Effects of Whey Peptides on the Quality of Pork Ball Preprepared Dishes during Repeated Freezing-Thawing.

This study evaluated the effect of FI (Fraction I, molecular weight < 1 kDa), which is separated from natural whey protein, on the antioxidant activity, sensory quality, color, texture characteristics, and microbial growth of pork balls during repeated freeze-thaw cycles (F-T cycles). The results indicated that pork balls mixed with FI significantly improved in quality after repeating the F-T cycle, especially with the addition of 10% FI. The quality was improved significantly after repeated F-T cycles by adding 10% FI, and the antioxidant activity after seven F-T cycles decreased by 40.78%, a similar result to that obtained with the addition of 0.02% BHA. In addition, FI effectively reduced the sensory damage of pork balls caused by repeated freezing-thawing and also significantly inhibited the growth of microorganisms. In summary, FI not only has excellent antioxidant capacity under repeated freeze-thaw conditions but also has significant antibacterial and quality preservation effects and is expected to be quantified as a kind of natural food additive with antibacterial and antioxidant properties. This paper not only explores the effect of FI on the quality characteristics of frozen and thawed pork balls in prepared dishes but also provides a theoretical basis for the application of whey polypeptides in prepared meat.

A Minispidroin Guides the Molecular Design for Cellular Condensation Mechanisms in S. cerevisiae.

Structural engineering of molecules for condensation is an emerging technique within synthetic biology. Liquid-liquid phase separation of biomolecules leading to condensation is a central step in the assembly of biological materials into their functional forms. Intracellular condensates can also function within cells in a regulatory manner to facilitate reaction pathways and to compartmentalize interactions. We need to develop a strong understanding of how to design molecules for condensates and how their in vivo-in vitro properties are related. The spider silk protein NT2RepCT undergoes condensation during its fiber-forming process. Using parallel in vivo and in vitro characterization, in this study, we mapped the effects of intracellular conditions for NT2RepCT and its several structural variants. We found that intracellular conditions may suppress to some extent condensation whereas molecular crowding affects both condensate properties and their formation. Intracellular characterization of protein condensation allowed experiments on pH effects and solubilization to be performed within yeast cells. The growth of intracellular NT2RepCT condensates was restricted, and Ostwald ripening was not observed in yeast cells, in contrast to earlier observations in E. coli. Our results lead the way to using intracellular condensation to screen for properties of molecular assembly. For characterizing different structural variants, intracellular functional characterization can eliminate the need for time-consuming batch purification and in vitro condensation. Therefore, we suggest that the in vivo-in vitro understanding will become useful in, e.g., high-throughput screening for molecular functions and in strategies for designing tunable intracellular condensates.

Recombinant protein condensation inside E. coli enables the development of building blocks for bioinspired materials engineering - Biomimetic spider silk protein as a case study.

Recombinant expression of proteins destined to form biological materials often results in poor production yields or loss of their function due to premature aggregation. Recently, liquid-liquid phase separation has been proposed as a mechanism to control protein solubility during expression and accumulation in the cytoplasm. Here, we investigate this process in vivo during the recombinant overexpression of the mimetic spider silk mini-spidroin NT2RepCT in Escherichia coli. The protein forms intracellular liquid-like condensates that shift to a solid-like state triggered by a decrease in their microenvironmental pH. These features are also maintained in the purified sample in vitro both in the presence of a molecular crowding agent mimicking the bacterial intracellular environment, and during a biomimetic extrusion process leading to fiber formation. Overall, we demonstrate that characterization of protein condensates inside E. coli could be used as a basis for selecting proteins for both materials applications and their fundamental structure-function studies.

Major yolk protein from sea cucumber (Stichopus japonicus) attenuates acute colitis via regulation of microbial dysbiosis and inflammatory responses.

Inflammatory bowel disease afflicted individuals and most medications have adverse effects. The objection of this study is to investigate whether the major yolk protein (MYP) could aid in the remission of colitis. The function of MYP on acute colitis was assessed through a dextran sulfate sodium -induced colitis mice model. Compared to the model group, the anti-inflammatory cytokines increased significantly in the MYP group, whereas the pro-inflammatory cytokines were not significantly different between the model and treatment group. The results also showed that supplementation of MYP improved the shift in microbial community composition of mice with colitis induced by DSS. In addition, MYP supplementation enriched the contents of fecal short-chain fatty acids. The alleviation of MYP on the colitis was probably related to repair the dysbiosis state of colonic microbiota, which thus induced an increase in short-chain fatty acids level and secrete anti-inflammatory cytokines (IL-4 and IL-10). In sum, oral MYP may be a potential candidate for the attenuating of acute colitis.

The Optimal Second-Line Systemic Treatment Model for Recurrent and/or Metastatic Head and Neck Squamous Cell Carcinoma: A Bayesian Network Meta-Analysis.

Background: The optimal second-line systemic treatment model for recurrent and/or metastatic head and neck squamous cell carcinoma (R/M HNSCC) remains controversial. A Bayesian network meta-analysis (NMA) was performed to address this issue with regard to efficacy and toxicity. Methods: By searching MEDLINE (via PubMed), Embase, the Cochrane Central Register of Controlled Trials and Web of Science, we extracted eligible studies. Efficacy, represented as overall survival (OS) and progression-free survival (PFS), and overall toxicity, represented as ≥ grade 3 severe acute events (sAE), were assessed to compare the following 7 treatment models through an NMA: standard-of-care therapy (SoC), single targeted therapy different from SoC (ST), double targeted therapy (DT), targeted therapy combined with chemotherapy (T+C), single immune checkpoint inhibitor therapy (SI), double immune checkpoint inhibitor therapy (DI) and single chemotherapy different from SoC (SC). Rank probabilities according to the values of the surface under the cumulative ranking curve (SUCRA) were separately determined for efficacy and toxicity. Results: In total, 5285 patients from 24 eligible studies were ultimately screened, with 5184, 4532 and 4026 involved in the NMA of OS, PFS and sAE, respectively. All qualifying studies were absent from first-line immune checkpoint inhibitor therapy. In terms of OS, SI was superior to the other treatments, followed by DI, ST, T+C, SoC, DT and SC. Other than SI and SC, all treatments tended to be consistent, with hazard ratios (HRs) close to 1 between groups. For PFS, ST ranked first, while DT ranked last. For the toxicity profiles, compared with the other models, SI resulted in the lowest incidences of sAE, with statistical significance over SoC (odds ratio [OR] 0.31, 95% credible interval [CrI] 0.11 to 0.90), ST (OR 0.23, 95% CrI 0.06 to 0.86) and DT (OR 0.11, 95% CrI 0.02 to 0.53), while DT was the worst. When the SUCRA values of OS and sAE were combined, a cluster plot illustrated the superiority of SI, which demonstrated the best OS and tolerability toward sAE. Conclusion: For R/M HNSCC patients without immune checkpoint inhibitors in the first-line setting, SI may serve as the optimal second-line systemic treatment model, demonstrating the best OS and least sAE.

Effect of geographic variation on the proteome of sea cucumber (Stichopus japonicus).

Sea cucumber is a sensitive organism that is easily challenged by environmental change. The aim of this study was to characterize the proteome of sea cucumbers from 5 main Chinese origins, including Xiamen (XM), Dalian (DL), Weihai (WH), Yantai (YT) and Qingdao (QD). In this work, a tandem mass tag (TMT) labeling proteomic approach was applied to identify and quantify the proteome of sea cucumber. A total of 5051 proteins were identified in the body wall; among those proteins, 1594 proteins (31.6%) were identified as enzyme proteins, and 33 proteins belonged to collagen. In addition, the 10 most highly abundant proteins were further discussed. Among all quantified proteins, 2266 were significantly differentially expressed proteins (SDEPs) across the 5 origins. These SDEPs were related to pigmentation (5 proteins), antioxidant activity (13 proteins), and immune system processes (29 proteins). Based on SDEPs, DL differed the most from QD and XM, as well as WH and YT, as shown in principal component analysis (PCA) and hierarchical clustering. In conclusion, one-fourth of the significantly different proteins found in the sea cucumber body wall among the 5 main Chinese locations indicated the sensitivity of sea cucumber to variations in temperature, environment, and feeding.

Effect of egg albumen protein addition on physicochemical properties and nanostructure of gelatin from fish skin.

The physicochemical properties and nanostructure of mixtures of egg albumen protein (EAP) and gelatin from under-utilised grass carp (Ctenopharyngodon idella) skins were studied. The gelatin with 1% EAP had an acceptable gel strength. The addition of 5% EAP significantly increased the melting and gelling temperatures of gelatin gels. Additionally, the colour turned white and the crystallinity was higher in gelatin gels with gradient concentrations of EAP (1, 3, and 5%). Gelatin with 5% EAP had the highest G' values while gelatin with 1% EAP had the lowest G' values. Atomic force microscopy showed the heterogeneous nanostructure of fish gelatin, and a simple coacervate with a homogeneous distribution was only observed with the addition of 1% EAP, indicating interaction between gelatin and EAP. These results showed that EAP effect fish gelatin's physicochemical and nanostructure properties and has potential applications in foods and pharmaceuticals.

Symmetric key structural residues in symmetric proteins with beta-trefoil fold.

To understand how symmetric structures of many proteins are formed from asymmetric sequences, the proteins with two repeated beta-trefoil domains in Plant Cytotoxin B-chain family and all presently known beta-trefoil proteins are analyzed by structure-based multi-sequence alignments. The results show that all these proteins have similar key structural residues that are distributed symmetrically in their structures. These symmetric key structural residues are further analyzed in terms of inter-residues interaction numbers and B-factors. It is found that they can be distinguished from other residues and have significant propensities for structural framework. This indicates that these key structural residues may conduct the formation of symmetric structures although the sequences are asymmetric.