Class III peroxidases (PRXs) link apoplastic redox regulation with cell-wall remodeling, yet their coordinated transcript- and protein-level responses to cadmium (Cd) remain poorly resolved in woody plants. We identified 80 high-confidence class III PRX genes in Salix variegata and characterized their genomic organization, evolutionary relationships, protein properties, and predicted regulatory features. Root RNA-seq and data-independent acquisition proteomic datasets from plants exposed to 0 or 100 μM CdCl2 for 10 d were then integrated to prioritize members supported across molecular layers. Segmental and dispersed duplication were the predominant expansion modes, and most SvaPRX proteins showed sequence features compatible with extracellular localization. RNA-seq identified 42 upregulated and two downregulated SvaPRX genes. The DIA dataset comprised 22,883 peptides and 6,068 proteins, with 4,849 proteins quantified in all six samples; biological replicates showed high within-treatment correlations and separation of CK and Cd100 in principal component analysis. Forty SvaPRX proteins were quantified, including 11 upregulated and six downregulated proteins. Cross-layer integration reduced the broad transcript-responsive set to seven concordantly upregulated candidates: SvPRX12, SvPRX23, SvPRX41, SvPRX47, SvPRX61, SvPRX62, and SvPRX66. Targeted qRT-PCR provided additional directional support for Cd-induced expression of SvPRX61 and SvPRX62, with response directions consistent across RNA-seq, proteomic, and qRT-PCR datasets. These results establish a reproducible genome-to-proteome framework for prioritizing Cd-responsive class III peroxidases and identify focused candidates for subsequent biochemical and genetic investigation in woody plants.

Recent advances in genetic engineering have substantially reduced the major carbohydrate barriers to pig-to-human xenotransplantation. Deletion of GGTA1, CMAH, and B4GALNT2 decreases αGal, Neu5Gc, and B4GALNT2-dependent Sda-like xenoantigens and thereby attenuates preformed antibody binding and complement-mediated injury. However, removal of these major xenoantigens does not necessarily generate a human-compatible cell-surface glycome. In particular, CMAH knockout eliminates Neu5Gc but leaves a Neu5Ac-dominant sialome whose linkage, chemical modification, glycoconjugate carrier, density, and spatial presentation are not necessarily human-like. This review examines sialylation and glycan-dependent recognition as an underappreciated regulatory layer linking innate immunity, blood-cell homeostasis, complement regulation, and thromboinflammation in xenotransplantation. Two conceptual distinctions are emphasized. First, the direction and evidence depth of glycan-receptor recognition must be explicitly considered. The best-established Siglec-1-dependent xenogeneic interaction is a graft-versus-host pathway, in which donor porcine Siglec-1-positive macrophages capture recipient human erythrocytes. Conversely, direct receptor-level binding of human Siglec-1 to CMAH-deficient gene-edited pig erythrocytes has recently been demonstrated, providing evidence for the reciprocal host-versus-graft direction; however, whether this interaction drives recipient macrophage phagocytosis or in vivo clearance remains unresolved. Donor ASGR1-dependent uptake of human platelets provides another direct example of glycan-dependent recipient blood-cell clearance. Second, α2,3-linked Neu5Ac illustrates a fundamental functional trade-off in sialome engineering. Neu5Acα2,3Gal-containing glycans can serve as preferred ligands for Siglec-1-mediated capture, whereas α2,3-linked sialoglycans also contribute to Factor H-dependent recognition of self-associated surfaces and alternative complement regulation. Conversely, increasing α2,6-linked sialylation may reduce macrophage xenoreactivity and enhance inhibitory Siglec engagement but decrease α2,3-dependent Factor H recognition; thus, ST6GAL1 engineering should not be considered uniformly protective or equivalent to glycan "humanization". Other pathways, including CD24-Siglec-10 signaling, Siglec-9-mediated regulation of neutrophil activation and NETosis, and platelet Siglec signaling, provide additional mechanistic frameworks but remain incompletely validated in solid-organ xenotransplantation. We therefore propose that post-editing glycan compatibility should be evaluated according to receptor directionality, evidence depth, and functional trade-offs, rather than glycan abundance or donor genotype alone. Organ-, cell-, carrier-, and receptor-specific analyses integrating quantitative glycomics/glycoproteomics, human receptor and immune-cell assays, functional crossmatching, and ex vivo perfusion are needed to link donor genotype to post-editing glycan phenotype and guide precision optimization of xenograft compatibility.

This study developed multifunctional sago starch active packaging films incorporating Abelmoschus esculentus extract (AE), green-synthesized ZnO nanoparticles (ZnONPs), and ZnO-MgO nanoparticles (ZnO-MgONPs). XRD confirmed the successful formation of crystalline nanoparticles, with average crystallite sizes of 22.4 ± 2.1 nm for ZnONPs and 18.7 ± 1.8 nm for ZnO-MgONPs. FESEM/TEM showed quasi-spherical nanoparticles with sizes of 25-35 nm and 20-30 nm, respectively. Compared with the AE-only film, the addition of ZnONPs and ZnO-MgONPs significantly improved film performance: tensile strength increased from 1.43 MPa to 2.12 MPa and 2.79 MPa, respectively, while elongation at break decreased from 60.39% to 46.12% and 32.33%. AE increased film moisture content from 6.98% to 8.78%, whereas nanoparticle incorporation improved barrier and structural properties. FTIR, XRD, FESEM, TGA, and UV-vis analyses confirmed good compatibility among the components, enhanced thermal stability, and stronger UV-shielding behavior in nanocomposite films, especially in the ZnO-MgO-containing system. The incorporation of AE enhanced the DPPH radical-scavenging activity of the films, indicating the contribution of AE-derived bioactive compounds to their antioxidant functionality. The nanocomposite films retained appreciable antioxidant activity while exhibiting improved mechanical, barrier, and antimicrobial properties. The films also exhibited antibacterial activity against Escherichia coli and Staphylococcus aureus, indicating their potential for active food packaging applications. Overall, the AE/ZnO-MgONPs film showed the best balance of mechanical strength, thermal stability, barrier performance, and antimicrobial function, making it a promising eco-friendly packaging material.

Resveratrol is stilbene phytoalexin that exhibits antimicrobial activity in diverse plant species. Although its direct antimicrobial effects have been investigated extensively, its role and regulatory mechanism in plant immune signaling remain unclear. In this study, we demonstrate that resveratrol enhances resistance to bacterial pathogens through activation of endogenous defense pathways in Arabidopsis thaliana. Resveratrol treatment reduced disease symptoms and bacterial growth caused by Pseudomonas syringae pv. tomato DC3000 (Pst DC3000) without exerting direct antibacterial activity and increased transcription of PATHOGENESIS-RELATED (PR) genes. Resveratrol promoted translocation of NONEXPRESSOR OF PATHOGENESIS-RELATED GENES 1 (NPR1) into the nuclear and failed to confer resistance in the npr1-1 mutant, which demonstrates that NPR1-mediated signaling is required for this response. Furthermore, resveratrol increased endogenous SA levels through transcriptional activation of SA biosynthesis genes. Accordingly, resveratrol-induced resistance and PR1 expression were abolished in the SA-deficient plants, which indicates that resveratrol-induces pathogen resistance depends on the SA signaling. In addition, resveratrol triggered rapid accumulation of reactive oxygen species (ROS), suggesting that ROS may contributes to this immune response. Collectively, these findings demonstrate that resveratrol functions as an immune-activating molecule that enhances bacterial resistance through NPR1-dependent SA signaling and ROS accumulation in Arabidopsis.

Medicinal plants provide an important source of structurally varied bioactive metabolites with medicinal potential; however, their effective extraction, characterization, and standardization present considerable analytical challenges. Counter-current separation (CCS) methods, such as centrifugal partition chromatography (CPC) and high-speed counter-current chromatography (HSCCC), represent advanced liquid-liquid chromatographic approaches that minimize irreversible adsorption of samples. A systematic literature search was performed in PubMed/MEDLINE, Scopus, Web of Science Core Collection, ScienceDirect and Google Scholar using pre-defined search terms related to counter-current separation, six omics domains, artificial intelligence/machine learning, molecular networking, network pharmacology, regulatory systems, and industrial translation. We screened relevant studies using pre-defined eligibility criteria and thematically synthesized them according to methodological development, analytical integration, evidence maturity, regulatory relevance, and translational potential. These methods enable the effective purification of bioactive natural compounds, ensuring high recovery rates. Simultaneously, the development of metabolomics, transcriptomics, proteomics, genomics, lipidomics, and glycomics has significantly advanced the research on natural products, enabling detailed molecular profiling and systems-level biological interpretation. This review critically considers the use of CCS in combination with multi-omics technologies for the discovery of bioactive compounds from medicinal plants. Special attention is paid to solvent-system design, AI-aided process optimization, molecular networking, chemometric analysis, and bioactivity-guided fractionation strategies. In addition, the use of CCS in quality control, phytopharmaceutical standardization, and industrial-scale production is discussed. Integration of CCS with omics technologies, artificial intelligence (AI), and network-based analytical approaches offers a complementary platform for the enhancement of the discovery, prioritization, isolation, and characterization of bioactive natural products. These integrated workflows can facilitate candidate identification, structural characterization, mechanistic investigation and biological validation in early translational research. However, it should be kept in mind that the identification of a bioactive compound or fraction is not the same as drug discovery or therapeutic development. Further pharmacological, toxicological, pharmacokinetic, manufacturing, regulatory and clinical investigations are required for a purified natural product or botanical preparation to advance toward therapeutic development.

Plasma p-tau217 accurately reflects amyloid pathology, but dual-cutoff strategies leave a non-negligible grey zone requiring confirmatory testing. We evaluated whether integrating plasma Aβ measurements could reduce diagnostic uncertainty in patients with mild cognitive impairment (MCI).

The RAS-RAF-MEK-ERK signaling pathway plays a key regulatory role in the survival and proliferation of mammalian cells. Accordingly, deregulated activation of this cascade by a mutated K-RAS (mutK-RAS) is a major cause of cancer initiation and progression. These findings led to the development of synthetic inhibitors that target the mutK-RAS protein, as a therapeutic approach. However, the challenging design of synthetic compounds that effectively and selectively inhibit the mutK-RAS, and the high prevalence of acquired resistance to the mutK-RAS inhibitors, motivated also the development of synthetic inhibitors to downstream effectors of mutK-RAS, such as ERK1/2. Unfortunately, these putative anti-cancer drugs also exhibited limited efficacy, due to the reactivation of the inhibited ERK1/2 kinases by their deregulated upstream activators, RAF and MEK, an effect that led to an acquired resistance. Recently, several studies reported the targeting of novel, mutK-RAS-RAF-MEK-ERK sustaining proteins whose downregulation led to ERK1/2 inhibition, without subsequent ERK1/2 reactivation and without acquired resistance. The knockdown or inhibition of the intracellular tyrosine kinase FER, whose expression level is linked to poor prognosis of cancer patients, led to significant and persistent inhibition of ERK1/2, without subsequent reactivation. In an additional report, myotrophin (MTPN) was identified as a crucial mediator of ERK1/2 oncogenic activation. Accordingly, knockdown of MTPN blocked ERK1/2 activity without inducing later reactivation, or acquired resistance of these kinases. Here, we discuss these novel findings, which could pave the way for novel intervention approaches with deregulated ERK1/2 activation, with significant ramifications for the therapeutic targeting of the key K-RAS-ERK1/2 oncogenic pathway.