Functional significance of O-linked N-acetylglucosamine protein modification in regulating autophagy.

Autophagy is a core molecular pathway that preserves cellular and organismal homeostasis. Being susceptible to nutrient availability and stress, eukaryotic cells recycle or degrade internal components via membrane transport pathways to provide sustainable biological molecules and energy sources. The dysregulation of this highly conserved physiological process has been strongly linked to human disease. Post-translational modification, a mechanism that regulates protein function, plays a crucial role in autophagy regulation. O-linked N-acetylglucosamine protein modification (O-GlcNAcylation), a monosaccharide post-translational modification of intracellular proteins, is essential in nutritional and stress regulatory mechanisms. O-GlcNAcylation has emerged as an essential regulatory mechanism of autophagy. It regulates autophagy throughout its lifetime by targeting the core components of the autophagy regulatory network. This review provides an overview of the O-GlcNAcylation of autophagy-associated proteins and their regulation and function in the autophagy pathway. Therefore, this article may contribute to further understanding of the role of O-GlcNAc-regulated autophagy and provide new perspectives for the treatment of human diseases.

Connexin32 ameliorates epithelial-to-mesenchymal-transition in diabetic renal tubular via inhibiting NOX4.

Renal tubulointerstitial fibrosis (RIF), characterized by epithelial-to-mesenchymal transition (EMT) of renal tubular epithelial cells (TECs), is the main cause of diabetic renal fibrosis. Oxidative stress plays a pivotal role in the development of diabetic RIF. Connexin32 (Cx32), prominently expressed in renal TECs, has emerged as an important player in the regulation of oxidative stress. However, the role of Cx32 in diabetic RIF has not been explored yet. Here, we showed that adenovirus-mediated Cx32 overexpression suppressed EMT to ameliorate RIF and renal function in STZ-induced diabetic mice, while knockout (KO) of Cx32 exacerbated RIF in diabetic mice. Moreover, overexpression of Cx32 inhibited EMT and the production of extra cellular matrix (ECM) in high glucose (HG) induced NRK-52E cells, whereas knockdown of Cx32 showed the opposite effects. Furthermore, we showed that NOX4, the main source of ROS in renal tubular, was down-regulated by Cx32. Mechanistically, Cx32 down-regulated the expression of PKC alpha in a carboxyl-terminal-dependent manner, thereby inhibiting the phosphorylation at Thr147 of p22phox triggered by PKC alpha, which ultimately repressed the formation of the p22phox-NOX4 complex to reduce the protein level of NOX4. Thus, we establish Cx32 as a novel target and confirm the protection mechanism in RIF.

Roles of the NLRP3 inflammasome in the pathogenesis of diabetic nephropathy.

Diabetic nephropathy (DN) is a serious complication of diabetes mellitus, and persistent inflammation in circulatory and renal tissues is an important pathophysiological basis for DN. The essence of the microinflammatory state is the innate immune response, which is central to the occurrence and development of DN. Members of the inflammasome family, including both "receptors" and "regulators", are key to the inflammatory immune response. Nucleotide binding and oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3) and other inflammasome components are able to detect endogenous danger signals, resulting in activation of caspase-1 as well as interleukin (IL)-1ß, IL-18 and other cytokines; these events stimulate the inflammatory cascade reaction, which is crucial for DN. Hyperglycaemia, hyperlipidaemia and hyperuricaemia can activate the NLRP3 inflammasome, which then mediates the occurrence and development of DN through the K+ channel model, the lysosomal damage model and the active oxygen cluster model. In this review, we survey the involvement of the NLRP3 inflammasome in various signalling pathways and highlight different aspects of their influence on DN. We also explore the important effects of the NLRP3 inflammasome on kidney function and structural changes that occur during DN development and progression. It is becoming more evident that NLRP3 inflammasome targeting has therapeutic potential for the treatment of DN.

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.

A new non-enzymatic biosensor for the determination of bisphenol-A.

In this study, a new non-enzymatic carbon paste biosensor was developed for the determination of Bisphenol-A (BPA) based on Multiwalled Carbon Nanotube (MWCNT) modified Myoglobin (Mb). The measurement principle of the biosensor was developed based on the inhibition effect of BPA on the heme group of myoglobin in the presence of hydrogen peroxide. With the designed biosensor, measurements were taken in the potential range of (-0.15 V & +0.65 V) using the differential pulse voltammetry (DPV) method in the medium containing K4[Fe(CN)6]. The linear range for BPA was determined to be 100-1000 µM. Response time was calculated as 16 s. The limit of detection was set at 89 µM. As a result, it has been proven that MWCNT modified myoglobin based biosensor is an alternative method that can be used for BPA determination, giving very sensitive and fast results.

The structures of two glucomannans from Bletilla formosana and their protective effect on inflammation via inhibiting NF-κB pathway.

Bletilla formosana is a traditional Chinese herbal medicine and is widely consumed as foods and medicines in China. However, the chemical structure and bioactivity of its polysaccharides remain unknown. Herein, two new polysaccharides, BFP60 and BFP80, with molecular weights of 3.99 kDa and 10.07 kDa, respectively, were isolated and purified from dried tuber of B. formosana. Structural analysis suggested that BFP60 and BFP80 may have backbone consisted of →4)-ß-d-Man-(1→,→4)-ß-d-Glc-(1→,→4)-2-O-acetyl-ß-d-Man-(1→, and →4)-3-O-acetyl-ß-d-Man-(1→. Inflammation assay in LPS-induced RAW264.7 cells showed that the productions of NO and pro-inflammatory cytokines including IL-6, IL-1ß, TNF-α, and IFN-γ were significantly reduced, and the expression of iNOS, COX-2, and target proteins in the NF-κB pathway were suppressed after BFP60 and BFP80 pretreatment. These findings indicated that this novel polysaccharide had significant inflammatory protective effects in vitro.

Preparation and structural properties of selenium modified heteropolysaccharide from the fruits of Akebia quinata and in vitro and in vivo antitumor activity.

Cancer is a complex disease, and blocking tumor angiogenesis has become one of the most promising approaches in cancer therapy. Here, an exopoly heteropolysaccharide (AQP70-2B) was firstly isolated from Akebia quinata. Monosaccharide composition indicated that the AQP70-2B was composed of rhamnose, glucose, galactose, and arabinose. The backbone of AQP70-2B consisted of →1)-l-Araf, →3)-l-Araf-(1→, →5)-l-Araf-(1→, →3,5)-l-Araf-(1→, →2,5)-l-Araf-(1→, →4)-d-Glcp-(1→, →6)-d-Galp-(1→, and →1)-d-Rhap residues. Based on the close relationship between selenium and anti-tumor activity, AQP70-2B was modified with selenium to obtain selenized polysaccharide Se-AQP70-2B. Then, a series of methods for analysis and characterization, especially scanning electron microscopy coupled with energy dispersive spectrometry (SEM-EDS), indicated that Se-AQP70-2B was successfully synthesized. Furthermore, zebrafish xenografts and anti-angiogenesis experiments indicated that selenization could improve the antitumor activity by inhibiting tumor cell proliferation and migration and blocking angiogenesis.

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.

Comparison of pyrazines formation in methionine/glucose and corresponding Amadori rearrangement product model.

The generation of pyrazines in a binary methionine/glucose (Met/Glc) mixture and corresponding methionine/glucose-derived Amadori rearrangement product (MG-ARP) was studied. Quantitative analyses of pyrazines and methional revealed that MG-ARP generated more methional compared to Met/Glc, whereas lower content and fewer species of pyrazines were observed in the MG-ARP model. Comparing the availability of α-dicarbonyl compounds generated from the Met/Glc model, methylglyoxal (MGO) was a considerably effective α-dicarbonyl compound for the formation of pyrazines during MG-ARP degradation, but glyoxal (GO) produced from MG-ARP did not effectively participate in the corresponding formation of pyrazines due to the asynchrony on the formation of GO and recovered Met. Diacetyl (DA) content was not high enough to form corresponding pyrazines in the MG-ARP model. The insufficient interaction of precursors and rapid drops in pH limited the formation of pyrazines during MG-ARP degradation. Increasing reaction temperature could reduce the negative inhibitory effect by promoting the content of precursors.

Reducing end thiol-modified nanocellulose: Bottom-up enzymatic synthesis and use for templated assembly of silver nanoparticles into biocidal composite material.

Nanoparticle-polymer composites are important functional materials but structural control of their assembly is challenging. Owing to its crystalline internal structure and tunable nanoscale morphology, cellulose is promising polymer scaffold for templating such composite materials. Here, we show bottom-up synthesis of reducing end thiol-modified cellulose chains by iterative bi-enzymatic ß-1,4-glycosylation of 1-thio-ß-d-glucose (10 mM), to a degree of polymerization of ∼8 and in a yield of ∼41% on the donor substrate (α-d-glucose 1-phosphate, 100 mM). Synthetic cellulose oligomers self-assemble into highly ordered crystalline (cellulose allomorph II) material showing long (micrometers) and thin nanosheet-like morphologies, with thickness of 5-7 nm. Silver nanoparticles were attached selectively and well dispersed on the surface of the thiol-modified cellulose, in excellent yield (≥ 95%) and high loading efficiency (∼2.2 g silver/g thiol-cellulose). Examined against Escherichia coli and Staphylococcus aureus, surface-patterned nanoparticles show excellent biocidal activity. Bottom-up approach by chemical design to a functional cellulose nanocomposite is presented. Synthetic thiol-containing nanocellulose can expand the scope of top-down produced cellulose materials.

Composition and potential as a prebiotic functional food of a Giant Willow Aphid (Tuberolachnus salignus) honeydew honey produced in New Zealand.

The Giant Willow Aphid (Tuberolachnus salignus, GWA) is an invasive pest insect in New Zealand, which excretes honeydew. European honeybees collect this honeydew and make it into a type of honey that crystallizes in the comb, representing a significant loss to apiarists. This crystallization has been ascribed to high concentrations of oligosaccharides, particularly melezitose. In this research, the first carbohydrate profile of GWA honeydew honey, a sample of GWA honeydew honey was found to contain 37.8% total oligosaccharides of which 27.4% was melezitose, and 2.5% gluconic acid (higher than typical honeydew honeys); 41.2% monosaccharides (lower than typical honeydew honeys); and 0.054% salicylic acid (higher than previous estimates). Melezitose extracted from GWA honeydew honey was not significantly hydrolyzed in crude human-stomach and human-small-intestine simulations and may therefore meet the prebiotic criterion of human indigestibility.

Structures of fructan and galactan from Polygonatum cyrtonema and their utilization by probiotic bacteria.

Polygonatum cyrtonema is a known tonic herb in Chinese Materia Medica, extensively consumed in China, but the structure and activity of its polysaccharide components remain to be clarified. Herein, two new polysaccharides (a fructan and a galactan) were purified from the dried and the processed P. cyrtonema rhizome, respectively. Structural analysis suggested that the fructan consisted of a (2 â†’ 6) linked ß-d-Fruf residues backbone with an internal α-d-Glcp residue and two (2 â†’ 1) linked ß-d-Fruf residues branches, and that the galactan was a (1 â†’ 4)-ß-d-galactan branched with a single ß-d-galactose at C-6 at about every nine residues in its main chain. The bioactive assay showed that the fructan and the galactan remarkably promoted growth of Bifidobacterium and Lactobacillus strains, indicating that they possess prebiotic activity. These findings may help expand the application of the polysaccharides from the tonic herb P. cyrtonema as functional ingredients in food products.

Combining cobalt ferrite and graphite with cellulose nanocrystals for magnetically active and electrically conducting mesoporous nanohybrids.

Free-standing mesoporous membranes based on cellulose nanocrystals (CNCs) are fabricated upon the incorporation of cobalt ferrite (CoFe2O4) and graphite nanoparticles at concentrations up to 20 wt % through a soft-templating process. Scanning electron microscopy (SEM) and N2 adsorption-desorption isotherms reveal the development of highly-porous interconnected random 3D structure with surface areas up to 193.9 m2 g-1. Thermogravimetric analysis (TGA) shows an enhanced thermal stability thanks to the formation of a tortuous network limiting the hindrance of degradation by-products. Vibrating sample magnetometer (VSM) reveals a maximum magnetization saturation of 8.77 emu·g-1 with materials having either ferromagnetic or diamagnetic behaviour upon the incorporation of CoFe2O4 and graphite, respectively. Four-point-probe measurements display a maximum electrical conductivity of 9.26 ±â€¯0.04 S·m-1 when graphite is incorporated into CNCs. A proof of concept for the applicability of synthesized nanohybrids for environmental remediation is provided, presenting the advantage of their easy recovery using external magnetic fields.

Kinetic study on the digestibility of lactose and lactulose using small intestinal glycosidases.

Lactose is mostly hydrolysed in the small intestine, whereas lactulose, recognised prebiotic carbohydrate, reaches the colon to be fermented by the intestinal microbiota. Digestibility of these substrates was investigated by an in vitro digestion model using a Rat Small Intestine Extract (RSIE). A kinetic study of lactose digestion showed levels of hydrolysis (82.8%) at 0.2 mg*mL-1 and the highest hydrolysis rate constant (kobt). Considering these conditions, lactulose showed high resistance to intestinal digestion by RSIE, resulting in low hydrolysis degrees (20.4%) after 5 h reaction. These results underline the suitability of these intestinal extracts under the studied conditions, as a reliable tool to evaluate carbohydrate digestion and support the evidences towards the higher resistance of galactosyl-fructose linkages during its intestinal degradation.

Physicochemical, bioactive and rheological properties of an exopolysaccharide produced by a probiotic Pediococcus pentosaceus M41.

This study aimed to investigate the physicochemical properties, health-promoting benefits and rheological properties of an EPS produced by a novel probiotic Pediococcus pentosaceus M41 isolated from a marine source. P. pentosaceus M41 was able to produce an EPS with average molecular weight of 682.07 kDa. EPS-M41 consisted of arabinose, mannose, glucose and galactose with a molar ratio of 1.2:1.8:15.1:1.0. EPS-M41 structure could be proposed as →3)α-D-Glc(1→2)ß-D-Man(1→2)α-D-Glc(1→6)α-D-Glc(1→4)α-D-Glc(1→4)α-D-Gal(1→ with arabinose linked at the terminals. At concentration of 10 mg.ml-1, the antioxidant capacity was 76.5% and 48.9% for DPPH and ABTS, respectively. EPS-M41 inhibited 86.8% and 90.8% of the α-amylase and α-glucosidase activities, respectively, at 100 µg.ml-1. A 77.5% and 46.4% of antitumor inhibition occurred by EPS-M41 against Caco-2 and MCF-7 cells, respectively. The apparent viscosity (ƞ) of all EPS-M41 solutions decreased with shear rate increases. Salt type and pH value had an impact on the rheological properties of EPS-M41.

A fructan from Anemarrhena asphodeloides Bunge showing neuroprotective and immunoregulatory effects.

A novel polysaccharide, AAP70-1, was isolated from Anemarrhena asphodeloides for the first time. The primary structural analysis revealed that AAP70-1 was composed of glucose and fructose, had an absolute molecular weight of 2720 Da, and contained a (2→6)-linked ß-D-fructofuranose (Fruf) backbone and a (2→1,6)-linked ß-D-Fruf side chain with an internal α-D-glucopyranose (Glcp) in the form of a neokestose. To explore the potential factors responsible for the medicinally relevant bioactivities of A. asphodeloides, a biological assay was performed. Using flow cytometry analysis, AAP70-1 was experimentally shown to have neuroprotective effects, and it can prevent and ameliorate neurological damage via reducing apoptosis. The immunomodulation assay further revealed that AAP70-1 can significantly improve immune function by promoting phagocytic capacity and the secretion of cytokines (IL-6, IL-1ß and TNF-α) in RAW264.7 cells. These results suggest that AAP70-1 has potential as a therapeutic agent for central nervous system diseases or as an immunomodulatory agent.

A heteropolysaccharide purified from leaves of Ilex latifolia displaying immunomodulatory activity in vitro and in vivo.

A novel polysaccharide (ILP50-2) was extracted, isolated and purified from the leaves of Ilex latifolia Thunb. Its structure was characterized as a repeating unit consisting of α-L-Araf-(1→, →3)-α-L-Araf-(1→, →5)-α-L-Araf-(1→, →3,5)-α-L-Araf-(1→, →2)-α-L-Rhap-(1→, →2,4)-α-L-Rhap-(1→, ß-D-Galp-(1→, →4)-ß-D-Galp-(1→, →4)-ß-D-Glcp-(1→, →6)-α-D-Manp-(1→, and →3,6)-α-D-Galp-(1→. The absolute molecular weight of ILP50-2 was 1.49 × 105 g/mol, which adapted a compact coil conformation in 0.1 M NaCl solution with Rz of 25.4 nm. Furthermore, ILP50-2 exhibited immunoregulatory activity, mainly through enhancing the phagocytosis ability of macrophages and prompting the release of nitric oxide (NO) and cytokines tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1ß (IL-1ß). Simultaneously, ILP50-2 was found to significantly increase the release of ROS and NO in zebrafish embryos, showing immunoregulatory effects in vivo.

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.

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.

Hydration characteristics, structural effects and the taste quality of some polyhydroxy compounds in aqueous solutions of nicotinic acid (vitamin B3) at (288.15-318.15) K.

Physicochemical properties elucidating the hydration characteristics and structural effects of polyhydroxy compounds in mixed aqueous solutions offer significant information for the growth of pharmaceutical and food industries. Consequently, standard partial molar volumes and isentropic compressibilities at infinite dilution of saccharides, their derivatives and sugar alcohols in (0.01, 0.05, 0.09 and 0.13) mol kg-1 nicotinic acid(aq) (vitamin B3) solutions have been investigated by experimental density and ultrasonic velocity with respect to temperature. Their transfer values, compressibility hydration numbers, apparent massic volumes and isentropic compressibilities have also been determined. These parameters are important to study the taste behavior of polyhydroxy compounds and intermolecular interactions occurring in ternary mixtures. UV absorption spectra of the studied polyhydroxy compounds have been recorded in 1 × 10-4 mol kg-1 nicotinic acid(aq) solutions. The comparison of present results has been made with the studies reported earlier in l-ascorbic acid, thiamine HCl and pyridoxine HCl.