The continuously growing mouse incisor provides a useful model for investigating stem cell niche organization and the MSC-to-TAC transition because it contains spatially distinct mesenchymal stem/stromal cell (MSC)-enriched and transit-amplifying cell (TAC)-enriched compartments and abundant TAC populations. However, the metabolic programs associated with MSCs and their transition to TACs within the native tissue environment remain poorly understood. Here, we integrated region-resolved transcriptomics, proteomics, and metabolomics with single-cell RNA sequencing to characterize metabolic remodeling associated with the MSC-to-TAC transition in the mouse incisor pulp. We found that the MSC-enriched apical pulp was enriched for molecular features associated with fatty acid β-oxidation and ketone body metabolism, whereas the TAC-enriched mitotic pulp showed molecular signatures associated with the tricarboxylic acid cycle, oxidative phosphorylation, and nucleotide biosynthesis. Integrated transcriptomic and proteomic analyses revealed lower mRNA-protein concordance in the MSC-enriched apical pulp, consistent with a contribution of post-transcriptional regulation, whereas the TAC-enriched mitotic pulp showed tighter mRNA-protein coupling. Metabolite profiling further showed higher abundance of acyl-carnitines and ketone bodies in the MSC-enriched apical pulp and higher abundance of glutamine, aspartate, and purine intermediates in the TAC-enriched mitotic pulp. Pseudotime analysis suggested coordinated changes in transcriptional programs related to fatty acid oxidation, mitochondrial oxidative metabolism, and nucleotide synthesis along an MSC-to-TAC trajectory. Collectively, our study establishes a region-resolved multi-omics atlas of metabolic remodeling associated with the MSC-to-TAC transition in the mouse incisor and provides insights into the metabolic features associated with stem cell activation, progenitor expansion, and tissue homeostasis in the native dental stem cell niche.

Soybean cyst nematode (Heterodera glycines, SCN) severely threatens global soybean yields. Traditional genetic resistance relying on Rhg1 and Rhg4 loci is increasingly compromised by virulent SCN populations. This review synthesizes current knowledge on the molecular interplay among soybean (Glycine max), SCN, and rhizosphere microbiome with a specific focus on quorum sensing (QS) as a central regulatory mechanism enabling novel biotechnological interventions. Emphasis is placed on how soil molecular microbiology approaches can facilitate the transfer of disease-suppressive microbiomes from resistant to susceptible soybean genotypes through strategic inoculation. QS molecules, including N-acyl homoserine lactones (AHLs) and autoinducer-2 (AI-2), prime plant immune responses, while root exudates mediate bidirectional plant-microbe feedback loops. It is proposed that rationally designed synthetic microbial consortia (SynComs), microbiome-smart breeding, and optimized inoculation technologies offer complementary strategies to conventional genetic resistance for sustainable SCN management.

The natural flavonoid chrysin exhibits broad-spectrum bioactivity. However, its clinical utility is severely constrained by suboptimal potency and unfavorable pharmacokinetic properties. Rational chemical modifications, including site-specific derivatization, molecular hybridization, and metal complexation, have yielded numerous chrysin derivatives with enhanced activity and selectivity. These derivatives engage diverse molecular targets, demonstrating considerable potential in various therapeutic areas, including anti-tumor, anti‑inflammatory, anti-diabetic, neuroprotective, and antimicrobial applications. This review systematically summarizes key site‑based modification strategies for chrysin, categorizes and discusses the derivatives according to their target profiles, and provides a detailed analysis of the design, bioactivity, and mechanisms of action of compounds targeting critical proteins such as epidermal growth factor receptor (EGFR), topoisomerase II (Topo II), poly (ADP-ribose) polymerase 1 (PARP1), cyclooxygenase-2 (COX-2), β-ketoacyl-acyl carrier protein synthase III (FabH), α-glucosidase, peroxisome proliferator-activated receptor γ (PPARγ) and cholinesterases (ChE). Recent advances in multi‑target derivatives and metal complexes are also highlighted. Finally, current challenges and future directions are discussed, emphasizing the role of target‑driven design and emerging technologies in guiding the development of next‑generation chrysin‑based therapeutics.

Medium- and long-chain triglycerides (MLCTs) are triacylglycerols (TAGs) that have medium-chain fatty acids (MCFAs) and long-chain fatty acids (LCFAs) linked together on a single glycerol backbone. The positioning of the components, such as the MCFA-LCFA-MCFA (MLM-type) positioning, can affect lipase-mediated hydrolysis, release of fatty acids (FAs), absorption and metabolic behavior, however, structure-function relationships requires experimental validation. This review presents a comparison between chemical interesterification, lipase-catalyzed interesterification and fermentation-assisted biosynthesis of MLCT. Chemical interesterification will redistribute fatty acyl groups, but offers limited positional control, while lipase-catalyzed synthesis provides more positional selectivity but is susceptible to acyl migration. Fermentation- assisted biosynthesis is a prospective approach that connects FA chain-length control, acyl-CoA supply and TAG assembly more strongly. However, there is currently more evidence that strongly supports the production of tailored FAs by microbes than demonstrated positional control of MLCT molecular species. Structural validation requires intact TAG molecular species analysis, positional analysis, and stereospecific analysis. An integrated framework combining applications of structural analysis, selective biocatalysis, fermentation optimization, metabolic engineering and functional evaluation is proposed.

Sodium alginate (ALG)/Gellan Gum (GG), myofibrillar protein (MF), and ovalbumin (OVA) were used to fabricate an interpenetrating polymer network (IPN) soft gel. The Ca[2+]-induced ALG network formed the primary matrix, followed by a heat-induced MF-OVA protein network. Optimal ALG concentration (MA-1) maximized network interpenetration and suppressed protein self-aggregation, yielding high moisture content (97%) and water-holding capacity (∼95%). The polysaccharide network facilitated ordered protein distribution and enhanced acyl bond interactions, enabling stable gelation under minimum gelation concentration. The resulting IPN gel balanced high cohesion with soft texture, attributed to dual-network interpenetration and physical entanglement. This study offers a novel structural design strategy for developing soft, morphologically stable protein gels for individuals with swallowing disorders.

Cervical cancer treatment faces challenges from chemotherapy resistance. Inducing ferroptosis is a promising strategy. Calycosin (Cal), an active component of Astragalus membranaceus, exhibits antitumor activity, but its role in cervical cancer, particularly regarding epithelial-mesenchymal transition (EMT) and ferroptosis, remains unclear. Cell Counting Kit-8, colony formation and Transwell assays assessed Cal's effects on cell viability, migration and invasion. Ferroptosis was evaluated by detecting reactive oxygen species (ROS), malondialdehyde, glutathione and ferrous iron (Fe[2+]) levels, mitochondrial morphology and expression of ferroptosis-related proteins. EMT markers and signaling pathways (TGF-β/Smad and Hippo/YAP) were analyzed using western blot. TGF-β1 knockdown was used to investigate the functional link between EMT and ferroptosis in vitro and in vivo. Cal inhibited proliferation and induced ferroptosis in cervical cancer cells, evidenced by mitochondrial shrinkage, lipid peroxidation and altered glutathione peroxidase 4/solute carrier family 7 member 11/acyl-CoA synthetase long-chain family member 4 expression. However, Cal also promoted migration and invasion, upregulated mesenchymal markers, and activated TGF-β/Smad while inhibiting Hippo/YAP signaling. Importantly, TGF-β1 knockdown suppressed both Cal-induced EMT and ferroptosis, indicating a synergistic relationship. In vivo studies confirmed these findings, showing Cal inhibited primary tumor growth but promoted metastasis, effects attenuated by TGF-β1 knockdown. Cal exerts dual effects in cervical cancer: Inducing ferroptosis to inhibit growth while promoting EMT-associated metastasis via TGF-β/Smad and Hippo/YAP pathways. Crucially, Cal-induced EMT synergistically enhances ferroptosis sensitivity. These findings highlight the complex pharmacology of Cal and suggest that combining with EMT inhibitors may optimize its therapeutic potential while mitigating pro-metastatic risks.

Cell culture relies heavily on serum supplementation, but serum composition makes it difficult to ensure reproducibility and consistency of experimental results. Consequently, research10 groups must test multiple batches to ensure the functionality and reproducibility of their models. The commonly used serums are fetal bovine serum (FBS) and calf serum (CS), which have been recognized as crucial in modulating cellular processes. While having been utilized for decades, little information is known about their respective lipid mediator contents. This study explored the presence of several major bioactive lipids involved in the regulation of inflammation, metabolism, differentiation, immune response, neuroprotection, and vascular homeostasis. These included polyunsaturated fatty acids, monoacylglycerols (MAGs), N-acyl-ethanolamines (NAEs), and oxylipins. As compared to FBS, CS samples were enriched in most fatty acids except for arachidonic acid. The levels of the endocannabinoids 2-arachidonoyl-glycerol (2-AG) and N-arachidonoyl-ethanolamine (AEA) followed the same pattern as arachidonic acid. On the other hand, most lipoxygenase-derived mediators, including, leukotriene B4, showed higher abundance in CS. Accordingly, CS activated the random migration of human neutrophils to a much greater extent than FBS, an effect attenuated by the BLT1 receptor antagonist CP 105,696. Cytokines/chemokines levels were also different between FBS and CS samples. These findings highlight that bovine serum lipid composition might be a major determinant modulating cellular responses and might thus impact experimental reproducibility. The data presented herein will help obtaining insights beyond conventional cell culture optimization and might represent key features to consider when planning in cellulo experiments across numerous fields, also beyond immunology.

In plant cells, protein homeostasis is tightly regulated to prevent imbalances between protein synthesis and degradation that can lead to proteotoxic stress. Protein turnover is largely maintained by the ubiquitin-proteasome system (UPS), with the AAA+ ATPase CDC48 facilitating the extraction and processing of ubiquitinated substrates. Previous studies identified redox-related proteins as plant CDC48 partners. Here, using tobacco cell lines overexpressing CDC48, we identified the peroxisomal catalase CAT1 as a substrate of CDC48-dependent turnover. CDC48 overexpression markedly reduced CAT1 abundance and activity, indicating post-translational regulation. CAT1 undergoes extensive polyubiquitination, and was rapidly degraded in a CDC48-dependent manner. Inhibition of deubiquitinating enzymes or CDC48 ATPase activity both led to the accumulation of ubiquitinated CAT1. Finally, CDC48 overexpression, which leads to reduced CAT1 levels, correlated with impaired peroxisomal function. These findings reveal a role for CDC48 in regulating peroxisomal protein turnover in tobacco.

Anaerobic digestion (AD) represents a mature and extensively adopted technique to dispose organic waste, yet ammonia toxicity remains a major bottleneck for stable AD operation. For activating methanogenesis-related enzymes, regulating the microbial community structure, and optimizing the reaction efficiency of AD process, supplementation of trace elements has been validated as an effective approach to counteract ammonia inhibition. However, mechanisms of how different trace elements modulate the biosynthesis and release of N-acyl homoserine lactones (AHLs), as well as the subsequent relieving ammonia inhibition are still unknown. Therefore, the current study investigated the effects of different trace elements on the secretion of AHLs under ammonia stress. The results showed that optimal concentrations of Fe, Co and Ni could enhance the secretion of AHLs, mainly N-butyryl-DL-homoserine lactone (C4-HSL) and N-hexanoyl-L-homoserine lactone (C6-HSL), and adjust microbial social behaviors under high ammonia levels. In addition, microbial community changes (enriched Bacteroidetes_vadinHA17, Syntrophomonas and Methanosarcina) were closely associated with increased methane production and stimulated AHLs accumulation by trace elements. Furthermore, increased AHLs signals correlated with intercellular communication could regulate critical genes transcription (glk, pgk, and yhdR) and enzymes synthesis (glucokinase, acetate kinase, and Coenzyme-B sulfoethylthiotransferase) to alleviate ammonia inhibition during the whole AD process. Trace elements could serve for multiple enzymes, which could be beneficial for AHL secreting hydrolytic-acidogenic consortia. The current study could provide primary insights for applying trace elements to relieve ammonia stress via AHLs regulation.

Nanofiltration (NF) membranes are indispensable for removing hazardous contaminants such as dyes and salts from industrial wastewater, yet their performance is often bottlenecked by the inherent permeability-selectivity trade-off and membrane fouling. Zwitterionic surface modification offers a means to overcome these limitations by promoting hydrophilicity and introducing charge-mediated transport. Here we synthesized a pre-formed zwitterionic piperazine (ZW-PIP) via a one-pot ring-opening of piperazine with 1,3-propanesultone, and directly grafted it onto polyamide films through residual acyl chloride groups. Combined low-field nuclear magnetic resonance (LF NMR) and molecular dynamics (MD) simulations demonstrated that the grafted zwitterionic moieties stabilized a hydration layer and facilitated water and chloride-ion diffusion (>90%), while sterically excluding sulfate. The resulting membrane exhibited a pure water permeance of 29.2 L∙m[-2]∙h[-1]∙bar[-1]. Crucially, this enhanced permeability was achieved while maintaining the exceptional salt-rejection performance for Na2SO4 (>97.0%). This high selectivity persisted even under challenging conditions of elevated salt concentration (8.0 g/L) and operating pressure (8.0 bar). The membrane exhibited robust rejection of various dyes (>96%), sustained stability, and potent antifouling (BSA, FRR 89%) and antibacterial (>99.97%) properties. Collectively, this work offers a mechanistic basis for designing zwitterionic NF membranes with high permeability, selectivity, and fouling resistance for treating complex hazardous wastewater.

This study reports the design and synthesis of novel quinolin-2-one derivatives as bioisosteric analogues for native N-acyl homoserine lactones (AHLs) to fight multidrug-resistant (MDR) Pseudomonas aeruginosa. Among the synthesized compounds, the Schiff base derivative (6g) emerged as a potent lead, exhibiting a Minimum Inhibitory Concentration (MIC) of 1.25 μg/mL against MDR clinical isolate. Phenotypic assays demonstrated that sub-MIC concentrations of 6g reduced biofilm biomass by 92.30%. Time-kill analysis demonstrated concentration-dependent antibacterial activity, with 2× MIC producing 4.2-log10 CFU/mL reduction at 24 h, whereas the 1/2 MIC concentration used for the anti-virulence studies did not produce extensive bacterial killing. Quantitative RT-PCR revealed significant downregulation of the core QS genes lasR (94.72%), rhlR (91.65%), lasI (84.84%), and rhlI (76.46%). Importantly, functional evaluation using LasR-dependent β-galactosidase reporter assay demonstrated concentration-dependent LasR inhibition by 6g, with an IC50 of 0.62 ± 0.05 μM, providing experimental support for LasR-mediated QS interference. Molecular docking supported favorable accommodation of the active derivatives within the RhlR, RhlI, LasR, and LasI binding pockets offering a theoretical rationale for potential broad quorum-sensing interference, while SwissADME analysis computationally predicted limited blood-brain barrier penetration and a preliminary predicted ADME profile for 6g. Together with its low mammalian cytotoxicity and high selectivity index (SI ≈ 267), these findings identify 6g as a promising quinolin-2-one lead displaying combined antibacterial and QS-associated anti-virulence activity that warrants further biophysical target validation and in vivo investigation.

Nanopore-based technologies are revolutionizing nucleic acid sequencing and molecular sensing, yet the structural basis by which pore-forming proteins interact with lipid membranes remains poorly defined. The mycobacterial porin MspA is a benchmark biological nanopore owing to its exceptional stability and uniquely narrow constriction, but accurate structural information is limited to protein-only models. Here, we present a 2.5-angstrom resolution cryo-electron microscopy structure of the M2 mutant of MspA embedded in a phospholipid nanodisc, providing a high-resolution experimental observation of MspA in a lipid environment. The structure reveals an annular arrangement of extended lipid acyl chains surrounding the MspA oligomer and contacting the transmembrane barrel. Their resolved densities indicate a substantially more ordered annular lipid environment than would be expected from a largely heterogeneous and nonspecific protein-lipid interface. The structure also demonstrates how specific M2 MspA mutations modulate pore polarity, influencing ion selectivity and transport. By determining the membrane-embedded structure of MspA and revealing an ordered arrangement of acyl chains forming extensive hydrophobic interactions with the protein, this study demonstrates the importance of the surrounding lipid environment in the design of protein nanopores.

A metal- and photosensitizer-free photoinduced C-H acylation of azauracils with both aryl and alkyl aldehydes has been developed using H2O2 or TBHP as the sole additive under light-emitting diode irradiation. This protocol accommodates a broad range of substrates, including various N-substituted azauracil derivatives as well as aryl, heteroaryl, and alkyl aldehydes, affording the corresponding acylated products in moderate to good yields. Notably, this transformation proceeds efficiently without transition metals or external photocatalysts, representing a distinct advantage over conventional protocols. The synthetic utility of this method is further demonstrated by scalable continuous-flow synthesis, late-stage functionalization of bioactive molecules, and diverse product derivatizations. Mechanistic studies, including radical trapping experiments and high-resolution mass spectrometry analysis of the corresponding adducts, support a radical pathway involving acyl radical intermediates and sequential hydrogen atom transfer processes.