Search Results (3,151)

A nanoliter-scale fabrication method was applied to construct a colorimetric lateral flow strip for urea detection (Urea-CLFS). The device involves two main papers: a nitrocellulose membrane (NC-Mb) for urease enzyme immobilization and chromatography paper (CH-PP) containing a phenol red indicator. Urea-CLFS is a tool for detecting urea that is based on enzyme catalysis and the change in color of phenol red when urea is present. The Urea-CLFS fabrication was made possible by the minimal amount of nanoliters used in reagent consumption. The use of small arrays of phenol red dots provides a higher response result compared to single dots applied on CH-PP. To find the most effective design, it analyzed how urease was aligned on NC-Mb horizontally and vertically. According to our findings, the vertical alignment of the urease enzyme on NC-Mb leads to a prolonged reaction time, which leads to higher product production. The optimization process included optimizing various parameters, including the layer number of phenol red on CH-PP, phenol red concentration, urease concentration, reaction time, and sample volume. Under optimal conditions, the Urea-CLFS provided a linear range of 0.25–8.0 mmol L⁻¹ with an LOD of 0.34 mmol L⁻¹, which is sufficient for human health diagnostics. The accuracy of the Urea-CLFS was demonstrated by the recovery of the human urine sample between 95 ± 3% and 103 ± 3% (n = 3). Full article

Glucosylceramide (GlcCer), a central glycosphingolipid derived from ceramide, is increasingly recognized as a bioactive lipid that intersects with key metabolic, inflammatory, and oncogenic pathways. While its dysregulation has long been associated with lysosomal storage disorders such as Gaucher disease (GD), growing evidence implicates GlcCer in cancer initiation and progression, particularly within tumor-predisposing conditions. GlcCer modulates membrane microdomains, intracellular trafficking, and cell signaling, counteracting ceramide-induced apoptosis and promoting cellular survival. In cancer, aberrant upregulation of UDP-glucose ceramide glucosyltransferase (UGCG), the enzyme responsible for GlcCer synthesis, drives tumor growth, metastasis, and multidrug resistance through activation of pathways such as phosphoinositide 3-kinase/protein kinase B (PI3K/Akt), mitogen-activated protein kinase (MAPK), canonical Wnt pathway (Wnt/β-catenin), and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathways. Specific GlcCer species (e.g., C16:0, C18:0, C24:1) display tissue-dependent functions, adding structural specificity to their oncogenic potential. Moreover, emerging links between GlcCer metabolism and chronic inflammation, oxidative stress, and altered glucose utilization highlight its role as a metabolic node bridging inherited metabolic disorders and malignancy. This review integrates recent advances in GlcCer biology, emphasizing its roles in tumor-predisposing diseases and exploring its potential as a biomarker and therapeutic target in oncology. Full article

Antimicrobial resistance (AMR) is a growing global threat, exacerbated by the adaptive mechanisms of Gram-negative ESKAPE pathogens, which include biofilm formation and outer membrane vesicle (OMV) production. Biofilms create robust protective barriers that shield bacterial communities from immune responses and antibiotic treatments, while OMVs contribute to both defense and offense by carrying antibiotic-degrading enzymes and delivering virulence factors to host cells. These mechanisms not only enhance bacterial survival but also increase the virulence and persistence of infections, making them a significant concern in clinical settings. This review explores the molecular processes that drive biofilm and OMV formation, emphasizing their critical roles in the development of AMR. By understanding these mechanisms, new therapeutic strategies can be developed to disrupt these defenses, potentially improving the efficacy of existing antibiotics and slowing the spread of resistance. Additionally, the use of OMVs in vaccine development and drug delivery offers promising avenues for future research. Addressing these challenges requires a comprehensive approach, combining advanced research with innovative therapies to combat the escalating threat of AMR and improve patient outcomes. Full article

Sugarcane smut, caused by Sporisorium scitamineum, is a devastating fungal disease of sugarcane. Sexual mating/filamentation of opposite mating types is a key step in the infection and pathogenicity of S. scitamineum, yet its regulation remains unclear. In this study, we identified a cytochrome P450 enzyme-encoding gene, SsCYP64, which plays an important role in oxidative stress and maintaining cell membrane stability in S. scitamineum. Further investigations revealed that deletion of SsCYP64 leads to a decrease in the transcriptional level of SsPRF1, a key transcription factor regulating the sexual mating of S. scitamineum. Subsequently, the constitutive expression of SsPRF1 restored the defect in sexual mating/filamentation of the SsCYP64 deletion mutant, indicating that SsCyp64 regulates the sexual reproduction of S. scitamineum by mediating the transcriptional level of SsPRF1. In addition, metabolomic analysis revealed that the fatty alcohol metabolite γ-linolenyl alcohol significantly decreased in the SsCYP64 deletion mutant, whereas exogenous supplementation with γ-linolenyl alcohol increased the transcriptional level of SsPRF1 and partially restored the sexual mating/filamentation of the SsCYP64 deletion mutant. In conclusion, our results indicated that SsCyp64 mediated the transcription of SsPRF1 by modulating γ-linolenyl alcohol levels, thereby regulating the formation of dikaryotic hyphae in S. scitamineum. These findings provide new insights into the role of cytochrome P450 enzymes in the pathogenic process of plant pathogenic fungi. Full article

Background/Objectives: Melanoma cells enhance glycolysis and expand lysosomes to support energy metabolism, proliferation, and metastasis. However, lysosomal membrane permeabilization (LMP) causes cathepsin leakage into cytosol triggering cytotoxicity. This study investigated the antimelanoma effect of 2-deoxy-D-glucose (2DG), an inhibitor of glycolytic enzyme hexokinase-2, in combination with cathepsin C-dependent LMP inducer L-leucyl-L-leucine methyl ester (LLOMe) and cathepsin C-independent LMP-inducers mefloquine and siramesine. Methods: The viability of A375 and B16 melanoma cells and primary fibroblasts was measured by crystal violet. Apoptosis, necrosis, and LMP were assessed by flow cytometry; caspase activation, mitochondrial depolarization, superoxide production, and energy metabolism were analyzed by fluorimetry, and expression of cathepsins and hexokinase-2 was evaluated by immunoblot. Appropriate inhibitors, antioxidant, and energy boosters were used to confirm cell death type and mechanism. Results: LLOMe triggered LMP, mitochondrial depolarization, and mitochondrial superoxide production, while suppressing oxidative phosphorylation. 2DG suppressed glycolysis and, together with LLOMe, synergized in ATP depletion, caspase activation, and mixed apoptosis and necrosis in A375 cells. Inhibitors of lysosomal acidification, cysteine cathepsins, and caspases, as well as antioxidant and energy boosters, reduced 2DG+LLOMe-induced toxicity. Cathepsins B, C, and D were lower, while hexokinase-2 was higher in A375 cells than fibroblasts. Accordingly, 2DG exhibited lower while LLOMe exhibited higher toxicity against fibroblasts than A375 and B16 cells. However, mefloquine and siramesine induced stronger LMP in A375 cells than in fibroblasts and showed melanoma-selective toxicity when combined with 2DG. Conclusions: 2DG-mediated glycolysis inhibition in combination with lysosomal destabilization induced by mefloquine and siramesine, but not with non-selectively toxic LLOMe, may be promising antimelanoma strategy. Full article

Salt stress is a significant environmental factor that inhibits maize growth and development, severely affecting yield formation. Interestingly, nanomaterials, particularly ZnONPs, can enhance resistance to various stresses and support healthy crop growth. However, the effects of ZnONPs on maize under salt stress remain unclear. This study investigates the effect of foliar and seed exposure to zinc oxide nanoparticles (ZnONPs) on reducing NaCl-induced salt stress in two maize inbred lines (NKY298-1 and NKY211). Over a period of seven days, under 120 mM NaCl, we measured growth, reactive oxygen species (ROS), malondialdehyde (MDA), membrane stability index (MSI), water status (relative water content, RWC), photosynthetic pigments and parameters, selected photosynthetic enzymes, and antioxidant enzyme activities. Then, we propose four composite indices, including stress improvement index (SII), alleviation capacity index (ACI), comprehensive improvement effects (CIE), and comprehensive alleviation capacity (CAC), to rank the effectiveness of ZnONP doses. The findings suggested that 50–100 μM ZnONPs significantly mitigate salt damage, with optimal doses varying by genotype (50 μM for NKY211 and 100 μM for NKY298-1). Notably, the study’s originality lies in its side-by-side composite scoring across 26 traits in two maize genotypes’ seedlings. In conclusion, the findings will provide a new idea for research on the molecular mechanism by which exogenous ZnONPs application improves the salt tolerance of maize seedlings. Full article

Cadmium (Cd), a pervasive and highly phytotoxic metal pollutant, poses severe threats to agricultural productivity, ecosystem stability, and human health through its entry into the food chain. Plants have evolved intricate defense mechanisms, among which the strategic manipulation of nutrient elements emerges as a critical physiological and biochemical strategy for mitigating Cd stress. This comprehensive review delves deeply into the multifaceted roles of essential macronutrient elements (nitrogen, phosphorus, potassium, calcium, magnesium, sulfur), essential micronutrient elements (zinc, iron, manganese, copper) and non-essential beneficial elements (silicon, selenium) in modulating plant responses to Cd toxicity. We meticulously dissect the physiological, biochemical, and molecular underpinnings of how these nutrients influence Cd bioavailability in the rhizosphere, Cd uptake and translocation pathways, sequestration and compartmentalization within plant tissues, and the activation of antioxidant defense systems. Nutrient elements exert their influence through diverse mechanisms: competing with Cd for root uptake transporters, promoting the synthesis of complexes that reduce Cd mobility, stabilizing cell walls and plasma membranes to restrict apoplastic flow and symplastic influx, modulating redox homeostasis by enhancing antioxidant enzyme activities and non-enzymatic antioxidant pools, regulating signal transduction pathways, and influencing gene expression profiles related to metal transport, chelation, and detoxification. The complex interactions between nutrients themselves further shape the plant’s capacity to withstand Cd stress. Recent advances elucidating nutrient-mediated epigenetic regulation, microRNA involvement, and the role of nutrient-sensing signaling hubs in Cd responses are critically evaluated. Furthermore, we synthesize the practical implications of nutrient management strategies, including optimized fertilization regimes, selection of nutrient-efficient genotypes, and utilization of nutrient-enriched amendments, for enhancing phytoremediation efficiency and developing low-Cd-accumulating crops, thereby contributing to safer food production and environmental restoration in Cd-contaminated soils. The intricate interplay between plant nutritional status and Cd stress resilience underscores the necessity for a holistic, nutrient-centric approach in managing Cd toxicity in agroecosystems. Full article

Schizochytrium is often added to feed to enhance the growth and health of farmed animals, yet research on its effects on amphibians remains relatively scarce. Here, this study investigated the effects of dietary Schizochytrium meal on growth, metamorphosis, intermediate metabolism, and intestinal health of bullfrogs. Six compound feeds (S0–S5) containing different gradients of Schizochytrium meal (0.00, 2.00, 5.00, 10.00, 15.00, and 20.00 g/kg diets) were formulated. After 90 days, the S4 group (15.00 g/kg) exhibited significantly superior growth performance, with the weight gain rate (WGR) increasing by up to 23.78% compared to the control (S0). Metamorphosis rate (MR) peaked at 23.33% in the S4 group. The enzyme activities of digestion (amylase (AMS), lipase (LPS), protease), brush border membrane (Na⁺, K⁺-ATPase, alkaline phosphatase (AKP), γ-glutamyl transferase (γ-GT), creatine kinase (CK), and antioxidation (superoxide dismutase (SOD), catalase (CAT)), as well as microvilli length and mucosal epithelial cell height in the intestine were the highest in the S4 group. Intestinal microbial diversity (Ace index) significantly increased by 41.28% in the S4 group, which also promoted beneficial bacteria. Key genes related to the GH-IGF-1 axis, metabolism, and intestinal barrier function were significantly upregulated with increasing Schizochytrium levels up to 15.00 g/kg, whereas pro-inflammatory genes showed an opposite trend. Overall, dietary supplementation with Schizochytrium meal at 15.00 g/kg promotes growth, metamorphosis, and intestinal health in bullfrog tadpoles by modulating the GH-IGF-1 axis, enhancing digestion and absorption, and improving intestinal integrity. Optimal Schizochytrium meal levels were identified as 13.27 g/kg. Full article

Fusarium wilt disease, caused by Fusarium oxysporum f. sp. cucumerinum (FOC), leads to widespread yield losses and quality deterioration in cucumber. Endophytes, as environmentally friendly control agents that enhance pathogen resistance in their host plants, may mitigate these problems. In this study, we isolated 14 endophytic bacteria from invasive Ambrosia artemisiifolia and screened the strain Bacillus subtilis TCX1, which exhibited significant antagonistic activity against FOC (inhibitory rate of 86.0%). TCX1 killed Fusarium oxysporum by being highly likely to produce lipopeptide and producing wall hydrolytic enzymes including protease, cellulase, and β-glucanase, thereby inhibiting mycelial growth and spore germination and causing peroxidation of FOC’s cytoplasmic membrane. In addition to its direct effects, TCX1 exerts indirect effects by inducing cucumber resistance to FOC. When cucumber seedlings were inoculated with TCX1, antioxidant enzymes related to disease resistance, including Superoxide dismutase (SOD), Peroxidase (POD), Polyphenol oxidase (PPO) and Phenylalanine ammonialyase (PAL) in cucumber, were significantly increased. The marker genes involved in induced systemic resistance and the salicylic acid signaling pathway, such as npr1, pr1a, pr2, pr9, lox1, and ctr1, were also dramatically upregulated, indicating these pathways played an important role in improving cucumber resistance. Notably, TCX1 can also promote cucumber growth through producing indole-3-acetic acid, solubilizing phosphate, and secreting siderophores. Given that TCX1 has dual functions as both a biological control agent and a biofertilizer, it offers an effective strategy for managing cucumber seedling blight while enhancing plant productivity. Full article

Leprosy is a chronic infectious disease that targets the peripheral nervous system, leading to peripheral neuropathy. Mycobacterium leprae primarily infects Schwann cells, adipocytes, and macrophages, altering their metabolism and gene expression. This study investigates the metabolic interaction between M. leprae and Schwann cells, with a focus on indoleamine 2,3-dioxygenase (IDO), a key enzyme in tryptophan catabolism via the kynurenine pathway. We found that M. leprae induces IDO expression in Schwann cells, suggesting a role in immune modulation and neuropathy. Inhibition of IDO with 1-methyl-L-tryptophan (1-MT) reduced Schwann cell viability and metabolic activity in response to M. leprae. After 24 h of infection, M. leprae impaired mitochondrial membrane potential, although no significant changes in autophagy or mitochondrial ultrastructure were observed by electron microscopy. Interestingly, IDO1 inhibition upregulated the expression of antioxidant genes, including GPX4, NFE2L2, and HMOX1. In conclusion, these findings highlight a central role for IDO in shaping the metabolic and immunological response of Schwann cells to M. leprae infection. IDO induction contributes to immune regulation and cellular stress, while its inhibition disrupts cell viability and promotes antioxidant gene expression. These results position IDO as a potential therapeutic target for modulating host–pathogen interactions and mitigating nerve damage in leprosy. Full article

Background/Objectives: Malaria remains the most critical parasitic disease globally, responsible for over 600.000 deaths annually. In sub-Saharan Africa, co-infections of Plasmodium falciparum with other pathogens, particularly Staphylococcus aureus, are common in children with severe malaria. Therefore, the design of new compounds targeting both pathogens appears to be an urgent priority. Methods: A small series of hybrid compounds was designed and synthesized by linking the pharmacophore of the antimalarial drug chloroquine with the phenothiazine core. These compounds were tested in vitro against a panel of microbial strains and further analyzed through in silico simulations to predict their physical-chemical properties. Results: Compounds 4b and 5b emerged the most potent candidates of the series, showing a sub-micromolar inhibitory activity on P. falciparum, and a promising micromolar potency on S. aureus alongside with a low toxicity on mammalian cells. Molecular docking followed by molecular dynamics (MD) simulations identified the respiratory membrane NDH-2 enzyme as common target in both pathogens. Conclusions: Both experimental and computational findings provide compelling evidence for the use of the designed compounds in a STOP strategy, i.e., Same-Target-Other-Pathogen, to treat malaria and bacterial infections concurrently. Full article

Ilex verticillata (winterberry) is a valuable ornamental shrub increasingly threatened by leaf blight, a disease that compromises its aesthetic and economic value. While fungal pathogens like Alternaria alternata are known to cause leaf blight in horticultural crops, their role in I. verticillata and the host’s defense mechanisms have not been fully characterized. Our study investigated the pathogen-host interaction by identifying the causal agent and examining the physiological and molecular defense mechanisms of I. verticillata. Through morphological and multi-locus molecular analyses (ITS, TEF1-α, G3PDH, RPB2), A. alternata was confirmed as the primary pathogen, fulfilling Koch’s postulates. Pathogenicity assays revealed distinct disease progression stages, from necrotic lesions to tissue degradation. Transcriptomic profiling uncovered dynamic host responses, with early upregulation of pattern recognition receptors (PRRs) and transcripts encoding antioxidant enzymes (SOD, CAT), followed by downregulation of metabolic pathway genes. Phytohormone analysis highlighted intricate crosstalk, with salicylic acid (SA) peaking during mid-infection and jasmonic acid (JA) rebounding later, reflecting a coordinated defense strategy. Additionally, the oxidative stress marker malondialdehyde (MDA), an indicator of membrane lipid peroxidation, surged early, indicating membrane damage, while sustained induction of antioxidant enzymes suggested adaptive responses. The key finding was distinct phytohormone crosstalk, characterized by a mid-infection SA peak followed by a late JA rebound, alongside an early oxidative burst marked by MDA accumulation and sustained antioxidant enzyme activity. These findings provide a framework for understanding I. verticillata’s defense mechanisms and offer insights for developing targeted disease management strategies, such as resistant cultivar breeding or hormone-mediated interventions. Full article

Plant-based extracts and elicitors (signaling molecules that activate the fruit’s innate defense responses) have emerged as promising and sustainable alternatives to synthetic chemicals for preserving postharvest fruit quality and extending shelf life. This review provides a comprehensive analysis, uniquely complemented by a bibliometric assessment of the research landscape from 2005 to 2025, to identify key trends and effective solutions. This review systematically examined the efficacy of various natural compounds including essential oils (complex volatile compounds with potent antimicrobial activity such as lemongrass and thyme), phenolic-rich botanical extracts like neem and aloe vera, and plant-derived elicitors such as methyl jasmonate and salicylic acid. Their preservative mechanisms are multifaceted, involving direct antimicrobial activity by disrupting microbial membranes, potent antioxidant effects that scavenge free radicals, and the induction of a fruit’s innate defense systems, enhancing the activity of enzymes like superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD). Applications of edible coatings of chitosan or aloe vera gel, nano-emulsions, and pre- or postharvest treatments effectively reduce decay by Botrytis cinerea and Penicillium spp.), delay ripening by suppressing ethylene production, minimize water loss, and alleviate chilling injury. Despite their potential, challenges such as sensory changes, batch-to-batch variability, regulatory hurdles, and scaling production costs limit widespread commercialization. Future prospects hinge on innovative technologies like nano-encapsulation to improve stability and mask flavors, hurdle technology combining treatments synergistically, and optimizing elicitor application protocols. This review demonstrates the potential of continued research and advanced formulation to create plant-based preservatives, that can become integral components of an eco-friendly postharvest management strategy, effectively reducing losses and meeting consumer demands for safe, high-quality fruit. Full article

Biodegradable plastics are not a primary solution to plastic pollution, and empirical evidence on whether they are environmentally friendly remains lacking. In this study, we systematically compared the toxic effects of traditional microplastics (polypropylene, PP; polystyrene, PS) with biodegradable microplastics (polylactic acid, PLA; polyhydroxyalkanoates, PHA) on the haplic phaeozem ecosystem. Through mathematical modeling analysis, it was found that earthworms initially rely on antioxidant enzymes to resist stress, mid-term activation of detoxifying enzymes to repair damage, and maintaining physiological balance through metabolic regulation and immune enhancement in later stages. We elucidated their mechanism differences: PLA and PP caused severe damage to the antioxidant system and cell membrane, with PLA mainly relying on POD to clear peroxides and PP relying on GST. In addition, PLA and PS can induce early neurotoxicity (AChE), while PHA induces late neurotoxicity. Furthermore, this study provides direct evidence proving that biodegradable microplastics are not environmentally friendly by breaking through the one-way research framework of “microplastic biotoxicity” and innovatively constructing a path analysis model that links biological physiological responses with soil ecological functions. We also provide a scientific basis to evaluate the ecological risks of microplastic pollution in soil and the whether biodegradable plastics are truly environmentally friendly. Full article

Extracellular signal-regulated kinase 1/2 (ERK1/2) inhibitors show therapeutic potential in triple-negative breast cancer (TNBC), but resistance through compensatory signaling limits their efficacy. We previously identified the secretory carrier membrane protein 3 (SCAMP3) as a regulator of TNBC progression and ERK1/2 activation. Here, we investigated the role of SCAMP3 in ERK1/2 signaling and therapeutic response using TMT-based LC-MS/MS phosphoproteomics of wild-type (WT) and SCAMP3 knockout (SC3KO) SUM-149 cells under basal conditions, after epidermal growth factor (EGF) stimulation, and during ERK1/2 inhibition with MK-8353. A total of 4408 phosphosites were quantified, with 1093 significantly changed. SC3KO abolished residual ERK activity under MK-8353 and affected the compensatory activation of oncogenic pathways observed in WT cells. SC3KO reduced the phosphorylation of ERK feedback regulators RAF proto-oncogene serine/threonine-protein kinase Raf-1 (S43) and the dual-specificity mitogen-activated protein kinase kinase 2 (MEK2) (T394), affected other ERK targets, including nucleoporins, transcription factors, and metabolic enzymes triosephosphate isomerase (TPI1) (S21) and ATP-citrate lyase (ACLY) (S455). SCAMP3 loss also impaired the mammalian target of rapamycin complex I (mTORC1) signaling and disrupted autophagic flux, evidenced by elevated sequestosome-1 (SQSTM1/p62) and microtubule-associated protein light chain 3 (LC3B-II) with reduced levels of the autophagosome lysosome maturation marker, Rab7A. Beyond ERK substrates, SC3KO affected phosphorylation events mediated by other kinases. These findings position SCAMP3 as a central coordinator of ERK signaling and autophagy. Our results support SCAMP3 as a potential therapeutic target to enhance ERK1/2 inhibitor clinical efficacy and overcome adaptive resistance mechanisms in TNBC. Full article