Our understanding of trypsin, its zymogenicity and its inhibition is intimately intertwined with the history of biochemistry. Early structural studies revealed its close relationship to chymotrypsin, with its active site triad, oxyanion hole and N-terminus involved in a buried salt bridge near the active site. Its complex with basic pancreatic trypsin inhibitor provided a model for how peptide substrates bind to and are cleaved by the proteinase. Analysis of crystals of trypsinogen by Wolfram Bode and Robert Huber revealed that a large region of the zymogen is disordered prior to proteolytic activation, with an associated disruption of the oxyanion hole and substrate binding pockets, highlighting the importance of disorder in protein function. As archetype of numerous therapeutically important serine proteinases, trypsin can also serve as a surrogate for structure-based drug design. Trypsin variants designed for ligand binding studies resulted however in an unexpected plasticity of the mutant proteins that underlines the complexity of protein stability. A trypsin variant selected for peptide ligation (reverse proteolysis) was shown to possess zymogen-like characteristics that proved central to its application in modification of therapeutic proteins. The review pays homage to the seminal works of Bode and Huber and their influence on modern structural biology.

By using naturally abundant diatomite with unique particle and porosity structures as the silica precursor, a novel green-emitting diatomite-derived Zn2SiO4:Mn phosphor has been obtained as a dual functional sensor for developing latent fingerprints, serving as a fingerprint powder and a reagent for small particle reagents (SPRs). Parameters including calcination duration, calcination temperature, molar ratio of manganese salt to zinc and manganese salts, and molar ratio of zinc and manganese salts to diatomite have been adjusted to optimize luminescent property of this phosphor. Formation mechanism has been proposed based on the change of particle morphology and mesopore structure. With respect to material synthesis, high porosity of diatomite plays a critical part in ensuring uniform doping of Mn[2+], lowering reaction temperature, sparing inert or reducing atmosphere. Furthermore, surplus diatomite in the precursors results in basic retention of particle morphology which is favorable for latent fingerprint development. Efficient green emission centered at 525 nm of the diatomite-derived Zn2SiO4:Mn under 254 nm excitation originates from the [4]T1 → [6]A1 transition, making it a very capable fluorescent fingerprint powder that can suppress interferences from various surfaces, especially with the aid of 530 nm band-pass filter as an optical filter and green channel of RGB images as a digital filter. Given that the luminescence of diatomite-derived Zn2SiO4:Mn phosphor is not subject to external environment factors such as water or dissolved components, a fluorescent SPR prepared by adding a certain amount of this powder (10 mg mL[-1]) to an aqueous solution of sodium dodecyl sulfate (0.050 wt%) shows better performance compared with some commercially available SPRs. Latent fingerprints on glass sheets can be clearly visualized even if they have been immersed in water as long as 5 days. Therefore, the successful acquisition of green luminescent diatomite-derived Zn2SiO4:Mn contributes to both the synthesis of this popular green-emitting phosphor and the application of fingerprint development.

Biodegradable shape memory scaffolds have the unique potential to heal irregularly shaped craniomaxillofacial (CMF) defects through conformal 'self-fitting'. These have been previously prepared from poly(ε-caprolactone) diacrylate (PCL-DA), but the slow biodegradation rate of PCL is expected to limit neotissue formation. Subsequently, telechelic siloxane macromers polydimethylsiloxane-dimethacrylate (PDMS-DMA) and polymethylhydrosiloxane-DMA (PMHS-DMA) were combined with PCL-DA at varying weight (wt)% ratios, resulting in PCL/PDMS and PCL/PMHS co-network scaffolds with accelerated degradation rates owing to phase separation that increased water uptake. Still, these siloxane macromers lack a hydrolytically unstable backbone thus limiting degradation. Herein, poly(silyl fumarate) (PSF) was synthesized as a hybrid siloxane macromer with a hydrolytically unstable backbone as well as interchain crosslinkability. PCL/PSF scaffolds were prepared at 90:10, 75:25, 60:40, and 50:50 wt% of PCL-DA to PSF via solvent-casting particulate leaching (SCPL) with a fused salt template. Despite a reduction in PCL crystallinity (i.e., switching segments) with 40 and 50 wt% PSF, all scaffolds maintained excellent shape memory behavior. PCL/PSF scaffolds with 10 and 25 wt% PSF also maintained the modulus of the PCL-only scaffold as well as the corresponding PCL/PDMS and PCL/PMHS scaffolds. In vitro degradation under basic conditions revealed that PCL/PSF scaffolds with just 10 wt% PSF degraded faster than the PCL-only scaffold and further increased with 25 wt% PSF to surpass that of the corresponding PCL/PDMS scaffold. A lack of phases separation was observed, and thus indicated that faster degradation was achieved by the hydrolytic instability of the PSF.

Salt stress severely restricts plant growth and reduces crop yield. Strigolactones (SLs) are carotenoid-derived phytohormones involved in the regulation of plant salt tolerance. Salt stress can modulate the expression of SL biosynthetic and signaling genes, thereby affecting SL accumulation and signaling responses. SLs also interact with abscisic acid (ABA), reactive oxygen species (ROS), and other signaling molecules to coordinate downstream stress responses. At the physiological level, SLs alleviate salt stress by maintaining Na+/K+ homeostasis, enhancing osmotic adjustment and antioxidant defense, and reducing damage to the photosynthetic system. In addition, SLs can enhance plant resource acquisition and adaptive capacity under salt stress by regulating root architecture and promoting hyphal branching of arbuscular mycorrhizal fungi (AMF). This review focuses on SL-mediated regulation of plant salt tolerance at the molecular and physiological levels and further summarizes exogenous SL application strategies for alleviating salt stress, as well as research progress on key genes in the SL pathway for the genetic improvement of salt tolerance. Clarifying the potential of SLs in regulating plant responses to salt stress could provide new insights into sustainable crop production in saline-alkali environments.

Ulva prolifera is the main causative species of marine green tides and exhibits extreme tolerance to intertidal abiotic stress. However, the underlying molecular mechanisms remain largely unclear. In this study, we cloned and characterized an MYB-related transcription factor, UpMYB-PHL, from U. prolifera. Expression analysis showed that UpMYB-PHL is rapidly and significantly induced by high-salt stress. Furthermore, heterologous overexpression of UpMYB-PHL in the model microalga Chlamydomonas reinhardtii significantly improved its salt tolerance and biomass. By yeast one-hybrid and dual-luciferase assays, we demonstrated that UpMYB-PHL directly binds to and activates the promoter of the phosphate transporter gene UpPHT1, which is a typical target gene of phosphate response (PHR) transcription factor and participates in salt stress responses of plants. Interestingly, yeast two-hybrid assays revealed that UpMYB-PHL physically interacts with UpGAPDH, a core enzyme in energy metabolism. Taken together, our findings reveal a novel regulatory network in which UpMYB-PHL coordinates phosphorus transporter and energy metabolism in response to salt stress in U. prolifera. This study provides a vital molecular explanation for the rapid adaptation and massive growth of U. prolifera under severe intertidal salt stress.

Basic-region leucine zipper (bZIP) transcription factors are crucial for plant stress responses, but their characterization in the wild species Juglans mandshurica remains limited. Here, we identified 80 bZIP genes in the J. mandshurica genome and classified them into 13 subgroups, with notable enrichment in subgroups S, A, D, and I. All subgroup D members contain both bZIP and DELAY OF GERMINATION 1 (DOG1) domains, forming characteristic dual-module fusion proteins. Evolutionary analysis detected three orthologous gene pairs under positive selection since divergence from Juglans regia. Promoter cis-elements, especially MYB and MYC motifs, are abundant in JmbZIP genes. Protein-protein interaction networks suggest potential functional specialization and coordination among JmbZIP members. Expression profiling revealed distinct patterns across subgroups, with S, A, and D showing high activity across various physiological processes and light stress responses. qRT-PCR validated the dynamic expression of six ABA pathway marker genes, the ABRE-rich JmbZIP41 and JmbZIP42 genes, together with the highly expressed JmbZIP12 gene under salt and drought stress. Our genome-wide analysis enabled the functional screening of bZIP members across subgroups. The key genes identified in this study provide valuable genetic resources for stress-resistance breeding in forest trees, with JmABI5 (JmbZIP40) and JmbZIP42 serving as prime candidates for enhancing tree stress tolerance.

Background/Objectives: Chronic kidney disease (CKD) remains underdiagnosed until advanced stages, motivating reliable, clinically deployable screening models that pair high discrimination with an explicit operating threshold and transparent explanations. Methods: In this study, we propose a CKD detection framework that integrates structured preprocessing, class imbalance handling, stratified 10-fold cross-validation with out-of-fold (OOF) prediction, and clinically oriented threshold selection via the Youden index, followed by explainability using SHAP and LIME. Experiments were conducted on two datasets. Across a broad panel of ten machine learning models, gradient boosting methods consistently dominated. Results: LightGBM achieved the best overall clinical composite performance on both datasets. On Dataset 1, LightGBM delivered near-ceiling OOF discrimination (ROC-AUC = 99.98, PR-AUC = 99.98) and an excellent clinically balanced performance at the best Youden threshold (0.41), reaching sensitivity = 99.20, specificity = 99.60, accuracy = 99.40, F1 = 99.40, and MCC = 98.80, with robust cross-validation stability (CV AUC = 99.99 ± 0.04; CV sensitivity = 99.10 ± 1.81; CV specificity = 99.46 ± 1.42; CV MCC = 98.59 ± 2.19), strong calibration (Brier = 0.006), and fast training (0.078 ± 0.019 s/fold). On Dataset 2, LightGBM maintained high generalization (ROC-AUC = 99.72, PR-AUC = 99.64) and clinically deployable balance at the best Youden threshold (0.35), achieving sensitivity = 98.10, specificity = 98.03, accuracy = 98.06, F1 = 98.06, and MCC = 96.13, with consistent fold-wise performance (CV AUC = 99.69 ± 0.25; CV sensitivity = 97.25 ± 1.25; CV specificity = 98.11 ± 1.02; CV MCC = 95.37 ± 1.56), acceptable calibration (Brier = 0.0173), and practical training time (0.742 ± 0.144 s/fold). Conclusions: Finally, SHAP and LIME explanations confirmed that model decisions align with clinically meaningful renal function and symptom/biomarker patterns at both population and patient levels, supporting safer translation of the proposed framework into CKD screening and decision-support workflows.

Salinity is an escalating agricultural challenge, yet plant microbiomes offer a promising avenue for improving salt tolerance. Nevertheless, most naturally occurring microbes remain unevaluated for plant growth-promoting function, and systematic approaches to uncover salt-tolerance-enhancing plant growth-promoting rhizobacteria (PGPR) are limited. Here, using soybean as a model, we implement a quantitative framework to characterize rhizosphere microbial networks and nominate novel taxa functionally associated with plant salt tolerance. We introduced a salt tolerance index (STI) to quantify plant salt tolerance and normalize performance across heterogeneous natural soil salinity. Metagenomic sequencing and co-occurrence analysis revealed distinct rhizosphere microbiota structures between tolerant and susceptible soybeans. In tolerant soybeans, Pseudomonas dominated as the hub of a highly interconnected network, whereas susceptible accessions showed a fragmented network dominated by Acinetobacter. Correlation analyses identified bacterial taxa positively associated with STI, including documented salt-tolerant PGPR and novel candidates. Greenhouse experiments showed that one candidate, Thalassospira xiamenensis, enhances soybean salt tolerance and reshapes host ion-transport and oxidative-stress gene expression under salinity, validating our screening strategy. Our culture-independent metagenomic association approach reveals host genotype-structured rhizosphere microbial networks underlying salt tolerance and provides an efficient, labor-saving means for high-throughput identification of salt-tolerant PGPR.

The rational integration of antagonistic catalytic sites within robust porous architectures remains a critical challenge for sustainable multicomponent reactions to synthesize pharmacologically active molecules. Herein, we report a thermo-chemically stable, mixed-ligand Ni(II)-based pillar-layered metal-organic framework (MOF), featuring activation-induced open metal sites and pore-confined Lewis basic centres. This bifunctional framework acts as an efficient heterogeneous catalyst for one-pot Knoevenagel-Michael multicomponent reactions, enabling the rapid synthesis of structurally diverse 2-amino-3-cyano-4H-pyrans under mild conditions with low catalyst loading. The synthesis of 4H-pyrans is promoted by the synergistic interplay between Lewis acidic metal centres and framework-integrated basic sites, as corroborated by control experiments and analyte-responsive emission modulation. The protocol accommodates twenty-six electronically diverse substrates and affords bioactive pyran derivatives in excellent yields. Notably, the confined pore environment imparts distinct size-selectivity, restricting bulky substrates and demonstrates molecular-dimension-controlled catalysis. The framework exhibits remarkable recyclability, structural robustness, and negligible metal leaching, outperforming many reported catalytic systems. Importantly, eight 4H-pyrans were structurally confirmed in their purest form by single-crystal X-ray diffraction, alongside other spectroscopic characterizations. This work highlights a rational platform for designing bifunctional MOF catalysts to promote green organic synthesis through the synergistic cooperation of antagonistic functionalities confined within porous architectures.

Coagulopathy refers to any medical condition which affects the ability of the blood to clot. It can be caused due to genetic conditions like haemophilia, von Willebrand disease, or it can be caused through liver disease or deficiency of Vitamin K. It also involves a broad range of diseases affecting hemostasis as an unbalanced and even bidirectional relationship between thrombosis and bleeding. Coagulopathy can also be caused by thromboinflammation, as seen in VHFs like Ebola, Dengue, Marburg, Crimean-Congo Hemorrhagic Fever, Yellow Fever, and Hantavirus infection. The immune response and coagulation system are intricately linked in such cases. Infections from VHFs cause endothelial cell dysfunction through the immune response, monocytes/macrophages activation, and increased expression of tissue factor (TF), which in turn causes excessive thrombin production and fibrin formation. These conditions result in microvascular thrombosis, organ dysfunction, consumption of platelets and coagulation factors, causing a balanced but fragile state of hemostasis that could tip over towards either thrombosis or bleeding. New therapies have been developed that interfere with these processes, such as interference with the TF pathway (for instance, rNAPc2) and regulation of fibrinolysis (tranexamic acid). The recognition of the double-edged sword of coagulopathy is critical for the development of treatment strategies targeting coagulation disorders. This literature review discusses the molecular basis of immunothrombosis and endothelial dysfunction in VHFs.

Osteogenic phenotypic transition of vascular smooth muscle cells (VSMCs) is the core pathological event underlying the initiation and progression of vascular calcification (VC). Runt-related transcription factor 2 (Runx2), a master osteogenic transcription factor, induces the transdifferentiation of VSMCs toward an osteoblast-like phenotype, which subsequently facilitates calcium salt deposition and calcified plaque formation in the vascular wall. However, its upstream and downstream regulatory networks remain incompletely defined. This study found that lactate levels were increased in calcified aortic tissues and osteogenic VSMCs. Additionally, it demonstrated that global protein lactylation and histone H3 lysine 18 lactylation (H3K18la) were concurrently elevated. In VSMCS, lactate consistently improved H3K18la modification and the osteogenic phenotypic transition, while the glycolytic inhibitor 2-deoxy-d-glucose (2-DG) reversed these phenotypic alterations. Consistent with the above phenotypic changes, we identified AlkB homolog 5 (ALKBH5) as a potential transcriptional target of H3K18la through CUT&Tag combined with proteomic screening and analysis. H3K18la is enriched at the ALKBH5 promoter, which is required to activate its transcription. Mechanistic investigations revealed that the Runx2 serves as a downstream target of ALKBH5-mediated m[6]A demethylation. ALKBH5 positively regulates Runx2 expression by erasing m[6]A modifications. Conversely, ALKBH5 knockdown leads to excessive m[6]A accumulation and subsequently accelerates Runx2 mRNA degradation. Additionally, Runx2 directly transcriptionally regulates lactate dehydrogenase A (LDHA), thereby increasing lactate production and H3K18la levels and establishing the H3K18la-ALKBH5-Runx2-LDHA positive feedback loop that persistently drives osteogenic phenotypic transition. This study identifies a novel synergistic regulatory mechanism between histone lactylation and RNA m[6]A methylation modifications in VC pathogenesis.

Microbially induced struvite precipitation is widely used for treating eutrophic water. Nevertheless, the treatment of eutrophic saline water is considerably restricted due to the scarcity of halophilic bacteria and their relatively low salt tolerance. In this study, Virgibacillus salexigens XYD, with an optimal 5% (w/v) sodium chloride concentration, was used to treat eutrophic high-salt wastewater. In wastewater treatment, this bacterium not only increases the pH but also degrades organic matter, releasing ammonium and phosphate ions. The increased contents of acidic and basic amino acids and organic acids in extracellular polymeric substances enhanced the adsorption of ammonium, magnesium, and phosphate ions, thereby promoting struvite nucleation. Orthogonal experiments and response surface analysis were used to optimize the immobilization conditions for strain XYD. Under the optimized conditions, the immobilized bacteria achieved higher removal efficiencies for ammonium, phosphate, and magnesium than the suspended bacteria, reaching 97.41%, 99.07%, and 97.55%, respectively, by day 14. In the third reuse cycle, the immobilized bacteria still exhibited higher efficiency than the suspended bacteria. The conversion of these ions into struvite was confirmed by X-ray diffraction and Rietveld refinement. Biotic struvite formed larger prismatic crystals and contained complex organic functional groups (e.g., C-O-PO3, S-S) and protein secondary structures, compared to inorganic abiotic struvite. The struvite induced by immobilized bacteria contained more organic functional groups and exhibited higher crystallinity than that induced by suspended bacteria. This study advances our understanding of bacterial-induced struvite formation and paves the way for the sustainable recovery of nitrogen and phosphorus from eutrophic saline waters.

The basic helix-loop-helix (bHLH) gene family, a major class of transcription factors in plants, plays critical role in growth, development, and responses to environmental stresses. Despite its importance, knowledge of bHLH gene functions in cowpea remains limited. Here, we conducted a comprehensive genome-wide in silico analysis of the cowpea bHLH family. A total of 60 VubHLH genes were identified and found to be non-uniformly distributed across the 11 chromosomes. Phylogenetic analysis grouped 60 VubHLH and 127 AtbHLH genes into six distinct subgroups. Gene structural characteristics were conserved within subgroups but diversed among subgroups. Synteny analysis indicated that segmental duplication, rather than tandem duplication, was the primary driver of VubHLH family expansion. Promoter analysis revealed diverse cis-acting elements associated with plant growth, developmental processes, phytohormone regulation, and stress responses. Tissue-specific expression suggested functional diversification of VubHLH genes during evolution. qRT-PCR analysis showed that VubHLH1.1 and VubHLH11.1 genes were significantly upregulated under ABA, MeJA, and salt treatments, whereas VubHLH3.7, VubHLH5.1, and VubHLH6.3 were downregulated. Collectively, these results provide valuable insights into the evolutionary relationships and functional characteristics of the bHLH gene family and lay a foundation for future studies of bHLH gene function and improving the cowpea salt tolerance.