Search Results (29,129)

Assemblysomes are ribosome-nascent chain condensates that regulate co-translational processes through liquid–liquid phase separation, yet the sequence determinants underlying their formation remain incompletely understood. Previous studies identified the DNA helicase Sgs1 as an assemblysome-associated protein; however, whether its N-terminal region possesses intrinsic phase separation propensity has not been experimentally examined. Here, we investigated the first 135 amino acids of Sgs1 using a light-inducible optoDroplet assay. A mCherry–Cry2–Sgs1_1–135 fusion construct was compared with the established positive control FUS–mCherry–Cry2 and the negative control mCherry–Cry2 in live HEK293T cells. Following blue-light activation, Sgs1_1–135 reproducibly formed reversible condensates, indicating intrinsic phase separation propensity. Quantitative image analysis revealed light-dependent increases in condensate number, average condensate area, and integrated condensate fluorescence intensity. Compared with FUS, Sgs1_1–135 formed slightly fewer and smaller condensates but displayed reproducible light-dependent condensate formation. These findings indicate that the Sgs1 N-terminal region exhibits intrinsic phase separation propensity in a validated optogenetic assay. Although this proof-of-principle study does not establish the molecular mechanism of assemblysome formation, the results are consistent with the hypothesis that the Sgs1 N-terminus may contribute to the multivalent interactions underlying assemblysome organization. Full article

The development of structurally stable and nutritionally enhanced gluten-free bread is a major technological challenge due to the absence of a gluten network. This study investigates the role of zein, a hydrophobic corn prolamin, in green buckwheat dough systems and its impact on bread properties. Zein was extracted from corn gluten and incorporated at 0, 10, 20, and 30%. Zein incorporation significantly modified starch–protein interactions, promoting a composite matrix with enhanced thermomechanical stability. The 20% zein formulation showed optimal performance, with increased peak viscosity, higher C2 and C3 torque values, improved starch gelatinization, and a more cohesive microstructure. The scanning electron microscopy analysis confirmed a continuous protein–starch network, with the highest elasticity and cohesiveness (0.85 and 0.57, respectively). The optimized formulation maintained functional value and reduced the conditional glycemic index. In the 20% zein formulated sample, the rapidly digestible starch fraction decreased from 48.1% to 41.6%, while slowly digestible and resistant starch increased from 22.4% and 7.8% to 26.3% and 10.4%, respectively. Overall, zein enrichment effectively compensates for the absence of gluten, resulting in improved bread quality and controlled starch digestibility. These findings provide new insights into protein–starch interactions in gluten-free systems and support the application of zein as a functional structuring agent in pseudocereal-based bakery products. Full article

Bigels combine the advantages of hydrogel and oleogel matrices, enabling simultaneous encapsulation of both hydrophilic and lipophilic compounds with good ductility and water retention. Bigels were prepared from large yellow croaker myofibrillar protein (MP)–soybean oil oleogels and κ-carrageenan hydrogels. All the bigels exhibited a milky white, solid-like self-supporting structure. Increasing the oleogel fraction enhanced droplet interactions and uniformity. Hardness peaked at 11.67 g at a 5:5 (w/w) oleogel-to-hydrogel ratio. The rheological tests revealed shear-thinning behavior across all samples, with apparent viscosity increasing progressively with oleogel content. The highest thixotropic recovery (71.07%) was observed at an oleogel-to-hydrogel ratio of 6:4 (w/w). All bigels underwent structural destabilization above 40 °C and complete liquefaction above 60 °C, indicating thermal reversibility. This study elucidates how varying the oleogel-to-hydrogel ratio modulates the physicochemical properties of bigels, providing a theoretical basis for developing high-value products from large yellow croaker. Full article

Background: Cashmere goat coat types determine cashmere quality and economic value. Elucidating the molecular regulatory mechanisms is crucial for targeted breeding and the development of high-quality cashmere products. Methods: Skin tissues from Jinlan cashmere goats with two distinct coat types (CHSC and CHLC) were collected at three hair follicle developmental stages—anagen (AN), catagen (CA), and telogen (TE)—for RNA-seq. Results: A total of 178 differentially expressed (DE) lncRNAs and 267 DE mRNAs were screened in the AN phase, 62 DE lncRNAs and 93 DE mRNAs in the CA phase, and 65 DE lncRNAs and 158 DE mRNAs in the TE phase. GO and KEGG functional enrichment analyses showed that DE target mRNAs at the AN stage were mainly enriched in processes of material and energy metabolism and in protein glycosylation modification pathways. In the CA stage, DE target mRNAs were primarily enriched in immune-related GO terms and signaling pathways, including immune response, cytokine activity, cytokine–cytokine receptor interaction, IL-17 signaling pathway, and NF-kappa B signaling pathway. The Wnt signaling-related GO terms and pathways were enriched in the TE phase, which is closely associated with the maintenance of hair follicle quiescence and follicle cycle transition. Furthermore, ceRNA network analysis identified several key regulatory axes corresponding to different developmental stages, including the lncRNAs-chi-miR-671-5p-ADIPOQ, XR_001918825.1-chi-miR-128-5p-RFX2, and TCONS_00062482-chi-miR-874-3p-PGLYRP1. Conclusions: These findings advance our understanding of hair follicle biology and provide potential molecular targets for improving cashmere production through genetic selection. Full article

The intestinal mucosa is the primary site of Toxoplasma gondii (T. gondii) infection and interaction with the host. While the intestinal responses have been characterized through transcriptomic and histopathological studies, a comprehensive, system-wide analysis of the functional proteome remodeling by acute T. gondii infection is required to elucidate the underlying mechanisms. This study presents a temporal, quantitative proteomic profiling of T. gondii-infected C57BL/6J mouse intestine to define the protein-centric host response. Global analysis reveals profound reprogramming, characterized by upregulated acute-phase reactants and interferon-stimulated effectors and downregulated epithelial barrier and digestive function proteins. Systems-level bioinformatics analysis uncovers a coordinated host strategy of cellular resource reallocation, evidenced by the simultaneous and specific amplification of ribosome biogenesis and proteasomal degradation pathways. Subsequent protein–protein interaction network analysis substantiates this strategic investment in core protein homeostasis infrastructure, identifying the ribosome biogenesis machinery as the topological core of the response interactome. These findings support a model of defense-priority resource reallocation. That is, the host redirects cellular resources to optimize its biosynthetic and catabolic capacity, facilitating a high-output immune response at the potential expense of tissue homeostasis. This work provides an integrated proteomic atlas of the intestinal immunopathology mechanisms during toxoplasmosis and establishes a foundation for future host-directed therapeutic exploration. Full article

Background/Objectives: Osteoarthritis (OA) is a whole-joint disease, and cellular senescence is one of several processes associated with cartilage degeneration. This study aimed to identify senescence-associated differentially expressed genes in established OA and to examine checkpoint-related candidates without assuming a causal checkpoint-imbalance mechanism. Methods: Five Gene Expression Omnibus (GEO) datasets spanning articular-cartilage tissue and primary cartilage-derived chondrocytes (GSE57218, GSE117999, GSE114007, GSE246425, and GSE169077) were integrated as a training cohort; the meniscus dataset GSE98918 was reserved as an independent cross-tissue validation cohort. OA-associated differentially expressed genes (DEGs) were intersected with CELLAGE genes. Enrichment, protein–protein interaction, transcription-factor, competing endogenous RNA, drug-enrichment, and molecular-docking analyses were performed. CDK6 and WEE1 expression was evaluated in IL-1β-treated human C28/I2 chondrocytes by RT-qPCR and representative Western blotting. Results: Forty-one senescence-associated DEGs were identified, and seven network-central genes (CDKN1A, CDK6, WEE1, NFKB2, ID1, RBL2, and IGFBP7) were prioritized. CDKN1A, CDK6, and WEE1 were reduced in OA-associated meniscal samples in GSE98918; within-dataset ROC analyses yielded AUCs of 0.931, 0.882, and 0.792, respectively. In IL-1β-treated C28/I2 cells, WEE1 mRNA decreased whereas CDK6 mRNA increased; representative immunoblots showed concordant qualitative trends. Berberine- and folic acid-related docking findings were computational only. Conclusions: Checkpoint-related gene expression is associated with senescence-linked transcriptomic changes in established OA. The discordant CDK6 results between clinical tissue datasets and an acute inflammatory cell model do not establish stage-dependent regulation. The present data also do not demonstrate p53-mediated CDKN1A activation, checkpoint failure, cell-cycle arrest, or cellular senescence; these hypotheses require dedicated functional experiments. Full article

The shortage of protein-rich foods is a major challenge due to the rapid growth of the world’s population. For this reason, efforts are being made to achieve sustainability in the food system to produce nutritious foods with better qualities. The present study aimed to utilize a combination of widely consumed legumes with nutritional properties—such as lentils, chickpeas, and beans—considering their physicochemical and functional characteristics, to produce tempeh as a model. A simple mixture design was employed to develop a legume-based product through the production of fermented flour using Rhizopus oligosporus ATCC 22959. For this purpose, proximate composition, water absorption index (WAI), pH, phenolic content, and antioxidant capacity, via the DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)) radicals, were determined in the raw legumes. Optimal fermentation conditions were determined through digital image analysis, and fermentations were carried out according to the design. The crude and soluble protein content, phenolic content, and DPPH of the fermented samples were determined, and a statistical optimization was performed by maximizing each variable. Through optimization, it was found that a formulation of 80.81% lentil and 19.19% chickpea presented the best desirability (D = 0.75) according to the criteria mentioned above. These results were also compared with those obtained from the preparation of an original soy tempeh using the microorganism Rhizopus oligosporus; it was found that the protein differences between the original tempeh and the one made from the legume blend were 19 g/100 g dry matter for the original tempeh and 27.5 g/100 g dry matter for the one made with the legume blend, demonstrating that the combination of legumes exerts a favorable interaction within the mixture model on the physicochemical properties of tempeh and could represent significant potential for the production of flours applicable to the development of food products as part of alternative protein sources. Full article

Collagen functional fragments retain selected activities of parent collagens while allowing greater flexibility in sequence design and precision fermentation. Although recent reviews have covered recombinant collagen production technologies, expression platforms, purification strategies, quality control, and biomedical applications, fragment selection, host–process matching, production, and characterization have received less integrated attention. This review focuses primarily on collagen-derived functional fragments, while collagen-mimetic peptides and collagen-like proteins are discussed as related design systems. The biological basis for fragmentation includes receptor-recognition motifs, matrikines and matricryptins, and basement membrane-derived fragments. The review further examines how motif context, Gly-X-Y organization, stabilizing sequence features, protease susceptibility, post-translational modification requirements, and host compatibility influence fragment stability, expression performance, production feasibility, and product integrity. Microbial production using Escherichia coli, Komagataella phaffii, and Saccharomyces cerevisiae is discussed from the perspectives of construct–host matching, secretory or intracellular production, prolyl 4-hydroxylase configuration, fermentation optimization and scale-up, and product characterization. Finally, we discuss AI-assisted, quality-guided design-build-test-learn workflows that integrate computational prediction, curated structural, extracellular-matrix, interaction, and protease resources, two-tier candidate evaluation, and format-appropriate experimental testing to support iterative sequence, host, and process optimization. The development of collagen functional fragments therefore depends on coordinated optimization of biological function, molecular design, microbial host performance, fermentation processes, and product characterization. Full article

Bacterial diseases caused by Flavobacterium covae (Fc), Vibrio harveyi (Vh), Vibrio vulnificus (Vv) and Photobacterium damselae (Pd) seriously constrain Asian seabass aquaculture. Here we dissect the mucosal immune mechanisms engaged by a five-month sequential vaccination strategy that combines nanoemulsion immersion priming with multivalent oral hydrogel boosting. Juvenile seabass were vaccinated, then challenged with F. covae by freshwater immersion and with a Vibrio–Photobacterium (Vh/Vv/Pd) mix by immersion or intraperitoneal injection. Gills were sampled after immersion challenges and intestine after injection, and profiled by RNA sequencing and label-free quantitative proteomics, with selected genes validated by RT-qPCR. Principal component analysis showed clear separation of vaccinated and control fish in all tissues and challenges, indicating a strong and coherent transcriptional reprogramming. Vaccination markedly increased the number of upregulated genes, with Gene Ontology enrichment revealing dominant signatures of ribosome biogenesis, RNA processing, lysosomal organization and immune response. KEGG analysis highlighted cytokine receptor interaction; NOD and Toll-like receptor signaling; oxidative phosphorylation; and phagosome, lysosome and cell adhesion molecule pathways, consistent with heightened antimicrobial readiness. Volcano plots and focused heatmaps showed strong induction of interferon-stimulated genes, cytokines and chemokine receptors, complement components, macrophage mannose receptor, epithelial barrier mediators and numerous immunoglobulin transcripts, with tissue- and challenge-specific patterns. Proteomics corroborated these trends, demonstrating a higher abundance of immunoglobulin heavy chains, complement proteins, cathepsins, heat shock and redox chaperones, ribosomal proteins and cytoskeletal and adhesion regulators in vaccinated mucosae. Integrated pathway mapping linked endothelial adhesion molecules and leukocyte integrins with T cell costimulation networks and an intestinal immune network for immunoglobulin production, including enhanced pIgR-mediated transcytosis. Overall, the sequential vaccination regimen was associated with coordinated transcriptomic and proteomic signatures related to epithelial responses, innate immunity, and humoral immune functions across gill and intestinal tissues. These molecular patterns were accompanied by improved survival following bacterial challenge; however, the present data do not directly demonstrate the functional activity of the inferred immune mechanisms in Asian seabass. Full article

Integrated nutrient management reduces reliance on chemical fertilizers by combining organic and inorganic inputs. A field experiment was conducted under rainfed conditions in Hungary from 2022 to 2024 to evaluate organic, inorganic and biological fertilizers applied to the high-oleic sunflower hybrid ES Emeric. Seven treatments were compared, namely, an unfertilized control, potassium (K), combined organic and inorganic nitrogen (GOIM), effective microorganisms (EM-1), and the combinations K+GOIM, K+EM-1 and GOIM+EM-1. Seed oil, crude protein and moisture content were determined, together with the fatty acid profile of the oil. Growing season influenced every measured variable far more strongly than fertilization, and all treatment responses were expressed as year-by-treatment interactions. Mean oleic acid content was 69.9% in the dry season of 2022 and 85.3% in 2024, but only 28.9% in the cooler and wetter season of 2023, when linoleic acid reached 59.8%. In 2022, K and K+EM-1 gave the numerically highest oil contents, 48.7% and 48.0%, less than one percentage point above the control, while GOIM+EM-1, GOIM and K+GOIM gave significantly higher protein contents than the remaining treatments. In the same season, EM-1 and K+GOIM raised linoleic and alpha-linolenic acids and, therefore, total polyunsaturated fatty acids, whereas GOIM+EM-1 and K increased oleic acid and total monounsaturated fatty acids. Integrated fertilization can therefore be used to shift the balance between monounsaturated and polyunsaturated fatty acids in sunflower oil, but the size and direction of the shift are governed by the conditions of the growing season. Full article

Background: Colorectal cancer (CRC) remains one of the leading causes of cancer-related morbidity and mortality worldwide. Increasing evidence indicates that alterations in glycosylation and glycan-binding proteins, such as galectins, contribute substantially to colorectal carcinogenesis. We review the current knowledge on the role of known human galectins in CRC to provide up-to-date summaries, focusing on their involvement in tumor initiation, progression, invasion, angiogenesis, immune evasion, and treatment resistance, integrating molecular mechanisms with diagnostic, prognostic, and therapeutic perspectives. Methods: We searched for “colorectal cancer” and “Galectin”, “Galectin-X” and “colorectal cancer”, and “Gal-X” and “colorectal cancer” (X denotes the number of the respective galectin) in PubMed, ScienceDirect, Scopus, and Web of Science databases. All records were screened for relevance by the authors. Results: We summarize the expression patterns of individual galectins in colorectal tissues and circulation, their involvement in oncogenic signaling pathways, and their interactions with the tumor microenvironment. Furthermore, we critically evaluate the available evidence regarding their diagnostic, prognostic, and therapeutic potential. Conclusions: Although galectin-1 and galectin-3 are the most extensively investigated galectins, accumulating evidence suggests that galectin-4, galectin-8, and galectin-9 also play important context-dependent roles in CRC progression and immune regulation. However, methodological heterogeneity and limited clinical validation currently preclude the implementation of galectins as standalone biomarkers in routine practice. Future research should focus on standardized multicenter studies, clarification of context-dependent galectin functions, and the development of galectin-targeted therapies. A deeper understanding of galectin biology may facilitate the development of novel multimarker diagnostic approaches and personalized therapies for CRC patients. Full article

Fusarium sacchari is one of the major pathogenic fungi that cause sugarcane Pokkah Boeng disease (PBD). Effectors play pivotal roles in F. sacchari–sugarcane interaction; thus, characterizing these effectors is essential for elucidating the molecular mechanisms underlying F. sacchari pathogenicity and for developing effective strategies to control PBD. However, only a limited number of effectors have been functionally validated to date. Here, we report FsRGAE1, a candidate effector protein from F. sacchari predicted to encode a rhamnogalacturonan acetylesterase (RGAE). FsRGAE1 exhibits high expression during the early stages of infection and maintains relatively elevated expression levels throughout the F. sacchari–sugarcane interaction. Targeted deletion of the FsRGAE1 gene in F. sacchari had no discernible impact on mycelial growth, conidiation, or carbon-source utilization, yet it significantly attenuated fungal virulence. FsRGAE1 possesses both a signal peptide conferring secretory capacity and a transit peptide enabling its translocation into the host cytoplasm and nucleus. Using the Agrobacterium tumefaciens-mediated transient expression system in Nicotiana benthamiana, FsRGAE1 was confirmed to suppress cell death induced by Bcl-2-associated X protein (BAX), as well as ROS accumulation and callose deposition, and its nuclear localization is indispensable for this immunosuppressive activity. Collectively, these findings indicate that FsRGAE1 promotes F. sacchari virulence by suppressing host immune responses in a nuclear localization-dependent manner, providing new insights into effector-mediated F. sacchari pathogenesis and potential target for resistance breeding in sugarcane. Full article

The biological fate of lipid-based nanocarriers (LBNs) is shaped at the interface. Whereas core architecture governs cargo loading, protection, and baseline release, surface architecture mediates the carrier’s initial interactions with proteins, cells, extracellular matrices, and tissue barriers, thereby influencing colloidal stability, immune recognition, targeting, biodistribution, barrier transport, and release initiation. Layer-by-layer (LbL) engineering provides a modular strategy for programming this interface through sequentially assembled coatings in which functional components are spatially separated yet mechanistically coordinated. By integrating polymers, biomolecules—including peptides and nucleic acids—and stimuli-responsive materials, LbL systems can decouple functions that are difficult to regulate independently within conventional single-layer or compositionally mixed surface architectures. This review examines recent advances in LbL-engineered LBNs (LbL-LBNs), focusing on how multilayer surface architecture reshapes physicochemical properties, cargo localization and release, biological identity, cellular interactions, and transport across physiological barriers. Particular attention is given to the multilayer interface as a dynamic biointerfacial bridge between a cargo-specific core architecture and the surrounding biological environment, including its capacity for stimuli-responsive switching in pathological microenvironments. The discussion further extends to biomimetic hybrid interfaces and establishes a framework for translating hierarchical surface architectures into reproducible, clinically tractable platforms for precision therapeutic delivery. Full article

Signalling between the ER and mitochondria regulates a number of key cellular functions that are damaged in frontotemporal dementia and related amyotrophic lateral sclerosis (FTD/ALS). This signalling involves close physical contacts between the two organelles that are mediated by the VAPB-PTPIP51 ER–mitochondria “tethering” proteins. A number of studies have shown that mutant genes which cause familial FTD/ALS disrupt the VAPB-PTPIP51 tethers and that this involves activation of GSK3β. TDP43 is one such mutant and altered TDP43 metabolism is central to FTD/ALS pathogenesis. Loss of Sigma-1 receptor function is also seen in FTD/ALS and there is evidence that Sigma-1 receptor agonists can repair damaged ER–mitochondria signalling. However, the underlying mechanisms are not properly understood. In this study, we show that the reference Sigma-1 receptor agonist PRE-084 stimulates VAPB-PTPIP51 binding and rescues FTD/ALS mutant TDP43-induced disruption to the VAPB-PTPIP51 interaction and linked ER–mitochondria Ca²⁺ delivery. We also show that these effects involve inhibition of the kinase GSK3β, a known negative regulator of VAPB-PTPIP51 binding. Finally, we show that ANAVEX2-73, a further Sigma-1 receptor agonist which is in clinical trials for Alzheimer’s disease, also stimulates VAPB-PTPIP51 binding via GSK3β inhibition. Our findings provide novel insights into the mechanisms by which Sigma-1 receptor agonists influence defective ER–mitochondria signalling in FTD/ALS. Full article

Hydrophilic sucrose esters are widely used in whipped cream, but the relationship between their molecular structure, interfacial behavior, and whipping performance remains unclear. This study investigated three hydrophilic sucrose esters, S1170, S1570, and P1570, in sodium caseinate-stabilized cream systems. Interfacial measurements showed that sucrose ester concentration strongly affected the adsorption behavior and viscoelasticity of the oil/water interface. Low concentrations produced rheological responses consistent with the formation of a more elastic mixed interface, whereas excessive addition was associated with competitive adsorption, reduced interfacial protein coverage, and weakened interfacial viscoelasticity. The molecular structure of sucrose esters further influenced this process: S1170, with a higher polyester content, showed behavior consistent with stronger interfacial interactions, while P1570 showed a greater decrease in interfacial modulus. These interfacial differences were associated with changes in whipping behavior. At 0.10 wt%, sucrose esters slowed fat partial coalescence, prolonged the optimal whipping time, and increased overrun. At 0.50 wt%, they accelerated early-stage coalescence and increased serum loss, especially for P1570. These findings suggest that selecting appropriate sucrose ester type and dosage is essential for balancing interfacial stability, fat partial coalescence, and whipped cream quality. Full article