Identification of salty peptides from enzymolysis extract of oyster by peptidomics and virtual screening.

Salty peptide as an important sodium substitute, which could reduce the risk of cardiovascular disease caused by excessive sodium intake. In this study, novel salty peptides were prepared and identified from enzymolysis extract of oysters by peptitomic identification, virtual screening and solid phase synthesis. Additionally, molecular simulation was used to study the taste mechanism of salty peptides. 316 peptides were identified in the enzymatic hydrolysates of oysters. 6 peptides, selected through virtual screening, were synthesized using solid-phase synthesis, and EK, LFE, LEY and DR were confirmed to possess a pleasing salty taste through electronic tongue evaluation. Molecular docking results indicated that these 4 peptides could enter the binding pocket within the transmembrane channel-like 4 (TMC4) cavity, wherein salt bridges, hydrogen bonds and attractive charges were the main binding forces. This study provides a rapid screening method for salty peptides in sea food products but possibly applied for other sources.

Topological polymeric glucosyl nanoaggregates in scaffold enable high-density piscine muscle tissue.

Upon the pressure of conventional land agriculture and marine environment facing the future of human beings, the emerging of alternative proteins represented by cultured meat is expected with a breakthrough of efficient, safe and sustainable production. However, the cell proliferation efficiency and final myofiber density in current animal-derived scaffolds are still limited. Here, we incorporated five plant-derived edible polymeric glucosyl nanoparticles (GNPs) into gelatin/alginate hydrogels to spontaneously form nanoaggregates where nanotopographies were observed inside. The nanoscale topological morphology significantly enhances the adhesion and proliferation efficiencies of piscine satellite cells (PSCs) in the tailored extracellular matrix of as-prepared scaffold. Physically, the presence of GNP-induced nanoaggregate increases the interaction between ITG-A1 (membrane protein of PSCs) and hydrogel microenvironment, which activates the focal adhesion-integrin-cytoskeleton mechanotransduction signaling to promote cell proliferation. With a controlled diameter of hydrogel filament, these inner topological GNP nanoaggregates can also improve the density, alignment and differentiation efficiency of PSCs. When cultured in vitro for 15 days, the cell density, size and orientation of muscle fibers in the GNP-stimulated cultured fish fillet are very similar to the total cell mass in native fish muscle tissue.

Generative subgoal oriented multi-agent reinforcement learning through potential field.

Multi-agent reinforcement learning (MARL) effectively improves the learning speed of agents in sparse reward tasks with the guide of subgoals. However, existing works sever the consistency of the learning objectives of the subgoal generation and subgoal reached stages, thereby significantly inhibiting the effectiveness of subgoal learning. To address this problem, we propose a novel Potential field Subgoal-based Multi-Agent reinforcement learning (PSMA) method, which introduces the potential field (PF) to unify the two-stage learning objectives. Specifically, we design a state-to-PF representation model that describes agents' states as potential fields, allowing easy measurement of the interaction effect for both allied and enemy agents. With the PF representation, a subgoal selector is designed to automatically generate multiple subgoals for each agent, drawn from the experience replay buffer that contains both individual and total PF values. Based on the determined subgoals, we define an intrinsic reward function to guide the agent to reach their respective subgoals while maximizing the joint action-value. Experimental results show that our method outperforms the state-of-the-art MARL method on both StarCraft II micro-management (SMAC) and Google Research Football (GRF) tasks with sparse reward settings.

Co-encapsulation of hydrophilic and hydrophobic bioactives stabilized in nanostarch-assisted emulsion for inner core gel of coaxial 3D printing.

3D printing technology, especially coaxial 3D mode of multiple-component shaping, has great potential in the manufacture of personalized nutritional foods. However, integrating and stabilizing functional objectives of different natures remains a challenge for 3D customized foods. Here, we used starch nanoparticle (SNP) to assisted soy protein (SPI) emulsion to load hydrophilic and hydrophobic bioactives (anthocyanin, AC, and curcumin, Cur). The addition of SNP significantly improved the storage stability of the emulsion. Xanthan gum (XG) was also added to the SNP/SPI system to enhance its rheology and form an emulsion gel as inner core of coaxial 3D printing. Low field nuclear magnetic resonance and emulsification analyses showed that AC/Cur@SNP/SPI/XG functional inner core had a strong water binding state and good stability. After printing with outer layer, the SNP/SPI coaxial sample had the lowest deviation rate of 0.8 %. Also, SNP/SPI coaxial sample showed higher AC (90.2 %) and Cur (90.8 %) retention compared to pure starch (S), pure SNP, pure SPI, and S/SPI samples as well as SNP/SPI sample printed without outer layer. In summary, this study provides a new perspective for the manufacture of customized products as multifunctional foods, feeds and even potential delivery of drugs.

Exogenous microbubbles contribute to valorization of microalgal compounds by ultrasound-assisted extraction.

Ultrasound-assisted extraction (UAE) shows great potential in exploiting microalgal compounds. However, upgrading the extraction system lacks concerns. This study proposes a novel sono-reactor featuring a microbubble distributor for increasing bubble abundance and correspondingly improving microalgal compound extraction. Results indicate that protein concentrations increase with ultrasound powers and extraction time while an optimized gas flow rate exists. The optimal parameters by Box-Behnken design are power 646.0 W, nitrogen flow rate 25.0 mL/min, and time 40.0 min, with an optimal protein concentration of 249.1 mg/L - a substantial improvement over gas-free extraction. The strategic increase in bubble abundance enhances microalgal compound extraction efficiency and extraction kinetics. The system innovation will contribute to the advancement of bioresource utilization and sustainability.

Single-cell encapsulation systems for probiotic delivery: Armor probiotics.

Functional foods or drugs based on probiotics have gained unprecedented attention and development due to the increasingly clear relationship between probiotics and human health. Probiotics can regulate intestinal microbiota, dynamically participating in various physiological activities to directly affect human health. Some probiotic-based functional preparations have shown great potential in treating multiple refractory diseases. Currently, the survival and activity of probiotic cells in complex environments in vitro and in vivo have taken priority, and various encapsulation systems based on food-derived materials have been designed and constructed to protect and deliver probiotics. However, traditional encapsulation technology cannot achieve precise protection for a single probiotic, which makes it unable to have a significant effect after release. In this case, single-cell encapsulation systems can be assembled based on biological interfaces to protect and functionalize individual probiotic cells, maximizing their physiological activity. This review discussed the arduous challenges of probiotics in food processing, storage, human digestion, and the commonly used probiotic encapsulation system. Besides, a novel technology of probiotic encapsulation was introduced based on single-cell coating, namely, "armor probiotics". We focused on the classification, structural design, and functional characteristics of armor coatings, and emphasized the essential functional characteristics of armor probiotics in human health regulation, including regulating intestinal health and targeted bioimaging and treatment of diseased tissues. Subsequently, the benefits, limitations, potential challenges, as well as future direction of armor probiotics were put forward. We hope this review may provide new insights and ideas for developing a single-cell probiotics encapsulating system.

Stable cytoactivity of piscine satellite cells in rice bran-gelatin hydrogel scaffold of cultured meat.

In order to achieve high cell adhesion and growth efficiency on scaffolds for cultured meat, animal materials, especially gelatin, are necessary though the disadvantages of weak mechanical properties and poor stability of their hydrogel scaffolds are present during cell cultivation. Here, we use rice bran as a kind of filling and supporting materials to develop a composite scaffold with gelatin for fish cell cultivation, where rice bran is also inexpensive from high yield fibrous agricultural by-product. The rice bran (with a proportion of 1, 3, 5, 7, 10 to 3 of gelatin) could evenly distributed in the three-dimensional network composed of gelatin hydrogel. It contributed to delaying swelling and degradation rates, fixing water and improving elastic modulus. It is important that rice bran-gelatin hydrogel scaffolds (especially the hydrogel with 70 % rice bran, db) promoted piscine satellite cells (PSCs) proliferation effectively compared to the pure gelatin hydrogel, and the former could also support the differentiation of PSCs. Overall, this work showed a positive promotion to explore new source of scaffold materials like agricultural by-product for reducing the cost of cell cultured meat production.

Prognostic signature and immune efficacy of m1A-, m5C-, m6A-, m7G-, and DNA methylation-related regulators in hepatocellular carcinoma.

Background: Hepatocellular carcinoma (HCC) is the main type of primary liver cancer, and its related death ranks third worldwide. The curative methods and progress prediction markers of HCC are not sufficient enough. Nevertheless, little progress has been made in the signature of m1A-, m5C-, m6A-, m7G-, and DNA methylation of HCC. Results: We calibrated a risk gene signature model that can be used to categorize HCC patients based on univariate, multivariate, and LASSO Cox regression analysis. This gene signature classified the patients into high- and low-risk subgroups. Patients in the high-risk group showed significantly reduced overall survival (OS) compared with patients in the low-risk group. The gene set variation analysis (GSVA), immune infiltration, and immunotherapy response were analyzed. The results demonstrated that an immunosuppressive environment was exited and the high-risk group had higher sensitivity to 5-fluorouracil, cisplatin, sorafenib, tamoxifen, and epirubicin. These results indicated personalized therapy should be taken into consideration. Conclusions: Our findings enriched our understanding of the molecular heterogeneity, tumor microenvironment (TME), and drug susceptibility of HCC. m1A-, m5C-, m6A-, m7G-, and DNA methylation-related regulators may be promising biomarkers for future research.

Effect of Thermal Treatment on Gelling and Emulsifying Properties of Soy ß-Conglycinin and Glycinin.

This study investigated the impact of different preheat treatments on the emulsifying and gel textural properties of soy protein with varying 11S/7S ratios. A mixture of 7S and 11S globulins, obtained from defatted soybean meal, was prepared at different ratios. The mixed proteins were subjected to preheating (75 °C, 85 °C, and 95 °C for 5 min) or non-preheating, followed by spray drying or non-spray drying. The solubility of protein mixtures rich in the 7S fraction tended to decrease significantly after heating at 85 °C, while protein mixtures rich in the 11S fraction showed a significant decrease after heating at 95 °C. Surprisingly, the emulsion stability index (ESI) of protein mixtures rich in the 7S fraction significantly improved twofold during processing at 75 °C. This study revealed a negative correlation between the emulsifying ability of soy protein and the 11S/7S ratio. For protein mixtures rich in either the 7S or the 11S fractions, gelling proprieties as well as emulsion activity index (EAI) and ESI showed no significant changes after spray drying; however, surface hydrophobicity was significantly enhanced following heating at 85 °C post-spray drying treatment. These findings provide insights into the alterations in gelling and emulsifying properties during various heating processes, offering great potential for producing soy protein ingredients with enhanced emulsifying ability and gelling property. They also contribute to establishing a theoretical basis for the standardized production of soy protein isolate with specific functional characteristics.

Activating argumentation schema to write argumentatively and tactfully.

To argue tactfully is a goal in argumentative writing, which entails balanced argumentation schema. Although computer-supported collaborative learning (CSCL) has been widely acknowledged as language learning mediation, especially in writing, few studies investigate its effectiveness in activating the balanced argumentation schema. This study explores the effectiveness of QQ group discussion, a kind of CSCL mediation most popular in China, in argumentative writing by means of quasi-experiment and interview. Fifty-six second-year college students in an English Department participated in this study. The experimental group were asked to have a pre-writing QQ group discussion on a disputable topic while the control group had an in-class face-to-face discussion (a regular teaching method for English majors). Content analysis of the essays was made to investigate the use of Counterargument elements, Qualifier and Concession. The results show that the two groups had no difference in the use of Counterargument-claim and Rebuttal. However, the experimental group surpassed the control group in Counterargument-data, Concession and Qualifier, which signifies more argumentativeness and tactfulness. In the delayed post-test this group still performed better. The interview transcriptions were coded and analyzed by inductive content analysis with the functions of QQ discussion as the themes. The result not only supported that of the experiment, but also revealed why and how QQ mediation could help activate the balanced argumentation schema. It is suggested that CSCL mediation should be promoted in argumentative writing instruction so that the students could write argumentatively and tactfully.

Nanostarch-Stimulated Cell Adhesion in 3D Bioprinted Hydrogel Scaffolds for Cell Cultured Meat.

Three-dimensional (3D) bioprinting has great potential in the applications of tissue engineering, including cell culturing meat, because of its versatility and bioimitability. However, existing bio-inks used as edible scaffold materials lack high biocompatibility and mechanical strength to enable cell growth inside. Here, we added starch nanoparticles (SNPs) in a gelatin/sodium alginate (Gel/SA) hydrogel to enhance printing and supporting properties and created a microenvironment for adherent proliferation of piscine satellite cells (PSCs). We demonstrated the biocompatibility of SNPs for cells, with increasing 20.8% cell viability and 36.1% adhesion rate after 5 days of incubation. Transcriptomics analysis showed the mechanisms underlying the effects of SNPs on the adherent behavior of myoblasts. The 1% SNP group had a low gel point and viscosity for shaping with PSCs infusion and had a high cell number and myotube fusion index after cultivation. Furthermore, the formation of 3D muscle tissue with thicker myofibers was shown in the SNP-Gel/SA hydrogel by immunological staining.

Biocompatible hydrophobic cross-linked cyclodextrin-based metal-organic framework as quercetin nanocarrier for enhancing stability and controlled release.

Cyclodextrin-based metal-organic framework (CD-MOF) has been widely used in various delivery systems due to its excellent edibility and high drug loading capacity. However, its typically bulky size and high brittleness in aqueous solutions pose significant challenges for practical applications. Here, we proposed an ultrasonic-assisted method for rapid synthesis of uniformly-sized nanoscale CD-MOF, followed by its hydrophobic modification through ester bond cross-linking (Nano-CMOF). Proper ultrasound treatment effectively reduced particle size to nanoscale (393.14 nm). Notably, carbonate ester cross-linking method significantly improved water stability without altering its cubic shape and high porosity (1.3 cm3/g), resulting in a retention rate exceeding 90% in various media. Furthermore, the loading of quercetin did not disrupt cubic structure and showcased remarkable storage stability. Nano-CMOF achieved controlled release of quercetin in both aqueous environments and digestion. Additionally, Nano-CMOF demonstrated exceptional antioxidant (free radical scavenging 82.27%) and biocompatibility, indicating its significant potential as novel nutritional delivery systems in food and biomedical fields.

The Roles of CircRNAs in Mitochondria.

Mitochondria participate in varieties of cellular events. It is widely accepted that human mitochondrial genome encodes 13 proteins, 2 rRNAs, and 22 tRNAs. Gene variation derived from human nuclear genome cannot completely explain mitochondrial diseases. The advent of high-throughput sequencing coupled with novel bioinformatic analyses decode the complexity of mitochondria-derived transcripts. Recently, circular RNAs (circRNAs) from both human mitochondrial genome and nuclear genome have been found to be located at mitochondria. Studies about the roles and molecular mechanisms underlying trafficking of the nucleus encoded circRNAs to mitochondria and mitochondria encoded circRNAs to the nucleus or cytoplasm in mammals are only beginning to emerge. These circRNAs have been associated with a variety of diseases, especially cancers. Here, we discuss the emerging field of mitochondria-located circRNAs by reviewing their identification, expression patterns, regulatory roles, and functional mechanisms. Mitochondria-located circRNAs have regulatory roles in cellular physiology and pathology. We also highlight future perspectives and challenges in studying mitochondria-located circRNAs, as well as their potential biomedical applications.

An advance in novel intelligent sensory technologies: From an implicit-tracking perspective of food perception.

Food sensory evaluation mainly includes explicit and implicit measurement methods. Implicit measures of consumer perception are gaining significant attention in food sensory and consumer science as they provide effective, subconscious, objective analysis. A wide range of advanced technologies are now available for analyzing physiological and psychological responses, including facial analysis technology, neuroimaging technology, autonomic nervous system technology, and behavioral pattern measurement. However, researchers in the food field often lack systematic knowledge of these multidisciplinary technologies and struggle with interpreting their results. In order to bridge this gap, this review systematically describes the principles and highlights the applications in food sensory and consumer science of facial analysis technologies such as eye tracking, facial electromyography, and automatic facial expression analysis, as well as neuroimaging technologies like electroencephalography, magnetoencephalography, functional magnetic resonance imaging, and functional near-infrared spectroscopy. Furthermore, we critically compare and discuss these advanced implicit techniques in the context of food sensory research and then accordingly propose prospects. Ultimately, we conclude that implicit measures should be complemented by traditional explicit measures to capture responses beyond preference. Facial analysis technologies offer a more objective reflection of sensory perception and attitudes toward food, whereas neuroimaging techniques provide valuable insight into the implicit physiological responses during food consumption. To enhance the interpretability and generalizability of implicit measurement results, further sensory studies are needed. Looking ahead, the combination of different methodological techniques in real-life situations holds promise for consumer sensory science in the field of food research.

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.

Evaluation of multiscale mechanisms of ultrasound-assisted extraction from porous plant materials: Experiment and modeling on this intensified process.

Ultrasound-assisted extraction (UAE) is an intensified mass transfer process, which can utilize natural resources effectively, but still lacks detailed mechanisms for multiscale effects. This study investigates the mass transfer mechanisms of UAE combined with material's pore structure at multiscale. Porous material was prepared by roasting green coffee beans (GCB) at 120 °C (RCB120) and 180 °C (RCB180), and their UAE efficiency for phytochemicals (caffeine, trigonelline, chlorogenic acid, caffeic acid) were evaluated by experiment and modeling. Besides, the physicochemical properties, mass transfer kinetics, and multi-physical field simulation were studied. Results indicated that positive synergy effects on extraction existed between ultrasound and material's pore structure. Higher mass transfer coefficients of UAE (GCB 0.16 min-1, RCB120 0.38 min-1, RCB180 0.46 min-1) was achieved with higher total porosity (4.47 %, 9.17 %, 13.52 %) and connected porosity (0 %, 3.79 %, 5.98 %). Moreover, simulation results revealed that micro acoustic streaming and pressure difference around particles were the main driving force for enhancing mass transfer, and the velocity (0.29-0.36 m/s) increased with power density (0.64-1.01 W/mL). The microscale model proved that increased yield from UAE-RCB was attributed to internal convection diffusion within particles. This study exploited a novel benefit of ultrasound on extraction and inspired its future application in non-thermal food processing.

Shape-recovery of implanted shape-memory devices remotely triggered via image-guided ultrasound heating.

Shape-memory materials hold great potential to impart medical devices with functionalities useful during implantation, locomotion, drug delivery, and removal. However, their clinical translation is limited by a lack of non-invasive and precise methods to trigger and control the shape recovery, especially for devices implanted in deep tissues. In this study, the application of image-guided high-intensity focused ultrasound (HIFU) heating is tested. Magnetic resonance-guided HIFU triggered shape-recovery of a device made of polyurethane urea while monitoring its temperature by magnetic resonance thermometry. Deformation of the polyurethane urea in a live canine bladder (5 cm deep) is achieved with 8 seconds of ultrasound-guided HIFU with millimeter resolution energy focus. Tissue sections show no hyperthermic tissue injury. A conceptual application in ureteral stent shape-recovery reduces removal resistance. In conclusion, image-guided HIFU demonstrates deep energy penetration, safety and speed.

Optimizing the Solvent Selection of the Ultrasound-Assisted Extraction of Sea Buckthorn (Hippophae rhamnoides L.) Pomace: Phenolic Profiles and Antioxidant Activity.

Sea buckthorn pomace (SBP) is a by-product of sea buckthorn processing that is rich in bioactive compounds. In this study, different active ingredients were extracted by using different solvents (water, methanol, ethanol, glycerol, ethyl acetate, and petroleum ether) combined with an ultrasonic assisted method. The correlation between the active ingredients and antioxidant properties of the extract was studied, which provided a research basis for the comprehensive utilization of SBP. This study revealed that the 75% ethanol extract had the highest total phenolic content (TPC) of 42.86 ± 0.73 mg GAE/g, while the 75% glycerol extract had the highest total flavonoid content (TFC) of 25.52 ± 1.35 mg RTE/g. The ethanol extract exhibited the strongest antioxidant activity at the same concentration compared with other solvents. The antioxidant activity of the ethanol, methanol, and glycerol extracts increased in a concentration-dependent manner. Thirteen phenolic compounds were detected in the SBP extracts using UPLC-MS/MS analysis. Notably, the 75% glycerol extract contained the highest concentration of all identified phenolic compounds, with rutin (192.21 ± 8.19 µg/g), epigallocatechin (105.49 ± 0.69 µg/g), and protocatechuic acid (27.9 ± 2.38 µg/g) being the most abundant. Flavonols were found to be the main phenolic substances in SBP. A strong correlation was observed between TPC and the antioxidant activities of SBP extracts. In conclusion, the choice of solvent significantly influences the active compounds and antioxidant activities of SBP extracts. SBP extracts are a valuable source of natural phenolics and antioxidants.

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.

Chickpea-Derived Modified Antimicrobial Peptides KTA and KTR Inactivate Staphylococcus aureus via Disrupting Cell Membrane and Interfering with Peptidoglycan Synthesis.

The widespread bacterial contamination caused by foodborne pathogens has continuously driven the development of advanced and potent food antimicrobial agents. In this study, two novel antimicrobial peptides (AMPs) named KTA and KTR were obtained by modifying a natural AMP, Leg2, from chickpea storage protein legumin hydrolysates. They were further predicted to be stable hydrophobic cationic AMPs of α-helical structure with no hemolytic toxicity by several online servers. Moreover, the AMPs exerted superior antibacterial activity against two representative Staphylococcus aureus strains thanks to the increased hydrophobicity and positive charge, with minimum inhibition concentration value (4.74-7.41 µM) significantly lower than that of Leg2 (>1158.70 µM). Further, this study sought to elucidate the specific antimicrobial mechanism against Gram-positive bacteria. It was found that the electrostatic interactions of the AMPs with peptidoglycan were vital for peptide activity in combating Gram-positive bacteria. Subsequently, the cell membrane of S. aureus cells was irreversibly disrupted by increasing permeability and impairing membrane components, which led to the massive release of intracellular substances and eventual cell death. Overall, this work demonstrated that KTA and KTR were active against Gram-positive bacteria via peptidoglycan targeting and membrane-disruptive mechanisms and paved the way for expanding their application potential to alleviate food contamination.