Search Results (230)

Ants act as keystone species in terrestrial ecosystems, providing important ecosystem services. The large-scale production and application of GO constitute a predominant contributor to its inevitable environmental dispersion. Most GO toxicity studies have focused on plants, animals, and microorganisms, with limited research on ground-dwelling ants. In the study, we used Camponotus japonicus as a model to investigate the toxic effects of GO on ants by integrating physiological characteristics, gut microbiota and transcriptome profiling. Results showed that GO exposure induced mitochondrial dysfunction, as evidenced by mitochondrial ROS accumulation and elevated mitochondrial membrane permeability. Physiological assessments revealed that GO exposure induced oxidative stress. Specifically, GO treatment significantly suppressed superoxide dismutase (SOD) and catalase (CAT) activities, while enhancing peroxidase (POD) and carboxylesterase (CarE) activities and increasing the levels of malondialdehyde (MDA) and trehalose. Gut microbiota analyses showed that GO remarkably reduced the relative abundance of beneficial bacterial symbionts (e.g., Candidatus Blochmannia) and destabilized the whole community structure. Furthermore, transcriptome profiling revealed 680 differentially expressed genes (DEGs) in the ants after GO exposure, most of which were significantly enriched in pathways associated with oxidative phosphorylation. This study suggests that GO may compromise ant-mediated ecosystem function and provides a reference for understanding the environmental risks of GO. Our findings also offer new insights for protecting the ecosystem services of ants. Full article

Human carboxylesterases (CES) are enzymes that play a central role in the metabolism and biotransformation of diverse endogenous substances and xenobiotics. The two most relevant isoforms, CES1 and CES2, are crucial in clinical pharmacotherapy as they catalyze the hydrolysis of numerous approved drugs and prodrugs. Elucidating the structural basis of CES isoform substrate specificity is essential not only for understanding and anticipating the biological fate of administered drugs, but also for designing prodrugs with optimized site-specific bioactivation. Additionally, this knowledge is also important for the design of biomedically useful molecules such as subtype-targeted CES inhibitors and fluorescent probes. In this context, both experimental and computational methodologies have been used to explore the mechanistic and thermodynamic properties of CES-mediated catalysis. Experimental designs commonly employ recombinant CES or human tissue microsomes as enzyme sources, utilizing quantification methods such as spectrophotometry (UV and fluorescence) and mass spectrometry. Computational approaches fall into two categories: (1) modeling substrate: CES recognition and affinity (molecular docking, molecular dynamics simulation, and free-energy binding calculations), and (2) modeling substrate: CES reaction coordinates (hybrid QM/MM simulations). While experimental and theoretical approaches are highly synergistic in studying the catalytic properties of CES subtypes, they represent distinct technical and scientific fields. This review aims to provide an integrated discussion of the key concepts and the interplay between the most commonly used wet-lab and dry-lab strategies for investigating CES catalytic activity. We hope this report will serve as a concise resource for researchers exploring CES isoform specificity, enabling them to effectively utilize both experimental and computational methods. Full article

The widespread cultivation of transgenic Bt cotton has elevated Apolygus lucorum (Meyer-Dür) to a major pest in cotton agroecosystems. Its rapidly developing resistance to insecticides poses a serious challenge to sustainable agriculture. In this study, we assessed the susceptibility of a field-collected population from Anyang, Henan Province, in relation to a laboratory-susceptible strain, to elucidate the present status and molecular basis of resistance to beta-cypermethrin. First, the toxicity of beta-cypermethrin to A. lucorum was assessed through a diet-incorporation method. Subsequently, the enzymatic activities of carboxylesterase (CarE) and glutathione S-transferase (GST) were measured, and the expression levels of CarE1 and GST1 were quantified by quantitative real-time PCR (qRT-PCR). Finally, the function of candidate genes was confirmed using RNA interference (RNAi) technology. The bioassays results indicated that the median lethal concentration (LC₅₀) for the laboratory and Anyang field strain were 343.34 mg/L and 700.45 mg/L, respectively. Following 48 h of exposure to the LC₃₀ of the susceptible strain, the mortality rate of the field population (20.00%) was significantly lower than that of the laboratory population (33.33%), suggesting an increase in resistance. The field population of A. lucorum exhibited significantly higher activities of CarE (1.61-fold) and GST (1.71-fold) compared to the laboratory strain, accompanied by 3.63- and 4.23-fold overexpression of the corresponding genes CarE1 and GST1. Spatiotemporal expression profiling revealed that CarE1 expression was highest in 4th–5th instar nymphs and adults, with predominant localization in the midgut, while GST1 expression peaked in 4th–5th instar nymphs and was abundant in the midgut and fat body. RNAi-mediated knockdown of CarE1 and GST1 significantly enhanced susceptibility to beta-cypermethrin in field populations, resulting in elevated mortality 48 h post-treatment compared to controls. In conclusion, the field population of A. lucorum has developed considerable resistance to beta-cypermethrin, strongly correlated with overexpression of CarE1 and GST1. These results deepen our understanding of metabolic resistance mechanisms and offer valuable insights for developing targeted pest control strategies. Full article

Landfills represent increasingly common anthropogenic habitats that provide food resources but also expose wildlife to complex chemical mixtures. White Storks (Ciconia ciconia) have recently expanded breeding near such sites, yet little is known about the physiological consequences of landfill dependence across time. In 2025, we assessed biomarker responses in White Stork (Ciconia ciconia) nestlings from the Jakuševec landfill (Zagreb, Croatia), a post-remediated site still in partial operation, three years after the initial studies conducted in 2021 and 2022. Activities of acetylcholinesterase (AChE), carboxylesterase (CES), glutathione S-transferase (GST) and glutathione reductase (GR), as well as levels of reduced glutathione (GSH) and reactive oxygen species (ROS), were quantified in extracellular (plasma) and intracellular (post-mitochondrial S9) blood fractions. Neurotoxicity biomarkers (AChE, CES) showed small increases in 2022, followed by significant declines in 2025, indicating potential changes in exposure to neuroactive compounds. Oxidative-stress biomarkers displayed contrasting patterns: GST and GR decreased progressively, whereas ROS rose and GSH shifted in opposite directions between fractions, together suggesting rising oxidative challenge and altered redox balance. The combined biomarker response suggests continuing low-level exposure to neurotoxic and redox-active compounds despite landfill remediation. Our findings highlight that urban landfills, even in post-closure phases, remain physiologically active systems influencing wildlife health and should be incorporated into long-term ecotoxicological and conservation monitoring frameworks. While independent long-term monitoring shows that the Jakuševec White Stork colony has continued to grow over the past decade, the physiological responses detected in nestlings highlight the importance of assessing how chronic low-level exposure might influence population health in the long term. Full article

Chinese wolfberry (Lycium barbarum L.), a specialty crop with ecological, medical, and economic value in Ningxia province of China, is subject to severe damage from Aphis gossypii Glover. Currently, A. gossypii populations show extremely high-level resistance to beta-cypermethrin in the major wolfberry planting areas in Ningxia. The specific resistance mechanisms, however, are still not known. In this work, we collected a field A. gossypii strain (HSP) from a wolfberry orchard in Ningxia in 2021 using a single-time sampling method, and its resistance to beta-cypermethrin was determined to be extremely high (994.74-fold) as compared with that of a susceptible strain (SS). Then we explored the potential resistance mechanisms from two aspects, namely, metabolic detoxification and target-site alterations. Bioassays of beta-cypermethrin with or without a synergist showed that piperonyl butoxide (PBO) significantly increased the toxicity of beta-cypermethrin (4.72-fold) to the HSP strain, while triphenyl phosphate (TPP) and diethyl maleate (DEM) exhibited no significant synergistic effects. Correspondingly, the O-demethylase activity of cytochrome P450s in the HSP strain was 1.68-fold higher than that in the susceptive strain (SS), whereas changes in carboxylesterases and glutathione S-transferases activities were unremarkable. Also, fifteen upregulated P450 genes were identified by both RNA-Seq and qRT-PCR technologies, containing eleven CYP6 genes, three CYP4 genes, and one CYP380 gene. Especially, five CYP6 genes with high relative expression levels (>3.00-fold) were intensively expressed by beta-cypermethrin induction in the HSP aphids. These metabolism-related results indicate the key role of P450-mediated metabolic detoxification in HSP resistance to beta-cypermethrin. Sequencing of voltage-gated sodium channel (VGSC) genes identified a prevalent M918L mutation and a new G1012D mutation in HSP A. gossypii. Moreover, heterozygous 918 M/L and 918 M/L + G1012D mutations were the dominant genotypes with frequencies of 60.00% and 36.67% in the HSP population, respectively. Overall, VGSC mutations along with P450-mediated metabolic resistance contributed to the extremely high resistance of the HSP wolfberry aphids to beta-cypermethrin, providing support for A. gossypii control and resistance management in the wolfberry planting areas of Ningxia using insecticides with different modes of action. Full article

The high incidence of anthropogenic impacts in the Mediterranean basin raises concerns on the health and quality of commercial fish species. This study aims to evaluate the health status of the European hake, Merluccius merluccius, from three areas of the Catalan coast (NW Mediterranean Sea) with different anthropogenic impacts (i.e., chemical pollution, litter, …) and assess if hake could serve as a sentinel species. We measured biomarkers of chemical exposure including B-esterases, antioxidant enzymes (GST, GR, GPx, CAT), biotransformation markers (EROD), lipid peroxidation, and macro-parasite assemblages. Hake showed, generally, a good health status across all areas with homogeneous patterns for most parameters. Tissue-specific differences included elevated gonadal cholinesterases and higher brain and hepatic carboxylesterase activities in the south, and increased hepatic EROD but lower lipid peroxidation in the central Barcelona area. Parasite assemblages were dominated by Digenea, Cestoda, and Nematoda, with higher cestode prevalence in both central and south zones. In summary, despite a greater prevalence of environmental pollution in the central region, there was a homogeneous pattern in hake health indicators throughout the three studied fishing zones. These results establish a baseline for hake health in Mediterranean waters and suggest that the species’ high mobility and wide depth range may limit its utility to detect local-scale pollution impacts, though it may serve as a regional-scale bioindicator. Full article

Trace metals and metalloids pose persistent threats to freshwater ecosystems, yet their trophic transfer and sublethal effects across food webs remain poorly understood. We investigated bioaccumulation patterns and biomarker responses in a predator–prey system comprising Prussian carp (Carassius gibelio) and Great Cormorant (Phalacrocorax carbo) nestlings from the Danube floodplain wetland Kopački rit Nature Park (Croatia) during 2023–2024. Concentrations of arsenic (As), selenium (Se), cadmium (Cd), mercury (Hg) and lead (Pb) were determined in Prussian carp liver and in Great Cormorant whole blood. The activities of acetylcholinesterase (AChE), carboxylesterase (CES), glutathione S-transferase (GST) and the levels of reactive oxygen species (ROS) and reduced glutathione (GSH) were measured in brain, muscle and gill tissues of Prussian carp, as well as in plasma and S9 blood fractions of Great Cormorants. In addition, tissue-specific metal concentration ratios (TSMCR) were calculated to assess the relative magnitude of recent dietary exposure in the predator compared to the prey. Biomarker activity showed strong tissue- and fraction-specific variation, with temporal differences. Exposure–response modelling revealed significant associations between As, Cd, and Hg and specific biomarkers, particularly in gill and plasma. Cross-species comparisons indicated elevated TSMCR as a proxy for recent trophic exposure only for Hg in 2023, whereas As and Se exhibited lower TSMCR. These findings demonstrate that metal exposure in floodplain systems induces physiological responses and Hg poses the greatest prey-to-predator exposure risk, highlighting the value of integrating pollutant measurements with mechanistic biomarker endpoints to evaluate ecosystem-level impacts. Full article

Background/Objectives: This study focused on synthesizing novel alkenyl derivatives of azinylferrocenes and evaluating their potential as Alzheimer’s disease (AD) therapeutics. Methods: 1-Azinyl-1′-acetylferrocenes were obtained by regioselective acetylation of azinylferrocenes, followed by the Wittig reaction or reduction of 1-azinyl-1′-acetylferrocenes and subsequent dehydration of the resulting alcohols. The synthesized compounds underwent the following biological activity testing relevant to AD: inhibition of acetylcholinesterase (AChE), butyrylcholinesterase (BChE), and off-target carboxylesterase (CES); antioxidant capacity (ABTS and FRAP assays); inhibition of Aβ₄₂ self-aggregation (thioflavin method); blocking AChE-induced β-amyloid aggregation (propidium displacement); and cytotoxicity in SH-SY5Y and MSC-Neu cells (MTT assay). Results: Quinoline and bipyridine derivatives demonstrated effective cholinesterase inhibition, especially quinoline 7b (AChE IC₅₀ 3.32 μM; BChE IC₅₀ 3.68 μM), while acridine derivatives were poor inhibitors. Quantum chemical (QC) calculations predicted that acridine derivatives were especially prone to form stable dimers. Molecular docking into protein targets generated by an AlphaFold3 reproduction code showed that these dimers were too bulky to access enzyme active sites, yet they could bind to protein surfaces to inhibit Aβ₄₂ self-aggregation and displace propidium from the AChE peripheral anionic site. All compounds showed high antioxidant activity in ABTS and FRAP assays, with quinoline derivatives being 2–4 times more potent than Trolox. QC calculations supported these findings. Quinoline and bipyridine derivatives also exhibited low cytotoxicity and scant CES inhibition. Conclusions: Overall, the synthesized ferrocenes, particularly the quinoline and bipyridine derivatives, appear promising for further research as multifunctional therapeutic agents targeting AD due to their anticholinesterase, antiaggregating, and antioxidant activities combined with low toxicity. Full article

Background/Objectives: Metabolic enzymes are crucial for the detoxification of exogenously administered drugs, especially enzymes expressed in the intestine and the liver. Recent advancements in analytical methodologies enable sensitive and specific quantitative measurements of proteins, facilitating a more accurate evaluation of their expression and relative contribution to drug metabolism. Methods: The aim of the study was to characterize the protein expression levels of 16 Cytochrome P450s (CYP450s) and 2 carboxylesterases (CESs) in human liver and intestinal tissues using absolute quantification by HPLC-MS/MS. Human hepatocytes (HHEP) and human liver microsomes (HLM) were utilized, along with a novel intestinal preparation from cryopreserved human intestinal mucosa (CHIM), to perform proteomic analyses. Results: A comprehensive evaluation of 16 CYP450s and 2 CES enzyme expression in human liver and intestinal tissues is provided to reflect their relative abundance. Among the various in vitro systems evaluated, 14 of 16, 15/16, and 7/16 CYP450 of the isoforms analyzed were detected in HHEP, HLM, and CHIM, respectively. In hepatic systems, CYP2C9 exhibited the highest expression among CYP450 isoforms, a trend consistently observed in both HHEP and HLM. CYP3A4 was the most abundantly expressed isoform in CHIM preparations. Across all systems tested, CES1 and CES2 showed the highest overall protein expression levels, surpassing those of the CYP450s. Conclusions: Our findings demonstrate that the absolute quantification method employed is reliable, producing consistent results across two different in vitro hepatic systems (HHEP and HLM). This study supports the utility of absolute quantification approaches for accurately profiling drug-metabolizing enzymes and provides new, valuable insights to improve in vitro/in vivo extrapolation and more informed predictive pharmacokinetic modeling strategies. Full article

Dimethyl fumarate (DMF) has established a significant position among therapies for multiple sclerosis and psoriasis and is now being investigated for repurposing to many other non-malignant diseases. Despite decades of preclinical research, some issues about its pharmacology remain unresolved, with ongoing debate over which of the methyl esters of fumarate, whether DMF or monomethyl fumarate (MMF), is the active ingredient. It is generally accepted that DMF undergoes enzymatic hydrolysis to MMF and methanol. However, there is disagreement regarding its exact site, its extent, and the responsible enzyme(s). The enzymatic mechanisms, particularly the roles of carboxylesterases-1 and 2, vary across tissues and species, complicating the translation of in vitro and in vivo preclinical findings into clinical practice. In addition, the impact of DMF and MMF is often not clearly distinguishable and sometimes overlaps, making the true molecular mediators of therapeutic and side effects unclear. Thus, the interpretation of some results obtained in studies is inconsistent because of interchanging of in vitro and in vivo observed features of fumarate esters: while DMF demonstrates rapid and strong effects in cell culture studies, including nuclear factor erythroid 2-related factor 2 (NRF2) function activation and glutathione depletion, these observations may not exactly reflect systemic pharmacology and physiology dominated by MMF. Moreover, methanol, the co-product of DMF metabolism, may contribute to the observed DMF effects through increased production of reactive oxygen species, which could result in activation of NRF2-dependent mechanisms. This review highlights specific unresolved issues in DMF metabolism, which are sometimes overlooked. Full article

Polygala tenuifolia Willd., a widely used traditional Chinese medicine, has the function of coordinating heart and kidney and eliminating swelling. However, its renal-protective efficacy and possible material basis remain unknown. The aim of the study was to investigate the renal-protective effect of Polygala tenuifolia Willd. and identify the potential active substance and molecular mechanism. A gentamicin-induced kidney injury model was established to investigate efficacy. Secondly, potential active substances and molecular mechanisms were studied through integrated gut microbiota–drug interaction analysis and network pharmacology at a cellular level. Finally, 16S rRNA sequencing and untargeted metabolomics were used to elucidate the gut microbiota composition and metabolic profile change. Polygala tenuifolia Willd. extracts (PWE), with tenuifoliside A (TFSA) as the key compound, significantly reversed gentamicin-induced acute kidney injury in mice. The gut microbiota-derived carboxylesterase metabolized TFSA into four characteristic metabolites (M1–M4). Notably, both TFSA and M4 were detected in kidney and exerted protective effects via inhibiting TLR4–NF-κB pathway. Furthermore, metabolic pathways and gut microbiota composition change were identified. PWE treatment significantly increased the abundance of beneficial bacteria such as Akkermansia and Blautia, while reducing the abundance of harmful bacteria such as Oscillospira. Subsequently, PWE can reverse amino acid metabolic abnormalities by regulating the biosynthesis of phenylalanine, tyrosine, and tryptophan and ameliorating tryptophan metabolism disorder. This study was the first to verify the renal-protective effect of PWE and identify the effective substance basis (TFSA) and the molecular mechanism, providing a scientific foundation for the development of kidney drug treatment strategies targeting the intestinal flora. Full article

Schizaphis graminum is the most important pest of wheat, and its feeding harm not only affects photosynthesis but also causes diseases, posing a serious threat to wheat production. We measured the contact killing and fumigation activities, population changes, enzyme activities, and nanoemulsion physicochemical property of Eupatorium fortune essential oil (EFEO) and pyrethrin against S. graminum and its natural enemy (Harmonia axyridis). Toxic effects of EFEO and pyrethrin increased with increasing concentration. After 24 h of contact killing, the LD₅₀ of S. graminum were 9.23 and 0.02 μg/head, respectively. Harmonia axyridis were 36.71 and 0.07 μg/head. The fumigation of EFEO LD₅₀ on S. graminum was 9.779 mg/L (y = 2.88x − 2.852). EFEO contains 28 chemical components, including l-Caryophyllene, Lily aldehyde, α-Terpineol, and Cineole. EFEO LD₅₀ significantly reduced the adult longevity, nymph production, and nymph production duration of the F0 S. graminum, while there were no significant differences in various parameters of offspring. The activity of acetylcholinesterase, glutathione-S-transferase, and carboxylesterase is significantly affected. The population decline rate of EFEO nanoemulsion is significantly higher than that of EFEO. This research aims to clarify the toxic effect of EFEO on S. graminum, providing a scientific basis for developing new plant essential oil preparations and nano preparations. Full article

Castration-resistant prostate cancer (CRPC) remains difficult to treat with conventional chemotherapy. We evaluated a stem cell-based enzyme-prodrug strategy using hTERT-immortalized adipose-derived stem cells engineered to express rabbit carboxylesterase 2 (hTERT-ADSC.CE2) in combination with irinotecan (CPT-11). hTERT-ADSC.CE2 cells were generated via lentiviral transduction and confirmed to overexpress CE2. Their tumor-homing capacity toward PC3 prostate cancer cells was assessed, along with prodrug activation, apoptosis induction, and in vivo tumor suppression in a CRPC mouse model. hTERT-ADSC.CE2 cells demonstrated enhanced migration toward PC3 cells and higher expression of tumor-homing factors than the controls. Under CPT-11, they exhibited a strong “suicide” effect and induced selective killing of PC3 cells, with upregulation of BAX and cleaved caspase-3 and downregulation of BCL-2. By day 14, the combination arm showed significantly lower tumor burden than both the control and irinotecan-alone arms (p < 0.05). The pattern is consistent with intratumoral activation and localized SN-38 exposure. hTERT-ADSC.CE2 combined with irinotecan provides potent, tumor-targeted cytotoxicity and markedly suppresses CRPC progression. This cell-mediated prodrug activation system may represent a promising therapeutic approach for advanced prostate cancer. Full article

Mythimna separata (Walker) (Lepidoptera: Noctuidae) is a major migratory pest causing severe damage to cereal crops such as maize, wheat, and rice across Asia, and is also found in many parts of Oceania. With increasing insecticide resistance, botanical alternatives are urgently needed. This study evaluated the insecticidal potential of hypaconitine, a C19-diterpenoid alkaloid from Aconitum coreanum, against M. separata larvae. Hypaconitine exhibited significant stomach toxicity and strong antifeedant activity. It also caused pronounced growth inhibition, prolonged larval and pupal development, reduced pupation and adult emergence, induced morphological deformities, and significantly shortened adult longevity. Crucially, biochemical assays revealed sustained, time- and concentration-dependent upregulation of key detoxification enzymes—carboxylesterase (CarE), glutathione S-transferase (GST), and cytochrome P450 (CYP450)—over 72 h, indicating that hypaconitine imposes severe metabolic fitness costs rather than being readily detoxified. These effects collectively demonstrate that hypaconitine’s insecticidal efficacy arises not only from direct toxicity but also from exploiting the physiological trade-offs inherent in xenobiotic defense. Its multi-modal action—combining larvicidal, antifeedant, growth-regulatory, and metabolism-disrupting effects—presents a novel strategy for bioinsecticide development with a lower risk of resistance evolution. These findings highlight hypaconitine as a promising candidate for sustainable, integrated management of M. separata and other resistant lepidopteran pests. Full article

To provide a scientific basis for pest control, this study evaluated the insecticidal activity of novaluron against the fall armyworm (Spodoptera frugiperda) and diamondback moth (Plutella xylostella). The leaf-dip method determined the toxicity of novaluron to second-instar larvae, while corn leaves and cabbage treated with sublethal (LC₁₀) and median lethal concentrations (LC₅₀) of novaluron were used to feed the larvae. Enzyme-linked immunosorbent assays (ELISA) measured the activities of detoxifying enzymes [carboxylesterase (CarE), cytochrome P450 (P450), glutathione S-transferase (GST), and acetylcholinesterase (AChE)] and ecdysteroid (Ecd) levels in the pests after 24 and 48 h of treatment. Results indicated that after 24 h, AChE was involved in diamondback moth metabolism and CarE activity was inhibited. After 48 h, P450 and GST participated in fall armyworm detoxification, whereas P450 and GST were active in diamondback moth detoxification, with other enzyme activities normalizing. Novaluron also altered Ecd levels in both pests. These results demonstrate differing detoxification mechanisms in fall armyworm and diamondback moth, likely due to their unique physiological and ecological traits, and support the potential use of novaluron in pest management strategies. Full article