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Every fungal cell is encapsulated in a cell wall, essential for cell viability, morphogenesis, and pathogenesis. Most knowledge of the cell wall composition in fungi has focused on ascomycetes, especially human pathogens, but considerably less is known about early divergent fungal groups, such as species in the Zoopagomycota and Mucoromycota phyla. To shed light on evolutionary changes in the fungal cell wall, we studied the monosaccharide composition of the cell wall of 18 species including early diverging fungi and species in the Basidiomycota and Ascomycota phyla with a focus on those with pathogenic lifestyles and interactions with plants. Our data revealed that chitin is the most characteristic component of the fungal cell wall, and was found to be in a higher proportion in the early divergent groups. The Mucoromycota species possess few glucans, but instead have other monosaccharides such as fucose and glucuronic acid that are almost exclusively found in their cell walls. Additionally, we observed that hexoses (glucose, mannose and galactose) accumulate in much higher proportions in species belonging to Dikarya. Our data demonstrate a clear relationship between phylogenetic position and fungal cell wall carbohydrate composition and lay the foundation for a better understanding of their evolution and their role in plant interactions.

The cell wall integrity (CWI) signaling pathway regulates yeast cell wall biosynthesis, cell division, and responses to external stress. The cell wall, comprised of a dense network of chitin, β-1,3- and β-1,6- glucans, and mannoproteins, is very thin,

Invasive fungal pathogen Candida auris has become a public health threat causing outbreaks of high mortality infections. Drug resistance often limits treatment options. For Candida albicans, subinhibitory concentrations of echinocandins unmask immunostimulatory β-glucan, augmenting immunity. Here we analyze the impact of echinocandin treatment of C. auris on β-glucan exposure and human neutrophil interactions. We show subinhibitory concentrations lead to minimal glucan unmasking and only subtle influences on neutrophil functions for the isolates belonging to circulating clades. The data suggest that echinocandin treatment will not largely alter phagocytic responses. Glucan masking pathways appear to differ between C. auris and C. albicans. [ABSTRACT FROM AUTHOR]

An alkali-soluble β-glucan (AHEP-A-b, 20 kDa) purified from Hericium erinaceus fruiting bodies, was structurally characterized and examined for antioxidant activity. Methylation analysis and NMR spectroscopy show that the backbone of AHEP-A-b is composed of (1→6)-linked-D-β-glucopyran residues, branched at O-3 of glucopyranose (Glcp) residues with [→3)-β-D-Glcp-(1→] oligosaccharides or single unit of β-Glcp. Periodate oxidation analysis and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) indicate that the degree of polymerization (DP) of [→3)-β-D-Glcp-(1→] side chains is 2 to 8. Functionally, AHEP-A-b is a relatively strong antioxidant as demonstrated by using 2, 2′-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) free radical (ABTS·+), 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals, and hydroxyl radicals scavenging assays. The present study lays the foundation for further studies into structure-activity relationships of polysaccharides from H. erinaceus. [ABSTRACT FROM AUTHOR]

Abstract Phosphoesterification is the only naturally occurring covalent starch modification identified to date, and it has a major impact on overall starch metabolism. The incorporation of phosphate groups mediated by dikinases [α-glucan, water dikinase (GWD), EC 2.7.9.4; phosphoglucan, water dikinase (PWD), EC 2.7.9.5] massively alters the starch granule properties; however, previous studies did not determine whether the starch-related dikinases bind the phosphate to the glucosyl units within the amylopectin molecules in a specific pattern or randomly. In order to answer this challenging question, a number of approaches were initially pursued until a protocol could be established that enabled a massive step forward in the in vitro analysis of phosphorylated glucan chains obtained from starch. For this purpose, phosphorylation by GWD was investigated, including the final state of phosphorylation i.e., the state of substrate saturation when GWD lacks further free hydroxyl groups at OH-C6 for the catalysis of monophosphate esters. Since the separated phosphorylated glucan chains were required for the analysis, isoamylase digestion was performed to cleave the α-1,6-glycosidic bonds and to allow for the removal of the huge number of existing neutral chains by means of anion exchange chromatography. Via Matrix-Assisted Laser Desorption/Ionization–Time of Flight (MALDI-TOF) MS and MALDI-MS/MS, the phosphorylated α-glucan chains were analysed, and the position of the phosphate group within the chain in relation to the reducing end was determined. Here, we demonstrate a protocol that enables the analysis of phosphorylated oligosaccharides, even in small quantities.

Phosphoesterification is the only naturally occurring covalent starch modification identified to date, and it has a major impact on overall starch metabolism. The incorporation of phosphate groups mediated by dikinases [α-glucan, water dikinase (GWD), EC 2.7.9.4; phosphoglucan, water dikinase (PWD), EC 2.7.9.5] massively alters the starch granule properties; however, previous studies did not determine whether the starch-related dikinases bind the phosphate to the glucosyl units within the amylopectin molecules in a specific pattern or randomly. In order to answer this challenging question, a number of approaches were initially pursued until a protocol could be established that enabled a massive step forward in the in vitro analysis of phosphorylated glucan chains obtained from starch. For this purpose, phosphorylation by GWD was investigated, including the final state of phosphorylation i.e., the state of substrate saturation when GWD lacks further free hydroxyl groups at OH-C6 for the catalysis of monophosphate esters. Since the separated phosphorylated glucan chains were required for the analysis, isoamylase digestion was performed to cleave the α-1,6-glycosidic bonds and to allow for the removal of the huge number of existing neutral chains by means of anion exchange chromatography. Via Matrix-Assisted Laser Desorption/Ionization–Time of Flight (MALDI-TOF) MS and MALDI-MS/MS, the phosphorylated α-glucan chains were analysed, and the position of the phosphate group within the chain in relation to the reducing end was determined. Here, we demonstrate a protocol that enables the analysis of phosphorylated oligosaccharides, even in small quantities. [ABSTRACT FROM AUTHOR]

The serum (1,3)-beta- d -glucan (BDG) assay gives quicker results and has higher sensitivity than blood cultures, therefore it is advised for early diagnosis of invasive candidemia and/or discontinuation of empirical therapy. Its sensitivity may depend on different factors. The aim of our study was to analyse the in vitro and in vivo BDG levels in clinical isolates of three species of Candida responsible for candidemia. C. albicans, C. parapsilosis, and C. auris strains were collected from blood cultures of patients who had a concurrent (–1 to +3 days) serum BDG test (Fungitell assay). Supernatants of all strains were tested in quadruplicate for BDG levels. Twenty-two C. auris , 14 C. albicans, and ten C. parapsilosis strains were included. The median BDG levels in supernatants were 463 pg/mL (interquartile range [IQR] 379–648) for C. auris , 1080 pg/mL (IQR 830–1276) for C. albican s, and 755 pg/mL (IQR 511–930) for C. parapsilosis , with the significant difference among the species (p < 0.0001). Median serum BDG levels (IQR) were significantly lower in case C. auris and C. parapsilosis vs. C. albicans (p < 0.0001), respectively, 50 pg/mL (IQR 15–161) and 57 pg/mL (IQR 18–332), vs. 372 pg/mL (IQR 102–520). Sensitivity of serum BDG was 39% (95% confidence interval [CI], 18–64) in case of C. auris , 30% (95% CI, 8–65) C. parapsilosis and 78% (95% CI, 49–94) C. albicans candidemia. In our centre C. auris and C. parapsilosis strains have lower BDG content as compared with C. albicans , with a potential impact on serum BDG performance for the diagnosis of candidemia. [ABSTRACT FROM AUTHOR]

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Sacran is a supergiant cyanobacterial polysaccharide that forms mesogenic supercoil rods that exhibit liquid crystalline (LC) gels at deficient concentrations of around 0.5 wt%, and has several bioactive stimuli-responsive functions. Here, we attempted to form oriented microfibers of sacran by electrospinning, following structural analyses of the sacran rods. A heterogeneous acid-hydrolysis method using a protonated cation-exchange resin was adopted to examine the short-time exposition of concentrated acid to sacran rods. From the supernatant, the oligomeric fraction that was soluble in water and methanol was isolated. The oligomeric fraction had a main sugar ratio of α-Glc:β-Glc:α-Xyl:β-Xyl:α-Rha of 2:5:1.5:1.5:4 (Glc:Xyl:Rha = 7 (=4 + 3):3:4), and it was speculated that the sacran structure includes rhamnoglucan and xyloglucan (4:3), which are generally rigid enough to exhibit LC. To make oriented microfibers of LC sacran, solubility testing was performed on sacran to find good new solvents of polyhydroxy alcohols such as ethylene glycol, 1,2-propanediol, and glycerol. The oriented film was prepared from a sacran aqueous solution where calcium compound particles deposited on the film are different from polyhydroxy alcohol solutions. Although sacran could not form microfibers by itself, polymer composite microfibers of sacran with poly(vinyl alcohol) were prepared by electrospinning. Cross-polarizing microscopy revealed the molecular orientation of the microfibers. [ABSTRACT FROM AUTHOR]

Fungi are one of the most diverse organisms found in our surroundings. The heterotrophic lifestyle of fungi and the ever-changing external environmental factors pose numerous challenges for their survival. Despite all adversities, fungi continuously develop new survival strategies to secure nutrition and space from their host. During host–pathogen interaction, filamentous phytopathogens in particular, effectively infect their hosts by maintaining polarised growth at the tips of hyphae. The fungal cell wall, being the prime component of host contact, plays a crucial role in fortifying the intracellular environment against the harsh external environment. Structurally, the fungal cell wall is a highly dynamic yet rigid component, responsible for maintaining cellular morphology. Filamentous pathogens actively maintain their dynamic cell wall to compensate rapid growth on the host. Additionally, they secrete effectors to dampen the sophisticated mechanisms of plant defense and initiate various downstream signaling cascades to repair the damage inflicted by the host. Thus, the fungal cell wall serves as a key modulator of fungal pathogenicity. The fungal cell wall with their associated signaling mechanisms emerge as intriguing targets for host immunity. This review comprehensively examines and summarizes the multifaceted findings of various research groups regarding the dynamics of the cell wall in filamentous fungal pathogens during host invasion. [ABSTRACT FROM AUTHOR]

ABSTRACT The uridine derivatives UDP-glucose and UDP-N-acetylglucosamine are important for cell wall construction as they are the precursors for the synthesis of β-1,3-glucan and chitin, respectively. Previous studies have demonstrated attenuated virulence of uridine auxotrophs in mice, which has been attributed to insufficient uridine levels for growth in the host. We have discovered that uridine deprivation in the uridine auxotroph ura3ΔΔ disrupts cell wall architecture by increasing surface mannans, exposing β-1,3-glucan and chitin, and decreasing UDP-sugar levels. Cell wall architecture and UDP-sugars can be rescued with uridine supplementation. The cell wall architectural disruptions in the ura3ΔΔ mutant also impact immune activation since the mutant elicited greater TNFα secretion from RAW264.7 macrophages than wild type. To determine if cell wall defects contributed to decreased virulence in the ura3ΔΔ mutant, we used a murine model of systemic infection. Mice infected with the ura3ΔΔ mutant exhibited increased survival and reduced kidney fungal burden compared with mice infected with wild type. However, suppression of the immune response with cyclophosphamide did not rescue virulence in mice infected with the ura3ΔΔ mutant, indicating the attenuation in virulence of uridine auxotrophs can be attributed to decreased growth in the host but not increased exposure of β-1,3-glucan. Moreover, the ura3ΔΔ mutant is unable to grow on ex vivo kidney agar, which demonstrates its inability to colonize the kidneys due to poor growth. Thus, although uridine auxotrophy elicits changes to cell wall architecture that increase the exposure of immunogenic polymers, metabolic fitness costs more strongly drive the observed virulence attenuation.IMPORTANCECandida albicans is a common cause of bloodstream infections (candidemia). Treatment of these bloodstream infections is made difficult because of increasing antifungal resistance and drug toxicity. Thus, new tactics are needed for antifungal drug development, with immunotherapy being of particular interest. The cell wall of C. albicans is composed of highly immunogenic polymers, particularly β-1,3-glucan. However, β-1,3-glucan is naturally masked by an outer layer of mannoproteins, which hampers the detection of the fungus by the host immune system. Alteration in cell wall components has been shown to increase β-1,3-glucan exposure; however, it is unknown how the inability to synthesize precursors to cell wall components affects unmasking. Here, we demonstrate how cell wall architecture is altered in response to a deficit in precursors for cell wall synthesis and how uridine is a crucial component of these precursors.

Solid‐state nuclear magnetic resonance (ssNMR) measurements of intact cell walls and cellular samples often generate spectra that are difficult to interpret due to the presence of many coexisting glycans and the structural polymorphism observed in native conditions. To overcome this analytical challenge, we present a statistical approach for analyzing carbohydrate signals using high‐resolution ssNMR data indexed in a carbohydrate database. We generate simulated spectra to demonstrate the chemical shift dispersion and compare this with experimental data to facilitate the identification of important fungal and plant polysaccharides, such as chitin and glucans in fungi and cellulose, hemicellulose, and pectic polymers in plants. We also demonstrate that chemically distinct carbohydrates from different organisms may produce almost identical signals, highlighting the need for high‐resolution spectra and validation of resonance assignments. Our study provides a means to differentiate the characteristic signals of major carbohydrates and allows us to summarize currently undetected polysaccharides in plants and fungi, which may inspire future investigations. [ABSTRACT FROM AUTHOR]

Cold water fish skin gelatin (FSGltn) was chosen as the main material, wherein catechol groups (CAT) were linked through quantitative modification to prepare a catechol-modified gelatin (CAT-FSGltn). Then CAT-FSGltn and dextran dialdehyde (DDA) were mixed in situ to prepare a tissue adhesive (CAT-FSGltn/DDA) with good adhesion in a wet environment. It was found that the gelation time of CAT-FSGItn/DDA was within 60 s. The fully crosslinked CAT-FSGltn/DDA hydrogel showed minimal mass loss after being immersed in PBS solution for over 80 h. When adhered to a rubber film resembling the tissue membrane, the hydrogel CAT-FSGltn/DDA demonstrated an ability to withstand stretching up to twice its initial length without fracturing. An in vitro model was used to evaluate the compressive strength of the tissue adhesive. The compressive strength of CAT-FSGltn/DDA was observed to reach 105.7 mmHg, surpassing the human diastolic pressure. The results indicated that CAT-FSGltn/DDA possessed both strong cohesion and adhesion. Tissue sections showed that CAT-FSGltn/DDA cured for 30 min was closely attached to the casing surface, and the hydrogel/casing interface remained intact after the water flow impact. Cell experiments showed that the survival rate of L929 cells was more than 80%, CAT-FSGltn/DDA hydrogel had good biocompatibility. [ABSTRACT FROM AUTHOR]

The fungal cell wall plays a critical role in regulating cellular integrity and communication, and serves as a frontline defense against stress. It is also a prime target for the development of antifungal agents. The cell wall is comprised of diverse polysaccharides and proteins and poses a challenging target for high-resolution structural characterization. Recently, the solid-state nuclear magnetic resonance (ssNMR) analysis of intact Aspergillus fumigatus cells has provided atomic-level insights into the structural polymorphism and functional assembly principles of carbohydrate components within the cell wall. This physical perspective, alongside structural information from biochemical assays, offers a renewed understanding of the cell wall as a highly complex and dynamic organelle. Here, we summarize key conceptual advancements in the structural elucidation of A. fumigatus mycelial and conidial cell walls and their responses to stressors. We also highlight underexplored areas and discuss the opportunities facilitated by technical advancements in ssNMR spectroscopy. [ABSTRACT FROM AUTHOR]

Every fungal cell is encapsulated in a cell wall, essential for cell viability, morphogenesis, and pathogenesis. Most knowledge of the cell wall composition in fungi has focused on ascomycetes, especially human pathogens, but considerably less is known about early divergent fungal groups, such as species in the Zoopagomycota and Mucoromycota phyla. To shed light on evolutionary changes in the fungal cell wall, we studied the monosaccharide composition of the cell wall of 18 species including early diverging fungi and species in the Basidiomycota and Ascomycota phyla with a focus on those with pathogenic lifestyles and interactions with plants. Our data revealed that chitin is the most characteristic component of the fungal cell wall, and was found to be in a higher proportion in the early divergent groups. The Mucoromycota species possess few glucans, but instead have other monosaccharides such as fucose and glucuronic acid that are almost exclusively found in their cell walls. Additionally, we observed that hexoses (glucose, mannose and galactose) accumulate in much higher proportions in species belonging to Dikarya. Our data demonstrate a clear relationship between phylogenetic position and fungal cell wall carbohydrate composition and lay the foundation for a better understanding of their evolution and their role in plant interactions.

An alkali-soluble β-glucan (AHEP-A-b, 20 kDa) purified from Hericium erinaceus fruiting bodies, was structurally characterized and examined for antioxidant activity. Methylation analysis and NMR spectroscopy show that the backbone of AHEP-A-b is composed of (1→6)-linked-D-β-glucopyran residues, branched at O-3 of glucopyranose (Glcp) residues with [→3)-β-D-Glcp-(1→] oligosaccharides or single unit of β-Glcp. Periodate oxidation analysis and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) indicate that the degree of polymerization (DP) of [→3)-β-D-Glcp-(1→] side chains is 2 to 8. Functionally, AHEP-A-b is a relatively strong antioxidant as demonstrated by using 2, 2′-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) free radical (ABTS·+), 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals, and hydroxyl radicals scavenging assays. The present study lays the foundation for further studies into structure-activity relationships of polysaccharides from H. erinaceus.

AbstractFungal α/β-glucans have significant importance in cellular functions including cell wall structure, host-pathogen interactions and energy storage, and wide application in high-profile fields, including food, nutrition, and pharmaceuticals. Fungal species and their growth/developmental stages result in a diversity of glucan contents, structures and bioactivities. Substantial progresses have been made to elucidate the fine structures and functions, and reveal the potential molecular synthesis pathway of fungal α/β-glucans. Herein, we review the current knowledge about the biosynthetic machineries, including: precursor UDP-glucose synthesis, initiation, elongation/termination and remodeling of α/β-glucan chains, and molecular regulation to maximally produce glucans in edible fungi. This review would provide future perspectives to biosynthesize the targeted glucans and reveal the catalytic mechanism of enzymes associated with glucan synthesis, including: UDP-glucose pyrophosphate phosphorylases (UGP), glucan synthases, and glucanosyltransferases in edible fungi. [ABSTRACT FROM AUTHOR]

Candidemia is associated with a heavy burden of morbidity and mortality in hospitalized patients. The availability of blood culture results could require up to 48–72 h after blood draw; thus, early treatment decisions are made in the absence of a definite diagnosis. In this retrospective study, we assessed the performance of different supervised machine learning algorithms for the early differential diagnosis of candidemia and bacteremia in adult patients on a large dataset automatically extracted within the AUTO-CAND project. Overall, 12,483 episodes of candidemia (1275; 10%) or bacteremia (11,208; 90%) were included in the analysis. A random forest classifier achieved the best diagnostic performance for candidemia, with sensitivity 0.98 and specificity 0.65 on the training set (true skill statistic [TSS] = 0.63) and sensitivity 0.74 and specificity 0.57 on the test set (TSS = 0.31). Then, the random classifier was trained in the subgroup of patients with available serum β-D-glucan (BDG) and procalcitonin (PCT) values by exploiting the feature ranking learned in the entire dataset. Although no statistically significant differences were observed from the performance measures obtained by employing BDG and PCT alone, the performance measures of the classifier that included the features selected in the entire dataset, plus BDG and PCT, were the highest in most cases. Random forest classifiers trained on large datasets of automatically extracted data have the potential to improve current diagnostic algorithms for candidemia. However, further development through implementation of automatically extracted clinical features may be necessary to achieve crucial improvements. [ABSTRACT FROM AUTHOR]

The treatment of dermatophytoses, the most common human fungal infections, requires new alternatives. The aim of this study was to determine the antidermatophytic activity of the aqueous Azorean Black Tea extract (ABT), together with an approach to the mechanisms of action. The phytochemical analysis of ABT extract was performed by HPLC. The dermatophytes susceptibility was assessed using a broth microdilution assay; potential synergies with terbinafine and griseofulvin were evaluated by the checkerboard assay. The mechanism of action was appraised by the quantification of the fungal cell wall chitin and β-1,3-glucan, and by membrane ergosterol. The presence of ultrastructural modifications was studied by Transmission Electron Microscopy (TEM). The ABT extract contained organic and phenolic acids, flavonoids, theaflavins and alkaloids. It showed an antidermatophytic effect, with MIC values of 250 µg/mL for Trichophyton mentagrophytes, 125 µg/mL for Trichophyton rubrum and 500 µg/mL for Microsporum canis; at these concentrations, the extract was fungicidal. An additive effect of ABT in association to terbinafine on these three dermatophytes was observed. The ABT extract caused a significant reduction in β-1,3-glucan content, indicating the synthesis of this cell wall component as a possible target. The present study identifies the antidermatophytic activity of the ABT and highlights its potential to improve the effectiveness of conventional topical treatment currently used for the management of skin or mucosal fungal infections. [ABSTRACT FROM AUTHOR]

Structural analysis of macromolecular complexes within their natural cellular environment presents a significant challenge. Recent applications of solid-state NMR (ssNMR) techniques on living fungal cells and intact plant tissues have greatly enhanced our understanding of the structure of extracellular matrices. Here, we selectively highlight the most recent progress in this field. Specifically, we discuss how ssNMR can provide detailed insights into the chemical composition and conformational structure of pectin, and the consequential impact on polysaccharide interactions and cell wall organization. We elaborate on the use of ssNMR data to uncover the arrangement of the lignin-polysaccharide interface and the macrofibrillar structure in native plant stems or during degradation processes. We also comprehend the dynamic structure of fungal cell walls under various morphotypes and stress conditions. Finally, we assess how the combination of NMR with other techniques can enhance our capacity to address unresolved structural questions concerning these complex macromolecular assemblies. [Display omitted] The advance in solid-state NMR spectroscopy enables the analysis of polysaccharide and protein assemblies in living fungal cells and intact plant tissues at atomic-to-nanometer resolution. In this review, Fernando et al. discuss the most recent conceptual breakthroughs in cell wall structural biology. [ABSTRACT FROM AUTHOR]

In this study, a novel polysaccharide (CPP-80) was prepared from the leaves of Cyclocarya paliurus and its detailed structure, antioxidant and immunomodulatory effects were explored in vitro. The results showed that CPP-80 was a glucan and its main chain consisted of → 4)-α-D-Glcp-(1 → 4)-β-D-Glcp-(1→, while the end groups α-D-Glcp-(1 → 3)-α-D-Glcp-(1 → and α-D-Glcp-(1 → 6)-α-D-Glcp-(1 → were linked to the main chain by the O-3 of the → 3,4)-α-D-Glcp-(1 → and the O-6 of the → 4,6)-α-D-Glcp-(1 → bonds, respectively. In vitro, CPP-80 had a certain reducing ability and antioxidant activities with the scavenging ratios of 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2′-Azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS) radicals and hydroxyl radicals from about 50 % to more than 80 % at a concentration of 4 mg/mL. Additionally, CPP-80 could immunomodulate RAW264.7 macrophages by boosting phagocytosis and encouraging the release of nitric oxide (NO), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6). Thus, CPP-80 could be used as a promising antioxidant and immunomodulator for food and pharmaceutical applications.

Sacran is a supergiant cyanobacterial polysaccharide that forms mesogenic supercoil rods that exhibit liquid crystalline (LC) gels at deficient concentrations of around 0.5 wt%, and has several bioactive stimuli-responsive functions. Here, we attempted to form oriented microfibers of sacran by electrospinning, following structural analyses of the sacran rods. A heterogeneous acid-hydrolysis method using a protonated cation-exchange resin was adopted to examine the short-time exposition of concentrated acid to sacran rods. From the supernatant, the oligomeric fraction that was soluble in water and methanol was isolated. The oligomeric fraction had a main sugar ratio of α-Glc:β-Glc:α-Xyl:β-Xyl:α-Rha of 2:5:1.5:1.5:4 (Glc:Xyl:Rha = 7 (=4 + 3):3:4), and it was speculated that the sacran structure includes rhamnoglucan and xyloglucan (4:3), which are generally rigid enough to exhibit LC. To make oriented microfibers of LC sacran, solubility testing was performed on sacran to find good new solvents of polyhydroxy alcohols such as ethylene glycol, 1,2-propanediol, and glycerol. The oriented film was prepared from a sacran aqueous solution where calcium compound particles deposited on the film are different from polyhydroxy alcohol solutions. Although sacran could not form microfibers by itself, polymer composite microfibers of sacran with poly(vinyl alcohol) were prepared by electrospinning. Cross-polarizing microscopy revealed the molecular orientation of the microfibers.

Objectives: Dental caries is a prevalent chronic human infection worldwide and several plants have shown anticariogenic properties through antibacterial activity against oral pathogens. The present study aimed to assess anticariogenic activity of Verbascum speciosum, in search of novel agents for the prevention and treatment of dental caries.Methods: Hydro-alcoholic extracts from flowers and total aerial parts of the plant were prepared by maceration. Antibacterial activity of the extracts against Streptococcus mutans (ATCC 35668) and Streptococcus sobrinus (ATCC 27607) was investigated by agar diffusion and microdilution techniques. Inhibitory concentration-fifty values of the flowers’ extract against Streptococcus mutans glucosyltransferase enzymes were determined. The total flavonoid content of the extracts was determined using an aluminum chloride reaction.Results: Verbascum speciosum flowers’ extract showed significantly higher flavonoid content and antibacterial activity; with minimum inhibitory concentrations of 100 and 200 μg/mL for Streptococcus mutans and Streptococcus sobrinus , respectively. The extract inhibited the synthesis of glucan by cell-associated and extracellular glucosyltransferase enzymes in a dose-dependent manner with higher activity against the extracellular enzyme.Conclusion: This study indicated effective anticariogenic activity of Verbascum speciosum flowers extract. This extract can be considered as an alternative to current anticaries therapies or an additive to dental care products.

AbstractBackground Candidemia is associated with a heavy burden of morbidity and mortality in hospitalized patients. The availability of blood culture results could require up to 48–72 h after blood draw; thus, early treatment decisions are made in the absence of a definite diagnosis.Methods In this retrospective study, we assessed the performance of different supervised machine learning algorithms for the early differential diagnosis of candidemia and bacteremia in adult patients on a large dataset automatically extracted within the AUTO-CAND project.Results Overall, 12,483 episodes of candidemia (1275; 10%) or bacteremia (11,208; 90%) were included in the analysis. A random forest classifier achieved the best diagnostic performance for candidemia, with sensitivity 0.98 and specificity 0.65 on the training set (true skill statistic [TSS] = 0.63) and sensitivity 0.74 and specificity 0.57 on the test set (TSS = 0.31). Then, the random classifier was trained in the subgroup of patients with available serum β-D-glucan (BDG) and procalcitonin (PCT) values by exploiting the feature ranking learned in the entire dataset. Although no statistically significant differences were observed from the performance measures obtained by employing BDG and PCT alone, the performance measures of the classifier that included the features selected in the entire dataset, plus BDG and PCT, were the highest in most cases.Conclusions Random forest classifiers trained on large datasets of automatically extracted data have the potential to improve current diagnostic algorithms for candidemia. However, further development through implementation of automatically extracted clinical features may be necessary to achieve crucial improvements.

The major purpose of this study was to determine how varying doses of algaederived pure ß-glucan affected in vitro gas generation, volatile fatty acid (VFA) concentrations, methane production, and protozoa populations. Different doses of ß-glucan [i.e., 0, 50, 100, 150, and 200 mg/kg feed (DM basis)] were applied to corn silage as experimental treatments. After 6-96 hours of incubation, the dose of 200 mg/kg of DM ß-glucan reduced total gas production compared to control (P<0.01). The concentration of total VFA decreased quadratically (P<0.01) as the amount of ß-glucan inclusion decreased (except for 200 mg/kg DM) when compared to the control group. The total VFA concentration was found to be the lowest (P<0.01) at 50, 100, and 150 mg/kg DM ß-glucan than the other doses. Propionate and butyrate concentrations increased linearly (P<0.01) in the ß-glucan supplemented groups, except for the 50 mg/kg DM dosage. When compared to the control group, all doses of ß-glucans lowered acetate and the acetate: propionate ratio linearly and quadratically (P<0.01). The addition of ß-glucans reduced the number of protozoa linearly (except at the lowest dose) and reduced the methane generation linearly and quadratically (P<0.01). The concentration of NH3-N did not differ (Linear, P=0.12; Quadratic, P=0.19) between treatments. The key findings were that ß-glucan acted as a rumen modulator, and levels of more than 50 mg/kg of feed DM functioned as a potential methane regulator in the rumen due to reduced acetate and acetate to propionate ratio. [ABSTRACT FROM AUTHOR]

The aim of this study was to explore the current knowledge and practice of agrobiodiversity to improve nutrition and health. The study focused on wild mushrooms commonly consumed in Northeastern Thailand in terms of ecology, cooking and preservation methods, and folk remedies. The monosaccharide, sugar alcohol, glucan, and carbohydrate polymer content and the glycemic index of these wild mushrooms were determined using the enzymatic method. The mushrooms collected belonged to three biological groups and were mostly saprotrophic and symbiotic. The most abundant mushrooms were Amanita, Boletus, and Russula, followed by Calostoma sp., Astraeus asiaticus C. Phosri, and Astraeus odoratus C. Phosri. Wild edible mushrooms can be used for food and medicinal purposes. Cooking methods utilized in the area consist of steaming, boiling, and grilling. Glucose was the major monosaccharide detected in all mushroom samples. Xylitol and inositol were found in all mushroom samples, while some contained mannitol and arabitol. Glucan was present in all mushroom samples, ranging from 8.03 to 31.1 mg/g DW. All mushrooms were classified as having a low glycemic index. These findings provide important information to potentially enhance and promote the utilization of wild mushrooms to improve the accessibility, availability, and sustainability of nutritious food. [ABSTRACT FROM AUTHOR]

Brewer's spent yeast (BSY) is a prevalent by-product of the brewing industry currently used in animal feed as a cheap source of protein. In this study, chemical composition of BSY from two different beer production (American IPA, BSY1, and Imperial Stout, BSY2) was characterized, aiming for potential use as nutraceutical sources. Cytotoxicity of BSY extracts was also tested in freshly isolated peripheral blood mononuclear cells (PBMCs) and in tumoral (pancreatic cancer tumor cells, PANC-1, and colorectal adenocarcinoma tumor cells, CACO-2) and non-tumoral (umbilical vein endothelial cells, HUVEC) cell lineages. BSY samples showed similar lipid and ash content, but slight differences in the content of reducing sugars, total proteins, moisture and total proteins. BSY2 sample presented higher glucans and phenolic compound concentrations than BSY1. Main phenolic compounds identified were xanthohumol, ferulic acid, p-coumaric acid (BSY1 and BSY2), and gallic acid (BSY2). BSY1 and BSY2 extracts showed similar antioxidant capacity. Neither BSY1 nor BSY2 were cytotoxic to PBMCs, HUVEC and CACO-2, but were cytotoxic to PANC-1 cells, in a concentration-dependent manner. Our findings revealed that the residue of brewery can be a value-added functional food product with an adjuvant role against PANC-1 cell lines. [ABSTRACT FROM AUTHOR]

Background(1-3)-b-D-glucan (BDG) is a fungal cell wall component and, in the absence of invasive fungal infection, a novel biomarker for microbial translocation of endogenous fungal products from the gastrointestinal tract into systemic circulation. However, its value as a marker of fungal translocation is limited by a concern that plant BDG-rich food influences blood BDG levels.MethodsWe conducted a pilot clinical trial to evaluate the impact of a standardised oral BDG challenge on blood BDG levels in participants with and without elevated microbial translocation. We enrolled 14 participants including 8 with HIV infection, 2 with advanced liver cirrhosis, and 4 healthy controls. After obtaining a baseline blood sample, participants received a standardised milkshake containing high levels of BDG followed by serial blood samples up to 8 hours after intake.ResultsThe standardised oral BDG challenge approach did not change the blood BDG levels over time in all participants. We found consistently elevated blood BDG levels in one participant with advanced liver cirrhosis and a single person with HIV with a low CD4 count of 201 cells/mm3 .ConclusionOur findings indicate that BDG blood levels were not influenced by plant origin BDG-rich nutrition in PWH, people with advanced liver cirrhosis, or healthy controls. Future studies are needed to analyse gut mycobiota populations in individuals with elevated blood BDG levels.

Introduction: Polysaccharides, key components present in Grifola frondosa, can be divided into those derived from fruiting bodies, mycelium, and fermentation broth based on their source. The structure of G. frondosa fruiting body-derived polysaccharides has been fully characterized. However, the structure of G. frondosa mycelium-derived polysaccharides remains to be elucidated.Methods: In this study, we obtained mycelia from G. frondosa by liquid fermentation and extracted them with water and alkaline solution. Then, the mycelia were isolated and purified to obtain homogeneity and systematically characterized by methylation and FT infrared (FT-IR) and nuclear magnetic resonance (NMR) spectroscopy.Results and discussion: Structural analysis showed that two neutral fractions (WGFP-N-a and AGFP-N-a1) have a common backbone composed of α-1,6-D-Me-Galp and α-1,6-D-Galp that were substituted at O-2 by 1,2-Manp, α-1,3-L-Fucp, and α-T-D-Manp and thus are identified as fucomannogalactans. WGFP-A-a, AGFP-A-b, and AGFP-A-c are β-1,6-glucans with different molecular weights and are branched with β-1,3-D-Glcp and T-D-Glcp at the O-3 of Glc. Our results provide important structural information about G. frondosa mycelium-derived polysaccharides and provide the basis for their further development and application.

BACKGROUND: Frequent fungal diseases tend to lead to severe losses in rice production. As a main component of the fungal cell wall, glucan plays an important role in the growth and development of fungi. Glucanase can inhibit the growth of fungi by breaking glycosidic bonds, and may be a promising target for developing rice varieties with broad‐spectrum disease resistance. RESULTS: We transferred a codon‐optimized β‐1,6‐glucanase gene (GluM) from myxobacteria into the japonica rice variety Zhonghua11 (ZH11), and obtained a large number of individual transgenic plants with GluM overexpression. Based on molecular analysis, three single‐copy homozygous lines with GluM overexpression were selected for assessment of fungal disease resistance at the T3 generation. Compared with that of the recipient cultivar ZH11, the area of rice blast lesion in transgenic rice was reduced by 82.71%; that of sheath blight lesion was decreased by 35.76%–43.67%; the sheath blight resistance in the field was enhanced by an average of 0.75 grade over 3 years; and the incidence of diseased panicles due to rice false smut was decreased by 65.79%. More importantly, there was no obvious loss of yield (without a significant effect on agronomic traits). Furthermore, plants overexpressing a β‐1,6‐glucanase gene showed higher disease resistance than rice plants overexpressing a β‐1,3‐glucanase gene derived from tobacco. CONCLUSION: The β‐1,6‐glucanase gene GluM can confer broad‐spectrum disease resistance to rice, providing an environmentally friendly alternative way to effectively manage fungal pathogens in rice production. © 2023 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Inflammatory bowel diseases (IBD) and colorectal cancer (CRC) are difficult to cure, and available treatment is associated with troubling side effects. In addition, current therapies have limited efficacy and are characterized by high costs, and a large segment of the IBD and CRC patients are refractive to the treatment. Moreover, presently used anti-IBD therapies in the clinics are primarily aimed on the symptomatic control. That is why new agents with therapeutic potential against IBD and CRC are required. Currently, polyphenols have received great attention in the pharmaceutical industry and in medicine due to their health-promoting properties. They may exert anti-inflammatory, anti-oxidative, and anti-cancer activity, via inhibiting production of pro-inflammatory cytokines and enzymes or factors associated with carcinogenesis (e.g., matrix metalloproteinases, vascular endothelial growth factor), suggesting they may have therapeutic potential against IBD and CRC. However, their use is limited under both processing conditions or gastrointestinal interactions, reducing their stability and hence their bioaccessibility and bioavailability. Therefore, there is a need for more effective carriers that could be used for encapsulation of polyphenolic compounds. In recent years, natural polysaccharides have been proposed for creating carriers used in the synthesis of polyphenol encapsulates. Among these, hemicelluloses are particularly noteworthy, being characterized by good biocompatibility, biodegradation, low immunogenicity, and pro-health activity. They may also demonstrate synergy with the polyphenol payload. This review discusses the utility and potential of hemicellulose-based encapsulations of polyphenols as support for treatment of IBD and CRC. [ABSTRACT FROM AUTHOR]

Exopolysaccharides (EPSs) are novel functional additives for low-fat yogurt. Pharmaceutical, medical, and food industries are using more LAB-based EPSs. In this study, Leuconostoc spp. was used to produce ninth bacterial EPSs in a modified molasses medium. Production of EPSs was concentration-dependent on all stains and the highest yield was obtained from the S3 strain (55.23 g/l), followed by S6 (49.95 g/l), S8 (45.68 g/l), and S7 (44.23), respectively. HPLC and FTIR analysis showed that all purified EPSs from Leuconostoc citreum (S3) and Leuconstoc holzaapfelii (S8) were related to exopolysaccharide glucan. Anticancer activity of all EPSs samples (EPSs1-9) against Caco-2 cells and normal MCR-5 cells were investigated using MTT assay. The results revealed that Caco-2 cells were more sensitive than the normal MCR-5 cells. The highest anticancer activity against Caco-2 cancer cells was recorded for EPS8 (IC50 = 22.94 µg/ml, SI=3.73), followed by EPS3 (IC50 = 36.15 µg/ml, SI=8.72), EPS1 (IC50 = 50.01 µg/ml, SI=3.73), and EPS4 (IC50 = 94.90 µg/ml, SI=3.26), respectively. The lowest cytotoxicity was recorded for EPS5 (IC50 = 130.5 µg/ml). The most active EPSs (EPS3 and EPS8) were used as fat replacements and stabilizers in low-fat set yogurt at non-toxic concentrations (0.4, 0.8, and 1.2%). EPS3 and EPS8 improved the low-fat yogurt’s organoleptic and rheological properties. EPS8 had the highest water holding capacity (77.26%), viscosity (3660 CP), and lowest syneresis (22.95%) and whey off (0.6 ml). Low-fat set yogurt enhanced with EPS3 and EPS8 recorded the highest sensory evaluation values with overall acceptability, especially EPS3b, EPS3c, EPS8c, and EPS8b; the total score point of 97.50, 97.43, 96.51, and 96.36, respectively in fresh age compared to control yogurt (92.64). In conclusion, Leuconostoc EPSs, especially EPS8, can be explored for anti-cancer effects on Caco-2 colorectal cancer cells. It could also improve the rheological and organoleptic qualities of low-fat set yogurt.

In recent years, invasive fungal infections have emerged as a common source of infections in immunosuppressed patients. All fungal cells are surrounded by a cell wall that is essential for cell integrity and survival. It prevents cell death and lysis resulting from high internal turgor pressure. Since the cell wall is not present in animal cells, it is an ideal target for selective invasive fungal infection treatments. The antifungal family known as echinocandins, which specifically inhibit the synthesis of the cell wall β(13)glucan, has been established as an alternative treatment for mycoses. To explore the mechanism of action of these antifungals, we analyzed the cell morphology and glucan synthases localization in Schizosaccharomyces pombe cells during the initial times of growth in the presence of the echinocandin drug caspofungin. S. pombe are rod-shaped cells that grow at the poles and divide by a central division septum. The cell wall and septum are formed by different glucans, which are synthesized by four essential glucan synthases: Bgs1, Bgs3, Bgs4, and Ags1. Thus, S. pombe is not only a perfect model for studying the synthesis of the fungal β(1-3)glucan, but also it is ideal for examining the mechanisms of action and resistance of cell wall antifungals. Herein, we examined the cells in a drug susceptibility test in the presence of either lethal or sublethal concentrations of caspofungin, finding that exposure to the drug for long periods at high concentrations (>10 µg/mL) induced cell growth arrest and the formation of rounded, swollen, and dead cells, whereas low concentrations (<10 µg/mL) permitted cell growth with a mild effect on cell morphology. Interestingly, short-term treatments with either high or low concentrations of the drug induced effects contrary to those observed in the susceptibility tests. Thus, low drug concentrations induced a cell death phenotype that was not observed at high drug concentrations, which caused transient fungistatic cell growth arrest. After 3 h, high concentrations of the drug caused the following: (i) a decrease in the GFP-Bgs1 fluorescence level; (ii) altered locations of Bgs3, Bgs4, and Ags1; and (iii) a simultaneous accumulation of cells with calcofluor-stained incomplete septa, which at longer times resulted in septation uncoupling from plasma membrane ingression. The incomplete septa revealed with calcofluor were found to be complete when observed via the membrane-associated GFP-Bgs or Ags1-GFP. Finally, we found that the accumulation of incomplete septa depended on Pmk1, the last kinase of the cell wall integrity pathway. [ABSTRACT FROM AUTHOR]

Market competition in a globalized society requires increasing quality requirements in the production process, which is reflected in the final products. The main measure of the effects of introducing quality is the customer and the position of the product on the market. The requirements of industrial production in the time of Industry 4.0 era in competition require the introduction of innovation in the field of production as a means of increasing the competitiveness and performance of the company. In the 4.0 industrial era, competition between companies is fiercer, forcing many to produce to seek innovation to improve the competitiveness and performance of their company. The companies are implementing an integrated management system ISO 9001: 2015 Quality Management System and ISO 22000: 2018 Food Safety Management. In this paper we deal with a case study where we prove that the introduction of quality systems and quality standards leads to an increase in overall product quality, labour productivity and at the same time to a better position of the company in the market, i.e. in the pharmaceutical industry. The study examines food and pharmaceutical products containing glucan, specifically those found on the Slovak market. The results are applied using statistical methods and briefly summarized in tables. The study is based on similar findings from abroad and proves that a unique product was produced in Slovakia. [ABSTRACT FROM AUTHOR]

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

Marcia hiantina (Mollusca, Bivalvia) (Lamarck, 1818), is an edible clam mainly distributed along the tropical coastal regions. Recent researches have demonstrated that clams can possess compounds, including polysaccharides, with a wide range of biological actions including antioxidant, immunomodulatory and antitumor activities. Here an α-glucan was isolated from M. hiantina by hot water, purified by anion exchange chromatography, and its structure was characterized by a combination of multiple nuclear magnetic resonance (NMR) methods (1D 1H, 1H-1H COSY, 1H-1H TOCSY, 1H-1H NOESY, 1H-13C HSQC and 1H-13C HSQC-NOESY spectra), gas chromatography-mass spectrometry, and high performance size exclusion chromatography (HPSEC). The analysis from NMR, monosaccharide composition, methylation analyses and HPSEC combined with multi-angle light scattering (MALS) of M. hiantina-derived α-glycan confirmed a branched polysaccharide exclusively composed of glucose (Glc), mostly 4-linked in its backbone, branched occasionally at 6-positions, and having a molecular weight of ~ 570 kDa. The mollusk α-glucan was subjected to four cell-based assays: (i) viability of three cell lines (RAW264.7, HaCaT, and HT-29), (ii) activity on lipopolysaccharide (LPS)-induced prostaglandin production in RAW264.7 cells, (iii) inhibitory activities of in H2O2- and LPS-induced reactive oxygen species (ROS) production in HMC3 cells, and (iv) HaCaT cell proliferation. Results have indicated no cytotoxicity, potent inhibition of both H2O2- and LPS-induced ROS, and potent cell proliferative activity. [ABSTRACT FROM AUTHOR]

Methyl cellulose and its derivatives are widely used in the food industry, cosmetics, and as construction materials. The properties of methyl celluloses (MC) strongly depend on their degrees and positions of substitution. In order to generate MCs with uncommon blocky substitution, we apply fully protected O-benzyl-O-methyl celluloses (BnMC). Such complex polysaccharide derivatives could not be deprotected completely and without shift of the composition by methods usually applied to mono- and oligosaccharides. Therefore, a facile debenzylation method was developed based on photo-initiated free-radical bromination in the presence of hydrobromic acid scavengers followed by alkaline treatment. The reaction proceeds under homogeneous conditions and without the aid of any catalyst. There is no need for expensive equipment, materials, anhydrous reagents, or running the reaction under anhydrous conditions. Reaction parameters were investigated and optimized for successful debenzylation of completely protected BnMC with degrees of methyl substitution (DSMe) around 1.9 (and DSBn around 1.1). Side-product-free and almost complete debenzylation was achieved when 1,2-epoxybutane (0.5 eq./eq. N-bromosuccinimide) and 2,6-di-tert-butylpyridine (0.5 eq./eq. N-bromosuccinimide) were used in the reaction. Furthermore, ATR-IR and 1H NMR spectroscopy confirmed the successful removal of benzyl ether groups. The method was developed to monitor the transglycosylation reaction of the BnMC with permethylated cellulose, for which the deprotection of many small samples in parallel is required. This comprises the determination of the methyl pattern in the glucosyl units by gas-liquid chromatography (GLC), as well as oligosaccharide analysis by liquid chromatography mass spectrometry (LC-MS) after perdeuteromethylation and partial hydrolysis to determine the methyl pattern in the chains. The unavoidable partial chain degradation during debenzylation does not interfere with this analytical application, but, most importantly, the DS and the methyl pattern were almost congruent for the debenzylated product and the original MC, indicating the full success of this approach The presented method provides an unprecedented opportunity for high throughput and parallel debenzylation of complicated glucans, such as BnMC (as a model compound), for analytical purposes. For comparison, debenzylation using Na/NH3 was applied to BnMC and resulted in a completely debenzylated product with a remarkably high recovery yield of 99 mol% and is, thus, the method of choice for synthetic applications, e.g., for the transglycosylation product prepared under the selected conditions in a preparative scale.

The presently reported study intended to examine the effect of the oral administration of an immunomodulator ß-glucan and ß-glucan-containing product (BGN-2) on the levels of tumor necrosis factor-alpha (TNF-a), interleukin-6 (IL-6), fish heat shock protein 70 (HSP-70), and fish growth hormone (GH) as well as growth performance of cultured sea trout juveniles. The sea trout is a migratory sea-run brown trout, Salmo trutta Linnaeus, 1758. The fish (total of 15 000) were divided into four experimental and one control group including control (consisting of basal diet) (D1); basal diet + 1 g kg-1 ß-glucan (D2); basal diet + 3 g kg-1 ß-glucan (D3); basal diet + 6 g kg-1 BGN-2 (D4); basal diet + 14 g kg-1 BGN-2 (D5). The fish fed D4 and D5 diets showed significantly higher IL-6, HSP-70, and GH expression levels than other treatments (P < 0.05). Sea trout fed D4 and D5 diets showed significant improvements in growth performance compared to the fish fed the control diet. In conclusion, our results suggest that dietary supplementation with the product BGN-2 provides great immunostimulation and could be used as an effective measure to improve growth performance in juvenile sea trout. [ABSTRACT FROM AUTHOR]

The role of sweet potato tubers in human nutrition is not new. The above-ground biomass of sweet potatoes is not used for nutritional purposes in most countries, but it has a high biological value. Therefore, the aim of the present study is to investigate the production of press fibre from above ground biomass by wet fractionation. Two sweet potato varieties (purple- and white-fleshed sweet potato) and two types of irrigation system were used: bubbling water flow system (BWS) and continuous water flow system (CWS). Glucan, xylan, arabinan were analysed by HPLC and elemental content was measured by ICP-OES. Our results show that the total carbohydrate content in the pressed fibre of the leaf blades (27.64-29.88% w/w) is lower than in the stem with petiole (51.14-57.36% w/w). No significant difference in glucan, xylan and arabinan content was observed in the leaf blade. In the stem with petiole, significant differences were observed for xylan and arabinan contents. For elemental content, generally higher values were measured in the leaf blade than in the stem with petiole. This information may be relevant for the selection of the appropriate variety and treatment, even for the production of functional food. [ABSTRACT FROM AUTHOR]

Exopolysaccharides (EPSs) are novel functional additives for low-fat yogurt. Pharmaceutical, medical, and food industries are using more LAB-based EPSs. in this study, leuconostoc spp. was used to produce ninth bacterial ePSs in a modified molasses medium. Production of EPSs was concentration-dependent on all stains and the highest yield was obtained from the S3 strain (55.23 g/l), followed by S6 (49.95 g/l), S8 (45.68 g/l), and S7 (44.23), respectively. HPLC and FTIR analysis showed that all purified EPSs from leuconostoc citreum (S3) and leuconstoc holzaapfelii (S8) were related to exopolysaccharide glucan. Anticancer activity of all EPSs samples (EPSS1-9) against Caco-2 cells and normal MCR-5 cells were investigated using Mtt assay. the results revealed that Caco-2 cells were more sensitive than the normal MCR-5 cells. the highest anticancer activity against Caco-2 cancer cells was recorded for EPS8 (IC50 = 22.94 µg/ml, Si=3.73), followed by EPS3 (IC50 = 36.15 µg/ml, SI=8.72), ePS1 (IC50 = 50.01 µg/ml, SI=3.73), and EPS4 (IC50 = 94.90 µg/ml, SI=3.26), respectively. the lowest cytotoxicity was recorded for ePS5 (IC50 = 130.5 µg/ml). the most active ePSs (EPS3 and EPS8) were used as fat replacements and stabilizers in low-fat set yogurt at non-toxic concentrations (0.4, 0.8, and 1.2%). EPS3 and EPS8 improved the low-fat yogurt's organoleptic and rheological properties. ePS8 had the highest water holding capacity (77.26%), viscosity (3660 CP), and lowest syneresis (22.95%) and whey off (0.6 ml). low-fat set yogurt enhanced with EPS3 and EPS8 recorded the highest sensory evaluation values with overall acceptability, especially EPS3b, EPS3c, EPS8c, and EPS8b; the total score point of 97.50, 97.43, 96.51, and 96.36, respectively in fresh age compared to control yogurt (92.64). in conclusion, leuconostoc EPSs, especially EPS8, can be explored for anti-cancer effects on Caco-2 colorectal cancer cells. it could also improve the rheological and organoleptic qualities of low-fat set yogurt. [ABSTRACT FROM AUTHOR]

Paotianxiong (PTX) is a processing product of Aconitum carmichaelii Debx., often used as a tonic food daily. However, the structure and activity of the polysaccharide component that plays a major role still need to be determined. In our work, two new polysaccharides were purified from PTX and named PTXP-1 and PTXP-2. Structural analysis showed that PTXP-1 is a glucan with a molecular weight of 915 Da and a structure of 4)-α-D-Glcp-(1 → as the main chain. PTXP-2 is a glucose arabinoglycan with 4)-α-D-Glcp-(1 → as the main chain, containing 8 glycosidic bonds attached, and a molecular weight of 57.9KDa. In vitro probiotic experiments demonstrated that PTXP-1 could significantly promote probiotic growth and acid production. In vivo experiments demonstrated that both PTXP-1 and PTXP-2 exhibited significant effectiveness in promoting the growth of intestinal probiotics. These findings help expand the application of polysaccharide components extracted from tonic herbs as functional food ingredients. [Display omitted] • PTXP-1 is a glucan with a molecular weight of 915 Da and a structure of 4)-α-D-Glcp-(1→ as the main chain. • PTXP-2 is a glucose arabinoglycan with 4)-α-D-Glcp-(1→ as the main chain, containing 8 glycosidic bonds attached. •.PTXP-1 promotes the four probiotics to produce short-chain fatty acids, especially for Bifidobacterium infantis. • These findings imply that PTXP-1 has the potential to be used as a prebiotic additive like oligofructose. [ABSTRACT FROM AUTHOR]

The polysaccharide glucan was extracted from Gastrodia elata Blume, and its structural characterizations and beneficial effects against acute dextran sulfate sodium (DSS)-induced ulcerative colitis were investigated. The results showed that a polysaccharide GP with a molecular weight of 811.0 kDa was isolated from G. elata Blume. It had a backbone of α-D-1,4-linked glucan with branches of α- d -glucose linked to the C-6 position. GP exhibited protective effects against DSS-induced ulcerative colitis, and reflected in ameliorating weight loss and pathological damages in mice, increasing colon length, inhibiting the expression of inflammatory cytokines tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β), decreasing the levels of inflammatory related proteins NLRP3 and ASC, and elevating the anti-inflammatory cytokine interleukin-10 (IL-10) level in mouse colon tissues. GP supplementation also reinforced the intestinal barrier by promoting the expression of ZO-1, Occludin, and MUC2 of colon tissues, and positively regulated intestinal microbiota. Thus, GP treatment possessed a significant improvement in ulcerative colitis in mice, and it was expected to be developed as a functional food. [ABSTRACT FROM AUTHOR]

Emerging and re-emerging diseases in fish cause drastic economic losses in the aquaculture sector. To combat the impact of disease outbreaks and prevent the emergence of infections in culture systems, understanding the advanced strategies for protecting fish against infections is inevitable in fish health research. Therefore, the present study aimed to evaluate the induction of trained immunity and its protective efficacy against Streptococcus agalactiae in tilapia. For this, Nile tilapia and the Tilapia head kidney macrophage primary culture were primed using β-glucan @200 μg/10 g body weight and 10 μg/mL respectively. Expression profiles of the markers of trained immunity and production of metabolites were monitored at different time points, post-priming and training, which depicted enhanced responsiveness. Higher lactate and lactate dehydrogenase (LDH) production in vitro suggests heightened glycolysis induced by priming of the cells using β-glucan. A survival rate of 60% was observed in β-glucan trained fish post challenge with virulent S. agalactiae at an LD 50 of 2.6 × 107 cfu/ml, providing valuable insights into promising strategies of trained immunity for combating infections in fish. • Innate immune training using β-glucan can elevate cytokine transcript levels. • Expression of trained immunity markers, mTOR and HIF1α elevated post immune training with β-glucan in tilapia. • Higher lactate and LDH production post β-glucan induction in the supernatant of tilapia head kidney macrophage culture. • Survival rate of 60% in Streptococcus agalactiae- challenged immune trained tilapia. [ABSTRACT FROM AUTHOR]

ABSTRACT: Here we show that selected nutritional feed additives reduce the adverse effects of transport stress on the immune system and hematology of tambaqui. We formulated a control diet to contain normal dietary levels of vitamin E (21.6 mg/ kg diet) and C (143 mg/kg), then we added supra levels of these vitamins (vit E – 264 mg/kg and vit C – 1000 mg/kg) to a second diet. Finally, a third diet was produced to contain similar levels of vitamins from diet 2 with 0.1% beta-glucan supplementation. Four hundred thirty-two tambaquis (20.91 g ± 0.27 g) were randomly assigned to 12 aquaria and fed the diets for 15 days; then, all fish were transported for five h and then returned to the aquaria. Blood samples were collected before and after the transport and at the end of the trial (60 days). Transportation significantly increased blood glucose that returned to baseline levels at the end of the trial. However, cortisol seemed to be unresponsive to the stress. Surprisingly, the stress significantly increased the immunoglobulin level after transport. Additionally, the transport markedly reduced the red blood cell count and leukocyte and lymphocytes counts while increasing the control group's neutrophil number. These effects lasted until the end of the trial in the control group. Supra levels of the vitamins and glucan supplementation prevented the decrease in red blood cell and leukocyte count after the stress. Additionally, beta-glucan supplementation induced lower cortisol levels in all the sampling points. However, the effect on the immune parameters was limited, increasing only the lysozyme activity and serum protein levels in the beta-glucan supplemented group and the group fed only the supra levels of vitamins, respectively. In sum, our results indicated that transport for five h induced a limited effect on stress biomarkers. The use of supra levels of antioxidant vitamins alone or in combination with beta-glucan could restore or prevent the adverse effects of stress on hematology and the immune system.

In this study, a bacterium with the ability to extracellularly produce a water-soluble polysaccharide (with high molecular mass of 743 kDa) was obtained from saline soils. This strain named as ZB01 was identified as Rhizobium punsense by 16S rRNA sequence analysis. The monomer composition and structure of extracellular polysaccharides were investigated by high-performance anion-exchange chromatography-pulsed amperometric detector, Fourier-transform infrared, methylation and nuclear magnetic resonance spectroscopy analysis. In addition, the morphological characterization of extracellular polysaccharides was performed by scanning electron microscopy analysis. As a result, the polysaccharide is found to be a novel water-soluble type of glucan most likely consisting of repeating two β- d -Glc p -(1 → 3) and one α- d -Glc p -(1 → 3). Furthermore, it showed pronounced antimicrobial effects against Escherichia coli and Bacillus subtilis , suggesting it has the potential to be used as a natural antibacterial agent in the future. [Display omitted] • A Rhizobium punsense strain with ability to produce exopolysaccharides (EPS) was obtained. • The EPS likely consists of repeating two β- d -Glcp-(1 → 3) and one α- d -Glcp-(1 → 3). • The discovered EPS displayed obvious antimicrobial activity. • This study provides a novel approach to produce water-soluble β-glucan. [ABSTRACT FROM AUTHOR]