Laminaria japonica polysaccharide alleviates type 2 diabetes by regulating the microbiota-gut-liver axis: A multi-omics mechanistic analysis.

The hypoglycemic effects of low-molecular-weight Laminaria japonica polysaccharide (LJO) were investigated in type 2 diabetes mellitus (T2DM) mice, focusing on its effect on the microbiome, metabolome, and transcriptome. The findings demonstrated that LJO significantly reduced fasting blood glucose levels, insulin levels, and inflammatory factors. Additionally, LJO induced changes in gut microbiota composition and increased the concentrations of cecal short-chain fatty acids. Analysis of transcriptomics and metabolomics data revealed that LJO primarily altered the endocrine and digestive systems, signal transduction, and lipid metabolism. It led to a decrease in palmitic acid levels and an increase in glutathione levels. Real-time quantitative polymerase chain reaction assay suggested that LJO upregulated Irs1 expression, consequently reducing insulin resistance. These findings strongly suggest that LJO holds promise in ameliorating T2DM and may serve as a potential dietary supplement for patients with T2DM.

Agrocybe aegerita Polysaccharide Combined with Bifidobacterium lactis Bb-12 Attenuates Aging-Related Oxidative Stress and Restores Gut Microbiota.

The objective of this study was to examine the impacts of the combing of Agrocybe aegerita polysaccharides (AAPS) with Bifidobacterium lactis Bb-12 (Bb-12) on antioxidant activity, anti-aging properties, and modulation of gut microbiota. The results demonstrated that the AAPS and Bb-12 complex significantly increased the average lifespan of male and female Drosophila melanogaster under natural aging conditions (p < 0.05), with an improvement of 8.42% and 9.79%, respectively. Additionally, the complex enhanced their climbing ability and increased antioxidant enzyme activity, protecting them from oxidative damage induced by H2O2. In D-galactose induced aging mice, the addition of AAPS and Bb-12 resulted in significantly increase in antioxidant enzyme activity, regulation of aging-related biomarker levels, changed gut microbiota diversity, restoration of microbial structure, and increased abundance of beneficial bacteria, particularly lactobacilli, in the intestines. These findings suggested that the complex of AAPS and Bb-12 had the potential to serve as a dietary supplement against organism aging and oxidative stress.

The Potential Hypoglycemic Competence of Low Molecular Weight Polysaccharides Obtained from Laminaria japonica.

This study aimed to assess the hypoglycemic efficacy of low molecular weight polysaccharides fractions obtained from Laminaria japonica (LJOO) in a model of type 2 diabetes mellitus (T2DM) constructed using mice. Biochemical parameters were measured after 4 weeks of continuous gavage, and fasting blood glucose (FBG) concentrations were analyzed. Pathological changes in tissues were assessed. The intestinal contents were obtained for 16S rDNA high-throughput sequencing analysis and detection of short-chain fatty acids (SCFAs). LJOO lowered FBG and insulin concentrations. It altered the gut microbiota composition, as evidenced by enriched probiotic bacteria, along with an increase in the Bacteroidetes/Firmicutes ratio and a decrease in the population of harmful bacteria. LJOO stimulated the growth of SCFA-producing bacteria, thereby increasing cecal SCFAs levels. LJOO can potentially aid in alleviating T2DM and related gut microbiota dysbiosis. LJOO may be used as a food supplement for patients with T2DM.

Hetero-Diels-Alder Cycloaddition Reaction of Vinyl Ethers Enables Selective Fluorescent Labeling of Plasmalogens in Human Plasma Lipids.

Plasmalogens (Pls) are vinyl ether-containing glycerophospholipids of broad biological interest. Their abnormal levels are associated with neurological disorders and cardiovascular diseases. The intricacy of analyzing Pls in lipid samples arises from the wide variety of other coexisting lipid species, which underscores the urgent need for a Pls-specific labeling reaction. To address this challenge, we report an efficient hetero-Diels-Alder cycloaddition reaction between nonterminal vinyl ethers of Pls and o-quinolinone quinone methide probes under mild conditions. On the basis of this mechanism, a selective fluorescent labeling method for Pls is developed. The application of this method permits the exclusive derivatization of Pls over other human plasma lipids. The process also imparts labeled Pls with distinct fluorescence emission and chromatographic retention properties. By integrating this method with high-performance liquid chromatography, we are able to identify individual chromatographic signatures of Pls from 10 different human plasma samples. This Pls signature analytical technique, empowered by the Pls-specific labeling reaction, is cost-effective and simple in terms of instrumentation, suggesting its promising potential for the early screening and diagnosis of diseases linked to Pls abnormalities.

Discovering covalent inhibitors of protein-protein interactions from trillions of sulfur(VI) fluoride exchange-modified oligonucleotides.

Molecules that covalently engage target proteins are widely used as activity-based probes and covalent drugs. The performance of these covalent inhibitors is, however, often compromised by the paradox of efficacy and risk, which demands a balance between reactivity and selectivity. The challenge is more evident when targeting protein-protein interactions owing to their low ligandability and undefined reactivity. Here we report sulfur(VI) fluoride exchange (SuFEx) in vitro selection, a general platform for high-throughput discovery of covalent inhibitors from trillions of SuFEx-modified oligonucleotides. With SuFEx in vitro selection, we identified covalent inhibitors that cross-link distinct residues of the SARS-CoV-2 spike protein at its protein-protein interaction interface with the human angiotensin-converting enzyme 2. A separate suite of covalent inhibitors was isolated for the human complement C5 protein. In both cases, we observed a clear disconnection between binding affinity and cross-linking reactivity, indicating that direct search for the aimed reactivity-as enabled by SuFEx in vitro selection-is vital for discovering covalent inhibitors of high selectivity and potency.

Modulation of gut microbiota and lipid metabolism in rats fed high-fat diets by Ganoderma lucidum triterpenoids.

Ganoderma lucidum triterpenoids (GP) have been reported to help prevent and improve hyperlipidemia. Modulation of the gut microbiota was proposed as underlying factor as well as a novel measure to prevent and treat hyperlipidemia. The effects of GP on high-fat diet (HFD)-induced hyperlipidemia and gut microbiota modulation were determined in rats. Ultra-performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry (UPLC-QTOF MS-MS) indicated that GP were enriched with ganoderic acids G, B, H, A, and F. After feeding with GP supplementation, serum lipid levels including total triglyceride, total cholesterol, and low-density-lipoprotein cholesterol were significantly decreased in hyperlipidemic rats. Furthermore, administration of GP also has reversed the HFD-induced gut microbiota dysbiosis, including a significant increase in Alloprevotella and reduced proportion of Blautia. The result above suggests that GP would be developed as a functional food to ameliorate lipid metabolic disorders and hyperlipidemia.

Anti-Aging Effect of Agrocybe aegerita Polysaccharide through Regulation of Oxidative Stress and Gut Microbiota.

Polysaccharides extracted from Agrocybe aegerita (AAPS) have various physiological effects. In this study, we used the naturally aging Drosophila melanogaster and D-galactose-induced aging mice as animal models to study the anti-aging effects of AAPS via the alleviation of oxidative stress and regulation of gut microbiota. Results showed that AAPS could significantly prolong lifespan and alleviate oxidative stress induced by H2O2 of Drosophila melanogaster. In addition, AAPS significantly increased the activities of antioxidant enzymes in Drosophila melanogaster and mice, and reduced the content of MDA. Furthermore, AAPS reshaped the disordered intestinal flora, increased the abundance ratio of Firmicutes to Bacteroidetes, and increased the abundance of beneficial bacteria Lactobacillus. Our results demonstrated that AAPS had good antioxidant and potential anti-aging effects in vivo.

Comparative transcriptomics discloses the regulatory impact of carbon/nitrogen fermentation on the biosynthesis of Monascus kaoliang pigments.

Carbon and nitrogen play a fundamental role in the production of Monascus pigments. However, their effects on pigment biosynthesis remain undetermined. In this study, we found that Monascus kaoliang produces pigments via liquid fermentation using glycerol and peptone as suitable carbon and nitrogen sources, respectively. Comparative transcriptomic profiling was performed using RNA sequencing. It indicated that the differentially expressed genes (DEGs) of carbon were enriched using amino acids and carbohydrates via the transport and metabolism pathways, respectively. DEGs of nitrogen were enriched only using general functional prediction pathways. These data provide a comprehensive interpretation of the linkage between primary and secondary metabolisms in M. kaoliang. Moreover, they provide insights into the effects of various substances involved in secondary metabolism.

Treponema primitia α1-2-fucosyltransferase-catalyzed one-pot multienzyme synthesis of fucosylated oligosaccharide lacto-N-fucopentaose I with antiviral activity against enterovirus 71.

Fucosylated oligosaccharides have important biological functions as well as an excellent antiviral activity. A novel α 1-2-fucosyltransferase (α 2FT) from Treponema primitia (Tp2FT) was cloned and expressed in Escherichia coli BL21(DE3) and purified as an N-His6-tagged fusion protein (His6-Tp2FT). Mass spectrometry was carried out to identify the products of enzymatic reaction. The Tp2FT exhibited strict acceptor substrate specificity for type 1 structure (Galß1-3GlcNAc)-containing glycans. It might be a promising emzyme for the chemo-enzymatic synthesis of lacto-N-fucopentaose I (LNFP I), which is one of the important fucosylated oligosaccharides. In this study, different in vitro experiments were used to study the biological activities of LNFP I. It could reduce the concentrations of inflammatory cytokines and effectively inhibit the synthesis of enterovirus 71 proliferation. LNFP I was an inhibitor of enterovirus 71 in the early stages of infection, it can used in infant nutrition and might provide a new drug for hand foot mouth disease.

In Situ Visualizing Nascent RNA by Exploring DNA-Templated Oxidative Amination of 4-Thiouridine.

Tracking and mapping the nascent RNA molecules in cells is essential for deciphering embryonic development and neuronal differentiation. Here, we utilized 4-thiouridine (s4U) as a metabolic tag to label nascent RNA and developed a fluorescence imaging method based on the DNA-templated oxidative amination (DTOA) reaction of s4U. The DTOA reaction occurred between amine-modified DNA and s4U-containing RNA with high sequence specificity and chemical selectivity. Target nascent mRNAs in HeLa cells, including those encoding green fluorescent proteins (GFPs) and endogenous BAG-1, were thus lit up selectively by DTOA-based fluorescence in situ hybridization (DTOA FISH). We believe the DTOA conjugation chemistry shown in this study could be generally applied to investigate the spatial distribution of nascent transcription dynamics in cellular processes.

A ratiometric solid AIE sensor for detection of acetone vapor.

Acetone serves as a routine solvent and synthetic intermediate in chemical factories and laboratories. Monitoring the level of acetone vapor in working environment is of great necessity to employee health due to its strong volatility and toxicity, but there is still in lack of simple and easy-to-use portable sensors. In this study, we report a portable and intuitive indicator for real-time displaying acetone vapor concentration in air, based on the ratiometric fluorescence response of the designed organic molecule, PhB-SSB, to acetone. As an aggregation-induced emission (AIE) fluorophore, PhB-SSB underwent specific reaction with acetone through the salicylaldehyde Schiff base and phenylboronate groups to realize ratiometric fluorescence change from green to red after acetone vapor treatment. The reaction mechanism was proposed as acetone-induced breakage of the imine bond in PhB-SSB. We further fabricated PhB-SSB into a film fluorescent sensor for acetone vapor with good sensitivity and selectivity. Taking advantage of its intuitive fluorescent color contrast, acetone-specific response and small size, our sensor is practical in real-time alarming the acetone vapor hazard in the workplaces.

Post-Synthetic Modification of Oligonucleotides Through Oxidative Amination of 4-Thio-2'-Deoxyuridine.

Functionalized oligonucleotides (ONs) are widely applied as target recognition molecules for biosensing and gene regulation. Herein, we describe a general method for post-synthetic modification of ONs based on the oxidative amination of 4-thio-2'-deoxyuridine (4SdU) with sodium periodate and several amines. Alkyne-/azide-, biotin-, and fluorophore-modified ONs were prepared by modifying 4SdU-containing ONs with the corresponding amines and characterized for their bioorthogonal reactivity, streptavidin-binding affinity, and fluorescence properties, respectively. We synthesized three fluorophore-modified ONs with and without the aromatic fluorophores conjugated to modified nucleobases and investigated their emission properties. © 2021 Wiley Periodicals LLC. Basic Protocol 1: Post-synthetic modification of ONs Supporting Protocol: Preparation of dansyl ethylenediamine Basic Protocol 2: Quantification of ON reaction yields Basic Protocol 3: Characterization of modified ONs.

Long-Term Dynamic Imaging of Cellular Processes Using an AIE Lipid Order Probe in the Dual-Color Mode.

Lipid order in the cytoplasm membrane of eukaryotic cells undergoes dynamic changes in almost all cellular processes. Dynamically monitoring these changes is of essential biological significance and remains challenging. This work provides the first aggregation-induced emission probe, TPNPDA-C15, with highly three-dimensional specificity to cell membranes for fluorescent imaging of lipid order of live cells. TPNPDA-C15 displays red fluorescence enhancement with the viscosity increase while emits yellow fluorescence when aggregates form. Imaging analyses of giant unilamellar vesicles and live cells under osmotic shock by the probe demonstrate its sensitive response to the degree of phospholipids packing on artificial and cell membranes. Taking advantage of its superior low photocytotoxicity and high photostability, TPNPDA-C15 is further applied for long-term dynamic imaging of entire live cell physiological processes including apoptosis, ferroptosis, and mitosis in the dual-color mode. With the analysis of fluorescence signal changes in the two fluorescence channels, TPNPDA-C15 serves as a robust fluorescent probe for the imaging study of cellular dynamics.

General Method for Post-Synthetic Modification of Oligonucleotides Based on Oxidative Amination of 4-Thio-2'-deoxyuridine.

Functionalized oligonucleotides (ONs) are widely applied as target binding molecules for biosensing and regulators for gene expression. Numerous efforts have been focused on developing facile methods for preparing these useful ONs carrying diverse modifications. Herein, we present a general method for postsynthetic modification of ONs via oxidative amination of 4-thio-2'-deoxyuridine (4SdU). 4SdU-containing ON can be derived by both alkyl and aromatic amines. Using this approach, ONs are successfully attached with alkyne/azide, biotin and dansylamide moieties, and these as-prepared ONs possess the expected biorthogonal reactivity, streptavidin affinity and fluorescent property, respectively. Furthermore, we also directly install fluorophores to the ON nucleobase based on oxidative amination of 4SdU, and these fluorophores exhibit distinct luminescence behaviors before and after conjugation. We believe our method will be a versatile strategy for constructing various functionalized ONs used in a wide range of nucleic acid applications.

Organic Nanoparticles with Persistent Luminescence for In Vivo Afterglow Imaging-Guided Photodynamic Therapy.

Optical imaging-guided photodynamic therapy (PDT), with precise localization and non-invasive treatment of tumors, is an emerging technique with great potential for cancer therapy. However, impaired by tissue auto-fluorescence that causes low signal-to-background ratio (SBR), most fluorescence imaging systems show poor sensitivity to tumors in vivo. In this study, we synthesized organic nanoparticles (ONPs) with persistent luminescence and good biocompatibility for afterglow imaging-guided PDT. The ONPs displayed near-infrared light emission with half-life time at minute level, which offered high SBR and good tissue penetration for in vivo afterglow tumor imaging. Taking advantage of their abundant singlet oxygen generation by NIR laser irradiation guided to the tumor sites, the ONPs also enabled imaging-guided PDT for efficient suppression of tumor growth in mice with minimal damage to major organs.

Simultaneously and Selectively Imaging a Cytoplasm Membrane and Mitochondria Using a Dual-Colored Aggregation-Induced Emission Probe.

Analysis of subcellular organelles (e.g., a cytoplasm membrane and mitochondria) during cellular processes can provide particularly useful information for our understanding of cell chemistry and biology. For this purpose, fluorescent probes capable of dynamically imaging multiple organelles in a simultaneous and selective manner are highly demanded, yet such probes are scarcely reported due to the challenges in molecular design. In this study, we developed a dual-colored aggregation-induced emission (AIE) probe TPNPDA-C12 with twisted intramolecular charge transfer (TICT) to visualize the membrane and mitochondria of the same cells through distinct fluorescence channels simultaneously. We also successfully used the probe to monitor and distinguish the dynamic changes of the organelles during cell apoptosis and necrosis induced by reactive oxygen species (ROS) and cytotoxins.

Ganoderma polysaccharide and chitosan synergistically ameliorate lipid metabolic disorders and modulate gut microbiota composition in high fat diet-fed golden hamsters.

High-fat diet (HFD) and sucrose intake can lead to hyperlipidemia, hypercholesterolemia, and nonalcoholic fatty liver disease (NAFLD) as well as disturbed gastrointestinal microbiota and dysfunctional intestinal barrier. In the present study, we showed that Ganoderma lucidum polysaccharide and chitosan (PC) significantly mitigated the hyperlipidemia in HFD-fed hamsters via lowering the contents of serum total triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), and aspartate aminotransferase (AST). Furthermore, PC changed the composition of gastrointestinal microbiota and elevated the relative abundances of beneficial bacteria, such as Prevotella, Oscillibacter, and SCFA-producers. Interestingly, we also found that the abundances of Prevotella, Alloprevotella, Bifidobacterium, and Alistipes were negatively associated with serum lipid profiles. Collectively, the above-mentioned findings indicated that PC could improve lipid metabolic disorders, at least in part, by modulating gastrointestinal microbiota, suggesting that PC could be used as a potential lipid-lowering ingredient in functional foods. PRACTICAL APPLICATIONS: PC could ameliorate lipid metabolism disorder, at least in part, by regulating specific gut microbiota, suggesting its potential as a novel lipid-lowering ingredient in functional foods. We believed that our findings could be of interest to the readers because they help others further understand the gut microbiota alterations that occurred after PC supplementation in the context of metabolic syndrome (MetS).

Selective and sensitive fluorescence "turn-on" detection of 4-thiouridine in nucleic acids via oxidative amination.

A fluorescence "turn-on" method for digestion-free analysis of 4-thiouridine (s4U) in nucleic acids was developed in this work based on the oxidative amination of s4U by fluoresceinamine (FAM-NH2) and periodate (IO4-). It was 125-fold more sensitive for s4U detection than the traditional UV330 absorption method, and showed excellent selectivity to s4U over 2-thiouridine (s2U) analogues and biological thiols.

Polyunsaturated fatty acids from microalgae Spirulina platensis modulates lipid metabolism disorders and gut microbiota in high-fat diet rats.

Effects of Spirulina platensis 55% ethanol extract (SPL55) on lipid metabolism in high-fat diet-induced hyperlipidaemic rats were investigated. Ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry indicated that SPL55 was enriched with polyunsaturated fatty acids. Meanwhile, serum and liver lipid levels, including total triglyceride, total cholesterol, and low-density-lipoprotein cholesterol, were significantly decreased in hyperlipidaemic rats of SPL55. Analysis of tissue sections showed that SPL55 treatment could markedly inhibit hepatic lipid accumulation and steatosis. Moreover, SPL55 regulated the mRNA and protein expression levels of SREBP-1c, HMG-CoA, PEPCK, ACC, and AMPK genes involved in lipid metabolism. Furthermore, SPL55 led to decrease the abundances of Turicibacter, Clostridium_XlVa, and Romboutsia, which were positive correlation with lipid metabolism indicators, and has also enriched Alloprevotella, Prevotella, Porphyromonadaceae, and Barnesiella. These results provided evidence that SPL55 might be developed as a functional food to ameliorate lipid metabolic disorders and hyperlipidaemia.