Applied Biological Chemistry最新文献

This paper aimed to design a biochar-based nanobiocomposite from bamboo to remove fifth-generation cephalosporin antibiotics, such as Ceftaroline, from pharmaceutical wastewater. Porous biochar was prepared by pyrolysis of bamboo biomass at 500 °C (BBC-A500) and 700 °C with KOH activation (BBC-Fe10), whereas iron nanoparticles were produced via a chemical reduction pathway and later linked to the biochar network. This was systematically characterized by a series of analyses, i.e., SEM, TEM, XRD, FTIR, BET, and XPS, which verified the presence of stable iron loading, a high specific surface area, and numerous surface functional groups. XPS analysis confirmed mixed Fe²⁺/Fe³⁺ surface states (Fe²⁺/Fe³⁺ ratio = 0.92), which are catalytically active for Fenton-like reactions. The degradation experiment was also conducted in batch mode to evaluate degradation performance as a function of contact time, pH, catalyst load, and oxidant concentration. This elimination was carried out in a two-step mechanism, in which preliminary adsorption concentrated Ceftaroline molecules on the biochar surface, and catalytic oxidation was conducted via Fenton-like oxidation by encapsulated iron nanoparticles. The resulting reactive species are generated to enable rapid and extensive degradation of antibiotics, and mechanistic probing indicated that hydroxyl radicals — both free and surface-bound — were the predominant oxidizing species, with minor contributions from superoxide and singlet oxygen. The composite achieved 98% removal within 60 min (k_app = 0.092 min⁻¹) and retained 82% activity after five cycles, with cumulative Fe leaching below 1.5%. Overall, the adsorption-oxidation synergy makes the bamboo biochar-iron nanocomposite a promising solution for treating pharmaceutical effluents.

Assessing the sublethal behavioral effects of pesticides on honey bees (Apis mellifera) is a key component of honey bee risk assessment. In this study, we aimed to refine experimental conditions for RFID-based tracking and to evaluate the sublethal effects of fenobucarb (FNC) and carbendazim (CBZ) on honey bee homing performance. Individually tagged forager bees were exposed to sublethal doses of FNC (0.00025 and 0.0025 µg/bee) or CBZ (0.005 and 0.05 µg/bee) and monitored for 24 h. Behavioral endpoints, including the return rate, first-return time, and frequency of out-of-hive trips, were quantified via RFID detection at the hive entrance. Control and solvent-control groups showed high (> 90%) and comparable return rates, indicating high baseline homing performance and no apparent effect of solvent exposure. FNC exposure at both doses did not significantly affect homing success, median return time, or frequency of out-of-hive trips relative to solvent exposure. At the lower dose, CBZ exposure significantly reduced the return rate; however, this effect was not observed at the higher dose and was not accompanied by changes in median return time or frequency of out-of-hive trips, suggesting the absence of a consistent or dose-dependent behavioral response. Considering the overall pattern across behavioral endpoints, no consistent adverse effects on homing-related behavior were observed for either substance under the study conditions. Overall, the refined RFID methodology provided consistent and reliable measurements of sublethal behavioral performance in honey bees.

Drought stress severely affects tomato (Solanum lycopersicum L.) growth, yield, and fruit quality. Although RNA-seq has identified drought-responsive genes in tomato, differences in experimental design and stress conditions can limit the reproducibility of DEG-based results. This study performed a meta-analysis of four independent public RNA-seq datasets comprising 51 tomato leaf samples to identify drought-responsive candidate genes supported across datasets. Differential gene expression data from individual datasets were integrated with heterogeneity assessment, identifying 5,681 heterogeneity-filtered drought-responsive meta-DEGs after study-coverage and heterogeneity-based filtering. Fixed- and random-effects pooled directions agreed for 5,647 genes (99.4%), and 3,294 genes remained significant at a random-effects FDR < 0.05. The prioritized candidate genes included those associated with MAPK signaling, auxin-responsive and redox regulation, phenylpropanoid, lipid, and cuticle metabolism, and proteolysis. GO enrichment of effect-size-significant meta-DEGs represented in at least three datasets identified nominally enriched terms related to protein processing, proteolysis, transport-related processes, photosynthesis, auxin response, and redox homeostasis. In ‘Micro-Tom’ plants subjected to soil-drying-induced drought stress, RT-qPCR analysis of ten representative genes revealed distinct temporal expression patterns. The drought-responsive marker genes SlPP2C3, SlAREB1, and SlNCED1 were also induced during progressive soil drying, providing molecular evidence of a response to irrigation withdrawal. These results demonstrate that RNA-seq-based meta-analysis combined with heterogeneity assessment can prioritize drought-responsive candidates in tomato and provide a candidate set for future functional studies.

Background

Intracranial aneurysms (IAs) are a significant cause of morbidity and mortality, often leading to subarachnoid hemorrhage and subsequent neurological deficits. The kinase insert domain receptor (KDR) plays a crucial role in vascular cell function during IA. However, the mechanism responsible for KDR’s role in IA remains unclear.

Methods

Human brain microvascular endothelial cells (hBMECs) were treated with angiotensin II (Ang II) to simulate an in vitro IA model. Quantitative real-time PCR (qRT-PCR) was used to analyze the mRNA expression levels of KDR and Wilms’ tumor 1-associating protein (WTAP). Western blotting was employed to detect the protein expression of KDR and WTAP. Cell viability was assessed using a cell counting kit-8 assay, while cell apoptosis and mitochondrial membrane potential were measured by flow cytometry. Fluorometric assay was conducted to evaluate reactive oxygen species (ROS) levels. Colorimetric assays were used to determine the levels of reduced glutathione (GSH) and malondialdehyde (MDA), as well as Fe²⁺ levels. Methylated RNA immunoprecipitation (MeRIP) assay and dual-luciferase reporter assay were performed to validate m6A modification of KDR mRNA and the regulatory effect of WTAP on KDR.

Results

Treatment with Ang II induced apoptosis, oxidative stress, and ferroptosis in hBMECs. KDR and WTAP expression were significantly reduced in the arterial wall tissues of IA patients and in Ang II-treated hBMECs. Overexpression of KDR attenuated Ang II-induced apoptosis, oxidative stress, and ferroptosis in hBMECs. In addition, WTAP was found to promote KDR expression in a m6A-dependent manner. Moreover, overexpression of WTAP protected hBMECs from Ang II-induced apoptosis, oxidative stress, and ferroptosis by upregulating KDR.

Conclusion

WTAP-induced upregulation of KDR mitigated Ang II-induced apoptosis, oxidative stress, and ferroptosis in hBMECs. These findings suggest that targeting the WTAP-KDR axis could have significant clinical implications for the prevention and treatment of IA.

Investigation of an antimicrobial and cytotoxic ethyl acetate and methanol extracts from the marine sponge-derived fungus Aspergillus versicolor QT21 led to the isolation of twelve metabolites, including 5α,6α-epoxy-ergosta-8(14),22-diene-3β,7α-diol (1); 5α,6α-epoxy-ergosta-8(9),22-diene-3β,7α-diol (2); 4,8-dihydroxy-3,4-dihydronaphthalen-1(2 H)-one (isosclerone) (3); 5-methylbenzene-1,3-diol (orcinol) (4); ergosterol (5); (22E, 24R)-24-methylcholesta-5,22-diene-3β,7β-diol (6); cyclo-(Pro-Gly) (7); cyclo-(Pro-Ala) (8); cyclo-(Tyr-Trp) (9); N-acetyltryptamine (10); uridine (11); adenosine (12). Of these, compound 3 was isolated for the first time from the species Aspergillus versicolor. Antimicrobial assays showed that most compounds exhibited selective antimicrobial activity against Gram-positive bacteria and yeast. Compound 5 demonstrated the most promising antimicrobial activity, with MIC values of 32 µg/mL against E. faecalis, 64 µg/mL against B. cereus, and 64 µg/mL against C. albicans. In addition, compounds 1, 4 and 5 exhibited cytotoxicity against all tested cell lines A549, MCF7, and HepG2 with IC₅₀ values of 13.07 ± 1.6 to 83.02 ± 1.65 µg/mL, whereas compound 2 had selective cytotoxicity against the A549 and MCF7 cell lines with IC₅₀ of 11.34 ± 1.56 and 10.91 ± 1.61 µg/mL, respectively. The structure–activity relationships of the isolated compounds were also discussed. This study highlights A. versicolor as a promising source of biologically active natural compounds.

Natural compounds are attracting increasing attention in cancer treatment due to their ability to regulate key pathways associated with cell proliferation, apoptosis, and chemotherapy resistance. Myo-inositol, a naturally occurring carbocyclic sugar, has demonstrated its potential in hormone-sensitive cancers, including breast cancer. This study investigates the synergistic effect of combining doxorubicin with myo-inositol on MCF7 breast cancer cells (ER+), and the possible involvement of the p38 MAPK signaling pathway. We hypothesized that myo-inositol enhances doxorubicin-induced cytotoxicity and promotes apoptosis in MCF7 cells, potentially by modulating stress-responsive Total p38 MAPK levels. Cell viability was assessed using the MTS assay, while apoptosis, p38 MAPK levels, and ROS were assessed by flow cytometry. ADME predictions, KEGG pathway analysis, and subcellular compartment analysis were performed to explore potential mechanistic links. The combination treatment significantly reduced MCF7 cell proliferation in a dose- and time-dependent manner, with minimal effect on normal MCF 10 A cells, suggesting selective cytotoxicity. The combined treatment induced 25.12% apoptotic cell death. Interestingly, Phospho-p38-MAPK levels were lower in the combination group (58.82%) compared to doxorubicin (90.17%) and myo-inositol (72.51%) alone, indicating a 1.53-fold (p < 0.01) and 1.23-fold (p < 0.05) reduction, respectively. These findings suggest that the combination of myo-inositol and doxorubicin enhances apoptotic cell death in MCF7 cells, potentially through modulation of Total p38 MAPK, with computational analyses supporting a mechanistic basis.

GenX (ammonium perfluoro(2-methyl-3-oxahexanoate)) is an HFPO-DA-related PFAS replacement with emerging health concerns, yet its effects on prostate stromal cells remain poorly understood. This study investigated the cytotoxic, apoptotic, and oxidative effects of GenX in WPMY-1 human prostate stromal cells and characterized the accompanying transcriptomic remodeling. WPMY-1 cells were exposed to 0–50 nM GenX for 24 h; viability was assessed by MTT assay, apoptotic and antioxidant markers (PARP, cleaved PARP, caspase-3, cleaved caspase-3, caspase-9, Bax, Bcl-2, SOD2, catalase) were evaluated by Western blot with Z-VAD-FMK and NAC rescue experiments, and intracellular ROS was measured by DCFDA imaging. Transcriptomic profiling via mRNA-seq (|fold change| ≥ 1.5) was followed by GO, KEGG, and Reactome enrichment analyses, and protein–protein interactions were analyzed using STRING with MCL clustering (inflation parameter = 2.0). GenX induced a dose-dependent viability decrease in WPMY-1 cells at 25 and 50 nM, increased the cleaved PARP/PARP and cleaved caspase-3/caspase-3 ratios, elevated Bax and reduced Bcl-2 at 50 nM, and increased intracellular ROS with concomitant SOD2 and catalase upregulation; Z-VAD-FMK attenuated apoptotic marker cleavage, and NAC attenuated ROS accumulation and antioxidant protein induction. Transcriptomic analysis identified 218 differentially expressed genes (113 upregulated, 105 downregulated), with enriched terms and pathways associated with stress-responsive transcription, MAPK and ER-stress signaling, interleukin-mediated inflammation, ECM organization and adhesion, growth factor/vascular signaling, and Wnt-related developmental signaling; 10 PPI modules were delineated. These findings indicate that nanomolar GenX elicits caspase-associated apoptotic response and oxidative stress in prostate stromal cells and provide a hypothesis-generating transcriptomic network map for targeted mechanistic follow-up of GenX-induced stromal toxicity.

Transcription factor (TF)-based whole-cell biosensors provide genetically encoded tools for detecting intracellular metabolites and have potential applications in monitoring microbial biosynthetic processes. TtgR, a multidrug-responsive transcriptional repressor from Pseudomonas putida, responds to several flavonoids, including resveratrol, and has previously been developed as a whole-cell biosensor for quantitative detection of flavonoids using authentic standards. In this study, we extended the application of the TtgR-based biosensor to the quantitative monitoring of resveratrol produced through a heterologous biosynthetic pathway in engineered Escherichia coli. Resveratrol production was monitored over time using the TtgR-based biosensor, and the estimated concentrations were compared with those determined by HPLC. The biosensor-derived concentrations showed relative agreement of 92.7–93.8% with the corresponding HPLC measurements, supporting the quantitative applicability of the biosensor for monitoring metabolite production in a microbial production system. In addition, ligand-response profiles of TtgR WT and selected engineered variants were independently evaluated in this study using resveratrol, quercetin, and naringenin. The engineered variants exhibited distinct ligand-response profiles, suggesting the potential extension of the TtgR-based sensing platform to other flavonoid production pathways. These results demonstrate the application of a previously established TtgR-based sensing platform beyond analytical characterization with authentic standards toward quantitative monitoring of biosynthetically produced metabolites in engineered microbial systems.

Diabetic nephropathy (DN) ranks as the primary contributor to end-stage renal disease across the globe, characterized by progressive glomerular mesangial proliferation and tubular interstitial fibrosis, which poses a severe threat to public health. Pinocembrin is a major flavonoid compound that has been reported to exhibit a variety of prominent biological effects. However, its dual regulatory effects on glomerular and tubular injuries in DN remain unclear. The present study aims to investigate the potential role of pinocembrin in alleviating the progression of DN, providing a combination therapeutic strategy. In vivo experiments were performed to verify its efficacy in improving renal injury and fibrosis of pinocembrin in Lep^db/Lep^db (db/db) mice. Pinocembrin-related targets were retrieved from Traditional Chinese Medicine Systems Pharmacology (TCMSP), Comparative Toxigenomics Database (CTD), Swiss Target Prediction, and Symmap databases. Differentially expressed genes in DN tubules and glomeruli were obtained from Gene Expression Omnibus (GEO) dataset GSE30122. Overlapping targets between pinocembrin and DN tissues were identified, followed by Protein-Protein Interaction (PPI) network construction, Gene Ontology (GO) annotation, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. Molecular docking predicted potential binding interactions between pinocembrin and key targets. In vitro experiments demonstrated that in HG-induced HK-2 cells, pinocembrin might target HSP90AB1 to suppress inflammatory response and epithelial-mesenchymal transition, thereby alleviating tubular fibrosis injury. In HG-treated SV40 cells, pinocembrin downregulated expression of VEGFR, thereby inhibiting cell proliferation and extracellular matrix deposition to improve glomerular pathological damage. This study clarifies the dual protective mechanism of pinocembrin on DN via regulatory pathways. These findings provide experimental foundation and theoretical insight for the application of pinocembrin as a functional food ingredient for the prevention and treatment of DN.