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The molecular composition of starch leachates from starch-based foods has been recently recognised as a crucial determinant of food properties. However, there is limited knowledge on the regulation of this composition through irradiative depolymerization of starch. This research investigates the leaching behaviour of maize starch depolymerized by electron beam irradiation, and the relationship between the composition of leached starch and structures of modified starch granules. The analysis using 1 H NMR spectroscopy confirmed a decrease in the degree of branching (from 4.4 % to 2.8 %), while size-exclusion chromatography identified a newly-derived amylopectin fraction of a smaller hydrodynamic radius (approximately 60-80 nm). The structural properties of the starch granules were also analysed, revealing an increased BET-area of granules and reduced total crystallinity after depolymerization. In the leachates of swollen granules, the bimodal distribution of starch molecules evolves into unimodal with the increase of the irradiative dosage, while modified starch leached more starch molecules with R h  < 10 nm. The results of principal component analysis and Pearson correlation analysis indicate that the degree of branching of degraded starch molecules, as well as the newly-derived amylopectin fraction, significantly correlates (p < 0.01) with the molecular size of leached starch molecules (R h  < 10 nm). It is thus proposed that the cleavage of α-1,6 linkage may be a critical factor in controlling the leaching process of irradiated starch granules. This study highlights the potential of irradiative degradation to control the molecular composition and structure of starch leachates, thereby optimizing the properties of starch-based foods., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

This study explored a new physical modification approach to regulate enzymatic resistance of high-amylose starch for potentially better nutritional outcomes. High-amylose maize starch (HAMS) was subjected to chain depolymerization by electron beam irradiation (EBI), followed by inducing ordered structure through annealing in palmitic acid solution (APAS). APAS treatment significantly promotes the formation of ordered structure. Starch after the combinative modification showed up to 5.2 % increase in total crystallinity and up to 1.2 % increase in V-type fraction. The EBI-APAS modification led to increased gelatinization temperature (from 66.1 to 87.6 °C) and reduced final digested percentage under in vitro stimulated digestion conditions. The moderate extent of depolymerization resulted in higher enzymatic resistance, indicating that the extent of depolymerization is crucial in EBI-APAS modification. Pearson analysis showed a significant correlation between gelatinization onset temperature and digestion kinetic parameter (k 1 , rate constant of fast-phase digestion). Overall, the result suggests that ordered structures of degraded molecules induced by the combinative modification contribute to the enzymatic resistance of starch. This study sheds lights on future applications of EBI-APAS approach to regulate multi-scale structures and nutritional values of high-amylose starch., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

This study explores a new strategy for manipulating the digestibility of high-amylose maize starch (HAMS) through combinative modifications, namely depolymerization via electron beam irradiation (EBI) followed by reorganizing glucan chains via heat moisture treatment (HMT). The results show that semi-crystalline structure, morphological features and thermal properties of HAMS remained similar. However, EBI increased branching degree of the starch at high irradiation dosage (20 kGy), resulting in more readily leached amylose during heating. HMT increased the relative crystallinity (3.9-5.4% increase) and V-type fraction (0.6-1.9% increase), without significant changes (p > 0.05) in gelatinization onset temperature, peak temperature and enthalpy. Under simulated gastrointestinal conditions, the combination of EBI and HMT either had no effect or negative effect on starch enzymatic resistance, depending on the irradiation dosage. These results suggest that the depolymerization by EBI predominantly affects the changes in enzyme resistance, rather than the growth and perfection of crystallites induced by HMT., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Irradiation can be used for the preservation of chickpea protein as it can destroy microorganisms, bacteria, virus, or insects that might be present. However, irradiation may provoke oxidative stress, and therefore modify the functionality and nutritional value of chickpea protein. In order to study the effects of irradiation on the physicochemical properties and digestion behaviour of chickpea protein, chickpea protein concentrate (CPC) was treated with electron beam irradiation (EBI) at doses of 5, 10, 15, and 20 kGy. After irradiation, protein solubility first increased at 10 kGy and 15 kGy, and then decreased at the higher dose of 20 kGy. This was supported by SDS-PAGE, where the intensity of major protein bands first increased and then decreased. Increased doses of EBI generally led to greater oxidative modification of proteins in CPC, indicated by reduced sulfhydryls and increased carbonyls. In addition, the protein structure was modified by EBI as shown by Fourier transform infrared spectroscopy analysis, where α-helix generally decreased, and β-sheet increased. Although the protein digestibility was not significantly affected by EBI, the peptidomic analysis of the digests revealed significant differences among CPC irradiated with varying doses. A total of 337 peptides were identified from CPC irradiated with 0 kGy, 10 kGy, and 20 kGy, with 18 overlapping peptides and 60, 29, and 40 peptides specific to the groups of 0, 10, and 20 kGy respectively. Theoretical calculation showed that the distribution of peptide length, hydrophobicity, net charge, and C-terminal residues were affected by irradiation. The 2, 2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activity showed a marginal decrease with an increasing dose of irradiation. In conclusion, EBI led to oxidative modification and structural changes in chickpea protein, which subsequently affected the physicochemical properties of peptides obtained from in-vitro digestion of CPC, despite similar digestibility.

In order to clarify the individual role of freezing and frozen storage on the quality of fish, fillets of large-mouth bass ( Micropterus salmoides ) were subjected to different freezing rates (freezing with -18 °C (A), -60 °C (B), and -60 °C with forced air circulation at 2 m/s (C), respectively) followed by frozen storage at -18 °C for 30 and 90 days. Another two groups were frozen at -60 °C, followed by storage at -40 °C (D) and -60 °C (E), respectively. Results showed that water-holding and TVBN were mainly affected by storage time. No significant changes were found in free thiol content among treatments. A greater freezing rate and lower storage temperature generally led to lower TBARS. GC × GC-TOFMS revealed a total of 66 volatile compounds, which were related to lipid oxidation. PLS-DA showed that fresh samples were separated from the frozen-thawed ones, and fillets in groups D and E were relatively close to fresh fillets in the composition of oxidation-related volatiles. In conclusion, freezing rate and storage temperature had a significant impact on lipid oxidation and protein denaturation in the fillets of large-mouth bass, while protein oxidation was more affected by freezing rate.

The objective of this study was to elucidate the influence of oxidative modifications of myofibrillar proteins (MPs) on their surface properties. Oxidative modifications (deamination, formation of disulfide bonds and Schiff bases), particle size, net surface charge, and binding ability of volatiles (2-enthylfuran, 1-octen-3-ol, hexanal, and octanal) of oxidized MPs was measured. Molecular docking of volatiles with actomyosin was performed using Qvina-W program and the specific oxidative modifications (monoxidation and deamination) of MPs were determined using LC-MS/MS. Results showed that oxidation of Cys (forming sulfinic, sulfonic, sulfenic acid, and disulfide bonds), monoxidation of Ala, Lys, Glu, and Asn, and deamination of Lys changed the surface properties of oxidized MPs including enhanced surface hydrophobicity and decreased affinity to volatile compounds and water. Overall, this study gives evidence of how protein oxidation affects the properties of MPs and therefore deteriorates fish meat quality., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Noninfectious liver injury, including the effects of chemical material, drugs and diet, is a major cause of liver diseases worldwide. In chemical and drugs-induced liver injury, innate inflammatory responses are mediated by extracellular danger signals. The S100 protein can act as danger signals, which can promote the migration and chemotaxis of immune cells, promote the release of various inflammatory cytokines, and regulate the body's inflammatory and immune responses. However, the role of S100A6 in inflammatory response in chemical and drugs-induced sterile liver injury remains unclear. We constructed the model of sterile liver injury induced by carbon tetrachloride (CCl 4 )/Paracetamol (APAP) and performed RNA sequencing (RNA-seq) on the liver tissues after injury (days 2 and 5). We analyzed inflammatory protein secretion in the liver tissue supernatant by enzyme-linked immunosorbent assay (ELISA), determined the inflammation response by bioinformatic analysis during sterile liver injury, and assessed mononuclear/macrophage infiltration by immunohistochemistry and flow cytometry. Immunohistochemistry was used to analyze the location of S100A6. We conducted inflammatory factor expression analysis and molecular mechanistic studies in Kupffer cells (KCs) induced by S100A6 using quantitative reverse transcription-polymerase chain reaction (qRT-PCR), ELISA, and western blot in vitro experiments. We performed chemokine CCL2 expression analysis and molecular mechanism studies using the same method. We used a Transwell assay to show the infiltration of mononuclear/macrophage. We here observed that aggravated inflammatory response was shown in CCl 4 and APAP-administrated mice, as evidenced by enhanced production of inflammatory cytokines (TNF-α, IL-1β), and elevated mononuclear/macrophage infiltration and activation of immunity. The expression of S100A6 was significantly increased on day 2 after sterile liver injury, which is primarily produced by injured liver cells. Mechanistic studies established that S100A6 activates Kupffer cells (KCs) via the p-P38, p-JNK and P65 pathways to induce inflammation in vitro. Furthermore, TNF-α can stimulate liver cells via the p-P38 and p-JNK pathways to produce CCL2 and promote the infiltration of mononuclear/macrophage. In summary, we showed that S100A6 plays an important role in regulating inflammation, thus influencing sterile liver injury. Our findings provide novel evidence that S100A6 can as a danger signal that contributes to pro-inflammatory activation through p-P38 and p-JNK pathways in CCl 4 and APAP-induced sterile liver injury in mice. In addition, the inflammatory factor TNF-α induces a large amount of CCL2 production in normal liver cells surrounding the injured area through a paracrine action, which is chemotactic for blood mononuclear/macrophage infiltration., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

In order to study the effect of oxidation on the dye-binding behavior of myofibrillar proteins, selected dyes with different charges (positively charged Sarfarin O (SO), neutral bromophenol blue (BPB), and negatively charged Orange G (OG)) were incubated with myofibrils oxidized by the Fenton system with H 2 O 2 (10 mM). Upon oxidation, loss of free thiols, formation of carbonyls, particle size, and hydrophobicity of myofibrillar proteins (MPs) increased. The absolute value of Zeta-potential increased by 14.48 % after oxidation, the myofibrillar proteins shifted to a more acidic isoelectric point (pI) upon oxidation. Oxidation decreased net positive charges of myofibrillar protein and the binding ability of MPs towards OG in the environment with pH less than pI and the affinity of MPs towards SO in the environment with pH more than pI were thus increased. Here we propose a hypothesis that oxidation-induced change in net charges is the driving force affecting the amount of protein-bound dye. This paper aims to examine the effect of oxidation on the net charges of myofibrillar proteins and to provide insight into the mechanism of oxidation-induced changes in protein-bound dyes., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The reduction of the salt content of foods is of interest for health reasons. The present study showed that low-salt surimi gels could be produced using a combination of l-arginine (Arg) and transglutaminase (TGase) under microwave (MW) irradiation. The low-salt surimi gels produced by MW had similar physicochemical properties as regular-salt surimi gels produced by conventional two-stage water bath heating. Compared to low-salt surimi gels containing TGase alone, Arg increased the water holding capacity and gel strength (P < 0.05) and promoted the formation of a more homogeneous and compact three-dimensional gel network. As a result, there was a significant increase in the proportion of immobilized water and decrease in the proportion of free water (P < 0.05). The nature of the binding sites between Arg and myosin heavy chain was predicted using the molecular docking simulations analysis. These results may be useful for the development of low-salt surimi products., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Plasma is a new technology used to modify myofibrillar proteins (MPs) structure and promote protein aggregation. In order to study the mechanism of plasma modifying MPs thus the effects on qualities of MP gels, MPs were extracted by 0.6 M NaCl solution prepared with plasma-activated water (PAW) at different treatment time (0 s, 30 s, 60 s, 120 s, 240 s). With the prolonged PAW treatment time from 0 to 240 s, the pH values of natural MP solutions decreased significantly from 5.91 to 2.61 ( P  < 0.05), the H 2 O 2 concentration in PAW increased from 0 to 70.82 μg/L ( P  < 0.05), and the net negative charges of MPs first decreased and then increased ( P  < 0.05). In addition, PAW caused significantly ( P  < 0.05) weakened ionic bonds and enhanced hydrophobic interactions, which promoted the aggregation and gelation of MPs thus forming MP gel with higher gel strength and a denser three-dimensional network. Furthermore, Raman spectra and intrinsic fluorescence suggested that PAW promoted the unfolding of MP structures and transformation from α-helixes and random coils to β-sheets and β-turns. Dynamic rheology indicated a gradually increased storage modulus and shortened degradation time of MPs with an increasing treatment time of PAW. Furthermore, PAW modification significantly improved the water holding capacity of MPs gels. These results demonstrated that the declined pH of MP solutions induced by PAW and increased H 2 O 2 in PAW altered the ζ-potential of MP solutions and promoted the unfolding and aggregation of MPs during heating via hydrophobic interactions, ultimately enhancing gelling properties of MPs. The present work suggested the potential use of PAW in preparing freshwater MP gels with high quality., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2022 The Authors.)

Ethnopharmacological Relevance: Honghua Qinggan 13 Flavor Pills (HHQG), whose Mongolian name is Guri Gumu-13, is a traditional Mongolian medicine, that was stated in the "Diagnosis and Treatment of Ming Medical Code". The HHQG has been included in the Mongolian Medicine Division of the Ministry of Health Drug Standards (1998 edition). Based on our clinical expertise, HHQG demonstrated satisfactory therapeutic effects in hepatitis and liver failure. However, the pharmacological effects and potential mechanisms of HHQG have not been investigated., Aim of the Study: In this study, we combined network pharmacology, transcriptomics, and molecular biology to detect the underlying mechanism for the effect of HHQG on acute liver injury in mice., Materials and Methods: Network pharmacology was used to explore the pathways involved in the protective effect HHQG in acute liver injury. This effect was further verified by injecting carbon tetrachloride (CCl 4 ; 10 mL/kg, i.p.) to induce acute liver injury in mice. Serum markers of liver injury, morphology, histology, and monocyte/macrophage infiltration in the liver tissue were investigated. Transcriptomics further defined the HHQG targets. Transwell analysis was performed to confirm that HHQG inhibited monocyte/macrophage RAW.264.7 infiltration. qPCR and Western blot were performed to explore the mechanism of action of HHQG., Results: Network pharmacology showed that HHQG exerted anti-oxidative and anti-inflammatory effects and promoted metabolic effects against acute liver injury. Pretreatment of mice with HHQG significantly maintained their body weight and decreased serum tumor necrosis factor-alpha (TNF-α) levels induced by CCl 4 treatment in vivo. Histopathological examination further confirmed that HHQG protected the liver cells from CCl 4 -induced damage. Importantly, HHQG significantly inhibited CCl 4 -induced monocyte/macrophage infiltration. Transcriptomic analysis revealed that HHQG significantly reduced the expression of chemokines and cell adhesion molecules. We determined that HHQG significantly downregulated the expression of the key chemokine (monocyte chemokine protein-1, CCL2) at the gene and protein levels. Further research showed that HHQG inhibited chemokine production in hepatocytes by inhibiting the p-P38 and p-JNK pathways, thereby reducing monocyte/macrophage infiltration., Conclusions: These combined data showed that HHQG alleviated acute liver injury in mice, and further verified that HHQG exerted protective effects by inhibiting the production of CCL2 and reducing the infiltration of monocyte/macrophage by inhibiting the p-P38 and p-JNK pathways., (Copyright © 2022 Elsevier B.V. All rights reserved.)

In this study, blueberry anthocyanins, gelatin and Fe 2+ were incorporated into zein matrix via electrospinning method to prepare colorimetric indicator films for monitoring milk freshness. Gelatin and Fe 2+ were incorporated into the film to improve visual discrimination of indicator films' color changes in milk with different freshness degrees and in solution with pH 3-7. Results of SEM, FT-IR and XRD showed that there were intermolecular hydrogen bonds among components, which associated with the larger color difference of indicator films. UV-vis spectral analysis showed that blueberry anthocyanin solutions containing both gelatin and Fe 2+ displayed the highest intensity absorption peaks. The optimal ability to distinguish the pH (3-7) of solutions was presented by the indicator film incorporating gelatin (1% (w/v)) and Fe 2+ (0.07 mg/mL). Gelatin and Fe 2+ increased the color-responsive sensitivity of the indicator film to pH. The film could be successfully used to detect the freshness of milk, whose color changes were visually perceivable: from purple black (fresh milk) to royal purple (spoiling milk) and then to violet red (spoiled milk). The color parameters ( L* , a* , R, G and B) of the film revealed a high correlation with the pH/acidity of the milk during storage. The successful application of the indicator film embedding gelatin and Fe 2+ for monitoring milk quality changes indicated that the addition of special substances could provide great potential for monitoring freshness and preparing intelligent packaging of food., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2022 The Authors. Published by Elsevier B.V.)

Background: Exploration of the genes with abnormal expression during the development of breast cancer is essential to provide a deeper understanding of the mechanisms involved. Transcriptome sequencing and bioinformatics analysis of invasive ductal carcinoma and paracancerous tissues from the same patient were performed to identify the key genes and signaling pathways related to breast cancer development., Methods: Samples of breast tumor tissue and paracancerous breast tissue were obtained from 6 patients. Sequencing used the Illumina HiSeq platform. All. Only perfectly matched clean reads were mapped to the reference genome database, further analyzed and annotated based on the reference genome information. Differentially expressed genes (DEGs) were identified using the DESeq R package (1.10.1) and DEGSeq R package (1.12.0). Using KOBAS software to execute the KEGG bioinformatics analyses, enriched signaling pathways of DEGs involved in the occurrence of breast cancer were determined. Subsequently, quantitative real time PCR was used to verify the accuracy of the expression profile of key DEGs from the RNA-seq result and to explore the expression patterns of novel cancer-related genes on 8 different clinical individuals., Results: The transcriptomic sequencing results showed 937 DEGs, including 487 upregulated and 450 downregulated genes in the breast cancer specimens. Further quantitative gene expression analysis was performed and captured 252 DEGs (201 downregulated and 51 upregulated) that showed the same differential expression pattern in all libraries. Finally, 6 upregulated DEGs (CST2, DRP2, CLEC5A, SCD, KIAA1211, DTL) and 6 downregulated DEGs (STAC2, BTNL9, CA4, CD300LG, GPIHBP1 and PIGR), were confirmed in a quantitative real time PCR comparison of breast cancer and paracancerous breast tissues from 8 clinical specimens. KEGG analysis revealed various pathway changes, including 20 upregulated and 21 downregulated gene enrichment pathways. The extracellular matrix-receptor (ECM-receptor) interaction pathway was the most enriched pathway: all genes in this pathway were DEGs, including the THBS family, collagen and fibronectin. These DEGs and the ECM-receptor interaction pathway may perform important roles in breast cancer., Conclusion: Several potential breast cancer-related genes and pathways were captured, including 7 novel upregulated genes and 76 novel downregulated genes that were not found in other studies. These genes are related to cell proliferation, movement and adhesion. They may be important for research into breast cancer mechanisms, particularly CST2 and CA4. A key signaling pathway, the ECM-receptor interaction signal pathway, was also identified as possibly involved in the development of breast cancer., Competing Interests: Competing interestsThe authors declare that they have no competing interests.

Protein oxidation readily occurs in postmortem muscle during storage and processing. Over the past decade new analytical methods have been developed and new aspects of protein oxidation in meat have been studied, such as the reaction mechanism, and impacts on eating quality and nutritional value. It is now evident that amino acid side chains in myofibrillar proteins undergoes modifications due to oxidative stress. In turn this will lead to formation of new protein-protein cross-links in structural proteins, however, also the overall level of fixed-charge groups attached to the peptide backbones is modified. Meat texture and water-holding are important quality attributes and they are affected by the oxidation of structural proteins. Different mechanisms have been suggested to explain the oxidation-induced quality changes, focusing mainly on reduced proteolysis and formation of cross-links. This review explores the current understanding of protein oxidation in fresh meat in relation to texture and water-holding. The consequences of protein oxidation at molecular level in relation to oxidation-induced cross-linking and changes in net charges of myofibrillar proteins, and the impacts on texture and water-holding are discussed.