N4-Allylcytidine: a new nucleoside analogue for RNA labelling and chemical sequencing.

RNA labelling has become indispensable in studying RNA biology. Nucleoside analogues with a chemical sequencing power represent desirable RNA labelling molecules because precise labelling information at base resolution can be obtained. Here, we report a new nucleoside analogue, N4-allylcytidine (a4C), which is able to tag RNA through both in vitro and in vivo pathways and further specifically reacts with iodine to form 3, N4-cyclized cytidine (cyc-C) in a catalyst-free, fast and complete manner. Full spectroscopic characterization concluded that cyc-C consisted of paired diastereoisomers with opposite chiral carbon centers in the fused 3, N4-five-membered ring. During RNA reverse transcription into complementary DNA, cyc-C induces base misincorporation due to the disruption of canonical hydrogen bonding by the cyclized structure and thus can be accurately identified by sequencing at single base resolution. With the chemical sequencing rationale of a4C, successful applications have been performed including pinpointing N4-methylcytidine methyltransferases' substrate modification sites, metabolically labelling mammalian cellular RNAs, and mapping active cellular RNA polymerase locations with the chromatin run-on RNA sequencing technique. Collectively, our work demonstrates that a4C is a promising molecule for RNA labelling and chemical sequencing and expands the toolkit for studying sophisticated RNA biology.

Comparative Site-Specific O-Glycosylation Analysis of the Milk Fat Globule Membrane Proteome in Donkey Colostrum and Mature Milk.

Donkey milk fat globule membrane (MFGM) proteins are a class of membrane-bound secreted proteins with broad-spectrum biofunctional activities; however, their site-specific O-glycosylation landscapes have not been systematically mapped. In this study, an in-depth MFGM O-glycoproteome profile of donkey milk during lactation was constructed based on an intact glycopeptide-centered, label-free glycoproteomics pipeline, with 2137 site-specific O-glycans from 1121 MFGM glycoproteins and 619 site-specific O-glycans from 217 MFGM glycoproteins identified in donkey colostrum and donkey mature milk, respectively. As lactation progressed, the number of site-specific O-glycans from three glycoproteins significantly increased, whereas that of 11 site-specific O-glycans from five glycoproteins significantly decreased. Furthermore, donkey MFGM O-glycoproteins with core-1 and core-2 core structures and Lewis and sialylated branch structures may be involved in regulating apoptosis. The findings of this study reveal the differences in the composition of donkey MFGM O-glycoproteins and their site-specific O-glycosylation modification dynamic change rules during lactation, providing a molecular basis for understanding the complexity and biological functions of donkey MFGM protein O-glycosylation.

Ultrasound Treatment Improves Fruit Quality of Postharvest Blood Oranges (Citrus sinensis L. Osbeck): Anthocyanin Enrichment and Its Biosynthesis.

This study was to investigate the effects of different nonthermal treatments on quality attributes, anthocyanin profiles, and gene expressions related to anthocyanin biosynthesis during low-temperature storage, including pulsed light (PL), magnetic energy (ME), and ultrasound (US). Among these treatments, 1 min US treatment was the most effective method for improving fruit quality and increasing total anthocyanin contents (by 29.89 ± 3.32%) as well as individual anthocyanins during low-temperature storage of 28 days. This treatment resulted in high color intensity, intact cellular architectures, and positive sensory evaluation. In contrast, PL and ME treatments displayed negative effects on quality improvement, leading to the destruction of cell architectures and inhibiting anthocyanin levels. Furthermore, qPCR analysis revealed that the structural genes (C4H, CHS1, CHS2, CHI, F3H, ANS, and GST) related to anthocyanin biosynthesis and transport were the target genes and upregulated in response to the cavitation effect of US treatment.

Inhibitory mechanisms of promising antimicrobials from plant byproducts: A review.

Plant byproducts and waste present enormous environmental challenges and an opportunity for valorization and industrial application. Due to consumer demands for natural compounds, the evident paucity of novel antimicrobial agents against foodborne pathogens, and the urgent need to improve the arsenal against infectious diseases and antimicrobial resistance (AMR), plant byproduct compounds have attracted significant research interest. Emerging research highlighted their promising antimicrobial activity, yet the inhibitory mechanisms remain largely unexplored. Therefore, this review summarizes the overall research on the antimicrobial activity and inhibitory mechanisms of plant byproduct compounds. A total of 315 natural antimicrobials from plant byproducts, totaling 1338 minimum inhibitory concentrations (MIC) (in µg/mL) against a broad spectrum of bacteria, were identified, and a particular emphasis was given to compounds with high or good antimicrobial activity (typically <100 µg/mL MIC). Moreover, the antimicrobial mechanisms, particularly against bacterial pathogens, were discussed in-depth, summarizing the latest research on using natural compounds to combat pathogenic microorganisms and AMR. Furthermore, safety concerns, relevant legislation, consumer perspective, and current gaps in the valorization of plant byproducts-derived compounds were comprehensively discussed. This comprehensive review covering up-to-date information on antimicrobial activity and mechanisms represents a powerful tool for screening and selecting the most promising plant byproduct compounds and sources for developing novel antimicrobial agents.

Fabrication of multifunctional materials based on chitosan/gelatin incorporating curcumin-clove oil emulsion for meat freshness monitoring and shelf-life extension.

A new multifunctional film with active and intelligent effects was developed by incorporating curcumin-clove oil emulsion into natural materials. The basic properties, functional characteristics, and pH/NH3-sensitivity of films were investigated, and then these films were applied to extend shelf-life and monitor freshness of meat. Curcumin solution and emulsion illustrated significant color variations at different pH values. The incorporation of emulsion improved the UV-vis barrier and water resistance properties of films, which blocked most of UV-light and its water contact angle reached 100.03°. Meanwhile, the films had stronger mechanical strength and higher thermal stability, with elongation at break reaching 79.18 % and the maximum degradation temperature rising to 316 °C. Moreover, emulsion made films have a slow-release effect on clove oil, which not only enhanced the antioxidant property but also significantly improved their antibacterial activity. Additionally, the multifunctional films presented a significant color response to acidic/alkaline environments over a short time interval and could be easily identified by naked eyes. Finally, the films effectively extended the shelf-life of fresh meat by 3 days at 4 °C and visually monitored freshness through color changes in real-time. This knowledge provides insights and ideas for the development of novel food packaging with both active and intelligent functions.

Conjugation of soy protein isolate (SPI) with pectin: effects of structural modification of the grafting polysaccharide.

Recently, there has been a great interest in enhancing the emulsifying properties of soy protein isolate (SPI) by Maillard reaction. As a commonly-used grafting polysaccharide, pectin has proved useful in modifying proteins. However, effects of its structural characteristics on conjugation are still not fully understood. To address this problem, we employed alkaline or/and enzymatic treatments to modify pectin and obtained three modified samples. Structural characteristics of pectin, including the molecular weight, degree of methoxylation and acetylation, and monosaccharide compositions were measured. When conjugated with SPI, pectin with lower molecular weight and less main chains induced higher conjugate yield. Fluorescence intensity and surface hydrophobicity of all conjugates markedly reduced compared to the original SPI, suggesting a more loosened protein structure after Maillard reaction. In this study, the enzymolysis pectin proved an optimum grafting polysaccharide considering the simple preparation procedures and the highest emulsifying properties of its resulting conjugates.

Whey protein isolate-gum Acacia Maillard conjugates as emulsifiers for nutraceutical emulsions: Impact of glycation methods on physicochemical stability and in vitro bioaccessibility of ß-carotene emulsions.

The physicochemical stability and in vitro bioaccessibility of ß-carotene emulsions stabilized by the whey protein isolate (WPI)-gum Acacia (GA) conjugates prepared by dry-heating (Con WPI (D)) and ultrasound (Con WPI (U)) Maillard reaction were investigated in this study. The conjugate-stabilized emulsions have a larger mean size than the WPI-stabilized emulsion. The stability of emulsions under freeze-thaw treatment, high ionic strength conditions, thermal treatment and pH conditions near the isoelectric point of WPI were improved by glycation. The chemical stability of ß-carotene in emulsions was also enhanced by WPI-GA conjugates. The WPI-GA conjugate-stabilized emulsions have better stability for droplet aggregation during in vitro simulated gastrointestinal digestion, which increased ß-carotene bioaccessibility. However, the Con WPI (U)-stabilized emulsion shows poor thermal stability and lower ß-carotene bioaccessibility than the Con WPI (D)-stabilized emulsion, which may be attributed to ultrasound-induced structural changes during the process of glycation.

Challenges of pectic polysaccharides as a prebiotic from the perspective of fermentation characteristics and anti-colitis activity.

Several studies are described that contribute to the systematic exploration of new aspects of digestion, fermentation, and biological activities of pectic polysaccharides (PPS) leading to a better understanding of prebiotics. Inflammatory bowel disease (IBD) is thought to be associated with the dysbacteriosis induced by different environmental agents in genetically susceptible persons. PPS are considered as an indispensable gut-microbiota-accessible carbohydrate that play a dominant role in maintaining gut microbiota balance and show a better effect in ameliorating IBD than some traditional prebiotics. The aim of this review is to summarize the fermentation characteristics of PPS, highlight its role in improving IBD, and propose a view that PPS may be a new and effective prebiotic.

Dietary pectic substances enhance gut health by its polycomponent: A review.

Pectic substances, one of the cell wall polysaccharides, exist widespread in vegetables and fruits. A surge of recent research has revealed that pectic substances can inhibit gut inflammation and relieve inflammatory bowel disease symptoms. However, physiological functions of pectins are strongly structure dependent. Pectic substances are essentially heteropolysaccharides composed of homogalacturonan and rhamnogalacturonan backbones substituted by various neutral sugar sidechains. Subtle changes in the architecture of pectic substances may remarkably influence the nutritional function of gut microbiota and the host homeostasis of immune system. In this context, developing a structure-function understanding of how pectic substances have an impact on an inflammatory bowel is of primary importance for diet therapy and new drugs. Therefore, the present review has summarized the polycomponent nature of pectic substances, the activities of different pectic polymers, the effects of molecular characteristics and the underlying mechanisms of pectic substances. The immunomodulated property of pectic substances depends on not only the chemical composition but also the physical structure characteristics, such as molecular weight (Mw ) and chain conformation. The potential mechanisms by which pectic substances exert their protective effects are mainly reversing the disordered gut microbiota, regulating immune cells, enhancing barrier function, and inhibiting pathogen adhesion. The manipulation of pectic substances on gut health is sophisticated, and the link between structural specificity of pectins and selective regulation needs further exploration.

Citrus pectin modified by microfluidization and ultrasonication: Improved emulsifying and encapsulation properties.

In this study, modified citrus pectin treated with a combination of microfluidization and ultrasonication was compared to the original and ultrasonication treated pectin on hydrodynamic diameter, molecular weight, polydispersity, zeta potential, apparent viscosity, Fourier-transform infrared spectroscopy (FTIR), 2,2-diphenyl-1-picryl hydrazyl (DPPH) radical scavenging capacity, scanning electron microscope (SEM), atomic force microscopy (AFM), their emulsifying properties and encapsulation properties. Modified pectin treated with a combination of microfluidization and moderate ultrasonication (MUB) was found to have lowest hydrodynamic diameter (418 nm), molecular weight (237.69 kDa) and polydispersity (0.12), and relatively low apparent viscosity among all pectin samples. Furthermore, it showed significantly higher DPPH radical scavenging capacity than the original pectin although only slightly higher than that of ultrasonication treated one (UB). MUB showed a thin fibrous morphology and decreased degree of branching from SEM and AFM. Emulsion stabilized by MUB had highest centrifugal and thermal stability compared to emulsions stabilized by UB and the original pectin. This could be attributed to higher interfacial loading of MUB (17.90 mg/m2) forming more compact interfacial layer observed by confocal laser scanning microscopy (CLSM). Moreover, both MUB and UB exhibited improved encapsulation functionality to protect cholecalciferol (vitamin D3) from UV degradation compared to the original pectin.

Inhibition effect of three common proanthocyanidins from grape seeds, peanut skins and pine barks on maize starch retrogradation.

The inhibition effect of three common proanthocyanidins (PA) on the retrogradation properties of maize starch was investigated (including grape seed proanthocyanidins (GSPA), peanut skin proanthocyanidins (PSPA), and pine bark proanthocyanidins (PBPA)). Differential scanning calorimetry (DSC) and X-ray diffraction (XRD) analysis showed that PA could significantly decrease the values of melting enthalpy of retrogradation (ΔHr) and the degree of relative crystallinity, suggesting that the starch re-crystallization was retarded by PA. Scanning electron microscope (SEM) characterizations illustrated that retrograded PA-starch samples formed a looser matrix with less appearance of continuous flakes during storage. Overall, 0.5 %-2.0 % of three PAs exhibited suppression of starch retrogradation after 21-day cooling storage, mainly resulting from the PA-starch interaction. Among them, PSPA showed the most substantial inhibition effect on starch retrogradation, which might be attributed to its structural features. This study suggested that PA could be a new type of inhibitor to suppress starch retrogradation.

Effect of Potato Pulp Pectic Polysaccharide on the Stability of Acidified Milk Drinks.

In order to broaden the application of potato pulp pectic polysaccharide (PPP) in stabilizing acidified milk drinks (AMDs) and investigate the stabilizing effect and physical properties of AMDs prepared with PPP, a comparative study was made among PPP, commercial high methoxyl pectin (HMP) and low methoxyl pectin (LMP). The zeta potential, rheology, particle size and serum separation of AMDs were evaluated after preparing with PPP, HMP and LMP, respectively. Results indicated that PPP led to lower serum separation than LMP (14.65% for AMDs prepared with 0.5% PPP compared to 25.05% for AMDs prepared with 0.5% LMP), but still higher than HMP (9.09% for AMDs prepared with 0.5% HMP). However, narrower particle size distribution and lower viscosity of AMDs was achieved by PPP than by LMP and HMP. PPP can electrostatically adsorb on the surface of casein and its abundant neutral sugar side chains would provide steric hindrance to prevent casein flocculation in AMDs. Our results might provide some new ideas for the application of PPP in improving the stability of AMDs.

Effect of anion type on enzymatic hydrolysis of starch-(thermostable α-amylase)-calcium system in a low-moisture solid microenvironment of bioextrusion.

The effect of six anions (Cl-, OH-, NO3-, SO42-, C6H10O62- and PO43-) on a starch (St)-enzyme (thermostable α-amylase, TαA)-calcium (Ca) system was investigated in a low-moisture solid state. Two levels of Ca salts (1 and 10 mmol/100 g St) added to potato starch with and without TαA were analyzed by FT-IR, DSC and SEM. The surface morphologies of the St-Ca complexes were different in the presence of various anions, and the residual Ca salts around the St granules might decrease the enzymatic action. For bioextrusion, TαA (0.5‰ and 1.5‰) were introduced for a relatively low Ca content (1 mmol/100 g). Significant differences in enzyme activity were observed, increasing the activity of TαA by SO42- (146.54 %) > C6H10O62- > Cl- > control > NO3- > OH- ≈ PO43- and C6H10O62- (123.20 %) ≈ Cl- ≈ SO42- > control > PO43 > OH- > NO3- for the low and high enzyme levels, respectively.

Physicochemical and Digestion Properties of Potato Starch Were Modified by Complexing with Grape Seed Proanthocyanidins.

Dietary intake of potato starch could induce a dramatic increase in blood glucose and is positively associated with chronic metabolic diseases (type II diabetes, cardiovascular disease, etc.). Grape seed proanthocyanidins (GSP) are known to decrease starch digestion by inhibiting digestive enzymes or changing the physicochemical properties of starch. In the present study, GSP were complexed with potato starch to prepare polyphenol-starch complexes. The physiochemical properties and digestibility of complexes were investigated by in vitro digestion model, X-ray diffraction, differential scanning calorimetry, rapid visco analyzer, Fourier transform infrared spectroscopy as well as texture profile analysis. Results indicated that the peak viscosity, breakdown, trough, and setback of the complexes disappeared, replaced by a special continuous increase in paste viscosity. The complexes showed a lower final viscosity and higher thermal stability with the increasing binding amount of GSP. GSP decreased the hardness of the complexes' gel significantly. FT-IR indicated that GSP might interact with potato starch through noncovalent forces. Additionally, the complexes also showed a higher content of slowly digestible starch and resistant starch than that of the native starch. Thus, we inferred that the addition of GSP could modify the digestibility of potato starch and be an optional way to modify the starch with lower digestion.

Ultrasound-assisted adsorption/desorption for the enrichment and purification of flavonoids from baobab (Adansonia digitata) fruit pulp.

This work described the purification and enrichment of flavonoids from baobab (Adansonia digitata) fruit pulp (BFP) by ultrasound-assisted adsorption/desorption procedure using macroporous resins. Four resins were tested and HPD-500 polar resin exhibited the best adsorption/desorption properties. Based on preliminary experiments and literature reports, the effects of various ultrasonic conditions including high power short time (HPST, 540 W for 5 min), medium power medium time (MPMT, 270 W for 15 min) and low power long time (LPLT, 45 W for 30 min) as well as different temperatures (T = 25-45 °C) on the adsorption of Total Flavonoids Content (TFC) were investigated in comparison with orbital shaking/no sonication (NS). Also, the effect of ultrasound on the desorption capacity and recovery of TFC was determined at different concentrations of ethanol (30-100%). Remarkably, ultrasonic treatment significantly increased the adsorption/desorption capacity and recovery and shortened the equilibrium time. The pseudo-second-order kinetic and Freundlich isotherm models better delineated the adsorption process under ultrasound. Moreover, the adsorption process was both spontaneous and thermodynamically favourable with physical adsorption and multilinear intraparticle diffusion being the predominant mechanisms of the whole process. HPST treatment at 25 °C with 80% ethanol as the desorption solvent most noticeably enhanced the adsorption/desorption of flavonoids and contributed to the highest recovery of TFC, Total Phenolic Content (TPC), and antioxidant capacity in addition to a 5-8-fold reduction in total sugar and acid contents when compared with NS treatment. Moreover, HPLC analysis revealed that the content of nine out of thirteen phenolic compounds from the HPST treatment was the highest, and the individual flavonoids content increased by 2-3-fold compared with the other treatments. Our analyses suggested that ultrasound can be employed as a practical approach to intensify the adsorption and desorption of functional compounds in BFP.

Manothermosonication (MTS) treatment by a continuous-flow system: Effects on the degradation kinetics and microstructural characteristics of citrus pectin.

Modified pectin (MP) was reported to have increased bioactivities compared with the original one. However, traditional modification methods such as using an acidic solvent with heating are not only costly but causing severe pollution as well. In this study, manothermosonication (MTS) with a continuous-flow system was utilized to modify citrus pectin. The citrus pectin (5 g/L) treated by MTS (3.23 W/mL, 400 kPa, 45 °C) exhibited lower molecular weight (Mw, 248.17 kDa) and PDI (2.76). The pectin treated by MTS (400 KPa, 45 °C, 5 min) exhibited a narrower Mw distribution and lowered more Mw (48.8%) than the ultrasound(US)-treated (23.8%). Pectin degradation data fitted well to kinetic model of 1/Mwt -1/Mw0 = kt (45-65 °C). A lower activation energy of 13.33 kJ/mol was observed in the MTS treatment compared with the US-treated (16.38 kJ/mol). The MTS-treated pectin lowered the degree of methoxylation (DM), mol% of rhamnose and galacturonic acid (GalA) while increased mol% of galactose (Gal), xylose (Xyl), and arabinose (Ara). The 1H and 13C nuclear magnetic resonance showed that MTS could not alter the primary structures of citrus pectin. However, an elevated (Gal + Ara)/Rha and reduced GalA/(Rha + Ara + Gal + Xyl) molar ratios after MTS suggested that MTS resulted in more significant degradation on the main chains and less on the side chains of pectin, in agreement with the result of atomic force microscope. Moreover, the MTS-treated pectin exhibited a higher 1,1-diphenyl-2picryl hydrazyl radical scavenging capacity compared with original pectin.

Ultrasonic modification of pectin for enhanced 2-furfurylthiol encapsulation: process optimization and mechanisms.

BACKGROUND: Pectin is an intriguing polymer, which is usually regarded as a byproduct from agricultural and biological processes. In previous studies, ultrasound treatment has been explored to improve the functionality of pectin but most of that work focused on aspects of molecular structure and the chemical properties of pectin. In this study, we utilized ultrasound treatment to modify the physiochemical properties of pectin. Using ultrasound treatment, we evaluated the emulsifying capability of pectin as a function of ultrasonic time and power density, using a response surface approach. A very potent yet unstable coffee-like aroma compound, 2-furfurylthiol, was also used for comparing the encapsulation feasibility of emulsion made with original pectin and ultrasound-treated pectin. RESULTS: Our results showed that the particle size of pectin was highly correlated with power density and ultrasound time. Approximately 370 nm of pectin particle size could be reached at a power density of 1.06 W mL-1 for 40 min. Ultrasound treatment increased emulsion droplet size but significantly improved emulsifying capacities, such as centrifugal stability and surface loading, although it was highly dependent upon the ultrasound treatment condition. When used as the encapsulation wall material, the ultrasound-modified pectin had significantly enhanced performance compared with the original, in terms of flavor retention over time at 45 °C and 65 °C. CONCLUSION: Ultrasound treatment was able to modify the physiochemical properties of pectin, which thus improved emulsification stability and encapsulation feasibility by forming a thicker layer at the oil / water interface to protect the core materials. © 2019 Society of Chemical Industry.

Comparison of citrus pectin and apple pectin in conjugation with soy protein isolate (SPI) under controlled dry-heating conditions.

In this study, we selected two most commonly-available commercial pectin, i.e. citrus pectin and apple pectin as the grafting polysaccharides to prepare soy protein isolate-pectin conjugates. Despite the similar degrees of methoxylation and acetylation for two pectin samples, apple pectin showed much more complex structures compared to citrus pectin, with a 2.20-fold higher molecular weight and large numbers of side chains. The conjugates were prepared under controlled dry-heating conditions and achieved the degree of graft of 25.00% and 21.85% for citrus and apple pectin, respectively. Formation of the conjugates was further confirmed by SDS-PAGE gel electrophoresis and IR spectra. Attributed to the strong steric-hindrance effect of pectin, the fluorescence intensity and surface hydrophobicity of the soy protein isolate were significantly decreased after Maillard reaction. However, both solubility and emulsifying properties of the conjugates were significantly improved. Results indicated that both pectin samples played favorable roles in protein modification.

Characterizing the phenolic constituents of baobab (Adansonia digitata) fruit shell by LC-MS/QTOF and their in vitro biological activities.

Baobab (Adansonia digitata) fruit is a part of the baobab tree, a revered multi-purpose tree native to Africa with a myriad of potentials in providing shelter, food, medicine, clothing and as a valuable source of natural antioxidants. A massive quantity of baobab fruit shells (BFS) is generated as a waste from the baobab fruit processing representing significant economic and environmental challenges at the same time an opportunity for its valorization and commercial utilization. The present study explored the potentials of BFS as a source of phenolic compounds. For this purpose, the phenolic constituents of BFS were identified by LC-MS/QTOF analysis. Also, the Total Phenolic Chromatographic Index (TPCI), TPC, TFC, and antioxidant capacity were compared with the baobab seeds and pulp. The antidiabetic potential through α-amylase and α-glucosidase inhibitory activities was also compared with that of acarbose. The LC-MS/QTOF analysis led to the identification of 45 compounds, including quercetin, kaempferol, proanthocyanidins, phenolic acids and their derivatives, several of which had never been reported in baobab fruit. Moreover, the BFS showed higher TPC, TFC, and antioxidant capacity than the baobab seeds and pulp and inhibited α-amylase and α-glucosidase enzymes activities with much higher potency than acarbose. This research demonstrated the promising potentials of BFS as a good source of phenolic compounds that can further be utilized for food and pharmaceutical applications.

Novel antibacterial modalities against methicillin resistant Staphylococcus aureus derived from plants.

Methicillin-resistant Staphylococcus aureus (MRSA) is a notorious bacterial pathogen that induces high mortality and morbidity. Due to the emergence of multiple resistance, antibiotic treatments are rapidly becoming ineffective for the related infections. Natural products, especially those derived from plants, have been proven to be effective agents with unique antibacterial properties through different mechanisms. This review interprets the resistance mechanisms of MRSA with the aim to conquer public health threat. Further, recent researches about plant antimicrobials that showed remarkable antibacterial activity against MRSA are recorded, including the crude plant extracts and purified plant-derived bioactive compounds. Novel anti-MRSA modalities of plant antimicrobials such as alteration in efflux pump, inhibition of pyruvate kinase, and disturbance of quorum sensing in MRSA are also summarized which may be promising alternatives to antibacterial drug development in future.