Characterization of an Aptamer Targeting Neu5Gc, as an Endogenous Pathogenic Factor Derived from Red Meat.

N-glycolylneuraminic acid (Neu5Gc), a sialic acid predominantly found in the non-neurohumoral fluids of hind-mouthed animals, is incapable of synthesizing Neu5Gc due to a deletion in the CMAH exon of the gene encoding human CMP-Neu5Gc hydroxylase. But consumption of animal-derived foods that contain Neu5Gc, such as red meat, can instigate an immune response in humans, as Neu5Gc is recognized as a foreign substance by the human immune system. This recognition leads to the production of anti-Neu5Gc antibodies, subsequently resulting in chronic inflammation. When Neu5Gc is consumed excessively or frequently, it may contribute to the development of heart disease and cancer. This makes Neu5Gc, an endogenous pathogenic factor derived from red meat, a new hot topic in red meat safety research. In this study, aptamers obtained by the magnetic bead SELEX technique were subjected to homology and secondary structure prediction analysis as well as affinity determination. The result indicated that the aptamer 2B.N2A9 exhibited a robust binding affinity, with an affinity constant (Ka) of 1.87 × 108 L/mol. This aptamer demonstrated optimal binding specificity within a pH range of 5.4 to 7.4. Molecular docking analysis further revealed that aptamer 2B.N2A9 formed stable binding interactions with the target Neu5Gc at specific sites, namely G-14, C-15, G-13, G-58, G-60, and C-59. An Enzyme-Linked Oligonucleotide Sorbent Assay (ELOSA) methodology was established to detect the endogenous pathogenic factor Neu5Gc present in red meat. This method demonstrated a limit of detection (LOD) of 0.71 ng/mL, along with an average recovery rate of 92.23%. The aptamer obtained in this study exhibited favorable binding properties to Neu5Gc. The assay was relatively convenient and demonstrated good sensitivity. Further investigation into the distribution of Neu5Gc in various red meats is of public health significance and scientific potential. A practical detection method should be provided to guide red meat diets and ensure the nutrition and safety of meat products.

Notch1 signaling activation alleviates coronary microvascular dysfunction through histone modification of Nrg-1 via the interaction between NICD and GCN5.

The Notch signaling pathway is related to endothelial dysfunction in coronary atherosclerosis. Our objective was to explore the role of Notch signaling in coronary microvascular dysfunction (CMD). CMD models were constructed by sodium laurate injection in vivo and homocysteine (Hcy) stimulation in vitro. The binding ability of Notch Intracellular Domain (NICD)/H3K9Ac/GCN5 (General Control Non-derepressible 5) to Neuregulin-1 (Nrg-1) promoter was examined by chromatin immunoprecipitation. Immunofluorescence staining was conducted to detect CD31 positive cells, NICD localization, and co-localization of NICD and GCN5. Flow cytometry and Tunel staining were conducted to identify the apoptosis. Hematoxylin and eosin staining, quantitative real-time PCR, western blot, immunohistochemical staining, co-immunoprecipitation, and double luciferase report analysis were also conducted. Notch signaling pathway-related protein levels were decreased, levels of Nrg-1 and the phosphorylation of ErbB2 and ErbB4 were enhanced in CMD models. Interference with Nrg-1 further increased the apoptosis in Hcy-induced cardiac microvascular endothelial cells (CMECs). Meanwhile, the activation of the Notch signaling pathway increased the levels of Nrg-1 and the phosphorylation of ErbB2 and ErbB4, as well as inhibited the apoptosis induced by Hcy. Furthermore, NICD and histone acetyltransferase enzyme GCN5 could regulate Nrg-1 promoter activity by affecting the expression of acetylation-modified protein H3K9Ac. In addition, NICD also interacted with GCN5. In vivo results also confirmed that the activation of the Notch signal alleviated CMD. Notch signaling pathway regulates Nrg-1 level through synergistic interaction with GCN5, thereby mitigating CMD.

The Gut-Organ-Axis Concept: Advances the Application of Gut-on-Chip Technology.

The intestine is considered to be a vital digestive organ to absorb nutrients and is the largest immune organ, while numerous microorganisms coexist with the host. It is well known that the complex interactions between the gut microbiota and the host's immune system inevitably affect the function of other organs, creating an "axis" between them. During the past few years, a new technique based mainly on microfluidics and cell biology has been developed to emulate the structure, function, and microenvironment of the human gut, called the "gut-on-chip". This microfluidic chip provides insight into key aspects of gut function in health and disease, such as the gut-brain axis, gut-liver axis, gut-kidney axis, and gut-lung axis. In this review, we first describe the basic theory of the gut axis and the various composition and parameter monitoring of the gut microarray systems, as well as summarize the development and emerging advances in the gut-organ-on-chip, with a focus on the host-gut flora and nutrient metabolism, and highlight their role in pathophysiological studies. In addition, this paper discusses the challenges and prospects for the current development and further use of the gut-organ-on-chip platform.

Weakened Contractile Performance and Mitochondrial Respiratory Complex Activity in Skeletal Muscle Improve during Interbout Arousal in Hibernating Daurian Ground Squirrel, Spermophilus dauricus.

Mammalian hibernation is composed of multiple episodes of torpor bout, separated by phases of interbout arousal. During torpor, the skeletal muscles of mammals are undoubtedly inactive, but it has been proven to mitigate disuse atrophy. While interbout arousal has been implicated in the prevention of muscle atrophy, the underlying mechanisms sustaining muscle contraction remain to be explored. In the present study, Daurian ground squirrels (Spermophilus dauricus) were divided into four groups: pre-hibernation (PRE), torpor (TOR), interbout arousal (IBA), and post-hibernation (POST). The contractile performance of slow-twitch soleus muscle (SOL) and fast-twitch extensor digitorum longus muscle (EDL) was detected both in situ and in vitro. Concurrently, mitochondrial respiratory chain complex activity in these muscles was quantified. Our findings revealed that in situ contractile properties of both muscles, including force, power output, time duration, and force development/relaxation rates of twitch contraction, and force and power output of tetanic contraction declined in the TOR group compared to the PRE group, but improved in the IBA and POST groups. Fatigue resistance of muscles, determined by the power output of repetitive tetanic contractions in situ, decreased in the TOR group but recovered in the IBA and POST groups. In vitro studies demonstrated that tetanic contraction power output in isolated muscles increased with muscle temperature in both TOR and IBA groups. However, at the same temperature, power output was consistently lower in the TOR group compared to the IBA group. Moreover, the activity of the mitochondrial respiratory chain complex, especially Complexes I and II, decreased in the TOR group but showed recovery in the IBA and POST groups. These findings suggest that both the contractile performance and fatigue resistance of mammalian skeletal muscle are compromised during torpor but can be improved during interbout arousal and post-hibernation. The rebound in body temperature and rise in mitochondrial respiratory chain complex activity in skeletal muscle are involved in enhancing contractile performance and fatigue resistance. This study suggests that interbout arousal functions as a vital temporal interval during which skeletal muscles can transition from the inactivity induced by torpor to a state of restored contractile functionality. Thus, interbout arousal serves as a behavioral safeguard against disuse-induced damage to skeletal muscles during hibernation.

Protective Effect of Astragaloside IV against Cadmium-Induced Damage on Mouse Renal Podocytes (MPC5).

In this study, we investigated the protective effect of Astragaloside IV (Ast) on mouse podocytes and its possible mechanism of action by constructing a cadmium-induced mouse renal podocytes model. We investigated the effects of cadmium (Cd) toxicity on cell number, morphology, the mitochondrial status of subcellular organelles, protein and gene levels, and the protective effects of Ast by constructing a model of Cd-induced damage to mouse renal podocytes (MPC5) and giving Ast protection at the same time. The results showed that exposure of MPC5 cells to CdCl2 culture medium containing 6.25 µM concentration acted with low cell mortality, but the mortality of MPC5 cells increased with the prolongation of cadmium exposure time. Given Ast, the death rate in the low dose group (12.5 µM) was significantly reduced, while the death rate in the medium dose group (25 µM) was extremely significantly reduced. In comparison to the control group, the Cd-exposed group exhibited a significant increase of 166.7% in malondialdehyde (MDA) content and a significant decrease of 17.1% in SOD activity. The mitochondrial membrane potential was also reduced to varying degrees. However, in the Ast-protected group compared to the Cd-exposed group, the MDA content significantly decreased by 20.8%, the SOD activity decreased by 7.14%, and the mitochondrial membrane potential showed a significant increase. Fluorescence staining of mitochondrial membrane potential indicated that Cd exposure caused mitochondrial apoptosis. In the 12-h cadmium-exposed group, the protein expression of Nephrin in mice significantly decreased by 33.4%. However, the expression of the Desmin protein significantly increased by 67.8%, and the expression of the autophagy protein LC3-II significantly increased by 55.5%. Meanwhile, the expression of PINK1, a mitochondrial autophagy pathway protein, was significantly increased in the 12 h and 24 h cadmium exposure groups. The mRNA level of PINK1 was significantly increased, and that of Parkin was decreased in the 48 h cadmium exposure group. Compared to the Cd-exposed group, the Ast group showed more significant improvements in the expression of podocyte structure, functional proteins, and mitochondrial autophagy pathway proteins. The immunological assay of mitochondrial autophagic pathway proteins further indicated that Cd-induced damage to MPC5 cells might be associated with the dysregulation of mitochondrial autophagy.

Recent advances in quality preservation of postharvest golden needle mushroom (Flammulina velutiper).

The golden needle mushroom (Flammulina velutiper) is one of the most productive mushrooms in the world. However, F. velutiper experiences continuous quality degradation in terms of changes in color and textural characteristics, loss of moisture, nutrition and flavor, and increased microbial populations due to its high respiratory activity during the postharvest phase. Postharvest preservation techniques, including physical, chemical and biological methods, play a vital role in maintaining postharvest quality and extending the shelf life of mushrooms. Therefore, in this study, the decay process of F. velutiper and the factors affecting its quality were comprehensively reviewed. Additionally, the preservation methods (e.g., low-temperature storage, packaging, plasma treatment, antimicrobial cleaning and 1-methylcyclopropene treatment) for F. velutiper used for the last 5 years were compared to provide an outlook on future research directions. Overall, this review aims to provide a reference for developing novel, green and safe preservation techniques for F. velutiper. © 2023 Society of Chemical Industry.

Isolation, Structural Characterization, and Hypoglycemic Activities In Vitro of Polysaccharides from Pleurotus eryngii.

Pleurotus eryngii (PE) is an edible mushroom with high nutritional value. Pleurotus eryngii polysaccharides (PEPs) are one of the main active ingredients and manifest a great variety of biological activities. This study mainly focused on the chemical characterization and biological activities of PEPs, which were separated into two fractions (named WPS and P-1). WPS is mainly dominated by ß-glycosidic bonds and contains α-glycosidic bonds, and P-1 only contains α-glycosidic bonds. The molecular weights of WPS and P-1 were 4.5 × 105 Da and 2.2 × 104 Da. The result of GC indicated that two the fractions were composed of rhamnose, arabinose, xylose, mannose, glucose, and galactose, with a ratio of 0.35:0.24:0.45:0.24:28.78:1.10 for WPS and 0.95:0.64:0.66:1.84:60.69:0.67 for P-1. The advanced structure studies indicated that the two fractions had no triple-helical structure, where WPS had a dense structure and P-1 had a loose structure. In addition, the antioxidant activity of WPS surpassed P-1, and the two fractions also exhibited a high hypoglycemic activity via inhibiting α-glycosidase activities and promoting the expression of PI3K-AKT signaling pathway based on in vitro assay and cell experiments.

Structural Characteristics, Antioxidant and Hypoglycemic Activities of Polysaccharide from Siraitia grosvenorii.

The structural characterization, the in vitro antioxidant activity, and the hypoglycemic activity of a polysaccharide (SGP-1-1) isolated from Siraitia grosvenorii (SG) were studied in this paper. SGP-1-1, whose molecular weight is 19.037 kDa, consisted of Gal:Man:Glc in the molar ratio of 1:2.56:4.90. According to the results of methylation analysis, GC-MS, and NMR, HSQC was interpreted as a glucomannan with a backbone composed of 4)-ß-D-Glcp-(1→4)-, α-D-Glcp-(1→4)-, and 4)-Manp-(1 residues. α-1,6 linked an α-D-Galp branch, and α-1,6 linked an α-D-Glcp branch. The study indirectly showed that SGP-1-1 has good in vitro hypoglycemic and antioxidant activities and that these activities may be related to the fact that the SGP-1-1's monosaccharide composition (a higher proportion of Gal and Man) is the glycosidic-bond type (α- and ß-glycosidic bonds). SGP-1-1 could be used as a potential antioxidant and hypoglycemic candidate for functional and nutritional food applications.

Advances in Chemical Composition, Extraction Techniques, Analytical Methods, and Biological Activity of Astragali Radix.

Astragali Radix (AR) is one of the well-known traditional Chinese medicines with a long history of medical use and a wide range of clinical applications. AR contains a variety of chemical constituents which can be classified into the following categories: polysaccharides, saponins, flavonoids, amino acids, and trace elements. There are several techniques to extract these constituents, of which microwave-assisted, enzymatic, aqueous, ultrasonic and reflux extraction are the most used. Several methods such as spectroscopy, capillary electrophoresis and various chromatographic methods have been developed to identify and analyze AR. Meanwhile, this paper also summarizes the biological activities of AR, such as anti-inflammatory, antioxidant, antitumor and antiviral activities. It is expected to provide theoretical support for the better development and utilization of AR.

Process optimization of vanillin production by conversion of ferulic acid by Bacillus megaterium.

BACKGROUND: Vanillin is an important flavoring and aromatic ingredient found mainly in the pods of the tropical plant vanilla and is widely used in the food industry. Attempts have been made to produce vanillin from ferulic acid esters in agricultural residues of wheat bran. RESULTS: The results showed that a strain with high tolerance to the substrate ferulic acid was isolated and screened from soil and identified as belonging to the genus Bacillus (Bacillus megaterium). The concentration of vanillin produced by this strain was 0.048 g L-1 , and the molar conversion of vanillin was 12.25%. The production of vanillin was optimized by orthogonal experiments. Beef pastes 6.0 g L-1 , soybean meal 5.0 g L-1 , magnesium sulfate heptahydrate 1.0 g L-1 , iron(II) sulfate heptahydrate 1.0 g L-1 , calcium chloride 1.0 g L-1 , dipotassium hydrogen phosphate trihydrate 1.0 g L-1 ; fermentation culture conditions were pH 7.0, inoculum level 5%, loading volume 20%, ferulic acid 1.0 g L-1 , fermentation culture temperature 35 °C. The concentration of vanillin obtained was 0.218 g L-1 . Finally, transcriptomic analysis of the strain samples before and after the optimization of the fermentation conditions was carried out to study the effect of the optimization of the fermentation conditions on the concentration of vanillin produced by the strain. CONCLUSION: This study provides a theoretical basis for further improving the yield of vanillin and gradually realizing efficient industrial production. © 2022 Society of Chemical Industry.

Machine Learning Driven Synthesis of Few-Layered WTe2 with Geometrical Control.

Reducing the lateral scale of two-dimensional (2D) materials to one-dimensional (1D) has attracted substantial research interest not only to achieve competitive electronic applications but also for the exploration of fundamental physical properties. Controllable synthesis of high-quality 1D nanoribbons (NRs) is thus highly desirable and essential for further study. Here, we report the implementation of supervised machine learning (ML) for the chemical vapor deposition (CVD) synthesis of high-quality quasi-1D few-layered WTe2 NRs. Feature importance analysis indicates that H2 gas flow rate has a profound influence on the formation of WTe2, and the source ratio governs the sample morphology. Notably, the growth mechanism of 1T' few-layered WTe2 NRs is further proposed, which provides new insights for the growth of intriguing 2D and 1D tellurides and may inspire the growth strategies for other 1D nanostructures. Our findings suggest the effectiveness and capability of ML in guiding the synthesis of 1D nanostructures, opening up new opportunities for intelligent materials development.

Two-step CVD synthesis of NiTe2-MoS2vertical junctions with improved MoS2transistors performance.

The primary challenge for widespread applications of two-dimensional electronics is to achieve satisfactory electrical contacts due to the difficulties in inevitable physical damages and selectively doping during traditional metal integration process. The two-dimensional (2D) metal-semiconductor junctions have attracted captivated attention for potential applications in future atomically thin electronics as perfect candidates for achieving reliable electrical contacts. Here we demonstrate the van der Waals epitaxial growth of 2D NiTe2-MoS2 metal-semiconductor vertical junctions which the upper NiTe2 selectively nucleate at the edge of underlying MoS2. Optical microscopy (OM), scanning electron microscopy (SEM), atomic force microscopy (AFM), and scanning transmission electron microscope (STEM) studies confirm that NiTe2-MoS2 metal-semiconductor vertical junctions are successfully synthesized. Electrical properties of the NiTe2-contacted MoS2 field-effect transistors (FETs) show higher field-effect mobilities (µFE) than those with deposited Cr/Au contacts. This study demonstrates an effective pathway to improved MoS2 transistors performance with metal-semiconductor junctions.

Two-step chemical vapor deposition synthesis of NiTe2-MoS2 vertical junctions with improved MoS2 transistor performance.

The primary challenge for the widespread application of two-dimensional (2D) electronics is to achieve satisfactory electrical contacts because, during the traditional metal integration process, difficulties arise due to inevitable physical damage and selective doping. Two-dimensional metal-semiconductor junctions have attracted attention for the potential application to achieve reliable electrical contacts in future atomically thin electronics. Here we demonstrate the van der Waals epitaxial growth of 2D NiTe2-MoS2 metal-semiconductor vertical junctions where the upper NiTe2 selectively nucleates at the edge of the underlying MoS2. Optical microscopy (OM), scanning electron microscopy (SEM), atomic force microscopy (AFM), and scanning transmission electron microscope (STEM) studies confirmed that NiTe2-MoS2 metal-semiconductor vertical junctions had been successfully synthesized. The electrical properties of the NiTe2-contacted MoS2 field-effect transistors (FETs) showed higher field-effect mobilities (µ FE) than those with deposited Cr/Au contacts. This study demonstrates an effective pathway to improved MoS2 transistor performance with metal-semiconductor junctions.

Molecular cloning and characterization of AlgL17, a new exo-oligoalginate lyase from Microbulbifer sp. ALW1.

A new alginate lyase gene of algl17 was cloned from an alginate-degrading marine bacterium Microbulbifer sp. ALW1. The gene contained 2220 bp and encoded a 739-amino acid protein classified into the PL-17 family. The recombinant alginate lyase AlgL17 was overexpressed and purified from Escherichia coli BL21 (DE3) with a molecular mass of 84.9 kDa. This enzyme showed activities towards sodium alginate, polyM and alginate oligosaccharide, but very low activity towards polyG. These results indicate that AlgL17 is a polyM-specific oligoalginate lyase. When sodium alginate was used as a substrate, the optimum reaction temperature and pH for the enzyme were 35 °C and pH 8.0, respectively. Recombinant AlgL17 was stable at 25 °C, but not stable at 30 °C and 35 °C. It showed good stability over a pH range of 5.0-8.0. The enzyme activity was increased to 1.7 times by adding NaCl to a final concentration of 0.7 M. The ability of the recombinant AlgL17 producing 4-deoxy-L-erythro-5-hexoseulose uronic acid (DEH) from sodium alginate and polyM block indicates that AlgL17 is an exo-type alginate lyase.

Machine-Driven Enzymatic Oligosaccharide Synthesis by Using a Peptide Synthesizer.

For decades, researchers have endeavored to develop a general automated system to synthesize oligosaccharides that is comparable to the preparation of oligonucleotides and oligopeptides by commercially available machines. Inspired by the success of automated oligosaccharide synthesis through chemical glycosylation, a fully automated system is reported for oligosaccharides synthesis through enzymatic glycosylation in aqueous solution. The designed system is based on the use of a thermosensitive polymer and a commercially available peptide synthesizer. This study represents a proof-of-concept demonstration that the enzymatic synthesis of oligosaccharides can be achieved in an automated manner using a commercially available peptide synthesizer.

Facile Chemoenzymatic Synthesis of O-Mannosyl Glycans.

O Mannosylation is a vital protein modification involved in brain and muscle development whereas the biological relevance of O-mannosyl glycans has remained largely unknown owing to the lack of structurally defined glycoforms. An efficient scaffold synthesis/enzymatic extension (SSEE) strategy was developed to prepare such structures by combining gram-scale convergent chemical syntheses of three scaffolds and strictly controlled sequential enzymatic extension catalyzed by glycosyltransferases. In total, 45 O-mannosyl glycans were obtained, covering the majority of identified mammalian structures. Subsequent glycan microarray analysis revealed fine specificities of glycan-binding proteins and specific antisera.

Cation exchange assisted binding-elution strategy for enzymatic synthesis of human milk oligosaccharides (HMOs).

A cation exchange assisted binding-elution (BE) strategy for enzymatic synthesis of human milk oligosaccharides (HMOs) was developed. An amino linker was used to provide the cation ion under acidic condition which can be readily bound to cation exchange resin and then eluted off by saturated ammonium bicarbonate. Ammonium bicarbonate in the collections was easily removed by vacuum evaporation. This strategy circumvented the incompatible issue between glycosyltransferases and solid support or large polymers, and no purification was needed for intermediate products. With current approach, polyLacNAc backbones of HMOs and fucosylated HMOs were synthesized smoothly.

An enzymatic strategy to asymmetrically branched N-glycans.

An enzymatic strategy was developed to generate asymmetrically branched N-glycans from natural sources by using a panel of glycosidases and glycosyltransferases. Briefly, LacZ ß-galactosidase was employed to selectively trim symmetrically branched N-glycans isolated from bovine fetuin. The yielding structures were then converted to asymmetrically branched core structures by robust glycosyltransferase for further extension.

Decoding glycan protein interactions by a new class of asymmetric N-glycans.

N-Glycans are normally involved in crucial physiological and disease processes by interactions with glycan-binding proteins. So far structurally defined N-glycans have been good candidates for glycan binding study. Herein, a class of homogeneous asymmetric N-glycans was synthesized by coupling glycan-oxazoline and N-glycans using EndoM N175Q catalyzed quick glycan extension. Branch-biased binding and spacial inhibition caused by the bulky group on the other branch of N-glycan were observed in glycan protein interactions involving lectins and these glycans by glycan microarray study. These new compounds are valuable for functional glycomic studies to better understand new functions of glycans and glycan-binding proteins.

Chemoenzymatic Synthesis of a Library of Human Milk Oligosaccharides.

Human milk oligosaccharides (HMOs) are a family of diverse unconjugated glycans that exist in human milk as one of the major components. Characterization, quantification, and biofunctional studies of HMOs remain a great challenge due to their diversity and complexity. The accessibility of a homogeneous HMO library is essential to solve these issues which have beset academia for several decades. In this study, an efficient chemoenzymatic strategy, namely core synthesis/enzymatic extension (CSEE), for rapid production of diverse HMOs was reported. On the basis of 3 versatile building blocks, 3 core structures were chemically synthesized via consistent use of oligosaccharyl thioether and oligosaccharyl bromide as glycosylation donors in a convergent fragment coupling strategy. Each of these core structures was then extended to up to 11 HMOs by 4 robust glycosyltransferases. A library of 31 HMOs were chemoenzymatically synthesized and characterized by MS and NMR. CSEE indeed provides a practical approach to harvest structurally defined HMOs for various applications.