.743, 77.78%, 74.28%, respectively. In metabolic pathway analysis of urine samples, tyrosine metabolism and fructose and mannose metabolism were found to be statistically significant (p

1,2-β-Mannobiose phosphorylases (1,2-β-MBPs) from glycoside hydrolase 130 (GH130) family are important bio-catalysts in glycochemistry applications owing to their ability in synthesizing oligomannans. Here, we report the crystal structure of a thermostable 1,2-β-MBP from Thermoanaerobacter sp. X-514 termed Teth514_1789 to reveal the molecular basis of its higher thermostability and mechanism of action. We also solved the enzyme complexes of mannose, mannose-1-phosphate (M1P) and 1,4-β-mannobiose to manifest the enzyme-substrate interaction networks of three main subsites. Notably, a Zn ion that should be derived from crystallization buffer was found in the active site and coordinates the phosphate moiety of M1P. Nonetheless, this Zn-coordination should reflect an inhibitory status as supplementing Zn severely impairs the enzyme activity. These results indicate that the effects of metal ions should be taken into consideration when applying Teth514_1789 and other related enzymes. Based on the structure, a reliable model of Teth514_1788 that shares 61.7% sequence identity to Teth514_1789 but displays a different substrate preference was built. Analyzing the structural features of these two closely related enzymes, we hypothesized that the length of a loop fragment that covers the entrance of the catalytic center might regulate the substrate selectivity. In conclusion, these information provide in-depth understanding of GH130 1,2-β-MBPs and should serve as an important guidance for enzyme engineering for further applications.

Graphical Abstract Non-starch polysaccharides derived from natural resources play a significant role in the field of food science and human health due to their extensive distribution in nature and less toxicity. In this order, the immunostimulatory activity of a non-starch polysaccharide (CQNP) from Chenopodium quinoa was examined before and after deproteination in murine macrophage RAW 264.7 cells. The chemical composition of CQNP and deproteinated-CQNP (D-CQNP) were spectrometrically analysed that revealed the presence of carbohydrate (22.7 ± 0.8% and 39.5 ± 0.8%), protein (41.4 ± 0.5% and 20.8 ± 0.5%) and uronic acid (8.7 ± 0.3% and 6.7 ± 0.2%). The monosaccharide composition results exposed that CQNP possesses a high amount of arabinose (34.5 ± 0.3) followed by galactose (26.5 ± 0.2), glucose (21.9 ± 0.3), rhamnose (7.0 ± 0.1), mannose (6.0 ± 0.1) and xylose (4.2 ± 0.2). However, after deproteination, a difference was found in the order of the monosaccharide components, with galactose (41.1 ± 0.5) as a major unit followed by arabinose (34.7 ± 0.5), rhamnose (10.9 ± 0.2), glucose (6.6 ± 0.2), mannose (3.4 ± 0.2) and xylose (3.2 ± 0.2). Further, D-CQNP potentially stimulate the RAW 264.7 cells through the production of nitric oxide (NO), upregulating inducible nitric oxide synthase (iNOS) and various pro-inflammatory cytokines including interleukin (IL)-1β, IL-6, IL-10, and tumor necrosis factor-alpha (TNF-α). Moreover, stimulation of RAW 264.7 cells by D-CQNP takes place along the NF-κB and the MAPKs signaling pathways through the expression of cluster of differentiation 40 (CD40). This results demonstrate that RAW 264.7 cells are effectively stimulated after removal of the protein content in C. quinoa non-starch polysaccharides, which could be useful for develop a new immunostimulant agent.

Introduction and Aim: In this study, the interaction between pathogenic Naegleria fowleri and pathogenic bacteria such as methicillin-resistant Staphylococcus aureus (MRSA), Enterococcus faecalis and Salmonella typhi was analyzed by a monosaccharide of mannose. Moreover, since the form of N. fowleri was found in diseases as cysts, the interaction between cysts and bacteria was analyzed. Materials and Methods: In order to analyze the role of a monosaccharide called mannose in bacterial interaction, the analysis of bacterial association, invasion, and survival for amoeba treated with mannose was performed. N. fowleri trophozoites or cysts were pre-treated with a mannose at a concentration of 10, 50 and 100 mM for 1 hr at 37°C. Results: The MRSA association was hardly suppressed until the concentration of mannose was 50 mM, but its association was reduced by about 168% to N. fowleri trophozoites by 100 mM mannose. Compared to the results for MRSA, the association of E. faecalis had little effect by mannose. Very interestingly, although S. typhi showed much higher invasion than the above MRSA and E. faecalis, it did not survive at all within N. fowleri trophozoites. Ten mM mannose showed a nearly similar 1% association with N. fowleri cyst treated, but not with 50 mM and 100 mM mannose treated N. fowleri cyst at all. Conclusion: The association and invasion of S. typhi was highest for N. fowleri trophozoites and cysts, but the three bacteria did not survive in N. fowleri trophozoites and cysts.

A bifunctional glycoside hydrolase GH78 from the ascomycete Xylaria polymorpha (XpoGH78) possesses catalytic versatility towards both glycosides and esters, which may be advantageous for the efficient degradation of the plant cell-wall complex that contains both diverse sugar residues and esterified structures. The contribution of XpoGH78 to the conversion of lignocellulosic materials without any chemical pretreatment to release the water-soluble aromatic fragments, carbohydrates, and methanol was studied. The disintegrating effect of enzymatic lignocellulose treatment can be significantly improved by using different kinds of hydrolases and phenoloxidases. The considerable changes in low (3 kDa), medium (30 kDa), and high (> 200 kDa) aromatic fragments were observed after the treatment with XpoGH78 alone or with this potent cocktail. Synergistic conversion of rape straw also resulted in a release of 17.3 mg of total carbohydrates (e.g., arabinose, galactose, glucose, mannose, xylose) per gram of substrate after incubating for 72 h. Moreover, the treatment of rape straw with XpoGH78 led to a marginal methanol release of approximately 17 μg/g and improved to 270 μg/g by cooperation with the above accessory enzymes. In the case of beech wood conversion, the combined catalysis by XpoGH78 and laccase caused an effect comparable with that of fungal strain X. polymorpha in woody cultures concerning the liberation of aromatic lignocellulose fragments.

Viruses can evade the host immune system by displaying numerous glycans on their surface "spike-proteins" that cover immune epitopes. We have developed an ultrasensitive "single-pot" method to assess glycan occupancy and the extent of glycan processing from high-mannose to complex forms at each N-glycosylation site. Though aimed at characterizing glycosylation of viral spike-proteins as potential vaccines, this method is applicable for the analysis of site-specific glycosylation of any glycoprotein.

Recently, Lanzhou lily has attracted more attention because of its bioactive components specifically polysaccharides. We studied in vitro the effects of Lactobacillus plantarum fermentation on the physicochemical properties, chemical structure, and antioxidant activity of the Lanzhou lily polysaccharide. The results showed that compared with the unfermented Lanzhou lily polysaccharide (LP-W), the molecular weight (Mw) of the fermented Lanzhou lily polysaccharide (LPF-W) decreased from 4334 to 1684 kDa, the particle size decreased from 300.8 ± 6.38 to 141.9 ± 4.96 nm, and the solubility increased from 72.33 ± 3.58 to 104.27 ± 2.91 mg/mL. In addition, after fermentation, the monosaccharide composition of LPF-W changed, and the alternation of mannose residues and glucose residues disappeared. The results of the analysis of the antioxidant activity in vitro showed that compared with LP-W, the fermented LPF-W had higher DPPH radical ability, superoxide anion radical scavenging ability, and reducing efficiency, but the hydroxyl radical scavenging ability decreased. These findings provide a reference for the potential application of the lily polysaccharide as a plant-derived antioxidant in functional foods.

Tremella fuciformis TFC6 mycelia was isolated from its fruiting body. T. fuciformis TFCUV5 was obtained from TFC6 mycelia by ultraviolet mutagenesis to improve extracellular-polysaccharide (EPS) yield. In addition, we investigated the physicochemical properties and functionality of EPS for industrial applications and the effect of EPS on enhancement of the moisturizing factor and antiwrinkle effect in skin cells. The EPS yield was improved upon fermentation by the mutant strain through optimization of the medium composition and culture conditions to a maximum of 8.4 g/L; the productivity was 1.9 g/L-day under optimal culture conditions (initial pH 7.0 at 25°C, 300 rpm, 2 vvm). EPS with a molecular weight of 1.79 × 106 (± 0.721%) comprised mannose, xylose, glucuronic acid, fucose, and glucose. EPS extract increased both moisture and the antiwrinkle factor in skin cells. Therefore, EPS from TFCUV5 might have potential to be used in the cosmetics industry.

Microbial adhesion to host cells represents the initial step in the infection process. Several methods have been explored to inhibit microbial adhesion including the use of glycopolymers based on mannose, galactose, sialic acid and glucose. These sugar receptors are, however, abundant in the body, and are not unique to bacteria. Trehalose, in contrast, is a unique disaccharide that is widely expressed by microbes. This carbohydrate has not yet been explored as an anti-adhesive agent. Herein, gold nanoparticles (AuNPs) coated with trehalose-based polymers were prepared and compared to glucose-functionalized AuNPs and examined for their ability to prevent binding to endothelial cells. Acting as anti-adhesive agents, trehalose-functionalized NPs decreased the binding of S. aureus to HUVECs, while outperforming the control NPs. Microscopy revealed that trehalose-coated NPs bound strongly to S. aureus compared to the controls. In conclusion, nanoparticles based on trehalose could be a non-toxic alternative to inhibit S. aureus infection.

Lipopolysaccharide biosynthesis metabolites, such as d-glycero-β-d-manno-heptopyranosyl 1,7-diphosphate, d-glycero-β-d-manno-heptopyranosyl phosphate, and adenosine 5′-(l-glycero-β-d-manno-heptopyranosyl)diphosphate, have been found to activate NF-κB via alpha-kinase 1 and TRAF-interacting protein with forkhead associated domain. This axis has been determined as a novel pathway of innate immunity yet to be targeted for immunomodulatory treatment approaches. Key in understanding this new axis has been the ability to synthesize these metabolites. The design of synthetic analogues and probes have also been published not only to design new drugs, but also to gain insight into the mechanism of action for these compounds. The focus of the present review is the synthesis of heptose phosphate metabolites­ as well as synthetic analogues and probes.1 Introduction2 Synthesis of d-glycero-d-manno-Heptose2.1 Using d-Mannose as Starting Material2.2 Using d-Ribose as Starting Material2.3 Using 2,2-Dimethyl-1,3-dioxan-5-one as Starting Material3 Synthesis of l-glycero-d-manno-Heptose3.1 Using d-Mannose as Starting Material3.2 Using 2,2-Dimethyl-1,3-dioxan-5-one as Starting Material3.3 Using l-Lyxose as Starting Material4 Synthesis of Heptose Phosphates4.1 Synthesis of d-glycero-β-d-manno-Heptose 1,7-Diphosphate4.2 Synthesis of Heptose Phosphate Derivatives4.2.1 Development of Scaffolds for Conjugation4.2.2 Development of Heptose Phosphates Derivatives for Cell Intake and Metabolic Stability5 Conclusion and Outlook

The regulatory mechanisms of hypertension in youth are incompletely understood. We aimed to identify potential serum metabolic alterations associated with hypertension in adolescents. A 1:1 age- and sex-matched case-control study including 30 hypertensive adolescents aged 12-17 years and 30 normotensive adolescents for the training set and 14 hypertensive adolescents and 14 normotensive adolescents for the test set was performed, which came from one cross-sectional study in Ningxia, China. Hypertension was defined based on blood pressure (BP) values measured on three different occasions according to the BP reference of Chinese children and adolescents. Untargeted ultra-high-performance liquid tandem chromatography quadrupole time of flight mass spectrometry was used to identify differential metabolites between hypertensive and normotensive adolescents. A total of 77 metabolites in positive mode and 101 in negative mode were identified (VIP > 1.0 and P < 0.05). After adjustment for the false discovery rate, 4 differential metabolites in positive mode and 10 in negative mode were found (Q value < 0.05). The logistic regression model adjusted for body mass index and lipid profile selected four significant metabolites (4-hydroxybutanoic acid, L-serine, acetone, and pterostilbene). The main metabolic pathways of amino acid metabolism, pantothenate and CoA biosynthesis, glyoxylate and dicarboxylate metabolism, fructose and mannose metabolism, and linoleic acid metabolism may contribute to the development of hypertension in Chinese adolescents. Based on the receiver operating characteristic plot, 4-hydroxybutanoic acid, L-serine, acetone, and pterostilbene may preliminarily help distinguish hypertension from normal BP in adolescents, with AUC values of 0.857 in the training set and 0.934 in the test set. The identified metabolites and pathways may foster a better understanding of hypertension pathogenesis in Chinese adolescents.

Rggs are a group of transcriptional regulators with diverse roles in metabolism and virulence. Here, we present work on the Rgg1518/SHP1518 quorum sensing system of Streptococcus pneumoniae. The activity of Rgg1518 is induced by its cognate peptide, SHP1518. In vitro analysis showed that the Rgg1518 system is active in conditions rich in galactose and mannose, key nutrients during nasopharyngeal colonization. Rgg1518 expression is highly induced in the presence of these sugars and its isogenic mutant is attenuated in growth on galactose and mannose. When compared with other Rgg systems, Rgg1518 has the largest regulon on galactose. On galactose it controls up- or downregulation of a functionally diverse set of genes involved in galactose metabolism, capsule biosynthesis, iron metabolism, protein translation, as well as other metabolic functions, acting mainly as a repressor of gene expression. Rgg1518 is a repressor of capsule biosynthesis, and binds directly to the capsule regulatory region. Comparison with other Rggs revealed inter-regulatory interactions among Rggs. Finally, the rgg1518 mutant is attenuated in colonization and virulence in a mouse model of colonization and pneumonia. We conclude that Rgg1518 is a virulence determinant that contributes to a regulatory network composed of multiple Rgg systems.

Zahra Alipour,1 Diana Agostini-Vulaj,2 Jennifer Findeis-Hosey,2 Lei Liu,3 Raul S Gonzalez,4 Michael G Drage,2 Hannah Krigman,1 Zhongren Zhou5 1Department of Pathology and Immunology, Washington University, St Louis, MO, 63110, USA; 2Department of Pathology and Laboratory Medicine, University of Rochester, Rochester, NY, 14642, USA; 3Division of Biostatistics, Washington University, St Louis, MO, 63110, USA; 4Department of Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, 02215, USA; 5Department of Pathology and Laboratory Medicine, Robert Wood Johnson Medical School, Rutgers University, New Brunswick, NJ, 08901, USACorrespondence: Zhongren ZhouChief of Gastrointestinal Pathology, Department of Pathology and Laboratory Medicine, Robert Wood Johnson Medical School, Rutgers University, 125 Paterson Street, New Brunswick, NJ, 08903, USATel +1 732-667-0497Fax +1 732-235-8124Email zz442@rwjms.rutgers.eduIntroduction: Pancreatic ductal adenocarcinoma (PDAC) is the fourth leading cause of cancer-related death in the United States. The need for increased patient survival has not been met for PDAC. The addition of mannose to conventional chemotherapy leads to accumulation of mannose metabolite in cancer cells and increases subsequent cell death. This susceptibility to mannose depends on the levels of phosphomannose isomerase (PMI). The cancer cells with lower levels of PMI are more sensitive to mannose than cells with higher levels. In this study, we investigated the association of PMI expression with clinical and pathological features of PDAC cases.Methods: PMI antibody immunohistochemistry (AbCam) was performed on tissue microarrays from 235 PDAC by a standard protocol on Ventana automated immunostainer. The PMI intensity was graded (0– 3) and the proportion of positivity was scored. Correlation of PMI expression with staging and survival was analyzed.Results: Of the 235 cases, 51.5% (n=121) cases demonstrated grade 2 intensity with 90.1% of these (n=109) showing positivity in ≥ 70% of tumor cells. Ninety-eight (41.7%) cases exhibited grade 3 intensity with 94.9% (n=93) of these cases showing ≥ 70% reactivity. Sixteen cases (6.8%) were nonreactive (intensity grade 0– 1). Intensity of PMI expression was associated with significantly better prognosis as assessed by median survival in months (M): grade 0– 1 intensity group: 11.2 M; grade 2 intensity group: 25.2 M; and grade 3 intensity group: 33.2 M (p=0.03). A minority (6.8%) of PDACs show non-high PMI expression with poorer prognosis.Discussion: Mannose may be a particularly useful adjunct with chemotherapy to treat this aggressive subgroup. PMI expression is also a potential biomarker to predict the prognosis of PDAC.Keywords: pancreatic ductal adenocarcinoma, phosphomannose isomerase, PMI, immunohistochemistry, overall survival, gene expression

Background Prosopis juliflora is a clinically relevant allergic sensitizer worldwide and shares cross-reactivity with allergens from several tree pollen and food. The present study aims to purify and immunobiochemically characterize a major allergen from Prosopis pollen. The allergen was further investigated for its cross-reactivity with legume allergens. Methods Prosopis extract was fractionated by Q Sepharose and Superdex 75 gel filtration column to purify the allergen. Specific IgE against purified protein was estimated via ELISA and immunoblot. The protein was subjected to mass spectrometric analysis. Glycan characterization was performed by Schiff staining and lectin binding assay followed by deglycosylation studies. The functional activity of the purified protein was evaluated by the basophil activation test. Cross-reactivity was assessed by inhibition studies with legume extracts. Results A 35 kDa protein was purified and showed 75% IgE reactivity with the patients’ sera by ELISA and immunoblot. Glycan characterization of protein demonstrated the presence of terminal glucose and mannose residues. A reduction of 40% and 27% in IgE binding was observed upon chemical and enzymatic deglycosylation of the protein, respectively. The glycoprotein allergen upregulates the expression of CD203c on basophils which was significantly reduced upon deglycosylation, signifying its biological ability to activate the effector cells. The identified protein shared significant homology with Lup an 1 from the lupine bean. Immunoblot inhibition studies of the purified allergen with legume extracts underlined high cross-reactive potential. Complete inhibition was observed with peanut and common bean, while up to 70% inhibition was demonstrated with soy, black gram, chickpea, and lima bean. Conclusion A 35 kDa vicilin-like major allergen was isolated from P. juliflora. The protein possesses glycan moieties crucial for IgE binding and basophil activation. Furthermore, the purified protein shows homology with Lup an 1 and exhibits cross-reactivity with common edible legume proteins.

Glycoproteins carrying O-linked N-acetylgalactosamine, N-acetylgluco-samine, mannose, fucose, glucose, and xylose are found in the nervous system. Lipids can be glycosylated as well. Membrane lipid, ceramide, is modified by the addition of either glucose or galactose to form glycosphingolipid, galactosylceramide, or glucosylceramide. Recent analyses have identified glucosylated lipids of cholesterol and phosphatidic acid. These O-linked carbohydrate residues are found primarily on the outer surface of the plasma membrane or in the extracellular space. Their expression is cell or tissue specific and developmentally regulated. Due to their structural diversity, they play important roles in a variety of biological processes such as membrane transport and cell–cell interactions.

A Chlorella sp. was isolated from a brackish water high-altitude lake of the Northern-Western Himalayas and selected on the basis of its ability to secrete substantial amount of exopolysaccharide (EPS). The purified EPS was estimated to have molecular weight (Mw) of approximately 1.52 × 105 Da with polydispersity of 2.315 and showed amorphous dense morphology. The production of the EPS was maximized to ≅ 460 mg L−1 through nutrient optimisation. The monosaccharide composition of EPS showed the presence of hexose (glucose, galactose mannose), pentose (xylose, arabinose), deoxy (rhamnose), amino (galactosamine) and acid (galacturonic acid and glucuronic) sugars. The one-dimensional proton NMR and FT-IR analysis confirmed the presence of aliphatic groups. The pseudo-plastic rheology and the antioxidant activity of the EPS suggested its suitability for industrial applications. Importantly, the isolated polysaccharide exhibits its potential for easy and single pot biosynthesis of polysaccharide-capped gold-nanoparticles (AuNPs). The polysaccharide-capped AuNPs displayed exceptional stability at extended pH range and high salinity and suggest that this algal EPS can be utilized for its biotechnological application.

Cell-wall polysaccharides are synthesized from nucleotide sugars by glycosyltransferases. However, in what way the level of nucleotide sugars affects the structure of the polysaccharides is not entirely clear. guanosine diphosphate (GDP)-mannose (GDP-Man) is one of the major nucleotide sugars in plants and serves as a substrate in the synthesis of mannan polysaccharides. GDP-Man is synthesized from mannose 1-phosphate and GTP by a GDP-Man pyrophosphorylase, VITAMIN C DEFECTIVE1 (VTC1), which is positively regulated by the interacting protein KONJAC1 (KJC1) in Arabidopsis. Since seed-coat mucilage can serve as a model of the plant cell wall, we examined the influence of vtc1 and kjc1 mutations on the synthesis of mucilage galactoglucomannan. Sugar composition analysis showed that mannose content in adherent mucilage of kjc1 and vtc1 mutants was only 42% and 11% of the wild-type, respectively, indicating a drastic decrease of galactoglucomannan. On the other hand, structural analysis based on specific oligosaccharides released by endo-β-1,4-mannanase indicated that galactoglucomannan had a patterned glucomannan backbone consisting of alternating residues of glucose and mannose and the frequency of α-galactosyl branches was also similar to the wild type structure. These results suggest that the structure of mucilage galactoglucomannan is mainly determined by properties of glycosyltransferases rather than the availability of nucleotide sugars.

Immersion vaccination of single-walled carbon nanotubes loaded with mannose-modified glycoprotein (SWCNTs-MG) vaccine has been proved to be effective in preventing spring viraemia of carp virus (SVCV). Immunization procedure has immense consequence on the immune effect of the immersion vaccine. However, immunization procedure optimization for SWCNTs-MG vaccine against SVCV has not been reported. In this study, accordingly, a full-factor experiment was designed to optimize the immunization procedure of SWCNTs-MG vaccine by three aspects of vaccine dose (30 mg/L, 40 mg/L and 50 mg/L), immunization density (8 fish L-1 , 24 fish L-1 and 48 fish L-1 ) and immunization time (6, 12 and 24 hr). Furthermore, we used the immunization group (A1B2C1, 30 mg/L, 24 fish L-1 and 6 hr) in the previous study as a positive control (PC) to evaluate the immunization effect optimized conditions from the expression of immune-related genes and relative percentage survival (RPS). At 28 days post-vaccination (DPV), common carps were intraperitoneal injected SVCV challenged test indicated that the A1B2C2 group (30 mg/L, 24 fish L-1 , 12 hr) displayed superiority of protective efficacy compare with other groups and the RPS with 77.9%, which was 15.6% higher than the PC group of RPS with 62.3%. Moreover, the expression of immune-related genes such as IL-10, CD4 and MHC-II was also significantly higher than PC group. The specific experimental flow chart is shown in Figure 1. Conclusively, these results demonstrated that vaccine dose, immunization density and immunization time are 30 mg/L, 24 fish L-1 and 12 hr, which is the more appropriate immunization programme with juvenile carp for SWCNTs-MG vaccine. This study provides a profitable reference for improving the immune efficiency of aquatic immersion vaccine. [Figure: see text].

Immune checkpoint antibodies have emerged as novel therapeutics, while many patients are refractory. Researchers had identified tumor-associated macrophages (TAMs) is the pivotal factor involved in immune resistance and that manipulation of TAMs functions would improve the immunotherapies effectively. NF-κB pathway was one of the master regulators in TAMs manipulation. Inhibition of NF-κB pathway could achieve both re-polarization M2 TAMs and downregulation the expression of programmed cell death protein 1 (PD-1) ligand 1 (PD-L1) on TAMs to improve the effect of immunotherapies. Here, IMD-0354, inhibitor of NF-κB pathway was loaded in mannose modified lipid nanoparticles (M-IMD-LNP). Then, PD-1 antibody and M-IMD-LNP were co-loaded in matrix metalloproteinase 2 (MMP2) responsive and tumor target nanogels (P/ML-NNG). P/ML-NNG could co-deliver drugs to tumor site, disintegrated by MMP2 and release drugs to different targets. Evaluation of PD-1 expression, inhibition of NF-κB pathway, expression of PD-L1 on M2 TAMs and M2 TAMs re-polarization demonstrated that P/ML-NNG could block the PD-1/PD-L1 and NF-κB pathways simultaneously. Evaluation of CD4 + T cells, CD8 + T cells, Tregs, cytokines and antitumor immunity confirmed that IMD-0354 could improve the immunotherapies effectively. Those results provided forceful references for tumor immunetherapy.

Russula virescens is an edible wild mushroom that is widely distributed in south of China. This research aimed to analyze the structure characterization and evaluate the hypoglycemic, anticancer and immunological activities of two water soluble polysaccharides RVP-1 and RVP-2 from R. virescens. The results showed RVP-1 and RVP-2 were non-triple helix structured hetero-polysaccharides with different weight-average molecular weight 14,883 and 13,301 Da, respectively. Both RVP-1 and RVP-2 were composed of galactose, glucose, mannose and fructose, and the sugar residues were mainly linked by 1,6→, 1,2→, 1→ and 1,3,6→ glycosidic bonds. Moreover, the antidiabetic, anticancer and immune activities of RVP-1 and RVP-2 were explored in vitro methods. The two polysaccharides have potential for inhibiting α-glucosidase and α-amylase activities, suppressing HepG-2, A549 and MCF-7 cancer cells proliferation, and activating macrophage RAW 264.7 cells to secret immune cytokines for mediating cellular immune response. These findings provided a scientific basis for further utilization of polysaccharide from R. virescens.

Dendrobium officinale Kimura et Migo (D. officinale) is used as herbal medicine and new food resource in China, which is nontoxic and harmless, and can be used as common food. Polysaccharide as one of the main bioactive components in D. officinale, mainly composed of glucose and mannose (Manp: Glcp = 2.01:1.00-8.82:1.00), along with galactose, xylose, arabinose, and rhamnose in different molar ratios and types of glycosidic bonds. Polysaccharides of D. officinale exhibit a variety of biological effects, including immunomodulatory, anti-tumor, gastro-protective, hypoglycemic, anti-inflammatory, hepatoprotective, and vasodilating effects. This paper presents the extraction, purification, structural characteristics, bioactivities, structure-activity relationships and analyzes gaps in the current research on D. officinale polysaccharides. In addition, based on in vitro and in vivo experiments, the possible mechanisms of bioactivities of D. officinale polysaccharides were summarized. We hope that this work may provide helpful references and promising directions for further study and development of D. officinale polysaccharides.

Acanthopanax giraldii Harms is commonly used in traditional Chinese medicine to treat rheumatism, improve joints, and strengthen muscles and bones. The polysaccharides present in A. giraldii Harms contain major bioactive substances, which have antioxidant, anticancer, and antiviral activities. In this study, the structural characterization of the homogeneous polysaccharide isolated from A. giraldii Harms, known as AHP-II, and its immunomodulatory effects in vivo will be studied. High-performance ion chromatography (HPIC) and high-performance gel permeation chromatography (HPGPC) based analyses revealed that AHP-II was composed of various monosaccharides, which included rhamnose, arabinose, galactose, glucose, mannose, galacturonic acid, and glucuronic acid in molar ratios of 29.5 : 24.6 : 23.8 : 4.4 : 5.7 : 8.8 : 3.1, respectively, and had a collective molecular weight of 80.21 × 103 Da. Fourier-transform infrared (FTIR) spectroscopy indicated the presence of a pyranose ring and β-type glycosidic linkages in AHP-II. In addition, immunomodulatory effect analyses of AHP-II that used a cyclophosphamide-induced immunosuppressive mouse model demonstrated that its treatment could significantly restore spleen and thymus indices, promote the proliferation of splenic lymphocytes, elevate CD4+ T lymphocyte percentage and CD4+ : CD8+ ratio in the spleen, promote macrophage phagocytosis, and restore cytokines (IL-6, TNF-α, IgM, and IgG) levels. These results suggested that AHP-II could potentially be used as natural immunomodulator and as an alternative treatment to reduce chemotherapy-induced immunosuppression.

New therapeutic strategies for personalized medicine need to involve innovative pharmaceutical tools, for example, modular nanoparticles designed for direct immunomodulatory properties. We synthesized mannose-functionalized poly(propyleneimine) glycodendrimers with a novel architecture, where freely accessible mannose moieties are presented on poly(ethylene glycol)-based linkers embedded within an open-shell maltose coating. This design enhanced glycodendrimer bioactivity and led to complex functional effects in myeloid cells, with specific induction of interleukin-8 expression by mannose glycodendrimers detected in HL-60 and THP-1 cells. We concentrated on explaining the molecular mechanism of this phenomenon, which turned out to be different in both investigated cell lines: in HL-60 cells, transcriptional activation via AP-1 binding to the promoter predominated, while in THP-1 cells (which initially expressed less IL-8), induction was mediated mainly by mRNA stabilization. The success of directed immunomodulation, with synthetic design guided by assumptions about mannose-modified dendrimers as exogenous regulators of pro-inflammatory chemokine levels, opens new possibilities for designing bioactive nanoparticles.

Mannan polysaccharides (MPs), which contain substituted O-acetyl groups in their backbone, are abundant in the medicinal plant Dendrobium officinale. Acetyl groups can influence the physiological and biochemical properties of polysaccharides, which mainly accumulate in the stems of D. officinale at four developmental stages (S1-S4), showing an increasing trend and a link with water-soluble polysaccharides (WSPs) and mannose. The genes coding for enzymes that catalyze O-acetyl groups to MPs are unknown in D. officinale. The TRICHOME BIREFRINGENCE-LIKE (TBL) gene family contains TBL and DUF231 domains that can transfer O-acetyl groups to various polysaccharides. Based on an established D. officinale genome database, 37 DoTBL genes were identified. Analysis of cis-elements in the promoter region showed that DoTBL genes might respond to different hormones and abiotic stresses. Most of the genes with MeJA-responsive elements were upregulated or downregulated after treatment with MeJA. qRT-PCR results demonstrated that DoTBL genes had significantly higher expression levels in stems and leaves than in roots. Eight DoTBL genes showed relatively higher expression at S2-S4 stages, which showed a link with the content of WSPs and O-acetyl groups. DoTBL35 and its homologous gene DoTBL34 displayed the higher mRNA level in different organs and developmental stages, which might participate in the acetylation of MPs in D. officinale. The subcellular localization of DoTBL34 and DoTBL35 reveals that the endoplasmic reticulum may play an important role in the acetylation of MPs.

Molecular mobility is important for interactions of biofunctional polymers with target molecules. Monomer structures for synthetic biofunctional polymers are usually selected based on their compatibility with polymerization systems, whereas the influence of monomer structures on the interaction with target molecules is hardly considered. In this report, we evaluate the correlation between the monomer structures of glycopolymers and their interactions with concanavalin A (ConA) with respect to the molecular mobility. Two types of glycopolymers bearing mannose are synthesized with acrylamide or acrylate monomers. Despite the similar structures, except for amide or ester bonds in the side chains, the acrylate-type glycopolymers exhibit stronger interaction with ConA both in the isothermal titration calorimetry measurement and in a hemagglutination inhibition assay. Characterization of the acrylate-type glycopolymers suggests that the higher binding constant arises from the higher molecular mobility of mannose units, which results from the rotational freedom of ester bonds in their side chains.

In this study, two degraded polysaccharides were obtained by H2O2-Vc-ultrasonic and H2O2-Fe2+-ultrasonic treatment from Codium cylindricum. The basic structure of polysaccharides was characterized and the relationship between structure and antioxidant activity was studied. FTIR spectrum indicated that the degraded polysaccharides had similar functional groups (OH, CH, CO group) with ordinary polysaccharides. LC-MS analysis showed that the degraded polysaccharides were composed of the same monosaccharide units (mannose, galactose, arabinose, glucose, ribose) with Codium cylindricum polysaccharides, but the molar ratio was different. Meanwhile, the molecular weight and morphological feature of polysaccharides had been changed after degradation. Additionally, the antioxidant activity assay revealed that two degraded polysaccharides with lower molecular weight possessed better antioxidant property than ordinary polysaccharides. These results suggested that the basic structure of polysaccharides had not been damaged by two degradation methods, while the antioxidant activity was significantly enhanced.

Xiaoling Yu,1,2 Lingyi Huang,1 Chen You,2 Liying Huang1 1School of Pharmacy, Fujian Medical University, Fuzhou, Fujian, People’s Republic of China; 2Department of Pharmacy, Mengchao Hepatobiliary Hospital of Fujian Medical University, Fuzhou, People’s Republic of ChinaCorrespondence: Liying HuangSchool of Pharmacy, Fujian Medical University, No.1 Xuefu North Road University Town, Fuzhou, Fujian, 350122, People’s Republic of ChinaTel +86 13860635139Fax +86 591 22862016Email fjmuhly88@sina.com.cnPurpose: The polysaccharide of Anoectochilus roxburghii (wall.) Lindl. (ARPS) is one of its important active ingredients. Hepatoprotective effects of ARPS on rat liver injury induced by CCl4 were studied.Methods: ARPS was extracted using the ultrasonic method and successfully purified by high-speed counter-current chromatography (HSCCC) with a two-phase aqueous system composed of 12.5% PEG 1000– 20% K2HPO4:KH2PO4 (1:1). The HSCCC conditions were optimized, and the structure of ARPS was characterized. The hepatoprotective effects of ARPS against CCl4-induced chronic hepatic injury in SD rats were evaluated.Results: The results showed that ARPS was a water-soluble polysaccharide with a molecular weight of 28,518 Da. It was composed of mannose, ribose, glucose, and arabian sugar; its monosaccharide molar ratio was glucose:ribose:arabinose:mannose = 54.24:13.20:1.09:1.00. The purity of ARPS was determined by HPLC to be 96.93%. The intervention effects of ARPS on CCl4-induced hepatic damage model in rats showed that ARPS could effectively reduce the activity of alanine amino transferase and aspartate amino transferase, decrease the content of malondialdehyde and nitric oxide synthesis, and increase the content of glutathione. Pathology revealed that liver plate order, liver cell degeneration, and edema were improved; inflammatory cell infiltration was not observed after ARPS intervention.Conclusion: ARPS had the function of antioxidant for protecting CCl4-induced injured liver, and the mechanisms were related to anti-lipid peroxidation, which could eliminate oxygen-free radicals and protect liver cells from attacks by free radicals.Keywords: Anoectochilus roxburghi, polysaccharide, CCl4-induced hepatic damage, hepatoprotective effects, rat

Lactobacilli are able to produce exopolysaccharides (EPSs) with a wide diversity in structure and composition. However, changes in EPS production under environmental challenges are largely unknown. Here, to examine our hypothesis that EPS biosynthesis in lactobacilli is modulated by environmental conditions, we quantified and characterized the EPSs produced by Lactobacillus plantarum VAL6 following thermal stress and increased carbon dioxide (CO2) concentration. We found that the EPS yield and composition of L. plantarum strain VAL6 were markedly affected by these treatment conditions. The highest EPS yield reached 17.32 g/L under CO2 exposure, 2.2-fold higher compared to the control. The monosaccharide composition of synthesized EPSs was also altered when L. plantarum VAL6 was exposed to thermal stress and CO2 concentration increase. Whereas proportion of mannose was lower, glucose and galactose contents were higher than those in non-stressed variants. Both increased CO2 concentration and stress at 47°C enhanced the proportion of rhamnose in EPSs. Moreover, thermal stress and increased CO2 concentration triggered the expression of the genes involved in EPS biosynthesis, e.g., glmU, pgmB1, epsE, epsF, wzx, and wzy. Furthermore, the expression of epsE and epsF resulted in an increase in the content of galactose and glucose in EPS monosaccharide composition.

The structure and moisture retention of Tremella polysaccharide fermented from GCMCC5.39 (FTP) were evaluated. After UV, infrared spectrum analysis, HPAEC-PAD, HPSEC and 1 D NMR analysis, the composition of the purified FTP was determined. Purified components of fermented Tremella polysaccharide (FTPS) was made of galactose, mannose, glucose, galactosmine, glucosamine, and contain a large amount of hydroxyl, carbonyl and amino groups. FTPS was α -neutral pyranose without uronic acid. FTPS-1 and FTPS-2 were obtained after purification by DEAE-Sepharose Fast Flow Column. The molecular weights of FTPS-1 and FTPS-2 were 25 722 and 177 263 Da. FTPS-2 had a better ability to prevent moisture loss, and the optimal moisture retention period was 0–4 h. FTPS-2 could significantly increase the moisture content of the skin epidermis and showed a dose-concentration relationship. The effect of FTPS-2 on the expression of different moisturizing genes was evaluated in a human skin keratinocyte model. The results showed that FTPS-2 has no cytotoxicity, and could significantly promote AQP3, TGM1, CASP14, HYAL2, FLG gene expression level in HaCaT cells. It has the most significant influence at HYAL2 protein expression on 50 μg/mL.

Backgound A polysaccharide was purified in this paper, which was acquired from the fermentation broth of Dendrobium officinale Kimura et Migo. This article aims to investigate the structural features and bioactivity of this polysaccharide. RESULTS: The polysaccharide was purified and the main polysaccharide fraction (i.e. DOP-1) was obtained. High-performance gel permeation chromatography (HPGPC) revealed that the molecular weight of DOP-1 is 447.48 KDa. There are galactose, glucose and mannose in DOP-1 via monosaccharide composition analysis, and their ratio was 1:1.79:6.71. Methylation and NMR spectroscopic analysis indicated that the backbone of DOP-1 is: →4)-α-D-Glcp-(1→4)-α-D-Manp-(1→4)-α-D-Manp-(1→4,6)-α-D-Manp-(1→, its repeating units were also preliminarily established. In vitro tests proved that DOP-1 not only protects RAW264.7 macrophages from the cytotoxic effect induced by lipopolysaccharide (LPS), but also inhibits cytokine (i.e. IL-6 and TNF-α) produced which are induced by LPS. DOP-1 demonstrated good scavenging activity in vitro toward 1, 1-diphenyl-2-picrylhydrazyl (DPPH) and hydroxyl radicals, as well as good metal chelating activity. Therefore, DOP-1 has potential applications in antioxidant. CONCLUSION: These findings strengthen that the structural characteristics of DOP-1 support its favourable biological activities and lay a strong foundation for further exploration of its structure-activity relationships and activity development and providing experimental data for the development and utilization of fermentation broth of D. officinale This article is protected by copyright. All rights reserved.

This study targeted the production of exopolysaccharide from Bacillus sp. NRC5 grown in Egyptian seawater to use it as natural antitumor therapy. The biological activities of selected exopolysaccharide (BEPS) as an antioxidant, anti-inflammatory and anticancer have been studied. BEPS was evaluated as an anti-inflammatory in vitro against cyclooxygenase (COX-1 and COX-2) and evaluated as an anticancer on human breast and prostate cancer cell lines (MCF-7 and PC3). In addition, BEPS antitumor activity was tested against the Ehrlich Ascites Carcinoma (EAC) model. The BEPS presented potent antioxidant activities, consisted of glucose, mannose, and mannuronic acid in a molar ratio of 1.0:1.7:0.8 with a molecular weight of 3.59 × 105 g/mol. BEPS showed a promising COX-2 inhibitory effect in comparison with the reference drug celecoxib. BEPS appeared efficient anticancer property, where it killed 64.20 and 70.20% of MCF-7 and PC3 cells at 100 µg/ml, respectively (IC50, 76.70, and 70.40 µg/ml, respectively). BEPS exhibited antitumor ability as it prolonged the lifespan of mice to reach 75 days instead of 20 days in the tumor control, reduced viable cancer cells count, tumor volume and weight, modulated blood components, and white blood cells differentiation. BEPS produced from Bacillus sp. NRC5 showed its antioxidant and anti-inflammatory abilities and antitumor abilities, which may all be attributed to its unique composition containing sulfated moieties and uronic acids.

Disruption of the mannose 6‐phosphate (M‐6‐P) pathway in HeLa cells by inactivation of the GNPTAB gene, which encodes the α/β subunits of GlcNAc‐1‐phosphotransferase, results in missorting of newly synthesized lysosomal acid hydrolases to the cell culture media instead of transport to the endolysosomal system. We previously demonstrated that the majority of the lysosomal aspartyl protease, cathepsin D, is secreted in these GNPTAB −/− HeLa cells. However, the intracellular content of cathepsin D in these cells was still greater than that of WT HeLa cells which retained most of the protease, indicating a marked elevation of cathepsin D expression in response to abrogation of the M‐6‐P pathway. Here, we demonstrate that HeLa cells lacking GlcNAc‐1‐phosphotransferase show a fivefold increase in cathepsin D mRNA expression over control cells, accounting for the increase in cathepsin D at the protein level. Further, we show that this increase at the mRNA level occurs independent of the transcription factors TFEB and TFE3. The intracellular cathepsin D can still be trafficked to lysosomes in the absence of the M‐6‐P pathway, but fails to undergo proteolytic processing into the fully mature heavy and light chains. Uptake experiments performed by feeding GNPTAB −/− HeLa cells with various phosphorylated cathepsins reveal that only cathepsin B is capable of partially restoring cleavage, providing evidence for a role for cathepsin B in the proteolytic processing of cathepsin D., Cathepsin D mRNA expression is increased fivefold in HeLa cells when the Man‐6‐P pathway is inactivated. This increase is independent of both TFEB and TFE3. These cells also lack cathepsin B, which is partly responsible for the failure of cathepsin D processing to occur. The effect is reversed by feeding the knockout HeLa cells with cathepsin B.

Aim: To design D-Mannose conjugated 5-Fluorouracil (5-FU) loaded Jackfruit Seed Starch Nanoparticles (JFSSNPs) for site-specific delivery. Background: Liver cancer is the third leading cause of death in the world and the fifth most often diagnosed cancer. It is a major global threat to public health. Treatment of liver cancer with conventional method bears several side effects, thus to undertake these side effects as a formulation challenge, it is necessary to develop novel target-specific drug delivery system for the effective and better localization of drug into the proximity of target with restricting the movement of the drug in normal tissues. Objective: To optimize and characterize the developed D-Mannose conjugated 5-Fluorouracil (5- FU) loaded Jackfruit Seed Starch Nanoparticles (JFSSNPs) for effective treatment of liver cancer. Materials and methods: 5-FU loaded JFSSNPs were prepared and optimized formulations having higher encapsulation efficiency were conjugated with D-Mannose. These formulations were characterized for size, morphology, zeta potential, X-Ray Diffraction, and Differential Scanning Calorimetry. The potential of NPs was studied using in vitro cytotoxicity assay, in vivo kinetic studies, and bio-distribution studies. Results and Discussion: 5-Fluorouracil loaded NPs had a particle size between 336 to 802 nm with drug entrapment efficiency between 64.2 to 82.3%. In XRD analysis, 5-FU peak was diminished in the diffractogram, which could be attributed to the successful incorporation of the drug in amorphous form. DSC study suggests there was no physical interaction between 5-FU and Polymer. NPs showed sustained in vitro 5-FU release up to 2 hours. In vivo, mannose conjugated NPs prolonged the plasma level of 5-FU and assisted in the selective accumulation of 5-FU in the liver (vs. other organs spleen, kidney, lungs, and heart) compared to unconjugated one and plain drug. Conclusion: In vivo, bio-distribution, and plasma profile studies resulted in a significantly higher concentration of 5-Fluorouracil liver, suggesting that these carriers are efficient, viable, and targeted carrier of 5-FU treatment of liver cancer.

Agaricus bisporus mannose-binding protein (Abmb) was discovered as part of mushroom tyrosinase (PPO3) complex. Apart from its presence, nothing is known about its function or activity in the mushroom. The protein is evolutionarily related to lectins with β-trefoil fold, which are glucose or galactose (and their derivatives) binding proteins. Abmb is also recently showed to display the typical agglutination activity of lectin when in complex with PPO3; this further supports Abmb similarity to its structural homologs from lectin with β-trefoil fold. However, Abmb has no affinity towards glucose or galactose but for mannose, thus its binding to the sugar may be different from its homologs. To date, the natural ligand of Abmb is unknown and the structure of Abmb in the presence of a ligand is not available. Therefore, the mannose-binding site of Abmb was predicted using molecular docking, which was consulted with the information from its structural homologs. This conservative approach would prevent over-speculation. The mannose-binding site of Abmb is likely located in the same region to that of Abmb structural homologs but with a shift in position due to the presence of additional surface loop. In addition, benefiting from the information from an in vitro study on Abmb sugar specificity, the mannose poses suggested that the sugar might interact with the side chains of Arg15, Thr45, Gln48, Asp49, Asp51 and Arg51. Most of these residues were equally present in Abmb structural homologs despite variation of their positions in the amino acid sequence. The variation probably originates from alteration of its amino acid sequence during evolution.

Biofilm formation, as adapting strategies, is the result of stressful conditions that Salmonella faces in hostile environments like surface water. We evaluated river water effect on Salmonella biofilm formation ability in terms of physical, morphological characteristics and chemical composition. A new morphotype SPAM (soft, pink and mucoid) was detected in Oranienburg strains S-76 and S-347 (environmental and clinical isolate). Oranienburg serotypes showed very marked behavior in adherence, pellicle liquid-air and resistance, being Oranienburg S-76 the strongest biofilm producer. All strains when exposed to river water presented an overlapping mucoid layer in the morphotype and increased their motility except Oranienburg S-347. The most motile was Typhimurium (control) and the least Infantis S-304 (clinical isolate). Mannose, glucose, galactose and ribose were the main biofilm sugar components; type and concentration of sugar suggest a morphotype/serotype dependent pattern. Strong morphotypes expressed in this study may be an effective protective strategy for Salmonella in hostile environments.

A new immunoregulatory polysaccharide (DCP-W4) with a relative molecular mass of 1.37 × 104 Da was separated and purified from the stems of Dendrobium chrysotoxum under the guidance of immunoregulatory activity assay based on the small intestinal Peyer’s patch-mediated proliferation of bone marrow cells. Analysis of physicochemical features showed that DCP-W4 was a glucomannan consisting of glucose and mannose in a molar ratio of 3.0 : 1.0. Oral administration of mice suggested that DCP-W4 could significantly influence the body’s immune activity, which was evidenced by its ability to change the indexes of immune organs including spleen and thymus, the proliferation of bone marrow cells mediated by the supernatant of Peyer’s patch cells, the balance of Th1/Th2 through effectively regulating the secretion and ratio of IFN-γ and IL-4, the proportion and differentiation status of different immune cells including T-cells, dendritic cells and macrophages, and the production of sIgA as well as β-defensins and mucin-2 in the small intestine or in the spleen of mice. These results indicated that DCP-W4 was a polysaccharide with a hopeful potential for the modulation of intestine immune system and systemic immune system.

Presence of the cell wall glycopolymer rhamnan was established for members of the genus Curtobacterium (family Microbacteriaceae) by using chemical and NMR spectroscopic methods. The rhamnan with the repeating unit, →2)-α-L-Rhap-(1→3)-α-L-Rhap-(1→, was found in C. flaccumfaciens pv. flaccumfaciens VKM Ас-1923T, C. flaccumfaciens pv. poinsettiae VKM Ас-1924T, C. pusillum VKM Ac-2099T, C. herbarum VKM Ac-2512T, and Curtobacterium sp. VKM Ac-1811. Each of the studied strains also contained a second, acidic phosphate-free polysaccharide. NMR spectra of these polysaccharides differed, indicating the structural differences. Cell wall hydrolysates of all studied strains were found to contain rhamnose, mannose, glucose, galactose, and glucosamine. Further research will facilitate more detailed characterization of the composition of cell wall glycopolymers in Curtobacterium species, as well as evaluation of their taxonomic significance. The data on the polymer structure will contribute to understanding of the mechanisms of colonization and infection of higher organisms by Curtobacterium species and bacterial interaction in microbial communities and with abiotic environmental objects.

Carboxymethyl pachymaran (CMP) was prepared from Poria cocos polysaccharide by carboxymethylation. Two types of CMP (CMP-1 and CMP-2) were further purified by DEAE-52 anion-exchange chromatography. The structure characteristics and immunomodulatory activities of CMP-1 and CMP-2 were investigated. CMP-1 was determined as β-(1 → 3)-d-glucan. A β-(1 → 3)-d-glucan backbone structure was also found in CMP-2, which was mainly consistent of mannose and glucose, with the mole ratio of 0.03:1. The molecular weight of CMP-1 was 126.1 kDa with a 30.4 nm irregular sphere in distilled water. However, the molecular weight of CMP-2 was 172.6 kDa in a 19.9 nm spherical structure in water solution. Both CMP-1 and CMP-2 had triple helical structure, which can promote the proliferation and the phagocytosis of macrophages. Moreover, CMP-1 and CMP-2 both could improve the secretions of NO, TNF-α and IL-6 by increasing the expression of iNOS, TNF-α and IL-6 mRNA, but CMP-1 exhibited a stronger immunomodulatory ability than that of CMP-2. Our results indicated that CMP-1 and CMP-2 can act as potential immunomodulatory agents.

Cationic glycopolymers with structures similar to those of typical poly(ionic liquid)s (PILs) were synthesized via the quaternization reaction of poly(4-vinyl pyridine) with halogen-functionalized d-mannose and tetraphenylethylene units. Such postpolymerization modification provided PILs with aggregation-induced emission effect as well as specific carbohydrate-protein recognition with lectins such as concanavalin A. The interactions between cationic glycopolymers and different microorganisms, including Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli, were used for the killing, imaging, and detection of bacteria. Besides, these sugar-containing PILs showed a relatively low hemolysis rate due to the presence of saccharide units, which may have potential application in the field of biomaterials.

The α1,6-fucosyltransferase, FUT8, is the sole enzyme catalyzing the core-fucosylation of N-glycoproteins in mammalian systems. Previous studies using free N-glycans as acceptor substrates indicated that a terminal β1,2-GlcNAc moiety on the Man-α1,3-Man arm of N-glycan substrates is required for efficient FUT8-catalyzed core-fucosylation. In contrast, we recently demonstrated that, in a proper protein context, FUT8 could also fucosylate Man5GlcNAc2 without a GlcNAc at the non-reducing end. We describe here a further study of the substrate specificity of FUT8 using a range of N-glycans containing different aglycones. We found that FUT8 could fucosylate most of high-mannose and complex-type N-glycans, including highly branched N-glycans from chicken ovalbumin, when the aglycone moiety is modified with a 9-fluorenylmethyloxycarbonyl (Fmoc) moiety or in a suitable peptide/protein context, even if they lack the terminal GlcNAc moiety on the Man-α1,3-Man arm. FUT8 could also fucosylate paucimannose structures when they are on glycoprotein substrates. Such core-fucosylated paucimannosylation is a prominent feature of lysosomal proteins of human neutrophils and several types of cancers. We also found that sialylation of N-glycans significantly reduced their activity as a substrate of FUT8. Kinetic analysis demonstrated that Fmoc aglycone modification could either improve the turnover rate or decrease the KM value depending on the nature of the substrates, thus significantly enhancing the overall efficiency of FUT8 catalyzed fucosylation. Our results indicate that an appropriate aglycone context of N-glycans could significantly broaden the acceptor substrate specificity of FUT8 beyond what has previously been thought.

Introduction. One of the key tasks in freeze‑drying of drug is to choose the optimal cryoprotector that provides a high‑quality lyophilized product.The aim of research. Selection of cryoprotectant and its concentration for the preparation of lyophilizate of the liposomal dosage form of the indolocarbazole derivative LHS‑1269.Materials and methods. Substance LHS‑1269 ≥99 % (N. N. Blokhin National Medical Research Center of Onco logy), egg phosphatidylcholine Е РС S (Lipoid, Germany), cholesterol ≥99 % (Sigma‑Aldrich, Japan), polyethylene glycol‑2000‑distearoyl phosphatidylethanolamine (Lipoid, Germany), mannose‑d (+) 99 % (Kaden, Germany), sucrose (Himmed, Russia), trehalose dihydrate (Himmed, Russia). To obtain LHS‑1269 liposomes, the Bangham method was used in modification for hydrophobic substances with subsequent extrusion of the dispersion of multilayer phos‑ pholipid vesicles. The prepared liposomal dispersion was dosed into vials of 6 ml and lyophilized in the freeze‑dryng chamber using the «step‑by‑step» freezing mode. LHS‑1269 liposomes were analyzed before and after freeze‑drying using laser scattering spectroscopy and determination of the electrophoretic mobility of particles.Results. To prevent the destruction of LHS‑1269 liposomes during lyophilization, substances from the carbohy‑ drates class – mannose, sucrose and trehalose – were studied in two concentrations. In the course of compara‑ tive evaluation of the obtained lyophilizates in terms of quality, appearance, rehydration, size and zeta potential of liposomes before and after sublimation, it was found that sucrose introduced into the liposomal dosage form LHS‑1269 in the molar ratio sucrose / egg phosphatidylcholine 5:1 has optimal cryoprotective properties.Conclusion. As a result of the study, the optimal cryoprotector and its concentration were selected, which ensure the production of high‑quality lyophilizate of the liposomal composition of the indolocarbazole derivative LHS‑1269.

Cancer cells increase their metabolic activity by enhancing glucose uptake through overexpression of hexose transporters (Gluts). Gluts also have the capacity to transport other molecules besides glucose, including fructose, mannose, and dehydroascorbic acid (DHA), the oxidized form of vitamin C. The majority of research studies in this field have focused on the role of glucose transport and metabolism in cancer, leaving a substantial gap in our knowledge of the contribution of other hexoses and DHA in cancer biology. Here, we summarize the most recent advances in understanding the role that the multifunctional transport capacity of Gluts plays in biological and clinical aspects of cancer, and how these characteristics can be exploited in the search for novel diagnostic and therapeutic strategies.

Heterologous endo-xanthanase (EX) from the thermophilic planktomycete Thermogutta terrifontis strain was obtained using Penicillium verruculosum 537 (ΔniaD) expression system with the cellobiohydrolase 1 gene promoter. Homogeneous EX with a molecular weight of 23.7 kDa (pI 6.5) was isolated using liquid chromatography methods. This xanthan degrading enzyme also possesses the enzymatic activity towards CM-cellulose, β-glucan, curdlan, lichenan, laminarin, galactomannan, xyloglucan but not towards p-nitrophenyl derivatives of β-D-glucose, mannose and cellobiose. The temperature and pH optima of EX were 55°C and 4.0, respectively; the enzyme exhibited 90% of its maximum activity in the temperature range 50-60°C and pH 3-5.