Search Results (80)

Neglected arboviruses dengue (DENV), Zika (ZIKV), yellow fever (YFV), Japanese encephalitis (JEV), and chikungunya collectively affect hundreds of millions of people annually, yet no specific antiviral drug has been approved for any of them. Alkaloids, nitrogen-containing specialized metabolites produced by diverse plant families, have emerged as a promising source of broad-spectrum antiviral scaffolds. This review compiles and critically analyzes over 100 alkaloid-virus pairs across several RNA virus families, providing a comparative mechanistic analysis. Lycorine, narciclasine, emetine, and berbamine, among others, exhibit potent activity against phylogenetically distant viruses, with the most potent activities reported against flaviviruses (narciclasine: EC₅₀ 0.02 µM against DENV, ZIKV, YFV, and JEV; pancratistatine: 0.0063 µM against ZIKV). Structure-activity analysis of multiple alkaloid classes identifies key pharmacophoric features, including the phenanthridine nucleus (lycorine derivatives) and the bis-benzylisoquinoline scaffold (tetrandrine, berbamine), as determinants of antiviral potency, selectivity, and broad-spectrum activity. Genetic resistance data in West Nile virus challenge the widely accepted model of lycorine as a direct nucleoside inhibitor, instead pointing toward the involvement of the membrane-associated NS4A-2K-NS4B replication complex, though direct validation remains limited for other flaviviruses. Converging structural, biochemical, and transcriptomic evidence suggests that ribosome-mediated translational stress may represent an additional host-directed antiviral mechanism for isoquinoline-type alkaloids, though the causal chain from ribosome binding to activation of the integrated stress response and to antiviral effect has not been established. The present analysis highlights that in vivo validation remains limited to a few alkaloid-virus pairs. Unbiased target deconvolution and formal testing of the ribosome/integrated stress response hypothesis stand out as essential research priorities. Full article

Background/Objectives: Endometrial cancer (EC) is the most prevalent gynecological malignancy in developed countries, yet no clinically validated noninvasive screening biomarker exists. Fatty acid metabolic reprogramming is a hallmark of cancer that produces measurable changes in systemic free fatty acid (FFA) profiles. We aimed to identify serum FFAs altered in EC and evaluate their potential as early diagnostic biomarkers. Methods: Gene expression of fatty acid metabolic enzymes in EC tissues was assessed using public databases. Serum levels of 19 FFAs were quantified by gas chromatography–mass spectrometry in 72 patients with EC and 84 healthy donors. A two-variable diagnostic model was constructed using logistic regression and validated in an independent dataset. Results: Multiple fatty acid metabolic enzymes involved in de novo synthesis were upregulated in EC tissues as early as stage I, while enzymes mediating fatty acid uptake and release were downregulated. Serum levels of palmitoleic acid, oleic acid and dihomo-γ-linolenic acid were significantly elevated, whereas arachidic acid and arachidonic acid were significantly decreased in patients with EC from stage I onward. A diagnostic model combining palmitoleic acid and arachidonic acid achieved an area under the receiver operating characteristic curve of 0.9674, with a sensitivity of 91.3% and specificity of 96.4% in the validation dataset, including for stage I disease, and was independent of body mass index, age, metastasis status, and family history. Conclusions: Serum FFA profiling represents a promising noninvasive strategy for early EC detection, with the potential to complement existing diagnostic methods. Full article

Laccases (EC 1.10.3.2, LAC) are copper-containing glycoproteins involved in lignin biosynthesis, and as such, they play important roles in plant development and stress responses. In this study, a genome-wide analysis of the LAC gene family was performed in Pinus massoniana (Chinese red pine), identifying 78 PmaLAC genes, all predicted to encode cell membrane-localized proteins. These genes were unevenly distributed across eight chromosomes, with notable clusters on chromosomes 7 and 8, indicating gene duplication-driven expansion in P. massoniana. Phylogenetic analysis revealed that PmaLAC genes are classified into five subfamilies, reflecting the lineage-specific expansion and evolutionary divergence of gymnosperm LAC genes. Conserved motif and gene structure analyses showed high conservation among PmaLAC proteins. Promoter analysis identified numerous cis-acting elements related to hormone signaling, stress, and light responses. RNA-seq analysis revealed distinct tissue-specific expression patterns for PmaLAC gene family members. Moreover, degradome analysis combined with dual-luciferase assays supported the interaction between miR397c-9 and PmaLAC31, suggesting that miR397c-9 negatively regulates PmaLAC31 and indicating a potentially conserved miRNA-mediated regulatory mechanism. Overall, this study provides a systematic overview of the composition, evolution, and potential regulation mechanisms of the PmaLAC gene family in P. massoniana, providing a useful resource for future functional characterization of PmaLAC genes. Full article

Alternaria brassicicola is a necrotrophic fungal pathogen causing black spot disease on cruciferous crops worldwide. In this study, we comprehensively characterized the pathogenic isolate Ab0920a from diseased broccoli in Shanghai using morphological, phylogenetic, host range, fungicide sensitivity, genomic, and functional analyses. Pathogenicity tests on 27 cruciferous varieties revealed a broad host range with varying resistance levels. Fungicide sensitivity assays showed that fluxapyroxad (EC₅₀ = 0.0695 µg/mL), prochloraz (0.0711 µg/mL), and difenoconazole (0.0863 µg/mL) were highly effective, whereas fluazinam was least effective. Genome-wide annotation identified 941 secreted proteins (8.95% of the proteome) and 237 candidate effectors, including 31 small cysteine-rich secreted proteins and 68 homologs of known virulence factors. Conserved effector-associated motifs (e.g., RXLR, [Y/F/W]xC) were detected, and carbohydrate-active enzyme CAZyme annotation revealed diverse families potentially involved in plant cell wall degradation. Functional validation using the pSUC2 yeast system confirmed that N-terminal signal peptides of tested effectors are competent for secretion. A PVX-based transient expression assay in Nicotiana benthamiana identified two effectors that suppress Bax-induced programmed cell death, suggesting their potential roles in modulating host immunity. Overall, this study provides comprehensive insights into the pathogen Ab0920a, offering resources for disease management and functional studies on necrotrophic fungal pathogenesis. Full article

Lipophorin receptors play a crucial role in the reproductive development of insects. However, their regulatory function in insect reproductive diapause remains poorly understood. In this study, a lipophorin receptor gene (GdLpR) was cloned from Galeruca daurica using RT-PCR. The open reading frame (ORF) of GdLpR is 2589 bp in length, encoding 862 amino acids, and possesses typical structural characteristics of the lipophorin receptor family. RT-qPCR analysis indicated that the expression of GdLpR is up-regulated in G. daurica adults before diapause, exhibits an expression pattern of initial down-regulation, subsequent up-regulation, and then further down-regulation during diapause, and is up-regulated again after diapause termination. After RNA interference of GdLpR, the expression levels of diapause-related genes ecdysone receptor (EcR), nuclear hormone receptor (HR3), and vitellogenin (Vg) were downregulated, while the expression level of fatty acid synthase (FAS) and total lipid content were upregulated, leading to premature diapause in adult G. daurica. These results indicate that GdLpR is involved in the regulation of reproductive diapause in G. daurica. Full article

Polyamines (PAs) play critical roles in plant growth, somatic embryogenesis (SE), etc. Previous studies have demonstrated that exogenous PAs could promote SE in plants. However, the effects of PAs on Ginkgo biloba L. SE are still unknown, especially in the switch from the initial callus (IC) to the embryogenic callus (EC) stage or to the globular embryo (GE) stage. This work identified 34 genes involved in PAs metabolism in G. biloba using genome-wide analyses. These genes were clustered into six families and found to be unevenly distributed across 11 of the 12 chromosomes on the plant. These families contain 539 cis-acting elements that mainly respond to phytohormones, abiotic stress, meristem expression, etc. RNAseq analysis revealed that the expression of GbADC2, GbSAMDC2, GbSPMS1, GbCuAO1 and 3, and GbPAO3, 8, 6 and 13 genes in G. biloba were higher in the GE stage than in the IC stage. In addition, 1.0 mg·L⁻¹ spermine (Spm3) could promote the conversion of IC to EC, while 0.01 mg·L⁻¹ putrescine (Put1) could facilitate the transition from IC to EC and then to GE. During the conversion of IC to EC or to GE, higher levels of abscisic acid (ABA), superoxide dismutase (SOD), and peroxidase (POD) and lower levels of indole-3-acetic acid (IAA), gibberellin (GA3), and zeatin (ZT) were observed; concurrently, the H₂O₂ level was also observed to be high. Gene expressions of GbSPMS2, GbCuAO3, and GbPAO6 and 8 were upregulated, while GbADC2 expression was downregulated in the EC or GE stages under Spm3- or Put1- treatment. These results illustrate that exogenous PAs might alter the levels of the endogenous polyamine pool and lead to H₂O₂ production, which caused a certain oxidative stress. However, SOD and POD balanced H₂O₂ production and maintained homeostasis. The PAs–H₂O₂–ABA module might coordinate the regulation of early in of G. biloba. These relations were discussed in this work. These findings provide a foundation for comprehending the roles of PAs gene families in the key nodes of early SE in G. biloba. Full article

Cinnamomum camphora (L.) Presl, a perennial evergreen tree of the Lauraceae family, exhibits diverse chemotypes and abundant essential oil constituents, which are widely utilized in pharmaceuticals, perfumery, and fine chemicals. Based on the variation in dominant volatile constituents, five chemotypes have been identified: Borneol Chemotype (BC), Camphor Chemotype (CC), Eucalyptol Chemotype (EC), Isoborneol Chemotype (IC), and Linalool Chemotype (LC). Their compositions of monoterpenoids (MT) and sesquiterpenes (SQT) differ significantly. This review systematically summarizes the research progress on MT and SQT in different chemotypes of C. camphora over the past five decades, with a total of 164 compounds identified (83 MT and 81 SQT), and compares the unique and shared constituents among chemotypes. Pharmacological studies indicate that C. camphora essential oils from different chemotypes exhibit strong antibacterial, anti-inflammatory, and antitumor activities, with linalool, camphor, and several SQT compounds showing remarkable biological effects in multiple bacterial, fungal, and tumor cell models. The underlying mechanisms may involve the disruption of cell membrane integrity, inhibition of key metabolic enzymes, interference with genetic transcription, and synergistic effects among multiple constituents. However, research on low-abundance bioactive components in different chemotypes remains limited, and their pharmacological mechanisms require further elucidation. This review provides a systematic reference for the future exploration of bioactive constituents, mechanistic studies, and efficient utilization of essential oils from different chemotypes of C. camphora, offering valuable insights for refined resource exploitation and industrial application. Full article

Appendage autotomy frequently occurs during the cultivation of Exopalaemon carinicauda, which severely impacts its survival and economic benefits. To investigate the molecular mechanism underlying appendage regeneration in E. carinicauda, this study presents a comparative transcriptome analysis on samples from different stages of appendage regeneration in individuals of the same family of E. carinicauda. A total of 6460 differentially expressed genes (DEGs) were identified between the samples collected at 0 h post-autotomy (D0) and those collected at 18 h post-autotomy (D18h). Additionally, 7740 DEGs were identified between D0 and 14 d post-autotomy (D14d), with 3382 DEGs identified between D18h and D14d. Among them, differentially expressed genes such as EcR, RXR, BMP1, and Smad4 are related to muscle growth or molting and may be involved in the regeneration process. qRT-PCR results revealed that EcBMPR2 was expressed at relatively high levels in the gonad and ventral nerve cord tissues and that the highest level of expression was detected in the regenerative basal tissue at 24 h post-autotomy. In situ hybridization results indicated strong signals of this gene in the cells at the wound site at 72 h post-autotomy. Following knockdown of EcBMPR2, the expression levels of both EcBMPR1B and EcSmad1 were significantly downregulated, and long-term interference with the EcBMPR2 gene resulted in a significantly slower appendage regeneration process compared to the control group. When the downstream transcription factor EcSmad1 was knocked down, the two receptor genes EcBMPR2 and EcBMPR1B were downregulated, whereas EcBMP7 was upregulated. After inhibiting the BMP signaling pathway, the degree of cell aggregation at the autotomy site in the experimental group was significantly lower than that in the control group, the wound healing rate was delayed, and the blastema regeneration time was prolonged from 5 d to 7 d. Collectively, these results indicate that the BMP signaling pathway plays a critical role in the early stages of appendage regeneration in E. carinicauda. This study provides important theoretical insights for understanding limb regeneration in crustaceans. Full article

Chlorfenapyr is a widely used insecticide known to harm non-target insects, but its effects on reproductive development in the silkworm (Bombyx mori L.) remain incompletely understood. In this study, we investigated the histopathological and transcriptional changes in the gonads (ovaries and testes) of newly molted fifth-instar silkworm larvae exposed to chlorfenapyr. Histopathological analysis revealed delayed gonadal development, a reduction in oogonia and oocytes in the ovaries, and decreased numbers of spermatocytes in the testes. Transcriptome analysis identified significant differentially expressed genes (DEGs), mainly enriched in pathways such as “Drug metabolism—cytochrome P450”, “Insect hormone biosynthesis”, and “Ribosome”. Key up-regulated genes included members of the cytochrome P450 family (CYP6B5, CYP9f2, CYP6B6), glutathione S-transferases (GSTT1, GST1), and juvenile hormone-related enzymes (JHAMT, JHEH), indicating active detoxification and hormonal regulation responses. Several transcription factor families, particularly C2H2, HB-other, and TRAF, exhibited altered expression, suggesting roles in stress adaptation. Protein–protein interaction (PPI) network analysis identified hub genes such as EcR, Kr-h1, and various ribosomal proteins, highlighting their potential involvement in reproductive development. Quantitative PCR (qPCR) validated the transcriptomic data, confirming the reliability of the results. Overall, these findings enhance our understanding of chlorfenapyr’s impact on silkworm reproductive development and the underlying molecular mechanisms, providing valuable insights for sustainable pest management and ecological risk assessment of insecticides. Full article

This study aims to evaluate the tolerance of an endophytic fungus isolated from the fibrous roots of Gentiana yunnanensis Franch. to arsenic (As) and elucidate the underlying physiological and molecular mechanisms. The filamentous fungus is identified as Cladosporium cladosporioides based on morphological characteristics and phylogenetic tree analysis, belonging to the family Moniliaceae and Phyla Hyphomycetes. The tolerance of C. cladosporioides to As(V) was assessed by measuring its biomass under varying concentrations of As(V). The fungus exhibited remarkable As(V) tolerance, with an EC₅₀ value of 2051.94 mg/L, and accumulated high concentrations of As in its mycelium. Subcellular distribution analysis revealed that As was predominantly localized in the cell wall fraction, with levels 4.06 times higher than those in the non-cell wall fraction. Notably, the concentrations of total organic As and As(III) in the mycelium were 852.75 μg/g and 24.94 μg/g, respectively, with conversion ratios of 76.64% and 2.24%. The organic As levels significantly surpassed both As(V) and As(III) concentrations in all cellular fractions (cell wall and non-cell wall components), demonstrating particularly efficient As transformation in C. cladosporioides. Under As(V) stress, the membrane antioxidant system, including superoxide dismutase (SOD), metallothionein (MT), glutathione (GSH), and melanin, was activated and significantly enhanced to mitigate oxidative damage. Transcriptomic analysis identified 4771 differentially expressed genes (DEGs; 2527 upregulated), including highly expressed As-responsive genes (CcArsH_1, CcARR_1, CcARR_3, CcGST_1, and CcGST_3). Strong correlations emerged between As speciation (total/organic/As(V)/As(III)), antioxidant levels, and DEG expression patterns. Taken together, these findings demonstrate that C. cladosporioides employs a multi-faceted As detoxification strategy involving subcellular distribution and reductive transformation (As(V) to As(III)/organic As), antioxidant system enhancement, transcriptomic adaptations, and integrated defense strategy. This work highlights C. cladosporioides potential for As bioremediation and elucidates As accumulation mechanisms in G. yunnanensis. Full article

Partnerships with parents in early childhood education and care services are a hallmark of quality education. Educators in Western countries work within a highly regulated environment, where government documents, such as frameworks, standards, and curricula, direct most of their work, time, and energy. Despite this, data from our mixed methods online survey from Australia, Canada, Denmark, Georgia, and Italy revealed a strong resistance to the homogeneity these documents prescribe. For the quantitative data, we used cross-tabulation and descriptive statistics. For the qualitative data, we used deductive thematic analysis using a parent–educator partnership framework. Educators described parents in their service as partners in their child’s education. This included efforts to share information, consult, negotiate, and build partnerships; problem solve; and monitor, report and manage the partnership. The educators talked about the uniqueness of their approaches to parents and families within their contextualised services. They then revealed how these unique features impacted their notions of quality and practice in these services. This will be of interest to policymakers, educators, and teacher educators. Full article

Laccase (LAC, EC 1.10.3.2) is integral to the formation of lignin synthesis, flavonoid production, and responses to both biotic and abiotic stresses. While recent studies have characterized numerous LAC gene families and their functions across various plants, information regarding LAC genes in woodland strawberry (Fragaria vesca) remains limited. In this study, we identified a total of 57 FvLAC genes in the Fragaria vesca genome, which were phylogenetically categorized into five distinct groups. Analysis of the gene structures revealed a uniformity in the exon–intron structure among the subgroups, while conserved motifs identified unique motifs specific to certain subgroups, suggesting functional variations. Chromosomal localization studies indicated that FvLACs are distributed across seven chromosomes, and collinearity analysis demonstrated that FvLACs exhibit collinearity within the species. Additionally, cis-acting element analysis suggested that FvLAC genes are involved in stress responses, hormone responses, light responses, and the growth and development of plants. qRT-PCR demonstrated that FvLACs responded to salt, drought, and hormone stresses, with the expression levels of FvLAC24, FvLAC32, and FvLAC51 continuously increasing under these stress conditions. Furthermore, transgenic yeast experiments revealed that FvLAC51 enhanced yeast tolerance to both salt and drought stresses, while FvLAC24 and FvLAC32 negatively regulated yeast tolerance under these same conditions. These findings provide a theoretical foundation for further investigation into the functions of FvLAC genes in woodland strawberry. Full article

The comparative analysis of homologous enzymes is a valuable approach for elucidating enzymes’ structure–function relationships. Glutathione transferases (GSTs, EC. 2.5.1.18) are crucial enzymes in maintaining the homeostatic stability of plant cells by performing various metabolic, regulatory, and detoxifying functions. They are promiscuous enzymes that catalyze a broad range of reactions that involve the nucleophilic attack of the activated thiolate of glutathione (GSH) to electrophilic compounds. In the present work, three highly homologous (96–98%) GSTs from ryegrass Lolium perenne (LpGSTs) were identified by in silico homology searches and their full-length cDNAs were isolated, cloned, and expressed in E. coli cells. The recombinant enzymes were purified by affinity chromatography and their substrate specificity and kinetic parameters were determined. LpGSTs belong to the tau class of the GST superfamily, and despite their high sequence homology, their substrate specificity displays remarkable differences. High catalytic activity was determined towards hydroxyperoxides and alkenals, suggesting a detoxification role towards oxidative stress metabolites. The prediction of the structure of the most active LpGST by molecular modeling allowed the identification of a non-conserved residue (Phe215) with key structural and functional roles. Site-saturation mutagenesis at position 215 and the characterization of eight mutant enzymes revealed that this site plays pleiotropic roles, affecting the affinity of the enzyme for the substrates, catalytic constant, and structural stability. The results of the work have improved our understanding of the GST family in L. perenne, a significant threat to agriculture, sustainable food production, and safety worldwide. Full article

In order to shed light on the usage of protein language model-based alignment procedures, we attempted the classification of Glutathione S-transferases (GST; EC 2.5.1.18) and compared our results with the ARBA/UNI rule-based annotation in UniProt. GST is a protein superfamily involved in cellular detoxification from harmful xenobiotics and endobiotics, widely distributed in prokaryotes and eukaryotes. What is particularly interesting is that the superfamily is characterized by different classes, comprising proteins from different taxa that can act in different cell locations (cytosolic, mitochondrial and microsomal compartments) with different folds and different levels of sequence identity with remote homologs. For this reason, GST functional annotation in a specific class is problematic: unless a structure is released, the protein can be classified only on the basis of sequence similarity, which excludes the annotation of remote homologs. Here, we adopt an embedding-based alignment to classify 15,061 GST proteins automatically annotated by the UniProt-ARBA/UNI rules. Embedding is based on the Meta ESM2-15b protein language. The embedding-based alignment reaches more than a 99% rate of perfect matching with the UniProt automatic procedure. Data analysis indicates that 46% of the UniProt automatically classified proteins do not conserve the typical length of canonical GSTs, whose structure is known. Therefore, 46% of the classified proteins do not conserve the template/s structure required for their family classification. Our approach finds that 41% of 64,207 GST UniProt proteins not yet assigned to any class can be classified consistently with the structural template length. Full article

β C-S lyases (β-CSLs; EC 4.4.1.8) are enzymes catalyzing the dissociation of β carbon–sulfur bonds of cysteine S-conjugates to produce odorant metabolites with a free thiol group. These enzymes are increasingly studied for their role in flavor generation in a variety of food products, whether these processes occur directly in plants, by microbial β-CSLs during fermentation, or in the mouth under the action of the oral microbiota. Microbial β-CSLs react with sulfur aroma precursors present in beverages, vegetables, fruits, or aromatic herbs like hop but also potentially with some precursors formed through Maillard reactions in cooked foods such as meat or coffee. β-CSLs from microorganisms like yeasts and lactic acid bacteria have been studied for their role in the release of polyfunctional thiols in wine and beer during fermentation. In addition, β-CSLs from microorganisms of the human oral cavity were shown to metabolize similar precursors and to produce aroma in the mouth with an impact on retro-olfaction. This review summarizes the current knowledge on β-CSLs involved in flavor generation with a focus on enzymes from microbial species present either in the fermentative processes or in the oral cavity. This paper highlights the importance of this enzyme family in the food continuum, from production to consumption, and offers new perspectives concerning the utilization of β-CSLs as a flavor enhancer. Full article