Effects of multi-frequency ultrasound-assisted immersion freezing processing on myofibrillar protein structure and lipid oxidation of large yellow croaker (Larimichthys crocea) during long-time frozen storage.

In this study, large yellow croaker (Larimichthys crocea) was frozen using multi-frequency ultrasound-assisted freezing (MUIF) with different powers (160 W, 175 W, and 190 W, respectively) and stored at -18 °C for ten months. The effect of different ultrasound powers on the myofibrillar protein (MP) structures and lipid oxidation of large yellow croaker was investigated. The results showed that MUIF significantly slowed down the oxidation of MP by inhibiting carbonyl formation and maintaining high sulfhydryl contents. These treatments also held a high activity of Ca2+-ATPase in the MP. MUIF maintained a higher ratio of α-helix to ß-sheet during frozen storage, thereby protecting the secondary structure of the tissue and stabilizing the tertiary structure. In addition, MUIF inhibited the production of thiobarbituric acid reactive substances value and the loss of unsaturated fatty acid content, indicating that MUIF could better inhibit lipid oxidation of large yellow croaker during long-time frozen storage.

CO2 electrocatalytic reduction to ethylene and its application outlook in food science.

The efficient conversion of CO2 is considered to be an important step toward carbon emissions peak and carbon neutrality. Presently, great efforts have been devoted to the study of efficient nanocatalysts, electrolytic cell, and electrolytes to achieve high reactivity and selectivity in the electrochemical reduction of CO2 to mono- and multi-carbon (C2+) compounds. However, there are very few reviews focusing on highly reactive and selective ethylene production and application in the field of electrochemical carbon dioxide reduction reaction (CO2RR). Ethylene is a class of multi-carbon compounds that are widely applied in industrial, ecological, and agricultural fields. This review focuses especially on the convertibility of CO2 reduction to generate ethylene technology in practical applications and provides a detailed summary of the latest technologies for the efficient production of ethylene by CO2RR and suggests the potential application of CO2RR systems in food science to further expand the application market of CO2RR for ethylene production.

Advances in biomimetic hydrogels for organoid culture.

An organoid is a 3-dimensional (3D) cell culture system that mimics the structural and functional characteristics of organs, and it has promising applications in regenerative medicine, precision drug screening and personalised therapy. However, current culture techniques of organoids usually use mouse tumour-derived scaffolds (Matrigel) or other animal-derived decellularised extracellular matrices as culture systems with poorly defined components and undefined chemical and physical properties, which limit the growth of organoids and the reproducibility of culture conditions. In contrast, some synthetic culture materials have emerged in recent years with well-defined compositions, and flexible adjustment and optimisation of physical and chemical properties, which can effectively support organoid growth and development and prolong survival time of organoid in vitro. In this review, we will introduce the challenge of animal-derived decellularised extracellular matrices in organoid culture, and summarise the categories of biomimetic hydrogels currently used for organoid culture, and then discuss the future opportunities and perspectives in the development of advanced hydrogels in organoids. We hope that this review can promote academic communication in the field of organoid research and provide some assistance in advancing the development of organoid cultivation technology.

Apoptotic Changes, Oxidative Stress and Immunomodulatory Effects in the Liver of Japanese Seabass (Lateolabrax japonicus) Induced by Ammonia-Nitrogen Stress during Keep-Live Transport.

This study investigated the effects of NH3-N on antioxidant responses, histoarchitecture, and immunity of Japanese seabass (Lateolabrax japonicus) during keep-live transport. The findings suggest that NH3-N stress transport alters the transcription of P53, Caspase 9, Bcl2, Caspase 3 and Bax genes, demonstrating that NH3-N stress can trigger the apoptotic pathway of P53-Bax-Bcl2 and Caspase and induce apoptosis. NH3-N stress transport also evoked transcriptional upregulation of inflammatory cytokines (tumor necrosis factor α (TNF-α), Toll-like receptor 3 (TLR-3), nuclear factor kappa ß (NF-κB), interleukin 6 (IL-6) and interleukin 1ß (IL-1ß)) and increased complement C3, C4, lysozyme (LZM) and immunoglobulin (IgM) levels, activating the innate immunological system during keep-live transport. In addition, NH3-N stress transport altered changes in the levels of superoxide dismutase (SOD), catalase (CAT), glutathione-related enzymes, and heat shock proteins 70 and 90 in the liver, indicating that the antioxidant system and Hsp protected the cells from NH3-N-induced oxidative stress. When excess ROS were not removed, they caused the body to respond with immunological and inflammatory responses, as well as apoptosis and tissue damage. This helps towards understanding the effect of NH3-N levels on sea bass during keep-live transport.

Effect of different stunning methods on antioxidant status, myofibrillar protein oxidation, and gelation properties of large yellow croaker during postmortem.

Post-mortem muscle biochemical processes play a crucial role on fish fillets quality and they are strictly linked to stunning methods. The improper stunning methods before slaughter could cause the fish to deteriorate more quickly during cold storage. This study aimed to investigate the effect of stunning methods (hit on the head, T1; gill cut, T2; immersion in ice/water slurry, T3; CO2 narcosis, T4; 40% CO2 + 30 % N2 + 30% O2, T5) on myofibrillar proteins (MPs) of large yellow croaker. The results indicated that T2 and T3 samples were significantly damaged compared with other samples, which reflected that the activities of total superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) were significantly damaged during cold storage in T2 and T3 samples. And the gill cut and immersion in ice/water slurry resulted in the generation of protein carbonyl, the decrease of Ca2+-ATPase, free ammonia and protein solubility, and the production of dityrosine during storage. In addition, MPs gel of T2 and T3 samples showed the decrease of water hold capacity (WHC) and whiteness, structure destruction, and water migration. The T4 samples had the least damage of MPs and gel structure during cold storage.

Effective and stable adsorptive removal of Cadmium(II) and Lead(II) using selenium nanoparticles modified by microbial SmtA metallothionein.

Metallothionein SmtA-modified selenium nanoparticles (SmtA-SeNPs), efficient adsorbents for Cd(II) and Pb(II), were synthesized in the present work. The ligand, microbial SmtA protein, was synthesized using an engineered strain Escherichia coli, posing the benefits of simplicity, safety, and high production. SmtA-SeNPs were spheres with diameters between 68.1 and 122.4 nm, containing amino, hydroxyl, and sulfhydryl functional groups with negatively charged (pH > 5). SmtA-SeNPs displayed better adsorption performance than dissociative SmtA and SeNPs. The adsorption of Cd(II) and Pb(II) mainly depends on the electrostatic attractions and the metal chelation of abundant functional groups. The maximum adsorption capacity was 506.3 mg/g for Cd(II) and 346.7 mg/g for Pb(II), which were higher than the values of most nanoparticles. In addition, SmtA-SeNPs were immobilized with a membrane filter to produce a SmtA-SeNPs filter, and the percentage removal of Cd(II) and Pb(II) increased from 26.75% to 98.13% for Cd(II) and from 9.95% to 99.20% compared with the blank filter. Moreover, the SmtA-SeNPs filter was regenerated using subacid deionized water, and the filter exhibited a stable removal ratio of Cd(II) and Pb(II) in ten continuous cycles of Cd(II)- or Pb(II)-containing wastewater treatment. The residual amounts of Cd and Pb met national standard levels of wastewater discharge.

Identification of Potential Biomarkers for Liver Cancer Through Gene Mutation and Clinical Characteristics.

Liver cancer is a common malignant tumor worldwide, which is a serious threat to the health of people. We try to investigate some mutations and clinical indicators as candidate markers for the development of liver cancer through targeted region capture technology combined with next-generation sequencing. We collected peripheral blood and liver cancer tissue samples from 32 liver patients concurrently. The SeqCap EZ Prime Choice Probe was used to perform the targeted enrichment; this probe captures 1,000 known cancer-associated genes. We calculated the tumor mutation burden (TMB) for each patient. The high-frequency mutations and these relative genes were identified. Eventually, survival analysis was performed based on the mutations and clinical indicators. In 32 liver patients, a total of 29 high-frequency mutations were investigated. They were located in 25 genes, which were enriched in 9 cellular components (CCs), 6 molecular functions (MFs), and 21 biological processes (BPs). Among them, EZH2 c.1544A>G and CCND1 c.839A>T had the highest mutation frequency (5/32). In the protein-protein interaction (PPI) network, EZH2-DNMT3A, NOTCH1-CCND1, and ABL1-CCND1 were the top three pairs. The survival analysis showed that there were significant differences in progression-free survival (PFS) and overall survival (OS) between the Karnofsky performance score (KPS) groups. The PFS and OS in the TMB high group were higher than those in the TMB low group. OS and tumor stage had a remarkable relationship. In conclusion, EZH2 c.1544A>G and CCND1 c.839A>T might be potential biomarkers of liver cancer. TMB might be used as a prognosis and survival indicator of liver cancer.

Rubusoside-assisted solubilization of poorly soluble C6-Ceramide for a pilot pharmacokinetic study.

Although C6-Ceramide has attracted much attention as a possible tumor suppressor, the delivery of C6-Ceramide is still challenging due to its inherent hydrophobicity and insolubility. In this study we explored the use of a natural compound rubusoside (RUB) as a solubilizer to enhance the solubility of a fluorescence-labeled C6-Ceramide (NBD C6-Ceramide) and to characterize its pharmacokinetics and tissue distribution in an animal model. RUB significantly enhanced the solubility of NBD C6-Ceramide by forming nanomicelles, and efficiently delivered NBD C6-Ceramide in rats by oral and intravenous administration. RUB loaded 1.96 % of NBD C6-Ceramide in the nanomicelles and solubilized it to a concentration of 3.6 mg/mL in water. NBD C6-Ceramide in nanomicelles remained stable in aqueous solutions, allowing intravenous administration without the use of any organic solvents or surfactants. After oral administration, NBD C6-Ceramide rapidly rose to peak plasma concentrations within the first 90 min, distributed to tissues, and remained in vivo for more than 24 h. Tissular levels of NBD C6-Ceramide from high to low were associated with heart, lung, cerebellum, testicle, spleen, liver, kidney, and brain. Altogether, our study demonstrated that RUB-assisted nanomicelles can serve as an efficient and convenient delivery system for short-chain C6-Ceramide and enable in vivo evaluation of potential new cancer treatments.

Incorporation of Fluorescence Ceramide-Based HPLC Assay for Rapidly and Efficiently Assessing Glucosylceramide Synthase In Vivo.

Glucosylceramide synthase (GCS) is a rate-limiting enzyme catalyzing ceramide glycosylation, thereby regulating cellular ceramide levels and the synthesis of glycosphingolipids (GSLs) in cellular membranes. Alterations of GCS not only affect membrane integrity, but also closely correlate with stem cell pluripotency, cancer drug resistance, GSL storage disorders and other diseases. Enzyme activities measured conventionally with currently available ex-vivo methods do not enable reliable assessment of the roles played by GCS in vivo. We report herein a substrate-incorporation method enabling rapid and efficient assessment of GCS in-vivo activity. Upon nanoparticle-based delivery, fluorescent NBD C6-ceramide was efficiently converted to NBD C6-glucosylceramide in live cells or in mouse tissues, whereupon an HPLC assay enabled detection and quantification of NBD C6-glucosylceramide in the low-femtomolar range. The enzyme kinetics of GCS in live cells and mouse liver were well-described by the Michaelis-Menten model. GCS activities were significantly higher in drug-resistant cancer cells and in tumors overexpressing GCS, but reduced after silencing GCS expression or inhibiting this enzyme. Our studies indicate that this rapid and efficient method provides a valuable means for accurately assessing the roles played by GCS in normal vs. pathological states, including ones involving cancer drug resistance.

Applications of bacillus Calmette-Guerin and recombinant bacillus Calmette-Guerin in vaccine development and tumor immunotherapy.

Bacillus Calmette-Guerin (BCG) vaccines are attenuated live strains of Mycobacterium bovis and are among the most widely used vaccines in the world. BCG is proven to be effective in preventing severe infant meningitis and miliary tuberculosis. Intravesical instillation of BCG is also a standard treatment for non-muscle invasive bladder cancer. In the past few decades, recombinant BCG (rBCG) technology had been extensively applied to develop vaccine candidates for a variety of infectious diseases, including bacterial, viral, and parasite infections, and to improve the efficacy of BCG in bladder cancer therapy. This review is intended to show the vast applications of BCG and recombinant BCG (rBCG) in the prevention of infectious diseases and cancer immunotherapy, with a special emphasis on recent approaches and trends on both pre-clinical and clinical levels.

Transarterial chemoembolization using docetaxel-loaded phytantriol cubic phase precursor for the treatment of hepatocellular carcinoma.

The purpose of this study was to evaluate the transarterial chemoembolic agent based on docetaxel-loaded phytantriol cubic phase precursor (DTX PCPP) by in vitro cytotoxicity study and in vivo evaluation of antitumor efficacy as well as the histological examination. The methythiazolyl tereazolium bromide assay in Hep G2 cell line revealed that DTX PCPP generated high cytotoxicity by causing cell apoptosis and G2/M phase arrest. In vivo studies conducted in rabbits bearing VX2 tumors, which were treated with DTX PCPP, used as a transarterial chemoembolic agent, showed a significant antitumor efficacy and prominent higher DTX concentrations in tumor and liver than those in other organs. The histology presented typical necrosis in tumor that demonstrated excellent therapeutic effect. In conclusion, the DTX PCPP could achieve an excellent antitumor effect with low systemic toxicity for the treatment of hepatocellular carcinoma and therefore implied the prospect of DTX PCPP for clinical applications.