Carrageenan-Based Compounds as Wound Healing Materials.

The following review is focused on carrageenan, a heteroglycan-based substance that is a very significant wound healing biomaterial. Every biomaterial has advantages and weaknesses of its own, but these drawbacks are typically outweighed by combining the material in various ways with other substances. Carrageenans' key benefits include their water solubility, which enables them to keep the wound and periwound damp and absorb the wound exudate. They have low cytotoxicity, antimicrobial and antioxidant qualities, do not stick to the wound bed, and hence do not cause pain when removed from the wounded region. When combined with other materials, they can aid in hemostasis. This review emphasizes the advantages of using carrageenan for wound healing, including the use of several mixes that improve its properties.

Chitin-rich heteroglycan from Sporothrix schenckii sensu stricto potentiates fungal clearance in a mouse model of sporotrichosis and promotes macrophages phagocytosis.

BACKGROUND: Fungal cell wall polysaccharides maintain the integrity of fungi and interact with host immune cells. The immunomodulation of fungal polysaccharides has been demonstrated in previous studies. However, the effect of chitin-rich heteroglycan extracted from Sporothrix schenckii sensu stricto on the immune response has not been investigated. RESULTS: In this study, chitin-rich heteroglycan was extracted from S. schenckii sensu stricto, and immunomodulation was investigated via histopathological analysis of skin lesions in a mouse model of sporotrichosis and evaluation of the phagocytic function and cytokine secretion of macrophages in vitro. The results showed that the skin lesions regressed and granulomatous inflammation was reduced in infected mice within 5 weeks. Moreover, heteroglycan promoted the fungal phagocytosis by macrophages and modulated the cytokine secretion. Heteroglycan upregulated TNF-α expression early at 24 h and IL-12 expression late at 72 h after incubation, which might result from moderate activation of macrophages and contribute to the subsequent adaptive immune response. CONCLUSIONS: Chitin-rich heteroglycan extracted from S. schenckii sensu stricto potentiated fungal clearance in a mouse model of sporotrichosis. Moreover, chitin-rich heteroglycan promoted fungus phagocytosis by macrophages and modulated cytokines secretion. These results might indicate that chitin-rich heteroglycan could be considered as an immunomodulator used in the treatment of sporotrichosis.

Molecular Mechanisms Associated With Particulate and Soluble Heteroglycan Mediated Immune Response.

Immune responses are outcomes of complex molecular machinery which occur inside the cells. Unravelling the cellular mechanisms induced by immune stimulating molecules such as glycans and determining their structure-function relationship are therefore important factors to be assessed. With this viewpoint, the present study identifies the functional receptor binding unit of a well characterized heteroglycan and also delineates the cellular and molecular processes that are induced upon heteroglycan binding to specific cell surface receptors in immune cells. The heteroglycan was acid hydrolysed and it was revealed that 10-30 kDa fractions served as the functional receptor binding unit of the molecule. Increasing the size of 10-30 kDa heteroglycan showed prominent immune activity. The whole soluble heteroglycan was also conjugated with hyperbranched dendrimers so as to generate a particulate form of the molecule. Dectin-1 and TLR2 were identified as the major receptors in macrophages that bind to particulate as well as soluble form of the heteroglycan and subsequently caused downstream signaling molecules such as NF-κß and MAPK to get activated. High levels of 1L-1ß and IL-10 mRNA were observed in particulate heteroglycan treated macrophages, signifying that increasing the size and availability of the heteroglycan to its specific receptors is pertinent to its biological functioning. Upregulated expression of PKC and iNOS were also noted in particulate heteroglycan treated RAW 264.7 cells than the soluble forms. Taken together, our results indicate that biological functions of immunomodulatory heteroglycan are dependent on their size and molecular weight. J. Cell. Biochem. 117: 1580-1593, 2016. © 2015 Wiley Periodicals, Inc.

Potential Use of Cardunculus Biomass on Pleurotus eryngii Production: Heteroglycans Content and Nutritional Properties (Preliminary Results).

The new perspective of using waste biomass to cultivate mushrooms as a source of protein for human nutrition, in line with the circular economy principles, is receiving increasing attention in the scientific community and represents great wealth in terms of environmental sustainability. Pleurotus eryngii is a mushroom also known as cardunculus mushroom due to its ability to grow on this plant. This study explores the potential intrinsic properties of cardunculus (for example, the presence of inulin in the roots) as raw material for the growth of cardunculus mushrooms, and the influence on heteroglycan content and nutrition parameters of the fruiting bodies. Both mycelium and fruiting bodies were used to determine the heteroglycan content in the presence of inulin or cardunculus roots rich in inulin. To produce heteroglycans from P. eryngii in greater quantities and shorter times without having to wait for the formation of the fruiting bodies, the mycelium could be used. The results showed that the presence of cardunculus biomass positively influences the heteroglycan content of P. eryngii. In terms of nutritional parameters, higher contents of polyphenols, flavonoids, anthocyanins, and antioxidant activity were detected in P. eryngii grown on the cardunculus stem and root substrate. In conclusion, recycling cardunculus biomass to generate growth blocks for edible mushrooms is a winning choice due to the opportunity to use this biomass waste, which is gaining more and more attention due to the increase in cultivated areas and the use of fruiting bodies of P. eryngii as a functional food and source of molecules with potential biological activities.

Structural Characterization and Anticoagulant Activity of a 3-O-Methylated Heteroglycan From Fruiting Bodies of Pleurotus placentodes.

Pleurotus placentodes, a fungus, belongs to the Pleurotaceae family. The aim of the present study was to characterize the structure of a novel polysaccharide from fruiting bodies of P. placentodes (PPp-W) and evaluate its anticoagulant activity in vitro. The high-performance liquid chromatography and GC-MS analysis indicated that PPp-W with a molecular weight of 27.4 kDa was mainly composed of mannose (17.56%), glucose (6.37%), galactose (44.89%), and fucose (1.22%) with a certain amount of 3-O-methyled galactose. SEM, XRD, and AFM combined with Congo red test revealed that PPp-W was an irregular curly sheet with triple-helix conformation. The FT-IR, methylation, and nuclear magnetic resonance analysis indicated that PPp-W contained→6)-α-D-Galp-(1→, →6)-3-O-Me-α-D-Galp-(1→and →2, 6)-α-D-Galp-(1→ as main chain, partially substituted at O-2 and O-6 by non-reducing ends of ß-D-Manp-(1→ and ß-L-Fucp-(1→ with a small amount of α-1,3-linked-Glcp in backbone. PPp-W could significantly prolong APTT (12.9 ± 0.42 s, p < 0.001) and thrombin time (39.9 ± 0.28 s, p < 0.01) compared with the control group (11.45 ± 0.071 s and 38.05 ± 0.21 s), which showed that PPp-W had anticoagulant activity. These studies suggested that PPp-W was a 3-O-methylated heteroglycan and might be suitable for functional foods and natural drugs as an anticoagulant ingredient, which provided a basis for the application of polysaccharides from P. placentodes.

Identification of Feronia-interacting proteins in Arabidopsis thaliana.

BACKGROUND: Plant growth and development are complex processes modulated by numerous genes, transcription factors, hormones, and peptides. Several reports implicate the membrane-localized Catharanthus roseus receptor-like kinase1 (CrRLK1L) protein, FERONIA (FER), involved in plant development. However, protein targets of FER remain poorly characterized. OBJECTIVE: FER recombinant proteins were analyzed, and FER-interacting proteins were identified, to better understand the function of the Arabidopsis thaliana FER (AtFER) gene in plant development. METHODS: AtFER-interacting proteins were identified through Yeast-Two Hybrid (Y2H) and validated by bimolecular fluorescence complementation (BiFC). Autophosphorylation activity was evaluated in AtFER site-directed and deletion mutants. RESULTS: AtFER cytoplasmic kinase domain (Flag-FER-CD) is autophosphorylated at the Thr residue (s), with T559 and T664 as important sites for AtFER kinase activity. In addition, the carboxy terminal region is essential for AtFER kinase activity. Y2H identified an Armadillo (ARM)-repeat protein (At4g16490) with tandem copies of a degenerate protein sequence motif, a U-BOX 9 (PUB9, At3g07360), IQ-DOMAIN 7 (IQD7, At1g17480), and heteroglycan glucosidase 1 (HGL1, At3g23640) as AtFER-interacting proteins. BiFC confirmed the in vivo interactions between these four proteins and AtFER in tobacco (Nicotiana benthamiana) leaf transient expression assays. The RAPID ALKALINIZATION FACTOR1 (RALF1) peptide, which is a FER ligand, induced the expression of genes encoding the four AtFER-interacting proteins. CONCLUSION: The AtFER-interacting proteins identified in this study are likely involved in FER-mediated intracellular signaling pathways that are essential in plant growth and development, and possibly plant immunity.

Structural characterization of a heteroglycan from an edible mushroom Termitomyces heimii.

A water soluble heteroglycan (THPS) of an average molecular weight ~1.98 × 105 Da was isolated from the aqueous extract of the fruit bodies of an edible mushroom Termitomyces heimii. Structural characterization of THPS was carried out using acid hydrolysis, methylation analysis, periodate oxidation, Smith degradation and 1D/2D NMR studies. Sugar analysis indicated the presence of glucose, mannose, galactose, and fucose in a molar ratio of nearly 6:2:2:1. The repeating unit of the THPS had a backbone consisting of four (1 â†’ 3)-ß-d-glucopyranosyl, one (1 â†’ 6)-ß-d-glucopyranosyl, two (1 â†’ 3)-α-D-manopyranosyl, and two (1 â†’ 6)-α-D-galactopyranosyl residues, out of which one (1 â†’ 3)-ß-d-glucopyranosyl residue was branched at O-6 position with terminal ß-d-glucopyranosyl residue and one (1 â†’ 6)-α-D-galactopyranosyl residue was branched at O-2 position with terminal α-L-fucopyranosyl residue.

Xylose rich heteroglycan from flaxseed gum mediates the immunostimulatory effects on macrophages via TLR2 activation.

Immunostimulatory activity of the flaxseed gum neutral fraction (NFG) was investigated. NFG was characterized as a xylose rich heteroglycan through monosaccharide composition analysis, FT-IR, methylation/GC-MS, and 1D/2D-NMR. NFG stimulated NO production and phagocytic activity of macrophages. Secretion of interleukin-6, interleukin-1ß, and tumor necrosis factor-α (254.7 pg/mL, 2.5 ng/mL, and 42.9 pg/mL, respectively) was significantly induced by NFG. Mitogen-activated protein kinases of JNK and P38 were activated by NFG with increased phosphorylation of JNK and P38, while NO production was reduced to 6.05 and 4.42 µM by JNK and P38 inhibitor, respectively. Nuclear factor-κB signaling pathway was also activated by NFG with the suppression of IκBα and up-regulation of phosphorylation of IκBα and nuclear factor-κB P65. Toll like receptor-2 was the molecular target of NFG and responsible for the activation of down-stream signaling pathways. Thus, NFG from flaxseed gum may potentially be used as a natural immunomodulator in functional foods.

Studies on structure and antioxidant properties of a heteroglycan isolated from wild edible mushroom Lentinus sajor-caju.

A water-soluble heteroglycan (PS-I) isolated from the aqueous extract of a wild edible mushroom Lentinus sajor-caju showed average molecular weight ∼1.79×105Da. The structure of the polysaccharide was determined using chemical and 1D/2D NMR experiments. Acid hydrolysis indicated the presence of d-glucose, d-galactose, d-mannose, and l-fucose in a molar ratio of nearly 4:4:1:1 respectively. The presence of terminal Fucp, terminal Galp, (1→3)-Glcp, (1→6)-Galp, (1→6)-Glcp, (1→4,6)-Galp, and (1→2,4)-Manp moieties were established from methylation analysis. The chemical and NMR analyses indicated that the PS-I was a heteroglycan composed of a repeating unit with backbone chain of three (1→6)-α-d-galactopyranosyl residues, two (1→6)-ß-d-glucopyranosyl residues, one (1→4)-α-d-mannopyranosyl residue, and two (1→3)-ß-d-glucopyranosyl residues where one (1→6)-α-d-galactopyranosyl residue was branched at O-4 position with terminal α-l-fucopyranosyl residue and (1→4)-α-d-mannopyranosyl residue was branched at O-2 position with terminal α-d-galactopyranosyl residue and the structure was proposed as; The PS-I is a moderate antioxidant compound which showed DPPH radical scavenging activity, hydroxyl radical scavenging activity, ABTS radical scavenging property, reducing power, and ferrous ion chelating ability.

Structural elucidation and immunostimulating property of a novel polysaccharide extracted from an edible mushroom Lentinus fusipes.

A water soluble heteroglycan (PS-II) with an average molecular weight∼60kDa was isolated from the hot aqueous extract of an edible mushroom Lentinus fusipes. The structural characterization of PS-II was carried out using total acid hydrolysis, methylation analyses, periodate oxidation, Smith degradation and 1D/2D NMR experiments. Total acid hydrolysis indicated the presence of D-galactose and D-glucose in a molar ratio of approximately 1:1. The chemical and NMR analyses revealed that the proposed repeating unit of the PS-II had a backbone chain consisting of three (1→6)-linked α-d-galactopyranosyl residue and two (1→6)-linked ß-d-glucopyranosyl residues, one of the ß-d-glucopyranosyl residue was branched at O-3 position with a terminal ß-d-glucopyranosyl. The PS-II exhibited significant in vitro splenocyte and macrophage activations with optimum dose of 20µg/ml and 80µg/ml respectively. Flow cytometry study revealed the protective role of the PS-II against nicotine stimulated lymphocytes. Moreover, the ROS scavenging property of PS-II was also established using DPPH radical scavenging assay.

Toll-like receptor 4-related immunostimulatory polysaccharides: Primary structure, activity relationships, and possible interaction models.

Toll-like receptor (TLR) 4 is an important polysaccharide receptor; however, the relationships between the structures and biological activities of TLR4 and polysaccharides remain unknown. Many recent findings have revealed the primary structure of TLR4/MD-2-related polysaccharides, and several three-dimensional structure models of polysaccharide-binding proteins have been reported; and these models provide insights into the mechanisms through which polysaccharides interact with TLR4. In this review, we first discuss the origins of polysaccharides related to TLR4, including polysaccharides from higher plants, fungi, bacteria, algae, and animals. We then briefly describe the glucosidic bond types of TLR4-related heteroglycans and homoglycans and describe the typical molecular weights of TLR4-related polysaccharides. The primary structures and activity relationships of polysaccharides with TLR4/MD-2 are also discussed. Finally, based on the existing interaction models of LPS with TLR4/MD-2 and linear polysaccharides with proteins, we provide insights into the possible interaction models of polysaccharide ligands with TLR4/MD-2. To our knowledge, this review is the first to summarize the primary structures and activity relationships of TLR4-related polysaccharides and the possible mechanisms of interaction for TLR4 and TLR4-related polysaccharides.

Heteroglycan of an edible mushroom Termitomyces clypeatus: structure elucidation and antioxidant properties.

A water-soluble heteroglycan (PS) of an average molecular weight ∼1.98 ×10(5) Da was isolated from the aqueous extract of an edible mushroom Termitomyces clypeatus (R. Heim). The structure of the polysaccharide (PS) was established using total hydrolysis, methylation analysis, Smith degradation, and 1D/2D NMR experiments. Total hydrolysis indicated the presence of d-glucose, d-galactose, d-mannose, and l-fucose in a molar ratio of 4.10:1.95:1.0:0.95, respectively. The chemical and NMR analysis indicated the presence of a repeating unit with a backbone consisting of one each of the residues (1→3)-α-d-galactopyranosyl, (1→3)-α-d-mannopyranosyl, (1→3)-α-d-glucopyranosyl, (1→3)-ß-d-glucopyranosyl, (1→6)-ß-d-glucopyranosyl, and (1→6)-α-d-galactopyranosyl, respectively. The (1→3)-α-d-mannopyranosyl residue was found branched at O-2 with terminal α-l-fucopyranosyl moiety and (1→3)-ß-d-glucopyranosyl residue was branched at O-6 with terminal α-d-glucopyranosyl residue. The PS exhibited antioxidant properties.

Heteroglycan of an edible mushroom Entoloma lividoalbum: structural characterization and study of its protective role for human lymphocytes.

A water soluble heteroglycan (PS-II) of an average molecular weight ∼5.2×10(4)Da was isolated from the alkaline extract of an edible mushroom Entoloma lividoalbum (Kühner & Romagn) Kubicka. Structural characterization of PS-II was carried out using sugar and methylation analysis, periodate oxidation study, and 1D/2D NMR experiments. Sugar analysis indicated the presence of glucose, mannose, galactose, and fucose in a molar ratio of nearly 5:1:2:1. The repeating unit of the PS-II had a backbone consisting of two (1→3)-ß-d-glucopyranosyl, one (1→6)-ß-d-glucopyranosyl, one (1→2)-α-L-fucopyranosyl, one (1→6)-α-d-glucopyranosyl, and two (1→6)-α-d-galactopyranosyl residues, out of which one (1→3)-ß-d-glucopyranosyl residue was branched at O-6 position with terminal ß-d-glucopyranosyl residue and one (1→6)-α-d-galactopyranosyl residue was branched at O-2 position with terminal ß-d-mannopyranosyl residue. PS-II showed ameliorative activities at different concentrations (50, 100, 200, 400µg/ml) and maintained the redox balance as well as reduced the lipid peroxidation to protect the cell destruction.