Isolation and Purification of Chitosan Oligosaccharides (Mw ≤ 1000) and Their Protective Effect on Acute Liver Injury Caused by CCl4.

Chitosan oligosaccharides are the degradation products of chitin obtained from the shell extracts of shrimps and crabs. Compared with chitosan, chitosan oligosaccharides have better solubility and a wider application range. In this study, high-molecular-weight chitosan oligosaccharides (COST, chitosan oligosaccharides, MW ≤ 1000) were isolated and purified by a GPC gel column, and the molecular weight range was further reduced to obtain high-purity and low-molecular-weight chitosan (COS46). Compared with COST, COS46 is better at inhibiting CCl4-induced cell death, improving cell morphology, reducing ALT content, and improving cell antioxidant capacity. The effects of COST and COS46 on CCl4-induced acute liver injury were further verified in mice. Both COS46 and COST improved the appearance of the liver induced by CCl4, decreased the levels of ALT and AST in serum, and decreased the oxidation/antioxidant index in the liver. From the liver pathological section, the effect of COS46 was better. In addition, some indicators of COS46 showed a dose-dependent effect. In conclusion, compared with COST, low-molecular-weight COS46 has better antioxidant capacity and a better therapeutic effect on CCl4-induced acute liver injury.

Effect of sugar reduction on sensory characteristics of dried fish mince product.

BACKGROUND: Excessive sugar consumption has been linked to type 2 diabetes, obesity and other diseases. Therefore, it is indispensable to reduce sugar of food. However, the sensory characteristics of food are affected after sugar reduction (SR). Currently, SR has been reported in drinks, jams, candies, and other fruit related or sweet foods; but salty or protein related foods have not been explored, therefore there is a big gap that needs to be filled. RESULTS: Sensory scores of initial sweetness and sweetness reduced by 0.26 and 0.12 in 10% SR dried fish mince product compared with control, and there was a small difference between 25% SR (3.33) and 40% SR (3.09) samples. It also showed that 10% SR sample had a small reduction in sweetness value and free sugar content by 3.5% (0.42/11.9) and 7.8% (2.12/27.06) compared with control; while values in 25% SR sample decreased sharply but were not much different from 40% SR sample. Electronic nose results showed that SR had a small effect on odor. Texture analysis showed that texture properties of 25% SR sample were significantly different from control. CONCLUSIONS: Dried fish mince product with below 10% SR had a small difference on the sensory characteristics and there was a big change when SR was more than 20%. Dried fish mince product with 25% SR and 40% SR had no significant difference. SR had a small effect on odor, but had a great effect on texture properties, especially over 20% SR. © 2023 Society of Chemical Industry.

Kinetics and mechanisms of thermal deterioration in silver carp (Hypophthalmichtys molitrix) surimi gel quality under high-temperature sterilization.

BACKGROUND: The gelation properties of surimi gel under various high temperatures (115, 118, and 121 °C) and sterilization intensities (F0 values of 3-7 min) were systematically investigated. A kinetic model detailed quality changes during heat treatment through mathematical analysis, elucidating mechanisms for gel quality degradation. RESULTS: Increased sterilization intensity significantly reduced the quality characteristics of surimi gel. Compared to the gel without sterilization treatment, when the sterilization intensity was increased to 7 min, the gel strength of the groups treated at 115 °C, 118 °C, and 121 °C decreased by 68.35%, 51.4%, and 51.71%, respectively, and the water-holding capacity decreased by 24.87%, 16.85%, and 22.5%, respectively. The hardness, chewiness, and whiteness of the gel also significantly decreased, and the changes in these indicators all conformed to a first-order kinetic model. Activation energy of 291.52 kJ mol-1 highlighted gel strength as the least heat-resistant. At equivalent sterilization intensities, 115 °C exhibited the poorest gel quality, followed by 121 °C, with 118 °C showing relatively better gel quality. Increased T22 and decreased PT22 suggested heightened water mobility and transition of immobilized water within the gel into free water. Protein degradation, weakened disulfide bonds and hydrophobic interaction, and protein conformation changes collectively led to a rough and incoherent gel network structure with large fissures, as verified by the results of scanning electron microscopy. Correlation analysis indicated potential for precise control over surimi gel quality by modulating physicochemical attributes. CONCLUSION: The outcomes may be beneficial to improve the production and quality control of ready-to-eat surimi-based products. © 2024 Society of Chemical Industry.

Study on the effect and mechanism of chicken breast on the gel properties of silver carp (Hypophthalmichtys molitrix) surimi.

BACKGROUND: Adding appropriate exogenous substances is an effective means to improve the quality of freshwater fish surimi. The present study investigated the effects of chicken breast on the gel properties of mixed minced meat products. RESULTS: With the increase in the proportion of chicken breast, the breaking force of mixed gels gradually increased. When the addition ratio was 30:70, the gel strength of mixed gels had the highest strength of 759.00 g cm-1 and also the highest water holding capacity of 87.36%. Compared with surimi gels (0:100), the hardness, adhesiveness and chewiness of mixed gels were significantly improved. The increase in the proportion of chicken breast increased the thermal stability of the mixed sol and improved the rheological properties of the mixed sol. When the proportion was 40:60, the area of immobile water (A22 ) in the mixed gel increased significantly, and the highest A22 was 3463.24. The hydrophobic interactions and disulfide bonds in the mixed gel were significantly increased as a result of the addition of chicken breast. The results of microstructure, electrophoresis and Raman spectroscopy indicated that the addition of chicken breast promoted the cross-linking of the proteins in mixed gels, which facilitated the transformation of the protein secondary structure from α-helical to ß-folded structure, thus forming a more uniform and orderly network structure. CONCLUSION: These results suggest that improving the gel properties of silver carp surimi by use of chicken breast has practical implications for the development of new blended products for surimi processing. © 2023 Society of Chemical Industry.

Physical exercise habits are related with reduced prevalence of falling among elderly women in China.

BACKGROUND: Although some studies have examined the association between exercise and falls, most have focused on specific exercises, and the results have been inconsistent. In addition, there is a lack of evidence on elderly Chinese women who have different living and exercise habits compared to those in other countries. Therefore, this study aimed to investigate whether physical exercise is associated with falls in elderly Chinese women. METHODS: This cross-sectional study included 1429 elderly Chinese women with a mean age of 69.2 years. Information on physical exercise habits and fall experiences was collected using a self-report questionnaire. Logistic regression models were used to analyze the association between physical exercise habits and falls. RESULTS: The results showed that 15% participants had a fall in the past year. After adjusting for confounding factors, the odd ratios (ORs) and 95% Confidence Intervals (CIs) for fall experiences across categories of exercise frequency were as follow: 1 (reference) for no exercise behavior, 0.50 (0.29, 0.85) for exercise 1 to 5 times a week, and 0.37 (0.25, 0.55) for exercise more than 6 times a week. Furthermore, the ORs (95% CIs) across categories of exercise insistence were 1 (reference) for less than 1 year, 0.78 (0.37, 1.65) for 1 to 3 years, and 0.38 (0.20, 0.74) for more than 3 years. In terms of exercise duration, the ORs (95% CIs) for < 1 h/day, 1-2 h/day, and > 2 h/day were 1 (reference), 0.85 (0.53, 1.36), and 2.80 (1.30, 6.05). Unlike other variables, longer exercise duration was associated unfavorably with falls. CONCLUSION: Physical exercise habits were associated with falls in elderly Chinese women. Keeping a proper exercise habit may contribute to lower risk of falling in elderly women.

Sulforaphane suppresses paraquat-induced oxidative damage in bovine in vitro-matured oocytes through Nrf2 transduction pathway.

Sulforaphane (SFN), a bioactive phytocompound extracted from cruciferous plants, has received increasing attention due to its vital cytoprotective role in eliminating oxidative free radical through activation of nuclear factor erythroid 2-related factor (Nrf2)-mediated signal transduction pathway. This study aims at a better insight into the protective benefit of SFN in attenuating paraquat (PQ)-caused impairment in bovine in vitro-matured oocytes and the possible mechanisms involved therein. Results showed that addition of 1 µM SFN during oocyte maturation obtained higher proportions of matured oocytes and in vitro-fertilized embryos. SFN application attenuated the toxicological effects of PQ on bovine oocytes, as manifested by enhanced extending capability of cumulus cell and increased extrusion proportion of first polar body. Following incubation with SFN, oocytes exposed to PQ exhibited reduced intracellular ROS and lipid accumulation levels, and elevated T-SOD and GSH contents. SFN also effectively inhibited PQ-mediated increase in BAX and CASPASE-3 protein expressions. Besides, SFN promoted the transcription of NRF2 and its downstream antioxidative-related genes GCLC, GCLM, HO-1, NQO-1, and TXN1 in a PQ-exposed environment, indicating that SFN prevents PQ-caused cytotoxicity through activation of Nrf2 signal transduction pathway. The mechanisms underlying the role of SFN against PQ-induced injury included the inhibition of TXNIP protein and restoration of the global O-GlcNAc level. Collectively, these findings provide novel evidence for the protective role of SFN in alleviating PQ-caused injury, and suggest that SFN application may be an efficacious intervention strategy against PQ cytotoxicity.

Research on apoptotic mechanism and related pathways involved in postmortem grass carp (Ctenopharyngodon idellus) muscle.

BACKGROUND: Apoptosis activation is an essential research to reveal the triggering mechanism of flesh quality deterioration. This study was aimed at explaining apoptotic mechanism of postmortem fish in terms of caspases activation, cytochrome c (cyt-c) release, B-cell lymphoma 2 (Bcl-2) and Bcl2-associated X (Bax) protein levels, transcriptional levels of its molecules, and apoptosis-inducing factor (AIF) translocation at 4 °C for 5 days. RESULTS: Activation of caspase-9, caspase-8, caspase-3 and the release of mitochondrial cyt-c were observed during storage. The decreased Bcl-2 protein levels, increased Bax protein expressions and Bax/Bcl-2 ratio were major steps for inducing apoptosis. Collectively, transcriptional regulation of Fas ligand (FasL), apoptotic protease activating factor-1 (Apaf-1), inhibitors of apoptosis proteins (IAPs), myeloid cell leukemia-1 (Mcl-1), c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (MAPK) indicated that extrinsic apoptotic pathways (FasL/caspase-8/caspase-3) and intrinsic pathway [(JNK and p38 MAPK)/(Bcl-2, Bax and Mcl-1)/cyt-c/Apaf-1/caspase-9/caspase-3] were involved in apoptotic process. Mitochondrial AIF translocation to nuclear indicated that AIF mediated caspase-independent pathway. CONCLUSION: Therefore, transcriptional and translational alterations of multiple signaling molecules acted important roles in regulating apoptosis activation in postmortem process. © 2022 Society of Chemical Industry.

The enhancer rare germline variation rs548071605 contributes to lung cancer development.

Rare germline variations contribute to the missing heritability of human complex diseases including cancers. Given their very low frequency, discovering and testing disease-causing rare germline variations remains challenging. The tag-single nucleotide polymorphism rs17728461 in 22q12.2 is highly associated with lung cancer risk. Here, we identified a functional rare germline variation rs548071605 (A>G) in a p65-responsive enhancer located within 22q12.2. The enhancer significantly promoted lung cancer cell proliferation in vitro and in a xenograft mouse model by upregulating the leukemia inhibitory factor (LIF) gene via the formation of a chromatin loop. Differential expression of LIF and its significant correlation with first progression survival time of patients further supported the lung cancer-driving effects of the 22q-Enh enhancer. Importantly, the rare variation was harbored in the p65 binding sequence and dramatically increased the enhancer activity by increasing responsiveness of the enhancer to p65 and B-cell lymphoma 3 protein, an oncoprotein that assisted the p65 binding. Our study revealed a regulatory rare germline variation with a potential lung cancer-driving role in the 22q12.2 risk region, providing intriguing clues for investigating the "missing heritability" of cancers, and also offered a useful experimental model for identifying causal rare variations.

Advances in the application of chitosan as a sustainable bioactive material in food preservation.

Chitosan is obtained from chitin and considered to be one of the most abundant natural polysaccharides. Due to its functional activity, chitosan has received intense and growing interest in terms of applications for food preservation over the last half-century. Compared with earlier studies, recent research has increasingly focused on the exploration of preservation mechanism as well as the targeted inhibition with higher efficiency, which is fueled by availability of more active composite ingredients and integration of more technologies, and gradually perceived as "chitosan-based biofilm preservation." In this Review, we comprehensively summarize the potential antimicrobial mechanisms or hypotheses of chitosan and its widely compounded ingredients, as well as their impacts on endogenous enzymes, oxidation and/or gas barriers. The strategies used for enhancing active function of the film-forming system and subsequent film fabrication processes including direct coating, bioactive packaging film and layer-by-layer assembly are introduced. Finally, future development of chitosan-based bioactive film is also proposed to broaden its application boundaries. Generally, our goal is that this Review is easily accessible and instructive for whose new to the field, as well as hope to advance to the filed forward.

The Microstructure, Antibacterial and Antitumor Activities of Chitosan Oligosaccharides and Derivatives.

Chitosan obtained from abundant marine resources has been proven to have a variety of biological activities. However, due to its poor water solubility, chitosan application is limited, and the degradation products of chitosan oligosaccharides are better than chitosan regarding performance. Chitosan oligosaccharides have two kinds of active groups, amino and hydroxyl groups, which can form a variety of derivatives, and the properties of these derivatives can be further improved. In this review, the key structures of chitosan oligosaccharides and recent studies on chitosan oligosaccharide derivatives, including their synthesis methods, are described. Finally, the antimicrobial and antitumor applications of chitosan oligosaccharides and their derivatives are discussed.

Knockdown of CLAUDIN-6 Inhibited Apoptosis and Induced Proliferation of Bovine Cumulus Cells.

This study aims to investigate the effects of CLAUDIN-6 (CLDN6) on cell apoptosis and proliferation of bovine cumulus cells (CCs). Immunofluorescence staining was used to localize CLDN6 protein in CCs. Three pairs of siRNA targeting CLDN6 and one pair of siRNA universal negative sequence as control were transfected into bovine CCs. Then, the effective siRNA was screened by real-time quantitative PCR (RT-qPCR) and Western blotting. The mRNA expression levels of apoptosis related genes (CASPASE-3, BAX and BCL-2) and proliferation related genes (PCNA, CDC42 and CCND2) were evaluated by RT-qPCR in CCs with CLDN6 knockdown. Cell proliferation, apoptosis and cell cycle were detected by flow cytometry with CCK-8 staining, Annexin V-FITC staining and propidium iodide staining, respectively. Results showed that the CLDN6 gene was expressed in bovine CCs and the protein was localized in cell membranes and cytoplasms. After CLDN6 was knocked down in CCs, the cell apoptosis rate significantly decreased and the pro-apoptotic genes BAX and CASPASE-3 were down-regulated significantly, whereas the anti-apoptotic gene BCL-2 was markedly up-regulated (p < 0.05). Additionally, CLDN6 knockdown significantly enhanced cell proliferation of CCs at 72 h after siRNA transfection. The mRNA levels of proliferation-related genes PCNA, CCND2 and CDC42 increased obviously in CCs with CLDN6 knockdown (p < 0.05). After CLDN6 was down-regulated, the percentage of CCs at S phase was significantly increased (p < 0.05). However, there was no remarkable difference in the percentages of cells at the G0/G1 phase and G2/M phase between CCs with or without CLDN6 knockdown (p > 0.05). Therefore, the expression of CLDN6 and its effects on cell proliferation, apoptosis and cell cycle of bovine CCs were first studied. CLDN6 low expression inhibited cell apoptosis, induced cell proliferation and cell cycle arrest of bovine CCs.

Effect of rainfall characteristics on the sewer sediment, hydrograph, and pollutant discharge of combined sewer overflow.

Significant management needs raised in urban sewer system to facilitate urban resilience to rainstorm. The work investigated the effects of temporal evolution of rainfall on hydrograph and pollutant discharge of CSO over an intensive observation period of 12 months, with special attention to differences in temporal scale for supporting management decision making. The characteristics of rainfall in different temporal scales helped overflow-risk identification and assessment. Prolonged dry seasons over 112 days in the CSO monitored year 2018 increased the sediment buildup in the pipes. The built sediment was mostly flushed out to overflow (and the treatment facility) by initial rainfall during 47 h. Following CSO hydraulics and pollutant discharge follows initial peak patterns which responded to rainfall patterns. Results of Redundancy analysis and network analysis showed that the combined effects of rainfall, urbanization, and sediments as "CSO troika" are the driving forces for CSO pollutants in the long-term. The improved characterization of CSO events and the associated pollutants has refined our understanding of how overflow hydrograph and pollutant discharge responds to rainfall temporally, which methodology supported decision making in the combining source/process control with terminal management for facilizing urban resilience.

FcRn is not the receptor mediating the transfer of serum IgG to colostrum in pigs.

In contrast to humans or rabbits, in which maternal IgG is transmitted to offspring prenatally via the placenta or the yolk sac, large domestic animals such as pigs, cows and sheep transmit IgG exclusively through colostrum feeding after delivery. The extremely high IgG content in colostrum is absorbed by newborns via the small intestine. Although it is widely accepted that the neonatal Fc receptor, FcRn, is the receptor mediating IgG transfer across both the placenta and small intestine, it remains unclear whether FcRn also mediates serum IgG transfer across the mammary barrier to colostrum/milk, especially in large domestic animals. In this study, using a FcRn knockout pig model generated with a CRISPR-Cas9-based approach, we clearly demonstrate that FcRn is not responsible for the IgG transfer from serum to colostrum in pigs, although like in other mammals, it is involved in IgG homeostasis and mediates IgG absorption in the small intestine of newborns.

Silencing of retrotransposon-derived imprinted gene RTL1 is the main cause for postimplantational failures in mammalian cloning.

Substantial rates of fetal loss plague all in vitro procedures involving embryo manipulations, including human-assisted reproduction, and are especially problematic for mammalian cloning where over 90% of reconstructed nuclear transfer embryos are typically lost during pregnancy. However, the epigenetic mechanism of these pregnancy failures has not been well described. Here we performed methylome and transcriptome analyses of pig induced pluripotent stem cells and associated cloned embryos, and revealed that aberrant silencing of imprinted genes, in particular the retrotransposon-derived RTL1 gene, is the principal epigenetic cause of pregnancy failure. Remarkably, restoration of RTL1 expression in pig induced pluripotent stem cells rescued fetal loss. Furthermore, in other mammals, including humans, low RTL1 levels appear to be the main epigenetic cause of pregnancy failure.

Recent advances in quality retention of non-frozen fish and fishery products: A review.

Fishery products are delicious, nutritional and healthy foods, and their popularity in today's market is growing. With the changes in consumer consumption expectations as well as the development of reliable cold-chains, fresh preprocessed fishery products such as fillets are more popular with consumers and producers because of their convenience for processing and cooking. However, fishery products after being removed from water have a short shelf life due to their intrinsic characteristics. Therefore, developing effective preservation technology to extend postmortem shelf life of fishery products has been an on-going area of study. This review summarizes the patterns of quality deterioration and external factors that affect postmortem quality of fishery products, and then updates the recent advances with preservation methods for fishery products including superchilling, modified atmosphere packaging, active packaging, edible coatings, irradiation and high hydrostatic pressure. Moreover, the advantages and disadvantages of each technology are discussed, and future trends for improving the preservation of fishery products are also discussed.

No imprinted XIST expression in pigs: biallelic XIST expression in early embryos and random X inactivation in placentas.

Dosage compensation, which is achieved by X-chromosome inactivation (XCI) in female mammals, ensures balanced X-linked gene expression levels between the sexes. Although eutherian mammals commonly display random XCI in embryonic and adult tissues, imprinted XCI has also been identified in extraembryonic tissues of mouse, rat, and cow. Little is known about XCI in pigs. Here, we sequenced the porcine XIST gene and identified an insertion/deletion mutation between Asian- and Western-origin pig breeds. Allele-specific analysis revealed biallelic XIST expression in porcine ICSI blastocysts. To investigate the XCI pattern in porcine placentas, we performed allele-specific RNA sequencing analysis on individuals from reciprocal crosses between Duroc and Rongchang pigs. Our results were the first to reveal that random XCI occurs in the placentas of pigs. Next, we investigated the H3K27me3 histone pattern in porcine blastocysts, showing that only 17-31.8% cells have attained XCI. The hypomethylation status of an important XIST DMR (differentially methylated region) in gametes and early embryos demonstrated that no methylation is pre-deposited on XIST in pigs. Our findings reveal that the XCI regulation mechanism in pigs is different from that in mice and highlight the importance of further study of the mechanisms regulating XCI during early porcine embryo development.

Correlations between microbiota succession and flavor formation during fermentation of Chinese low-salt fermented common carp (Cyprinus carpio L.) inoculated with mixed starter cultures.

The correlation between microbiota succession and flavor development in Chinese traditional fermented fish (Suanyu) inoculated with mixed starter cultures was studied. The results showed that 17 free amino acids, 22 free fatty acids and 9 organic acids were present. A total of 81 aroma compounds were detected during Suanyu fermentation. Aldehydes were the most abundant aroma compounds in the early stage of fermentation and esters contributed most to the aroma in the later stage of fermentation. The correlation analysis of microbial succession and flavor dynamics based on bidirectional orthogonal partial least squares (O2PLS) suggested that in addition to starter cultures, other microorganisms including 2 bacteria genera and 11 fungi genera were also responsible for the formation of flavor compounds. These results may help focus further research to improve the flavor quality of traditional fermented fish.

Exogenous glutathione improves intracellular glutathione synthesis via the γ-glutamyl cycle in bovine zygotes and cleavage embryos.

Excess reactive oxygen species (ROS) generated in embryos during in vitro culture damage cellular macromolecules and embryo development. Glutathione (GSH) scavenges ROS and optimizes the culture system. However, how exogenous GSH influences intracellular GSH and improves the embryo developmental rate is poorly understood. In this study, GSH or GSX (a stable GSH isotope) was added to the culture media of bovine in vitro fertilization embryos for 7 days. The cleavage rate, blastocyst rate, and total cell number of blastocysts were calculated. Similarly to GSH, GSX increased the in vitro development rate and embryo quality. We measured intracellular ROS, GSX, and GSH for 0-32-hr postinsemination (hpi) in embryos (including zygotes at G1, S, and G2 phases and cleaved embryos) cultured in medium containing GSX. Intracellular ROS significantly decreased with increasing intracellular GSH in S-stage zygotes (18 hpi) and cleaved embryos (32 hpi). γ-Glutamyltranspeptidase ( GGT) and glutathione synthetase ( GSS) messenger RNA expression increased in zygotes (18 hpi) and cleaved embryos treated with GSH, consistent with the tendency of overall GSH content. GGT activity increased significantly in 18 hpi zygotes. GGT and GCL enzyme inhibition with acivicin and buthionine sulfoximine, respectively, decreased cleavage rate, blastocyst rate, total cell number, and GSH and GSX content. All results indicated that exogenous GSH affects intracellular GSH levels through the γ-glutamyl cycle and improves early embryo development, enhancing our understanding of the redox regulation effects and transport of GSH during embryo culture in vitro.

Bio-based edible coatings for the preservation of fishery products: A Review.

The popularity of preprocessed fresh fishery products such as fillets and peeled shrimps is growing in today s market due to their convenience for subsequent processing and cooking. However, fishery products are highly perishable because of the combined actions of biochemical reactions and microbial metabolism. Various methods have been proposed to address this problem. Among these methods, bio-based edible coating has been highlighted as a promising solution. This review updates and summarizes the recent literature on the application of coatings for the preservation of fishery products including the aspects of coating carriers, composite natural preservatives and coating methods, and a discussion of the protective effects based on microbial, physicochemical and sensorial evaluations. Moreover, some challenges and future research directions regarding optimization of formulas and exploration of mechanisms of coating are also discussed. Given consumer demand for fresh fishery products with long shelf life, edible coatings that are environmentally friendly and effective alternative will be used to extend the shelf life of fishery products.

Editing porcine IGF2 regulatory element improved meat production in Chinese Bama pigs.

Insulin-like growth factor 2 (IGF2) is an important growth factor, which promotes growth and development in mammals during fetal and postnatal stages. Using CRISPR-Cas9 system, we generated multiple founder pigs containing 12 different mutant alleles around a regulatory element within the intron 3 of IGF2 gene. Crossing two male founders passed four mutant alleles onto F1 generation, and these mutations abolished repressor ZBED6 binding and rendered this regulatory element nonfunctional. Both founders and F1 animals showed significantly faster growth, without affecting meat quality. These results indicated that editing IGF2 intron 3-3072 site using CRISPR-Cas9 technology improved meat production in Bama pigs. This is the first demonstration that editing non-coding region can improve economic traits in livestock.