Identification of a Candidate Gene Regulating Intramuscular Fat Content in Pigs through the Integrative Analysis of Transcriptomics and Proteomics Data.

Pork is a widely consumed source of animal protein worldwide, and the intramuscular fat (IMF) content in pork plays a crucial role in determining its quality. In this study, we sought to identify candidate genes that regulate IMF deposition in pigs. We performed tandem mass tags (TMT)-based quantitative proteomics analysis using Longissimus dorsi (LD) muscle samples obtained from eight pigs with extremely high and low IMF content among a group of 28 Duroc pigs and identified 50 differentially abundant proteins (DAPs). Additionally, we compared the proteomics data with RNA-sequencing data obtained in our previous study and identified TUSC5 as a differentially expressed gene corresponding to the relevant DAP. To investigate the potential role of TUSC5 in adipogenesis, we suppressed TUSC5 expression in mouse 3T3-L1 preadipocytes using short hairpin RNA (shRNA) and observed a significant reduction in the differentiation of 3T3-L1 cells into adipocytes, as indicated by Oil Red O staining and triglyceride content. Moreover, we observed a reduction in the expression of genes associated with adipogenesis (PPARG, CEBPA, FABP4, and FASN) following TUSC5 suppression. Through an integrative analysis of transcriptomics and proteomics data, our study identified TUSC5 as a crucial candidate gene associated with the regulation of IMF content in pigs.

Bromodomain protein 4 is a key molecular driver of TGFß1-induced hepatic stellate cell activation.

Liver fibrosis is characterized by the excessive deposition of extracellular matrix in liver. Chronic liver injury induces the activation of hepatic stellate cell (HSCs), a key step in liver fibrogenesis. The activated HSC is the primary source of ECM and contributes significantly to liver fibrosis. TGFß1 is the most potent pro-fibrotic cytokine. Bromodomain protein 4 (BrD4), an epigenetic reader of histone acetylation marks, was crucial for profibrotic gene expression in HSCs. The present study aimed to investigate the roles of BRD4 in TGFß1-dependent HSC activation and liver fibrosis, focusing on TGFß1-induced alterations of the levels of the fibrotic-related important proteins in HSCs by employing the heterozygous TGFß1 knockout mice and BrD4 knockdown in vivo and in vitro. Results revealed that BrD4 protein level was significantly upregulated by TGFß1 and BrD4 knockdown reduced TGFß1-induced HSC activation and liver fibrosis. BrD4 was required for the influences of TGFß1 on PDGFß receptor and on the pathways of Smad3, Stat3, and Akt. BrD4 also mediated TGFß1-induced increases in histone acetyltransferase p300, the pivotal pro-inflammatory NFkB p65, and tissue inhibitor of metalloproteinase 1 whereas BrD4 reduced Caspase-3 protein levels in HSCs during liver injury, independent of TGFß1. Further experiments indicated the interaction between TGFß1-induced BrD4 and NFkB p65 in HSCs and in liver of TAA-induced liver injury. Human cirrhotic livers were demonstrated a parallel increase in the protein levels of BrD4 and NFkB p65 in HSCs. This study revealed that BrD4 was a key molecular driver of TGFß1-induced HSC activation and liver fibrosis.

Lactoferricin, an antimicrobial motif derived from lactoferrin with food preservation potential.

The growth of bacteria and fungi may cause disease inf human or spoilage of food. New antimicrobial substances need to be discovered. Lactoferricin (LFcin) is a group of antimicrobial peptides derived from the N-terminal region of the milk protein lactoferrin (LF). LFcin has antimicrobial ability against a variety of microorganisms, which is significantly better than that of its parent version. Here, we review the sequences, structures, and antimicrobial activities of this family and elucidated the motifs of structural and functional significance, as well as its application in food. Using sequence and structural similarity searches, we identified 43 new LFcins from the mammalian LFs deposited in the protein databases, which are grouped into six families according to their origins (Primates, Rodentia, Artiodactyla, Perissodactyla, Pholidota, and Carnivora). This work expands the LFcin family and will facilitate further characterization of novel peptides with antimicrobial potential. Considering the antimicrobial effect of LFcin on foodborne pathogens, we describe the application of these peptides from the prospective of food preservation.

Perspective of sodium reduction based on endogenous proteases via the strategy of sodium replacement in conjunction with mediated-curing.

NaCl is the main curing agent in dry-cured meat products, and a large amount of NaCl addition leads to high salt content of final products. Salt content and composition are important factors affecting the activity of endogenous proteases, which in turn could affect proteolysis as well as the quality of dry-cured meat products. With the increasing emphasis on the relationship between diet and health, reducing sodium content without sacrificing quality and safety of products is a great challenge for dry-cured meat industry. In this review, the change of endogenous proteases activity during processing, the potential relationship between sodium reduction strategy, endogenous proteases activity, and quality were summarized and discussed. The results showed that sodium replacement strategy and mediated-curing had a complementary advantage in influencing endogenous proteases activity. In addition, mediated-curing had the potential to salvage the negative effects of sodium substitution by affecting endogenous proteases. Based on the results, a sodium reduction strategy that sodium replacement in conjunction with mediated-curing based on endogenous proteases was proposed for the future perspective.

Meat food fraud risk in Chinese markets 2012-2021.

Food fraud is a major concern worldwide, and the majority of cases include meat adulteration or fraud. Many incidences of food fraud have been identified for meat products both in China and abroad over the last decade. We created a meat food fraud risk database compiled from 1987 pieces of information recorded by official circular information and media reports in China from 2012 to 2021. The data covered livestock, poultry, by-products, and various processed meat products. We conducted a summary analysis of meat food fraud incidents by researching fraud types, regional distribution, adulterants and categories involved, categories and sub-categories of foods, risk links and locations, etc. The findings can be used not only to analyze meat food safety situations and study the burden of food fraud but also help to promote the efficiency of detection and rapid screening, along with improving prevention and regulation of adulteration in the meat supply chain markets.

Identification and In Silico Simulation on Inhibitory Platelet-Activating Factor Acetyl Hydrolase Peptides from Dry-Cured Pork Coppa.

The unique processing technology of dry-cured meat products leads to strong proteolysis, which produces numerous peptides. The purpose of this investigation was the systematic isolation, purification, and identification of potentially cardioprotective bioactive peptides from dry-cured pork coppa during processing. According to the results of anti-platelet-activating factor acetyl hydrolase activity and radical scavenging ability in vitro, the inhibitory effect of M1F2 in purified fractions on cardiovascular inflammation was higher than that of M2F2. The peptide of M1F2 was identified by nano-liquid chromatography-tandem mass spectrometry. A total of 30 peptides were identified. Based on bioinformatics methods, including in silico analysis and molecular docking, LTDKPFL, VEAPPAKVP, KVPVPAPK, IPVPKK, and PIKRSP were identified as the most promising potential platelet-activating factor acetyl hydrolase inhibitory peptides. Overall, bioactive peptides produced during dry-cured pork coppa processing demonstrate positive effects on human health.

Insights into Feruloylated Oligosaccharide Impact on Gel Properties of Oxidized Myofibrillar Proteins Based on the Changes in Their Spatial Structure.

Polyphenolic compounds can protect against myofibrillar protein (MP) oxidation in meat products. In this study, the inhibitory effect of feruloyl oligosaccharides (FOs) on MP oxidation was investigated, and the gel properties of MPs were further studied. The results showed that 50-100 µmol/g protein of FOs could effectively inhibit damage to amino acid side chains by reducing carbonyl contents by 60.5% and increasing sulfhydryl and free amine contents by 89.5% and 66%, which may protect the secondary and tertiary structures of MPs. Additionally, FOs at 50 µmol/g protein had better effects on the crosslinking of MPs, leading to effective improvements in the gel properties, which can be seen in the rheology properties, scanning electron microscope (SEM) photographs, and the distribution of water in the MP gel. On the contrary, 150-200 µmol/g protein of FOs showed peroxidative effects on oxidatively stressed MPs, which were detrimental to MPs and contributed to their denaturation in the electrophoresis analysis and irregular aggregation in the SEM analysis. The concentration-dependent effects of FOs depended on MP-FOs interactions, indicating that an appropriate concentration of FOs has the potential to protect MPs from oxidation and enhance the gelation ability of pork meat during processing.

Determination of carrageenan and konjac gum in livestock meat and meat products by ultra performance liquid chromatography-tandem mass spectrometry.

At present, gum-injected meat appeared in the market, which has seriously damaged the legitimate rights and interests of consumers. Therefore, a method for the determination of carrageenan and konjac gum in livestock meat and meat products by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was established. The samples were hydrolyzed by hydrogen nitrate. After centrifugation and dilution, the supernatants were detected by the UPLC-MS/MS and the concentration of target compounds in samples were calibrated by the matrix calibration curves. A good linear relationship was observed in the concentration range of 5-100 µg/mL with correlation coefficients of more than 0.995. The limits of detection (LODs) and limits of quantification (LOQs) were found to be 20 and 50 mg/kg, respectively. The recoveries at three spiked levels (50, 100 and 500 mg/kg) in blank matrix were in the range of 84.8-108.6% with the relative standard deviations between 1.5% and 6.4%. The method has the advantages of convenient, accurate and efficient, and can be used as an effective method for detecting carrageenan and konjac gum in the different kinds of livestock meat and meat products.

Effects of Citrus Fiber on the Gel Properties of Mutton Myofibrillar Protein.

This work investigated the effects of different additions of water-soluble citrus fiber (SCF) and water-insoluble citrus fiber (ICF) on the gel properties of the mutton myofibrillar protein (MP). The key parameters of water-holding capacity (WHC), rheological properties, and microstructure were evaluated. The addition of 2.5-10% of SCF and ICF significantly improved (p < 0.05) the WHC and gel strength of mutton MP gel. The rheological results showed that the viscoelasticity of MP with 5% SCF was the best, and the T2 relaxation time of the gel was significantly shortened. SEM results showed SCF reduced the number of pores in the MP gel, forming a more compact network structure. ICF stabilized the MP gel network structure as a filler after water absorption and expansion. However, the gel lost moisture under the action of strong external force (freeze-drying), which left large pores. These data confirmed that SCF and ICF could effectively improve the gel properties of meat products.

Insights into the trace Sr2+ impact on the gel properties and spatial structure of mutton myofibrillar proteins.

Myofibrillar proteins (MPs) and the quality of meat strongly depend on the properties of MP gels, which in turn depend on several parameters that include the thermal history and the concentration of metal ions. Strontium element (Sr) widely exists in mineral water and is found as strontium ions (Sr2+), which is an essential trace element for humans. This study investigated the effects of trace Sr2+ on the structure-function relationship of mutton MPs, as well as their gels with water. Trace concentrations of Sr2+ were found to significantly alter the conformation of the MPs. An increase in Sr2+ concentration was associated with a reduction in the tightness and strength of the gel and a significant increase in its water-holding capacity As compared to the untreated control sample, the solubility, particle size, and the magnitude of the Zeta potential of the gels increased by 13.03 %, 12.62 %, and 19.73 %, respectively, whereas the water retention capacity and the gel strength increased by 23.13 % and 21.90 %, at a Sr2+ concentration of 5.0 mg/L. Molecular docking predicted an increase in ionic bonds and disulfide bonds because Sr2+ had a strong interaction with hydrophilic amino acids and acidic amino acids. The analysis of molecular forces further verified the significant facilitation of interactions between MP molecules with the induction of Sr2+. As compare to the untreated control group, the ionic and disulfide bonds increased by 141.17 % and 66.94 %, when treated with 5.0 mg/L Sr2+. These changes were likely due to the enhancement of protein-protein interactions caused by Sr2+, which could induce MP molecules to properly unfold and aggregate in gel formation. The results could provide a basis for improving the texture and the quality of meat and meat products.

Nisin: From a structural and meat preservation perspective.

Nisin is a posttranslationally modified antimicrobial peptide that is widely used as a food preservative. It contains five cyclic thioethers of varying sizes. Nisin activity and stability are closely related to its primary and three dimensional structures. It has nine reported natural variants. Nisin A is the most studied nisin as it was the first one purified. Here, we review the sequence feature of nisin A and its natural variants, and their biosynthesis pathway, mode of action and application as a meat preservative. We systematically illustrate the functional domains of the main enzymes (NisB, NisC, and NisP) involved in nisin synthesis. NisB was shown to dehydrate its substrate NisA via a tRNA associated glutamylation mechanism. NisC catalysed the cyclization of the didehydro amino acids with the neighboring cysteine residues. After cyclization, the leader peptide is removed by the protease NisP. According to multiple sequence alignments, we detected five conserved sites Dha5, Pro9, Gly14, Leu16, and Lys22. These residues are probably the structural and functional important ones that can be modified to produce peptides versions with enhanced antimicrobial activity. Through comparing various application methods of nisin in different meats, the antimicrobial effects of nisin used individually or in combination with other natural substances were clarified.

Development of meat analogs: Focus on the current status and challenges of regulatory legislation.

Population growth and the rising enthusiasm for meat consumption in developing countries have increased the global demand for animal protein. The limited increase in traditional meat production, which results in high resource consumption, greenhouse gas emissions, and zoonotic diseases, has affected the sustainable supply of meat protein. The technological development and commercialization of meat analogs derived from plant and microbial proteins provide a strategy for solving the abovementioned problems. However, before these innovative foods are marketed, they should comply with regulations and standards to ensure food safety and consumer rights. This review briefly summarizes the global development status and challenges of plant- and fungi-based meat analog products. It focuses on the current status, characteristics, and disputes in the regulations and standards worldwide for plant- and fungi-based meat analogs and proposes suggestions for perfecting the regulatory system from the perspective of ensuring safety and supporting innovation. Although plant- and fungi-based meat analogs have had a history of safe usage as foods for a certain period around the world, the nomenclature and product standards are uncertain, which affects product innovation and global sales. Regulatory authorities should promptly formulate and revise regulations or standards to clarify the naming of meat analogs and product standards, especially the use of animal-derived ingredients and limits of nutrients (e.g., protein, fat, vitamins, and minerals) to continuously introduce start-up products to the market.

Physicochemical and Antioxidant Properties of Nanoliposomes Loaded with Rosemary Oleoresin and Their Oxidative Stability Application in Dried Oysters.

Lipid and protein oxidation is a main problem related to the preservation of dried aquatic products. Rosemary oleoresin is widely used as an antioxidant, but its application is limited due to its instability and easy degradation. Nanoliposome encapsulation is a promising and rapidly emerging technology in which antioxidants are incorporated into the liposomes to provide the food high quality, safety and long shelf life. The objectives of this study were to prepare nanoliposome coatings of rosemary oleoresin to enhance the antioxidant stability, and to evaluate their potential application in inhibiting protein and lipid oxidation in dried oysters during storage. The nanoliposomes encapsulating rosemary oleoresin were applied with a thin-film evaporation method, and the optimal amount of encapsulated rosemary oleoresin was chosen based on changes in the dynamic light scattering, Zeta potential, and encapsulation efficiency of the nanoliposomes. The Fourier transform-infrared spectroscopy of rosemary oleoresin nanoliposomes showed no new characteristic peaks formed after rosemary oleoresin encapsulation, and the particle size of rosemary oleoresin nanoliposomes was 100-200 nm in transmission electron microscopy. The differential scanning calorimetry indicated that the nanoliposomes coated with rosemary oleoresin had better thermal stability. Rosemary oleoresin nanoliposomes presented good antioxidant stability, and still maintained 48% DPPH radical-scavenging activity and 45% ABTS radical-scavenging activity after 28 d of storage, which was 3.7 times and 2.8 times higher than that of empty nanoliposomes, respectively. Compared with the control, the dried oysters coated with rosemary oleoresin nanoliposomes showed significantly lower values of carbonyl, sulfhydryl content, thiobarbituric acid reactive substances, Peroxide value, and 4-Hydroxynonenal contents during 28 d of storage. The results provide a theoretical basis for developing an efficient and long-term antioxidant approach.

Effects of ultrasound-assisted thawing on lamb meat quality and oxidative stability during refrigerated storage using non-targeted metabolomics.

The aim of this study was to evaluate the changes of ultrasound-assisted thawing on lamb meat quality and differential metabolite profiles during refrigerated storage. Compared with flow water thawing (FW), pH, a*, C*, and sulfhydryl content of lamb were significantly increased, while L*, drip loss and cooking loss were significantly decreased after ultrasound-assisted thawing (UT). On day 1 (UT1 and FW1) and day 7 (UT7 and FW7) in the UT and FW groups, principal component analysis explained 42.22% and 39.25% of the total variance. In this study, 44 (UT1 and FW1) and 47 (UT7 and FW7) differentially expressed metabolites were identified, including amino acids, carbohydrates and their conjugates, nucleic acids, carbonyl compounds and others. The results of this study provide data to clarify the differences between UT and FW, and lay a foundation for the application of ultrasound-assisted thawing in the meat industry.

Modulating the structure of lamb myofibrillar protein gel influenced by psyllium husk powder at different NaCl concentrations: Effect of intermolecular interactions.

In this study, a strategy involving psyllium husk powder (PHP) was proposed to alleviate the textural deterioration of protein gels under low-sodium conditions. Results revealed that myofibrillar protein (MP) in 0.3 M NaCl could accommodate more PHP to achieve better gels properties compared with that of 0.6 M NaCl. The 3 % addition of PHP could lessen the textural deterioration of gels at 0.3 M NaCl because of the insertion of PHP into the hydrophobic cavity of MP. Consequently, the reduction in protein viscoelasticity and the thermal stability of the head and tail of myosin improved. α-Helix structures unfolded, intermolecular forces formed, and proteins aggregated. Molecular docking predicted hydrogen bonds and hydrophobic interactions as the main forces to stabilize the conformation of composites. Experiments further verified that hydrophobic interactions and disulfide bonds were the main forces that stabilized the structure of MP-PHP composite gels.

Simultaneous screening and analysis of 155 veterinary drugs in livestock foods using ultra-high performance liquid chromatography tandem quadrupole linear-ion-trap mass spectrometry.

Veterinary drugs are widely used to improve the health and growth of livestock. The supervision of these residues is necessary to ensure food safety. A high-throughput method based on Oasis PRiME HLB with solid phase extraction for simultaneous qualitative and quantitative analysis of 155 veterinary drugs in livestock foods was developed by the ultra-high performance liquid chromatography tandem quadrupole linear-ion-trap mass spectrometry (UHPLC-QTRAP-MS). The limits of detection and quantification ranged from 0.5 µg/kg to 5 µg/kg and 2 µg/kg to 20 µg/kg, respectively. For over 85% of the analytes, the recoveries were between 60% and 120%. The positive simulated samples perfectly matched with a purity fit value over 70% from the self-built library. The screening results of UHPLC-QTRAP-MS were almost consistent with UHPLC tandem quadrupole-exactive orbitrap mass spectrometry (UHPLC-Q-Exactive Orbitrap/MS). The evaluated UHPLC-QTRAP-MS method was powerful and reliable for the screening and quantification of veterinary drugs in real samples.

Physical optimization of cell proliferation and differentiation using spinner flask and microcarriers.

The traditional breeding industry has been increasingly saturated and caused environmental pollution, disease transmission, excessive resource use, and methane emission; however, it still cannot meet the needs of the growing population. To explore other alternatives, researchers focused on cell agriculture and cell-based meat, especially large-scale cell culture. As a prerequisite for production, large-scale culture technology has become an important bottleneck restricting cell-based meat industrialization. In this study, the single-factor variable method was adopted to examine the influence of Cytodex1 microcarrier pretreatment, spinner flask reaction vessel, cell culture medium, serum and cell incubation, and other influencing factors on large-scale cell cultures to identify the optimization parameters suitable for 3D culture environment. Collagen and 3D culture were also prospectively explored to promote myogenesis and cultivate tissue-like muscle fibers that contract spontaneously. This research lays a theoretical foundation and an exploratory practice for large-scale cell cultures and provides a study reference for the microenvironment of myoblast culture in vitro, a feasible direction for the cell therapy of muscular dystrophy, and prerequisites for the industrialized manufacturing of cell-based meat. Graphical summary: Research on large-scale myoblast culture using spinner flasks and microcarriers. For cell culture, the microcarriers were pretreated with UV and collagen. Cell seeding condition, spinner flask speed, resting time, and spinner flask culture microenvironment were then optimized. Finally, two culture systems were prepared: a culture system based on large-scale cell expansion and a culture system for myogenesis promotion and differentiation.

Changes in volatile profiles of a refrigerated-reheated xylose-cysteine-lecithin reaction model analyzed by GC×GC-MS and E-nose.

The changes in the volatile profiles of a xylose-cysteine-lecithin reaction model were investigated by using comprehensive two-dimensional gas chromatography-mass spectrometry (GC×GC-MS) in combination with headspace solid-phase microextraction (SPME) and electronic nose (E-nose) to evaluate the contribution of refrigerating and reheating treatment to warmed-over flavor (WOF). The volatile compound results and E-nose revealed that the contribution of refrigerating and reheating to the WOF was not consistent. After refrigerating, the level of furfuryl mercaptan increased, while that of 1-octene-3-ol, octanal, nonanal, and 2-decanone decreased, which affected the flavors. An increase in the level of 1-octene-3-ol, 2-pentyl-thiophene, and hexanoic acid and a decrease in the levels of furfural, 2-methyl-3-furanthiol, and 2-methyl-3-pentanethiol occurred during reheating. According to the odor activity value and sensory evaluation, the sulfur-like odor became more intense after refrigerating, while the rancid-like odor grew stronger, but the sulfur-like odor alleviated after reheating. Overall, the reaction between residual substances caused the WOF during refrigeration, also lead to the fatty acid oxidation increased after reheating. The overproduction of fatty acids oxidation products and decreased of volatile product of Maillard reaction leads to the WOF during reheating. PRACTICAL APPLICATION: This study provides theoretical guidance to reduce the off-flavors of meat products.

Rapid LC-MS/MS method for the detection of seven animal species in meat products.

The adulteration of meat products has been reported worldwide, and detection of specific peptides through mass spectrometry (MS) is a reliable method for meat species identification. However, the practical application of this method is limited by complicated steps and long reaction time of the traditional sample preparation. Therefore, this paper introduced a convenient and time-saving sample preparation by optimizing the steps of reduction, alkylation, digestion, and purification. With the rapid sample preparation, 35 species-specific peptides for seven species (pig, cattle, sheep, deer, chicken, duck, and turkey) were screened using high-resolution MS, and a rapid LC-MS/MS method was established. The method only takes 3 h from sample receipt to results. The meat species of 20 processed meat products were detected, and three samples were found potentially adulterated. The method is proved to have high sensitivity, specificity, practicability with respect to rapid identification of meat species in meat products.