Search Results (17,295)

In this study, the ultrasonic-assisted extraction of silkworm pupae protein (SPP) was optimized using response surface methodology. Subsequently, the effects of ultrasonic treatment on the structural and functional characteristics of SPP were systematically analyzed and verified through Pearson correlation analysis. The results showed that the optimal extraction parameters were an ultrasonic treatment time of 120 min, a power of 115 W, a temperature of 54 °C, pH of 10.5, and the average extraction yield was 68.087%. Compared to the control, ultrasonic treatment significantly improved the functional properties of SPP, including solubility (13.13 g/L), water holding capacity (0.18%), oil holding capacity (0.28%), foaming capacity (55.35%), foam stability (12.71%), emulsification activity (2.15 m²/g), emulsification stability (21.95%), gel water holding capacity (11.5%), gel hardness (1.02 N), and gel elasticity (0.49 mm). In addition, the adsorption ability of SPP for 2-octanone and aldehyde was enhanced after ultrasonic treatment. Furthermore, the absorption intensity and maximum wavelength of the SPP fluorescence spectrum extracted via ultrasonic treatment were enhanced, along with the increased surface hydrophobicity and more stable secondary structure which contributed to promoting the functional properties of SPP, proven by Pearson correlation analysis. This study provides a theoretical basis for the further utilization of SPP in the food industry. Full article

Selenium is an important trace element with certain antioxidant effects. Nano-selenium, as a novel selenium source, has the advantages of strong biological activity, high absorption efficiency, and low toxicity. The aim of the present study was to compare the protective effects of sodium selenite and nano-selenium on intestinal oxidative stress induced by hydrogen peroxide (H₂O₂) in mice. A total of 60 female mice were randomly divided into 6 groups with 10 replicates per group and 1 mouse per replicate (n = 10). The first three groups were as follows: the Control group (C), fed with basal diet; the sodium selenite group (SS), basal diet + 0.3 mg·kg⁻¹ sodium selenite; and the nano-selenium group (NS), basal diet + 0.3 mg·kg⁻¹ nano-selenium. The latter three groups (CH, SSH, NSH) were fed the same diet as the former three groups, but the last 10 days of the experiment were fed with drinking water containing 0.3% H₂O₂ to induce oxidative stress. The results showed that under normal conditions, the supplementation with sodium selenite or nano-selenium decreased the spleen index of mice; sodium selenate up-regulates GPX3 expression in the ileum, and increases T-SOD in the colon of mice; and nano-selenium up-regulated GPX1 expression but decreased T-AOC in the jejunum. After drinking water treated with H₂O₂, H₂O₂ increased the expression of intestinal inflammatory factors and selenium proteins, such as IL-1β and SOD in jejunum, IL-1β, NF-κB, IL-10, TXNRD1, TXNRD2, GPX1, GPX3, GPX4, and CAT in ileum, and IL-1β and SOD in colon. At the antioxidant level, H₂O₂ decreased T-AOC in the jejunum. In the H₂O₂ treatment, sodium selenite and nano-selenium increased the ratio of VH to CD (VH/CD) in jejunum; sodium selenite up-regulated the expression of TXNRD1 in jejunum, down-regulated the expression of GPX3 in ileum, at the antioxidant level, decreased the T-SOD and T-AOC in colon, and increased the content of MDA in ileum; and nano-selenium down-regulated the expression of TXNRD1 in colon. At the same time, the expression of IL-1β, NF-κB, IL-10, TXNRD1, TXNRD2, GPX1, GPX4, and CAT can be restored to normal levels by selenium supplementation. According to the results, drinking H₂O₂ induced intestinal oxidative stress in mice to a certain extent, and selenium supplementation mitigated the destructive effect of H₂O₂ on the intestinal morphology of mice jejunum and restored the level of related inflammatory factors, and had a positive effect on antioxidants. Full article

This study compared the effects of two C3 forages (Italian ryegrass [RG], Timothy grass [TG]) and two C4 forages (Klein grass [KG], Bermuda grass [BG]) on growth performance, nutrient digestibility, and nitrogen (N) balance in Korean crossbred black goats to evaluate C4 warm-season forages as alternatives under changing climate conditions. Sixteen castrated goats (10 months old) were allocated to four treatments using a randomized complete block design. Diets contained 40% treatment-specific forage and 60% commercial concentrate. After adaptation periods, a 5-day metabolism trial measured performance and digestibility parameters. No significant differences occurred among treatments for growth performance or digestibility of dry matter, crude protein, neutral detergent fiber, ash, and non-fiber carbohydrate. C4 grasses showed significantly lower acid detergent fiber and ether extract digestibility than C3 grasses, with KG having the lowest ether extract digestibility. The KG group had higher N intake and absorption than the TG group, while BG showed lower urinary and total N excretion than KG. No differences existed in retained N, utilization efficiency, or biological value among groups. Both C3 and C4 forages supported comparable goat growth performance, providing a reference for utilizing different photosynthetic pathway forages under changing climatic conditions. Full article

The environmental problems brought about by factory-based aquaculture have become increasingly prominent. Reducing nitrogen and phosphorus concentrations in tailwater has become the key to tailwater management. In order to assess the potential of microalgae in removing nitrogen and phosphorus ions from aquaculture wastewater, four microalgae species, i.e., Chlorella sp., Dicrateria zhanjiangensis, Nitzschia closterium minutissima, and Platymonas subcordiformis, were used in this study, and their growth and nitrogen and phosphorus removal rates in four nutrient concentrations of simulated aquaculture wastewater were systematically evaluated. After 15 days of cultivation, the cell counts of all four types of microalgae increased. Three species, i.e., Chlorella sp., N. closterium minutissima, and P. subcordiformis, grew best in high PO₄³⁻ and low NH₄⁺ medium, whereas D. zhanjiangensis possessed best growth in low PO₄³⁻ and high NH₄⁺ medium. The removal rate of PO₄³⁻, NH₄⁺, NO₃⁻, and NO₂⁻ by four microalgae species exceeded 82.64%, 89.06%, 59.27%, and 42.15%, respectively, even though the four microalgae had different performance in the removal of nitrogen and phosphorus. All microalgae in the low-phosphorus groups removed PO₄³⁻ at significantly lower rates than those in the high–phosphorus groups, while high NH₄⁺ removal rates were observed in all four microalgae groups. Moreover, in phosphorus-limited conditions, four microalgae exhibit lower removal rates of NO₃⁻ when nitrogen content was high. The chlorophyll a contents of microalgae in four culture media strictly corresponded to their final cell densities. P. subcordiformis exhibited the highest intracellular polysaccharide accumulation in high PO₄³⁻ and low NH₄⁺ type medium, whereas D. zhanjiangensis demonstrated the strongest protein synthesis capacity in high PO₄³⁻ and low NH₄⁺ medium. The activities of acid phosphatase in all microalgae were higher under phosphorus–deficient conditions than phosphorus-sufficient conditions. Our results might provide useful references for microalgae selection in the treatment of different aquaculture wastewater conditions. Full article

Bacillus cereus AS_3 and Bacillus cereus AS_5 are bacterial endophytes isolated from sterilized leaves of the medical plant Alectra sessiliflora, which were previously identified using 16S rRNA sequencing. Here, we present the whole-genome sequencing and annotation of strains AS_3 and AS_5, the first genome report of Bacillus cereus strains from A. sessiliflora. The genome of strain AS_3 has 59 contigs, 5 503 542 bp draft circular chromosome, an N₅₀ of 211,274 bp, and an average G+C content of 35.2%; whereas strain AS_5 has 38 contigs, 5,510,121 bp draft circular chromosome, an N₅₀ of 536,033 bp, and an average G+C content of 35.2%. A total of 5679 protein-coding genes, 62 genes coding for RNAs, and 122 pseudogenes in the strain AS_3 genome were identified by the National Center for Biotechnology Information Prokaryotic Annotation pipeline, whereas a total of 5688 gene protein-coding genes were identified in AS_5, with 60 genes coding for RNAs and 120 pseudogenes. Phenotypic analysis and whole-genome sequencing analysis showed that AS_3 and AS_5 share similar characteristics, including Gram-positive, motile, rod-shaped, and endospore-forming have shown a high sequence similarity with Bacillus cereus, type strain ATCC 14579^T. Strains AS_3 and AS_5 had genomic digital DNA–DNA hybridization (dDDH) with the type strain Bacillus cereus ATCC 14579^T of 85.8% and 86%, respectively, and average nucleotide identities (ANIs) of 98% and 98.01%, respectively. Phylogenomic analysis confirmed that strains AS_3 and AS_5 share very similar genomic and phenotypic characteristics, and are closely related to the type strain Bacillus cereus type strain ATCC 14579^T, supporting their classification within the Bacillus cereus species. A total of 10 secondary metabolite gene clusters, including siderophore type petrobactin, terpene type molybdenum cofactor, non-ribosomal peptide synthetase (NRPS) type bacillibactin, and β-lactone type fengycin, were predicted using AntiSMASH software (version 5.0). Putative genes potentially involved in bioremediation and endophytic lifestyle were identified in the genome analysis. Genome sequencing of Bacillus cereus AS_3 and Bacillus cereus AS_5 has provided genomic information and demonstrated potential biotechnological applications. Full article

Organic fertilisers release nutrients more slowly than mineral fertilisers, which is why combining organic and mineral fertilisation gives good results in crop cultivation. In the conducted pot experiment, the reaction of oats to compost fertilisation with or without additional nitrogen mineral fertilisation was examined. The following treatments were used: A, control (no fertilisation); B, compost (sewage sludge 80% + sawdust 20%); C, compost (garden and park waste 80% + sawdust 20%); D, compost (sewage sludge 40% + garden and park waste 40% + sawdust 20%); E, compost B with nitrogen fertilisation (30 N kg ha⁻¹); F, compost C with nitrogen fertilisation (30 N kg ha⁻¹); and G, compost D with nitrogen fertilisation (30 N kg ha⁻¹). The study results indicated that the composts used had an altering impact on the soil’s chemical composition by the end of the experiment. Overall, the lowest levels of nutrients were recorded in the control group, indicating that the composts increased soil fertility. Oat plants were better nourished (SPAD—soil–plant analysis development) after fertilisation with sewage sludge composts than garden and park waste composts. However, the most favourable results were obtained in the treatments where organic fertilisation (composts) was combined with mineral fertilisation (nitrogen). All fertilisation treatments significantly enhanced plant height and the number of panicles in the pot compared to the control. The highest values for the number of grains in the panicle, thousand-grain weight, grain mass from the pot, and protein content in the grain were observed after applying organic–mineral fertilisation. Therefore, fertilisation with composts, especially composts combined with mineral nitrogen, can be recommended for oat cultivation. Full article

Glycosylation of many proteins has been revealed to be closely related to food allergy, and screening and structural analysis of related glycoproteins are essential for studies in this important area. Gly m Bd 30K is one of the major allergens that exist in soybeans. N-Glycans of the Gly m Bd 30K influenced the immunoreactivity and antigen-presenting efficiency. In this paper, soybean allergen glycoprotein Gly m Bd 30K was used as the research object. It was separated and purified by the combination of isoelectric point and Sepharose CL-6B gel. The glycoprotein was analyzed and identified by SDS-PAGE and MALDI-TOF MS. The N-glycans of Gly m Bd 30K glycoprotein were released and labeled by a newly developed one-pot method, and qualitatively and quantitatively analyzed by ESI-MSⁿ and HILIC-UV-MS/MS. The results showed that the purity of Gly m Bd 30K glycoprotein was 95%, and the relative molecular mass was 33,923 Da. The Gly m Bd 30K glycoprotein contained a total of six kinds of glycans, including two types: oligo-mannose type (4.3%) and paucimannose type (95.7%). The paucimannose modified with core α-1,3-fucose and β-1,2-xylose accounted for 92.87%. This study provides quality-reliable materials for the follow-up study of glycan sensitization and also provides a theoretical basis for the in-depth study of the specificity and biological function of the antigenic determinant of Gly m Bd 30K glycoprotein. Full article

H5N1 Influenza A virus continues to pose a significant zoonotic threat, with increasing evidence of interspecies transmission and genetic adaptation. Previous studies primarily focused on avian or human isolates, with limited comprehensive analysis of H5N1 evolution across multiple mammalian hosts. Existing molecular surveillance often lags behind viral evolution; this study underscores the necessity for real-time monitoring of ongoing mutations affecting pathogenicity and transmissibility. Our goals are (1) to retrieve and analyze HA, NP and NA gene sequences of H5N1 Influenza A virus from diverse hosts, including humans, poultry and multiple mammalian species, to assess genetic diversity and evolutionary patterns and (2) to evaluate positive selection sites across the three major genes (HA, NP and NA) to determine adaptive mutations linked to host adaptation and viral survival. To achieve these goals, in this study, we considered (78 HA), (62 NP) and (61 NA) gene sequences from diverse hosts, including humans, poultry and multiple mammalian species, retrieved from the NCBI database. Phylogenetic analysis revealed distinct clade formations, indicating regional spread and cross-species transmission events, particularly from avian sources to mammals and humans. Selection pressure analysis identified positive selection across all three genes, suggesting adaptive mutations contributing to host adaptation and viral survival. Homology modeling and molecular dynamics simulations were performed to generate high-quality structural models of HA, NP and NA proteins, which were subsequently validated using multiple stereochemical parameters. Domain analysis confirmed conserved functional motifs, while protein–ligand docking demonstrated stable interactions at conserved binding sites, despite observed residue substitutions in recent isolates. Earlier research concentrated on HA alone; this study integrates HA, NP and NA genes for a broader understanding of viral evolution and adaptation. These findings highlight ongoing evolutionary changes in H5N1 genes that may enhance viral adaptability and pathogenicity, underscoring the need for continuous molecular surveillance and updated antiviral strategies. Full article

Salt–alkaline soil poses a significant challenge to soybean productivity. While plant growth-promoting rhizobacteria (PGPR) offer a sustainable strategy for stress mitigation, their field-level application remains underexplored. Here, a field experiment was conducted in the Yellow River Delta of Shandong, China, a typical salt–alkaline region. In this study, we evaluated the effectiveness of Bacillus velezensis 41S2 in enhancing soybean performance under salt–alkaline soil through integrated field trials and transcriptomic analysis. Inoculation with strain 41S2 significantly improved plant biomass, yield components, and seed yield under salt–alkaline soil, and notably increased seed protein and isoflavone contents. Physiological analyses revealed that strain 41S2 markedly reduced hydrogen peroxide (H₂O₂) accumulation, indicating alleviation of oxidative stress. Moreover, strain 41S2 modulated the levels of soluble sugars and amino acids, contributing to osmotic regulation and carbon–nitrogen (C-N) metabolic balance. Transcriptome profiling further indicated that strain 41S2 upregulated genes involved in antioxidant response, C–N metabolism, and phenylpropanoid biosynthesis, highlighting its role in coordinating multilayered stress response pathways. Overall, these findings highlight the potential of B. velezensis 41S2 as a multifunctional bioinoculant for improving salt tolerance and presents a promising tool for sustainable crop production and ecological restoration in salt–alkaline soil. Full article

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is a widespread enzymopathy affecting approximately 500 million individuals that represents a significant global health issue. Among the more than 230 identified mutations in the G6PD gene, six class A variants—G6PD Utrecht (Pro409Ser), G6PD Suwalki (Pro409Arg), G6PD Merlo (Pro409Gln), G6PD Kawasaki (Gly410Ala), G6PD Shinagawa (Gly410Asp), and G6PD Riverside (Gly410Cys)—are located in the beta-loop near the NADP+ binding site. These mutations are of particular interest due to their association with severe hematologic phenotypes, including chronic hemolytic anemia, as well as their proposed role in the allosteric regulation of G6PD multimerization. This study presents a comprehensive biochemical and functional characterization of these clinically relevant G6PD variants. The variant enzymes were cloned, expressed, and purified for characterization. Kinetic parameters and thermal stability assays, complemented by molecular dynamics simulations (MDS), were employed to elucidate the structural impacts of the mutations. Our results demonstrate that these mutations significantly impair protein function, characterized by reduced affinity for glucose-6-phosphate (G6P) and NADP⁺, as well as altered thermal stability compared with wild-type G6PD. MDS revealed that point mutations in the βN- and βM-sheets in the NADP⁺s region propagate subtle conformational changes, ultimately affecting the NADP⁺c region and the G6P binding cavity. Furthermore, secondary structure element analyses of the simulation data showed that Pro409 and Gly410 point mutations propagate several changes around residues 195–210 (G6P binding site) and 380–400 (NADP⁺s), explaining their effect on overall catalytic performance. These findings enhance our understanding of the molecular mechanisms underlying G6PD deficiency and its clinical implications, providing a foundation for future therapeutic strategies aimed at mitigating the effects of these variants. Full article

Tropaeolum tuberosum (mashua) is a native Andean tuber recognized for its high nutritional and bioactive compound content. Among the various morphotypes, the black and yellow variants show potential differences in composition and functionality. This study aimed to compare the thermo-energetic, nutritional, and physicochemical characteristics of two morphotypes (black and yellow) of Tropaeolum tuberosum flour from the Peruvian Andes. Flours were obtained from tubers harvested in Ayacucho, Peru, and analyzed using elemental analysis for carbon, hydrogen, nitrogen, and sulfur (CHNS), inductively coupled plasma optical emission spectrometry (ICP-OES), scanning electron microscopy (SEM), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), and bomb calorimetry. The empirical formula is CH_1.74O_0.91N_0.06S_0.005 for black mashua and CH_1.78O_0.92N_0.05S_0.005 for yellow mashua. Black flour exhibited higher protein (17.6% vs. 14.8%) and fat contents (8.0% vs. 6.7%), along with nearly double the iron content. Both flours showed similar starch granule morphology and gelatinization enthalpy (~2 J/g), but the black flour had higher gelatinization temperatures. Calorimetric analysis revealed a greater net calorific value (qNCV) in black mashua flour (4157 ± 22 kcal/kg) than in yellow flour (4022 ± 19 kcal/kg). The thermogravimetric profiles indicated good thermal stability with approximately 30% residual mass. These findings suggested that black mashua flour possesses superior nutritional and energy characteristics, supporting its application in functional food formulations and energy-rich gluten-free products. Full article

Influenza virus is a leading cause of global morbidity and mortality due to acute lower respiratory infection, even with the widespread use of multiple licensed influenza vaccines. However, antigenic drift during influenza replication can cause vaccine-induced antibodies to poorly neutralize influenza virus, thereby reducing vaccine effectiveness. To help overcome this problem, we leveraged a hydrogel platform with influenza hemagglutinin (HA) protein to induce prolonged antigen exposure. The hydrogel platform, Vaccine Self-Assembling Immune Matrix (VacSIM^®), in combination with recombinant influenza H1 or H3 HA protein antigens, increased antigen-specific antibody titers in vaccinated mice, which led to decreased disease severity after H1N1 infection for H1 HA-vaccinated mice and decreased lung viral titers after H3N2 challenge for H3 HA-vaccinated mice. Sera collected from mice immunized with VacSIM and HA also showed broader HAI activity, increasing by 1–3 log against a panel of influenza viruses. These results were consistent with the use of cocktail immunization, containing both an H1 and H3 HA, where mice immunized with VacSIM had an increase in antigen-specific antibody titers and decreased disease severity and lung viral titers against H1N1 and H3N2 influenza challenges, respectively. Finally, it was determined that a single immunization with VacSIM and H1 HA could provide protection against lethal H1N1 challenge compared to a group without VacSIM. In summary, we demonstrate that use of the slow-release platform VacSIM can improve the host immune response to vaccination and increase protection against influenza infection. Full article

The BRAF protein regulates cell growth and division through key signaling pathways. Mutations in BRAF, particularly the V600E variant, are frequently observed in colorectal cancer (CRC) and are associated with poor prognosis and therapeutic challenges. Tumors harboring certain BRAF mutations often exhibit primary resistance to BRAF inhibitor monotherapies. Over time, these tumors can also develop acquired resistance, further complicating treatment. In this study, we employed replica exchange molecular dynamics simulations combined with machine learning techniques to investigate the structural alterations induced by BRAF mutations and their contribution to drug resistance. Our analyses revealed that conformational changes in mutant BRAF proteins associated with dabrafenib residues psi494, phi600, phi644, phi663, psi675, and phi677 were sufficient for classifying drug-resistant vs. drug-sensitive variants. Similarly, for vemurafenib, residues psi450, phi484, phi495, phi518, psi622, and phi622 were the key residues that influence drug binding and resistance mechanisms. These residues are located in the N-lobe of CR3, which is responsible for ATP binding and the regulation of BRAF kinase activity. These findings offer deeper insights into the molecular basis of BRAF-driven resistance and provide predictive models for phenotypic outcomes of various BRAF mutations. The study underscores the importance of targeting specific BRAF variants for more effective, personalized therapeutic strategies in drug-resistant CRC patients. Full article

Microbial proteins offer a sustainable alternative for animal nutrition. Rossellomorea marisflavi NDS, a bacterium isolated from seawater, was previously identified as a promising candidate due to its high protein content. This study aimed to enhance its single cell protein production through systemic fermentation optimization. Single-factor optimization in shake flask determined the optimal conditions to be: a salinity of 20‰ NaCl, a temperature of 32 °C, and an initial pH of 7.3, and a medium composed of 1% (w/v) corn flour, 1% peptone, 0.3% beef extract, and 0.2% KCl. Scaling up to a 10 L bioreactor demonstrated that a two-stage agitation strategy (150 rpm for the first 20 h followed by 180 rpm for the remaining 12 h) enhanced single cell protein yield. Furthermore, allowing the pH to fluctuate freely was more beneficial for protein production than maintaining a constant pH of 7.3 ± 0.02. Under these optimized conditions, the biomass composition (wet weight) was determined to be 2.3767 ± 0.0205% crude ash, 15.6013 ± 0.0082% crude protein, 0.1023 ± 0.0026% crude lipid, and 2.6997 ± 0.0021% carbohydrates. Amino acid analysis revealed a rich profile, with lysine and glutamic acid being the predominant essential and non-essential amino acids, respectively. Fatty acids analysis indicated that C14:1n5 was the most dominant. These findings underscore the potential of R. marisflavi NDS as a high-quality dietary protein supplement and provide a solid foundation for its industrial-scale production. Full article

Mealworm (Tenebrio molitor, LINNAEUS, 1758) is a protein-rich edible insect. In this study, low-gluten wheat flour was formulated with mealworm protein powder at various concentrations (0%, 5%, 10%, 15%, and 20%) to investigate its influence on the pasting, farinographic, and extensographic properties of low-gluten wheat flour, as well as the changes in the overall quality of the resulting biscuits (soda crackers and cookies). The viscosity of the composite flour decreased with an increasing substitution level of mealworm protein powder, and the setback significantly decreased from 69.31 ± 0.16 RVU (M0) to 19.00 ± 0.71 RVU (M20), indicating enhanced resistance to starch retrogradation. Farinographic and extensographic analyses revealed that the addition of mealworm protein powder reduced dough water absorption, significantly prolonged dough development time and stability time, and enhanced overall dough stability. However, extensibility gradually decreased, with a further reduction observed as the proofing time increased. Concurrently, the baking expansion ratio and hardness of the biscuits decreased. Specifically, for soda crackers, the baking expansion ratio decreased from 198.96 ± 3.88% (M0) to 135.74 ± 1.28% (M20), and hardness dropped from 26.40 ± 1.53 N (M0) to 6.32 ± 0.08 N (M20). For cookies, the baking expansion ratio and hardness decreased from 93.77 ± 0.72% (M0) to 86.06 ± 1.08% (M20) and from 1.76 ± 0.06 N (M0) to 1.10 ± 0.16 N (M20), respectively. The impact of mealworm protein powder (5–20%) was relatively minor in cookies but more pronounced in soda crackers, likely due to differences in formulation and processing methods. Additionally, the crunchiness of soda crackers was 3.42 times greater than that of cookies, whereas resilience was only 0.15 times that of cookies under controlled conditions. Pearson correlation analysis and principal component analysis (PCA) further elucidated the relationships between the dough properties and final product quality. Furthermore, the substitution of mealworm protein powder affected the sensory properties of the product but significantly enhanced its nutritional value, confirming the feasibility of replacing low-gluten wheat flour with mealworm protein powder and offering a theoretical foundation for its development and application in diverse biscuit formulations. Full article