Investigation of the Alternations in the Muscle Quality of Swimming Crab (Ovalipes punctatus) during Cold-Chain Transportation Using Physicochemical and TMT-Based Quantitative Proteomic Analysis.

Physicochemical properties and protein alterations in Ovalipes punctatus during cold-chain transportation were examined via sensory scores, water-holding capacity (WHC), glucose (GLU) content, catalase (CAT) activity, urea nitrogen (UN) content, and tandem mass tag (TMT)-based proteomic analysis. The results revealed that sensory characteristics and texture of crab muscle deteriorated during transportation. Proteomic analysis revealed 442 and 470 different expressed proteins (DEPs) in crabs after 18 h (FC) and 36 h (DC) of transportation compared with live crabs (LC). Proteins related to muscle structure and amino acid metabolism significantly changed, as evidenced by the decreased WHC and sensory scores of crab muscle. Glycolysis, calcium signaling, and peroxisome pathways were upregulated in the FC/LC comparison, aligning with the changes in GLU content and CAT activity, revealing the stress response of energy metabolism and immune response in crabs during 0-18 h of transportation. The downregulated tricarboxylic acid (TCA) cycle and carcinogenesis-reactive oxygen species pathways were correlated with the decreasing trend in CAT activity, suggesting a gradual retardation in both energy and antioxidant metabolism in crabs during 18-36 h of transportation. Furthermore, the regulated purine nucleoside metabolic and nucleoside diphosphate-related processes, with the increasing changes in UN content, revealed the accumulation of metabolites in crabs.

Changes in the physicochemical characteristics and microbial community compositions of the abdomen and cheliped muscles in swimming crab (Portunus trituberculatus) during frozen storage.

The physicochemical indexes and microbial diversity were investigated to compare the altered quality properties of the abdomen and cheliped muscle in swimming crab (Portunus trituberculatus) during 100 days of frozen storage at -20℃. Over the extended duration of frozen storage, the sensory evaluation, moisture content, water activity (Aw), and water-holding capacity (WHC) in the abdomen and cheliped muscles of swimming crab decreased, while the pH, total volatile basic nitrogen (TVB-N), and trimethylamine (TMA) increased. The increase and decrease rates of these indicators were smaller in the abdomen than those in the cheliped muscle. High-throughput sequencing results indicated a reduction in the microbial richness and diversity in the abdomen and cheliped muscles of the swimming crab as frozen storage time extended. Proteobacteria, Actinobacteriota, and Firmicutes, Achromobacter, Kocuria, and Staphylococcus were the dominant phylum and genus in both muscle tissues, respectively. Furthermore, the correlation analysis between the composition of the microbiota and physiochemical properties revealed that the growths of Kocuria, Vibrio, Staphylococcus, and Aliiroseovarius were closely related to the physiochemical factors. The study provides a theoretical reference for quality deterioration and develops new products of different parts in the swimming crab during frozen storage.

Spectrophotometric- and LC/MS-Based Lipidomics Analyses Revealed Changes in Lipid Profiles of Pike Eel (Muraenesox cinereus) Treated with Stable Chlorine Dioxides and Vacuum-Packed during Chilled Storage.

Spectrophotometric- and liquid chromatography/mass spectrometry (LC/MS)-based lipidomics analyses were performed to explore the changes of lipid profiles in pike eel (Muraenesox cinereus) under stable chlorine dioxides (ClO2) and vacuum-packed treatment during chilled storage. The peroxide value (PV) and malondialdehyde (MDA) content in ClO2 treated and vacuum-packaged (VP) samples were significantly reduced compared to simple-packaged (SP) samples during whole chilled storage. The LC/MS-based lipidomics analyses identified 2182 lipid species in the pike eel muscle classified into 39 subclasses, including 712 triglycerides (TGs), 310 phosphatidylcholines (PCs), 153 phosphatidylethanolamines (PEs), and 147 diglycerides (DGs), among others. Further, in comparison with fresh pike eel (FE) muscle, 354 and 164 higher and 420 and 193 lower abundant levels of differentially abundant lipids (DALs) were identified in SP samples and VP samples, respectively. Compared with the VP batch, 396 higher and 404 lower abundant levels of DALs were identified in the SP batch. Among these, PCs, PEs, TGs, and DGs were more easily oxidized/hydrolyzed, which could be used as biomarkers to distinguish FE, SP, and VP samples. This research provides a reference for controlling lipid oxidation in fatty fish.

Cold-induced denaturation of muscle proteins in hairtail (Trichiurus lepturus) during storage: Physicochemical and label-free based proteomics analyses.

Physicochemical, proteomics, and bioinformatics analyses were conducted to investigate protein profiles in Trichiurus haumela under frozen (120 d) and chilled (6 d) storage. Springiness, chewiness, myofibrillar active sulfhydryl content, and Ca2+-ATPase activity significantly decreased, suggesting that cold stress altered muscle proteins. Compared with fresh hairtail (FH), 66 common differentially abundant proteins (DAPs) had lower abundances in chilled (3 d; CSH) and frozen (120 d; FSH) hairtail, including myosin binding proteins, filamins, actinin, troponin, and muscle-restricted coiled-coil protein. Gene Ontology (GO) annotation showed DAPs were mainly involved in cellular process, cellular anatomical entity, intracellular, and binding items. Eukaryotic orthologous group (KOG) analysis revealed that changes in cytoskeleton and energy production and conversion functions dominated during cold storage, degrading the myofibril and connective tissue structures and the physicochemical performance of muscle tissues. This study presents deep insights into the protein alternation mechanisms in hairtail muscle under cold stress.

Investigation of the changes in the lipid profiles in hairtail (Trichiurus haumela) muscle during frozen storage using chemical and LC/MS-based lipidomics analysis.

Chemical and LC/MS-based lipidomics strategies were performed to explore the alterations of lipid profiles in hairtail (Trichiurus haumela) muscle during 120 days of frozen storage. Chemical results indicated that the PV and TBARS values in hairtail muscle significantly increased during 120 days of frozen storage. Lipidomics results detected 1223 lipids in hairtail muscle assigned to 26 lipid categories, including 261 triglycerides (TGs), 251 phosphatidylcholines (PCs), 153 phosphatidylethanolamines (PEs), and 66 diglycerides (DGs). Totally, 153 and 67 differentially abundant lipids (DALs) accumulated at high and low levels, respectively, were detected in frozen hairtail (FSH) compared to in fresh (FH) samples. Among these, PEs, PCs, and TGs/DGs as predominant lipid components were vulnerable to oxidation/hydrolysis mainly due to their unsaturated properties. The apparent alterations between FSH and FH samples may result from lipid side-chain modifications, backbone cleavage, and/or decomposition of lipids during long-term storage. This study provides novel insight into the molecular mechanisms of lipid alternations in hairtail muscle during frozen storage.

Investigation of the activity of cathepsin B in red shrimp (Solenocera crassicornis) and its relation to the quality of muscle proteins during chilled and frozen storage.

Cathepsin B is a cysteine protease that has important effects on the quality of muscle products. In this study, the changes of cathepsin B activity and its relation to muscle proteins were investigated in intact and beheaded shrimp during chilled and frozen storage. The obtained results indicated that the water holding capacity (WHC), shear force, hardness, and myofibrillar protein (MP) content all significantly decreased in both the intact and beheaded shrimp samples with increasing storage period (p < 0.05). Specifically, beheading shrimp exhibited much more stable characteristics than intact shrimp samples during both chilled and frozen storage. The enzyme activity results suggested that cold temperature and storage induced the release of cathepsin B from the lysosomes to the mitochondria, sarcoplasm, and myofibrils in the muscle tissues. Furthermore, SDS-PAGE and transmission electron microscopy (TEM) analysis revealed that beheading the shrimp greatly inhibited the dissociation of shrimp muscle proteins during storage. The current findings suggest that cathepsin B located in the head of shrimp was likely transferred to the muscle through the first abdominal segment during storage, accelerating the dissociation of the muscle proteins. Therefore, beheading the shrimp was conducive to prolonging the shelf-life of stored shrimp products.

Phosphorylated Trehalose Suppresses the Denaturation of Myofibrillar Proteins in Peeled Shrimp (Litopenaeus vannamei) during Long-Term Frozen Storage.

The protective effects of phosphorylated trehalose on the quality and characteristics of peeled shrimp (Litopenaeus vannamei) were determined. Quality changes in treated samples were evaluated by assessing the physicochemical properties of myofibrillar proteins (MP) and compared to fresh water-, sodium tripolyphosphate-, and trehalose-treated samples during 12 weeks of frozen storage. The sensitivity of MP to oxidation and denaturation was increased during frozen storage. Phosphorylated trehalose significantly improved the quality of shrimp by increasing water-holding capacity. Further analysis showed that the addition of phosphorylated trehalose reduced the decrease in soluble MP content, Ca2+-ATPase activity, and total sulfhydryl contents and also effectively inhibited the increase in the surface hydrophobicity of MP. In addition, atomic force microscopy and hematoxylin and eosin staining revealed that phosphorylated trehalose preserved the integrity of the myofibril microstructure. Thermal stability results further confirmed that the denaturation temperature and denaturation enthalpy of MP were improved by phosphorylated trehalose. Overall, phosphorylated trehalose suppresses the denaturation of MP in peeled shrimp during long-term frozen storage.

Investigation of the changes in lipid profiles induced by hydroxyl radicals in whiteleg shrimp (Litopenaeus vannamei) muscle using LC/MS-based lipidomics analysis.

The oxidative effects of hydroxyl radical on the alterations of lipid profiles were investigated in shrimp muscle. Chemical results indicate peroxide value (PV) and thiobarbituric acid index (TBA-i) value in oxidation-treated shrimp significantly increased with oxidation time, and hydroxyl radical concentration increased, compared with those of in fresh samples. It was assumed that radical attack might induce lipid decomposition, backbone cleavage, and/or side-chain modifications. LC/MS-based lipidomics analysis revealed 835 lipids in shrimp assigned to 27 lipid classes, including 219 PCs and 98 CLs. In total, 86 and 34 differentially abundant lipids (DALs) accumulated at lower and higher levels, respectively, were identified in OS, compared with that in FS. This indicates hydroxyl radical attack altered the lipidomics profiles of shrimp muscle to a large extent. Furthermore, DALs, including CL 62:2, PC 38:3, and PE 34:9, could be considered as promising biomarkers to distinguish fresh and oxidation-treated shrimp products.

The differences of muscle proteins between neon flying squid (Ommastrephes bartramii) and jumbo squid (Dosidicus gigas) mantles via physicochemical and proteomic analyses.

Neon flying squid (OB) and jumbo squid (DG) mantles were evaluated to reveal the similarities and differences in their physicochemical features and protein abundances. Microstructural results indicated that the OB mantle exhibited numerous myofibril fragments and disordered microstructures after frozen storage compared with DG tissues. Chemical analysis suggested that freezing resulted in a rapid decrease in myofibrillar protein (MP) content, Ca2+-ATPase activity, and total sulfhydryl content, and promoted the increase in carbonyl content of MPs in both OB and DG. While, DG presented better MP stability than OB muscle after 120 days of frozen storage. Label-free proteomic analysis detected 24 down- and 33 up-regulated differentially abundant proteins (DAPs) in OB and DG mantles. Identified DAPs including isocitrate dehydrogenase and malic enzyme initiated a rapid decrease in the MP properties in OB samples. Moreover, DAPs were related to cytoskeleton function, including paramyosin, tropomyosin, and troponin C, which improved the stability of DG in response to freezing-induced changes.

Chrysin Stimulates Subcutaneous Fat Thermogenesis in Mice by Regulating PDGFRα and MicroRNA Expressions.

The activation of adipose tissue browning and thermogenesis provides a new strategy to counter obesity and associated metabolic diseases. Here, a natural flavonoid chrysin is used as the supplement of a high-fat diet (HFD). Dietary chrysin alleviates adiposity and insulin resistance in HFD-fed mice. Meanwhile, dietary chrysin elevates systemic energy expenditure and enhances the uncoupling protein-1 (UCP1) level in subcutaneous adipose tissue (SAT), which is accompanied by the increased thermogenic program, beige preadipocyte number, and angiogenesis in SAT. Dietary chrysin also induces the expression of SAT platelet-derived growth factor receptor α (PDGFRα), which commits adipose progenitor cells to differentiate into beige or white adipocytes in response to various environmental signals. Double immunofluorescent staining for UCP1 and PDGFRα reveals that chrysin elevates the number of UCP1+PDGFRα+ beige progenitors in SAT. Further, chrysin treatment reverses the effects of the specific PDGFRα inhibitor imatinib on browning differentiation of stromal vascular fraction cells from SAT. Finally, chrysin-induced adipocyte browning is correlated with the expressions of microRNAs as PDGFRα inhibitors or thermogenesis suppressors. In conclusion, dietary chrysin promotes subcutaneous adipocyte browning and systematic energy expenditure by regulating PDGFRα and microRNA expressions in HFD-fed mice.

Label-free proteomic analysis revealed the mechanisms of protein oxidation induced by hydroxyl radicals in whiteleg shrimp (Litopenaeus vannamei) muscle.

The oxidative effects of hydroxyl radicals derived from a FeCl3/ascorbic acid/H2O2 system on the stability of muscle proteins in peeled shrimp (Litopenaeus vannamei) were investigated. Physicochemical analysis indicated negative effects on the color (a* value), springiness, and pH of shrimp muscle, which appeared to be significantly exacerbated by higher concentrations of generated hydroxyl radicals when compared with the control. The microstructural results confirmed that a radical attack induced the incompact structure and disintegrated myofibers, thereby leading to weakened connective tissues and decreased stability of muscle proteins. Furthermore, label-free proteomic analysis revealed several differentially abundant proteins (DAPs) (i.e., ribosomal protein subunits, putative cytoskeleton proteins, and ion-binding proteins), which were detected and identified in oxidation-treated shrimp when compared with the control. The gene ontology (GO) and eukaryotic clusters of orthologous group (KOG) analyses further confirmed that the active hydroxyl radicals attacked vulnerable amino acids, modified peptide chains, and/or protein structures and/or conformations, which were responsible for a significant decrease in the muscle texture and stability of proteins in oxidation-treated shrimp. This study provides novel insight into the molecular mechanisms of muscle protein changes during oxidation development.

Insights into the similarities and differences of whiteleg shrimp pre-soaked with sodium tripolyphosphate and sodium trimetaphosphate during frozen storage.

In this study, similarities and differences of sodium tripolyphosphate (STPP) and sodium trimetaphosphate (STMP) pre-soaking on the stability of muscle proteins in shrimp were investigated during 12 weeks of frozen storage (-30 °C). The physicochemical analysis indicated significant improvements in the WHC, springiness, chewiness, and thermal stability of STPP and STMP pre-soaked samples when compared to the control. Interestingly, STMP pre-soaking showed better cryoprotective effects than the STPP treatment when the storage period reached the end of the 12 weeks. Furthermore, the label-free based proteomics results indicated that 62 upregulated differentially abundant proteins (DAPs) were detected in STMP when compared to STPP. These identified DAPs specifically included 40S ribosomal proteins, actin-related proteins, heat shock proteins, myosin heavy chain, and tubulin beta chain. Additionally, the gene ontology (GO) and eukaryotic clusters of orthologous group (KOG) analyses verified that the incorporation of STMP molecules enhanced the resistance of cytoskeleton proteins to cold-temperature stress.

Kappa-carrageenan and its oligosaccharides maintain the physicochemical properties of myofibrillar proteins in shrimp mud (Xia-Hua) during frozen storage.

As a popular hot-pot ingredient, Chinese-style shrimp mud (Xia-Hua) is usually transported and stored frozen. However, frozen storage leads to decreased quality of Xia-Hua products caused by the variations in physicochemical and functional properties of myofibrillar proteins (MPs). Κ-carrageenan and its oligosaccharides are reported as antioxidants and antifreeze and can stabilize proteins in whole shrimp, but their effects on MPs in Xia-Hua remain poorly understood. Compared to the control and Na4 P2 O7 treatments, the physicochemical properties of MPs in κ-carrageenan and its oligosaccharides-incorporated Xia-Hua were evaluated during 120 days of frozen storage. The results showed that cold stress increased the susceptibility of MPs to denaturation and oxidation during frozen storage. Carrageenan oligosaccharides maintained the turbidity, emulsifying activity, stability, and foaming capacity of MPs. Oxidation analysis showed that the incorporation of carrageenan oligosaccharides significantly retarded the rapid decrease of Ca2+ -ATPase activity, total sulphydryl and active sulphydryl contents, and also effectively inhibited the increases of carbonyl content and surface hydrophobicity of MPs. Thermal stability results confirmed that the oligosaccharides improved the denaturation temperature and enthalpy of MPs compared to the control, Na4 P2 O7 , and carrageenan treatments. This study suggests that κ-carrageenan and its oligosaccharides maintain the properties of MPs in Xia-Hua during frozen storage. PRACTICAL APPLICATION: The cryoprotection and antioxidant effects of carrageenan oligosaccharides on the stability of MPs in frozen shrimp mud can be used to extend the shelf-life and maintain the quality of frozen Xia-Hua products. Furthermore, it can drive the development of aquatic product health industry, improve the quality and safety of aquatic products, reduce the occurrence of public food safety incidents, and maintain social stability.

Small interfering RNA-loaded chitosan hydrochloride/carboxymethyl chitosan nanoparticles for ultrasound-triggered release to hamper colorectal cancer growth in vitro.

Development of nontoxic, targetable and potent small interfering RNAs (siRNA) delivery systems remains a predominant challenge for clinical application of siRNA therapy. The nanoparticles of carboxymethyl chitosan (CMC) and labeled fluorescein isothiocyanate (FITC)-chitosan hydrochloride (CHC) were fabricated as carriers for ultrasound-triggered drug delivery to treat colon cancer. The results showed the (FITC-CHC)-CMC nanoparticles could effectively encapsulate anti-ß-catenin siRNA through ionic gelation self-assembly to improve the stability of siRNA. The cumulative release ratio of siRNA from crosslinked (FITC-CHC)-CMC nanoparticles was merely 11.08% in pH 2.2 solution within 120 min, whereas about 70.07% of the loaded siRNA was released within 120 min in pH 5.5 solution after an 8-min ultrasonic treatment. It indicated that the (FITC-CHC)-CMC based pH-sensitive delivery system could fulfill a controlled release of siRNA through responding to external stimulus (ultrasound) under favorable pH condition. Fluorescence microscopy measurements clearly visualized the entry of fluorescently-labeled siRNA into HT-29 cells. Following the transfection of anti-ß-catenin siRNA for 48 h, the ß-catenin protein expression of the colon cancer cells was reduced to about 40.10%, indicating effective reduction of the protein that promotes colon cancer proliferation. Our results demonstrated that the siRNA-(FITC-CHC)-CMC delivery system hold substantial potential for RNAi therapeutical applications in diseased cells.

Effect of postharvest UV-B or UV-C irradiation on phenolic compounds and their transcription of phenolic biosynthetic genes of table grapes.

Ultraviolet (UV) irradiation has been related to the extension shelf-life and maintenance of postharvest quality in fruits. However, the comparison of UV-B and UV-C treatment on the biosynthesis of phenolic compounds of grape remain unclear. This study provides a comparison on the mechanism of phenolic secondary metabolism at the same dose of 3.6 kJ m-2 UV treatment. Total phenolic compounds, total flavonoid, total flavanol, and total anthocyanin content and antioxidant activities of grapes after UV-C treatments were higher than those of the control and UV-B treatment. Among the evaluated parameters of individual phenolic compounds, the content of trans-resveratrol showed the highest percentage increase after the UV application. The transcriptions of PAL, CHS, F3H, LAR, ANS and STS were higher in grapes treated by UV-C than in those treated by UV-B. The CHS, LAR, ANS and STS genes were more induced in UV-B treatment than in control group. The same applied dose of UV-B or UV-C irradiation have different impact on gene expression and phenolic metabolites synthesis. The UV-C irradiation stimulated a higher gene expression of the phenolic compounds biosynthesis and also induced a greater accumulation of these metabolites at the same applied dose.

Far-infrared therapy for cardiovascular, autoimmune, and other chronic health problems: A systematic review.

Physical therapy (physiotherapy), a complementary and alternative medicine therapy, has been widely applied in diagnosing and treating various diseases and defects. Increasing evidence suggests that convenient and non-invasive far-infrared (FIR) rays, a vital type of physiotherapy, improve the health of patients with cardiovascular disease, diabetes mellitus, and chronic kidney disease. Nevertheless, the molecular mechanisms by which FIR functions remain elusive. Hence, the purpose of this study was to review and summarize the results of previous investigations and to elaborate on the molecular mechanisms of FIR therapy in various types of disease. In conclusion, FIR therapy may be closely related to the increased expression of endothelial nitric oxide synthase as well as nitric oxide production and may modulate the profiles of some circulating miRNAs; thus, it may be a beneficial complement to treatments for some chronic diseases that yields no adverse effects.