Search Results (608)

Co-contamination with dibenzofuran (DBF) and cadmium (Cd(II)) poses a major challenge in environmental remediation. While Burkholderia sp. can degrade polycyclic aromatic hydrocarbons and tolerate heavy metals, the coordinated mechanism governing DBF degradation under high Cd(II) stress remains elusive. Here, we characterize Burkholderia sp. FM-2, which optimally degrades 600 mg/L DBF at pH 6.0 and 25 °C, achieving 91.8% removal within 48 h. FM-2 exhibits exceptional Cd(II) tolerance, with a minimum inhibitory concentration of 2000 mg/L. UPLC-MS/MS confirms DBF degradation via dioxygenase-mediated hydroxylation and sequential enzymatic reactions. Transcriptomics reveals, for the first time, concurrent upregulation of genes encoding RND efflux pumps, ABC transporters, P-type ATPases, and core DBF-degrading enzymes under high Cd(II) stress, enabling the synergistic maintenance of intracellular Cd(II) homeostasis and efficient DBF degradation. Collectively, FM-2 remediates DBF-Cd(II) co-contamination via coordinated transcriptional regulation of degradation and detoxification pathways, offering a promising strain resource and molecular basis for the bioremediation of co-contaminated environments. Full article

Lithocarpus litseifolius is a medicinal tea plant recognized for its sweet flavor and anti-diabetic properties, but its limited leaf yield under cultivation restricts its economic sustainability. Melatonin (MLT) is a multifunctional plant growth regulator, but its roles in leaf growth under normal conditions remain not fully understood. Herein, we investigated the effects and mechanisms of foliar-applied MLT on L. litseifolius seedlings, including growth indices, phytohormone profiles, photosynthetic characteristics, and transcriptome alterations. The 100 μM MLT treatment significantly enhanced leaf dry weight by 33.8% and leaf dry matter content by 22.2% compared to the control group. MLT decreased both free and bound abscisic acid (ABA), while increasing gibberellins (GAs), 5-deoxystrigol, auxins (e.g., IAM), and cytokinins (e.g., cZ9G). Additionally, exogenous MLT improved photosynthetic rate, stomatal conductance, chlorophyll content, and soluble sugars in leaves. RNA-seq revealed that MLT up-regulated DEGs involved in hormone biosynthesis and signaling (CYP707A, BAK1, D14, CCD1, and IAA6), photosynthesis (PsbC/B, PetH, PsaB, and ATPase β), and sugar metabolism (WAXY, glgC, and otsB). Our results demonstrate that MLT promotes leaf dry matter accumulation through coordinated phytohormone homeostasis, photosynthetic enhancement, and transcriptional regulation, offering a cost-effective strategy to improve leaf yield in L. litseifolius. Full article

Bradysia odoriphaga is a devastating soil pest of Allium tuberosum (Chinese chive), and current control relies heavily on chemical insecticides. Cry39Ab1 toxins from Bacillus thuringiensis (Bt), which are highly toxic to B. odoriphaga, offer an environmentally friendly alternative. However, its mechanism of action remains unclear. In this study, we identified the involvement of vacuolar H⁺-ATPase subunit A (V-ATPase A) in Cry39Ab1 insecticidal activities. The full-length cDNA sequences of BoV-ATPase A was contained 1659 bp open reading frame (ORF), encoding a protein of 552 amino acids with a calculated molecular weight of 59 kDa and an isoelectric point of 9.11. Successful expression and purification of BoV-ATPase A (with GST and His tags) and Cry39Ab1 (with GST and His tags) proteins were achieved. GST pull-down assays demonstrated a direct interaction between recombinant BoV-ATPase A and activated Cry39Ab1 toxin in vitro. Heterologous expression of BoV-ATPase A in Cry-insensitive Sf9 cells conferred susceptibility to Cry39Ab1, resulting in a significant increase in cytolysis compared with control cells. Finally, RNAi-mediated knockdown of BoV-ATPase A in larvae significantly decreased their susceptibility to Cry39Ab1, as evidenced by a marked decrease in mortality. This is the first report that BoV-ATPase A is a key protein required for Cry39Ab1 toxicity, revealing its insecticidal mechanism and establishing BoV-ATPase A as a potential target for pest control. Full article

This study aims to investigate the antioxidant, anti-inflammatory, and anti-ulcer properties of two newly isolated natural compounds (1–2) from the hydroalcoholic root extract of Tricholaena teneriffae L. Through bioassay-guided fractionation, two new natural compounds, xantenone and anthracenone, were isolated and characterized using 1D/2D NMR and mass spectrometry. While 6-hydroxy-3-methoxy-7H-benzo[de]anthracen-7-one (HK2) showed significant antioxidant activity, 11-hydroxy-12H-benzo[a]xanthen-12-one (HK1) exhibited only weak activity. However, both compounds demonstrated a gastroprotective effect in vivo, similar to that of omeprazole. In silico screening revealed that both molecules bind stably to the H⁺/K⁺-ATPase proton pump, suggesting a potential mechanism for inhibiting gastric acid secretion. In the ethanol-induced ulcer model, treatment with HK1 and HK2 significantly reduced inflammation, oxidative stress, gastric hypersecretion, and ulcer area, with HK2 showing greater efficacy. These results suggest that the active fraction of T. teneriffae root has anti-ulcer properties beyond its antioxidant effects, making it a promising natural source of gastroprotective agents. Full article

Research on mule and donkey muscle composition remains limited despite their global importance as working equids. The objective of this study is to identify Mammoth donkey jacks with higher percentage of fast twitch fibers for racing mule production. A total of 33 animals were biopsied; however, only 12 samples were suitable for analysis, including racing mules (n = 7) and male Mammoth donkeys (n = 5). Animals were sedated with detomidine (10 µg/kg body weight) and butorphanol (20 µg/kg body weight). Middle gluteal muscle biopsies were collected using a 6 mm Bergström biopsy needle at a site located 20 cm dorsocaudal to the tuber coxae at a 45° angle to the base of the tail. Collection depth was 7.5 cm in adult mules and 5 cm in donkeys. Samples were prepared aseptically, anesthetized subcutaneously with lidocaine hydrochloride, and frozen in liquid nitrogen. Histochemical analysis included myosin adenosine triphosphatase (ATPase) staining at pH 9.5, 4.6, and 4.3. Fibers were classified as Type I, Type IIA, or Type IIB, and CSA measurements were obtained using NIH ImageJ software. Statistical analysis included group contrasts, summarized as mean ± SD with 95% confidence intervals, while Bayesian ANOVA outputs were presented as exploratory evidence summaries. Type IIA fibers were greater in mules (47.84 ± 7.30%) than donkeys (38.47 ± 4.48%). Results suggest that differences in equid muscle architecture may be associated with variation in Type IIA fiber composition related to work or use. Full article

Nitrite is a key environmental challenge in intensive shrimp aquaculture, adversely affecting physiological regulation and survival. Although tolerant Penaeus vannamei families have been established by selective breeding, the basis of family-level variation in tolerance has yet to be clarified. In this study, nitrite-tolerant and nitrite-sensitive families were compared using survival analysis, transcriptomics, targeted qPCR validation, physiological assays, and RNA interference of representative transport-related genes. Under nitrite exposure, the tolerant family exhibited significantly higher survival and a distinct gill transcriptional response, characterized by stronger induction of acid–base and ion-transport genes, including carbonic anhydrase 2 (CA2), the Na⁺/K⁺-ATPase subunits ATP1A and ATP1B, as well as several V-type H⁺-ATPase-related genes. These transcriptional changes were accompanied by elevated ATP content and Na⁺/K⁺-ATPase activity, improved hemolymph pH stability, and reduced nitrite accumulation in both gill and hemolymph. RNAi-mediated knockdown of CA2 or ATP1B attenuated the nitrite-induced transport response, decreased ATP content and NKA activity, exacerbated hemolymph acidification, promoted internal nitrite accumulation, and ultimately reduced shrimp survival under nitrite stress. Family-based validation further showed that the tolerant family displayed higher survival than the sensitive family in the dsEGFP group, whereas this advantage was markedly reduced after CA2 or ATP1B knockdown under nitrite stress. These findings highlight that strengthened branchial ion transport and acid–base regulation represent key physiological mechanisms underlying nitrite tolerance in resistant shrimp families. Full article

Background: T-cell acute lymphoblastic leukemia (T-ALL) remains a challenging malignancy with limited targeted therapies. Natural phenanthrene derivatives represent a promising source of antileukemic agents. Objective: We screened a library of natural phenanthrene-type compounds to identify cytotoxic leads in Jurkat T-ALL cells and investigated the mechanisms underlying their activity, including potential synergy with the proteasome inhibitor bortezomib (BTZ). Methods: Jurkat cells were treated with thirteen natural compounds at 10 and 20 µM for 48 h; cell viability was assessed by WST-1 cell viability assay. Dose–response curves were generated to calculate IC₅₀ values. Apoptosis was evaluated by Hoechst 33342/PI staining and Annexin V/PI flow cytometry. Synergy with BTZ was analyzed using a fixed-ratio combination index (CI) approach and IC₅₀ shift analysis. ER stress signaling was characterized by Western blotting, quantitative RT-PCR of UPR genes (GRP78, ATF6), and immunoprecipitation of GRP78 followed by ubiquitin immunoblotting. Results: Among the compounds screened, Cypripedin showed the most potent cytotoxicity with an IC₅₀ of 6.52 µM. It induced a dose-dependent increase in apoptosis. Combination with BTZ yielded a CI < 0.5 and reduced BTZ IC₅₀ from 3.43 to 1.88 ng/mL. Cypripedin activated the unfolded protein response (UPR), modulated key ER stress markers including GRP78, p-PERK, p-eIF2α, p-JNK, and ATF6, downregulated UPR gene transcripts, and promoted GRP78 ubiquitination. Molecular docking predicted strong binding of Cypripedin to the GRP78 ATPase domain (Vina score −7.630 kcal/mol), supporting its mechanism of action. Conclusion: Cypripedin induces apoptosis in Jurkat T-ALL cells, synergizes with BTZ, and modulates ER stress through GRP78 ubiquitination. These findings support its further development as a potential T-ALL therapeutic. Full article

Anaerobic biodegradation is the most affordable method for the degradation of N,N-dimethylformamide. However, the degradation efficiency depends on the concentration. To elucidate the responses of microbial community to N,N-dimethylformamide load, microbial diversity, composition and functional changes at different concentrations of 100, 2000, and 3500 mg/L were analyzed. Results showed that as the N,N-dimethylformamide influent concentration increased from 100 to 2000 mg/L, the removal rate stabilized at 90%, whereas it decreased to ~75% at concentrations over 2000 mg/L. Microbial community diversity increased, and specialists were enriched at 3500 mg/L. Patescibacteria (42.88% and 42.90%), Bacillota (18.52% and 18.54%), and Pseudomonadota (7.13% and 7.09%) were the dominant phyla at 100 mg/L and 2000 mg/L, respectively, and Patescibacteria (16.88%) and Pseudomonadota (15.34%) were the dominant phyla at 3500 mg/L. Methylotrophic methanogeneic (Methanolobus and Methanomassiliicoccus) and syntrophic electron-donating bacteria (Clostridium and Trichococcus) were significantly enriched. DMF-degrading genes (fdh, rfA/nrfH, and ATPase) and methylotrophic methanogenesis genes (mcr, mta, and mtm) were significantly upregulated. Therefore, the degradation of N,N-dimethylformamide was characterized by a parallel carbon flux distribution, “methylamine-driven methanogenesis + further oxidation/integration of single-carbon intermediates”, and the nitrogen flux tended to enter a reductive nitrogen cycle characterized by retention and reuse. Full article

Tumour necrosis factor-alpha (TNF-α) disrupts bioenergetic homeostasis in skeletal muscle cells through the suppression of ion-dependent ATPase activities, mitochondrial depolarisation, and impairment of antioxidant defences. Carvacrol, a phenolic monoterpenoid constituent of thyme and oregano essential oil, has been shown to exert cytoprotective effects in TNF-α-challenged L6 rat myoblasts. The mechanistic basis of these effects, specifically the relationship between membrane-associated ATPase function, mitochondrial polarisation status, and transcriptional regulation of metabolic stress-response genes, has not been formally characterised. L6 rat myoblasts were exposed to TNF-α (10 ng/mL, 1 h), then treated with carvacrol (6.25 µg/mL, 24 h) in a post-inflammatory rescue paradigm. Cell viability (MTT), membrane integrity (LDH), ion-dependent ATPase activities (Na⁺/K⁺, Ca²⁺, Mg²⁺), antioxidant enzyme activities (catalase, SOD), mitochondrial membrane potential (Muse™ MitoPotential flow cytometry), and SIRT1/AMPK mRNA expression were quantified. TNF-α significantly suppressed Na⁺/K⁺, Ca²⁺, and Mg²⁺-dependent ATPase activities (all p < 0.001), consistent with impaired membrane-associated bioenergetic function. Post-TNF-α carvacrol treatment partially restored all three ATPase activities (p < 0.05) and reduced the proportion of mitochondrially depolarised cells from 31.65 ± 4.25% to 19.0 ± 2.6% (p < 0.05). LDH release, catalase activity, and SOD activity were also significantly modulated. At the transcriptional level, carvacrol increased SIRT1 mRNA by 1.6-fold and AMPK mRNA by 2.0-fold relative to TNF-α-treated cells. An integrative bioenergetic model is proposed in which carvacrol’s membrane-intercalating properties restore the phospholipid environment required for ATPase conformational cycling, attenuating the Ca²⁺ overload that drives mitochondrial permeability transition, and thereby partially preserving Δψm. Transcriptional upregulation of SIRT1 and AMPKα may represent an adaptive response to residual energetic stress. The mechanistic relationships among these endpoints and the causal contribution of SIRT1 and AMPK to observed bioenergetic changes require protein-level and pathway-specific experimental validation. Full article

(1) Background: Nitric oxide (NO) serves as a crucial signaling molecule in plant abiotic stress responses. Although its role in enhancing drought resistance in cotton has been recognized, the specific mechanisms underlying this physiological and molecular regulation remain largely unexplored. This study aims to elucidate the multi-layered mechanisms by which NO modulates drought resistance in cotton; (2) Methods: Cotton seedlings were subjected to drought stress with the application of the NO donor sodium nitroprusside (SNP). A combination of confocal laser scanning microscopy, transcriptional expression analysis, biochemical assay of enzyme activity, virus-induced gene silencing (VIGS), and in vitro protein modification assays was applied to characterize the effects of NO on the drought stress response in cotton; (3) Results: Exogenous NO significantly reinforced drought resistance in cotton seedlings by improving leaf water retention capacity and photosynthetic efficiency, eliminating excessive drought-induced reactive oxygen species (ROS), upregulating the transcription and enzymatic activity of antioxidant enzymes, and promoting stomatal closure. Mechanistically, NO triggered S-nitrosylation of the plasma membrane H⁺-ATPase isoform GhHA2, thereby enhancing its protein stability; (4) Conclusions: These findings reveal that exogenous NO orchestrates cotton drought tolerance via multiple interconnected physiological and molecular pathways, in which the activation of the antioxidant defense system and the modulation of stomatal closure serve as central regulatory mechanisms. Full article

Pomegranate peel is a food industry waste rich in polyphenols. To date, its effect in alleviating fatigue remains unclear. This study aimed to characterize the chemical composition of pomegranate peel polyphenols (PPPs), evaluate its antioxidant and anti-fatigue capacities, and investigate the underlying mechanism. In the current study, twenty main compounds, primarily flavonoids, phenolic acids, and anthocyanins, were identified from PPPs using LC-MS/MS. In H₂O₂-induced HepG2 cells, PPPs promoted cellular repair and reduced the production of intracellular malondialdehyde (MDA) and reactive oxygen species (ROS) via enhancing the activity of antioxidant enzymes (SOD, CAT, and GSH-Px). In the endurance swimming-induced fatigue mice model, PPPs prolonged mice exhaustion times, reduced accumulation of fatigue-related metabolites (BUN, LA, BA, LDH and CK), and alleviated liver and muscle tissue damage. Mechanistically, PPPs mitigated oxidative stress via activation of the Keap1/Nrf2 pathway, leading to increased expression of hemeoxygenase-1 (HO-1) and NAD(P)H quinone oxidoreductase 1 (NQO1). Furthermore, PPPs stimulated energy metabolism by activating the AMPK/PGC-1α/PPAR-α pathway, promoting mitochondrial biogenesis, enhancing glycogen storage, increasing ATPase activity (Na⁺-K⁺-ATPase, Ca²⁺-Mg²⁺-ATPase, and T-ATPase) and accelerating lipid β-oxidation. These findings suggest that PPPs is a promising anti-fatigue supplement and could be further utilized in the nutritional industry. Full article

Magnesium is an essential divalent cation required for adenosine triphosphate (ATP)-dependent reactions, nucleic acid metabolism, and ribosomal stability. Bacteria depend on specialized transport systems to maintain intracellular Mg²⁺ homeostasis as it cannot freely cross the phospholipid bilayer. During infection, host nutritional immunity restricts metal availability, and magnesium limitation within the phagosome compromises bacterial metabolism and stability. This review summarizes the major bacterial magnesium transport systems and their roles in survival and pathogenicity, with an emphasis on Salmonella and extension to clinically relevant ESKAPE pathogens. We focus on the PhoPQ-regulated MgtA, MgtB, and MgtC system, in which low magnesium, acidic pH, and other host-derived signals activate PhoPQ to induce mgt gene expression. MgtA and MgtB act as high-affinity P-type ATPases, whereas MgtC promotes bacterial survival within the intramacrophage environment by inhibiting bacterial F-type ATP synthase through specific interactions with subunit a. We also discuss CorA as a conserved channel for basal Mg²⁺ uptake and MgtE as a Mg²⁺-selective channel whose gating responds to intracellular Mg²⁺ and ATP. Finally, we consider the conservation and variation in these systems across pathogenic bacteria and their potential as therapeutic targets for antimicrobial development. Full article

Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb), remains a major cause of morbidity and mortality worldwide, particularly due to the emergence of drug-resistant strains. Membrane-active antimicrobial peptides (AMPs) represent attractive therapeutic candidates because they target bacterial envelope integrity and disrupt essential cellular processes. We evaluated two rationally designed LL-37-derived peptides: a truncated C-terminally amidated analog (LL37-1) and a modified variant incorporating N-terminal acetylation and a single D-amino acid substitution (D-LL37). Dose–response analysis demonstrated that D-LL37 exhibited greater antimycobacterial potency, with lower inhibitory concentrations of 90% (IC₉₀) and 50% (IC₅₀) values (18.40 ± 0.39 μM and 10.11 ± 0.60 μM, respectively) compared with LL37-1 (25.44 ± 0.36 μM and 15.45 ± 1.40 μM). Fluorescence-based permeability assays revealed partial membrane disruption (36% and 44% at IC₉₀ for LL37-1 and D-LL37, respectively), which was supported by ultrastructural alterations observed by scanning electron microscopy, including bacillary shortening, rough surface formation, cell clusters, and the presence of cellular debris, all of which are consistent with membrane damage. RT-qPCR analysis demonstrated significant upregulation of the P-type ATPase genes ctpF, ctpA, and ctpH following D-LL37 exposure. Collectively, these findings indicate that optimized LL-37-derived peptides exert antitubercular activity associated with envelope perturbation and coordinated activation of ion transport-related stress responses. Full article

Metabolic dysfunction, a hallmark of diet-induced obesity (DIO), is increasingly attributed to alterations in intestinal nutrient and electrolyte transport. Yet the mechanisms that drive obesity-associated functional alterations of intestinal transporters remain incompletely understood. In this context, the effects of a high-fat diet (HFD) induced obesity on sodium-dependent glucose co-transporter 1 (SGLT1), Na⁺/H⁺ exchanger 3 (NHE3), and Cl⁻/HCO₃⁻ exchangers (DRA/PAT1), the primary glucose, sodium, and chloride absorptive pathways in mice small intestinal villus cells, were investigated. SGLT1 activity significantly increased in intact villus cells and brush border membrane vesicles (BBMV) from HFD-fed mice. Kinetic analysis demonstrated reduced Km without a change in Vmax, indicating enhanced transporter affinity. Notably, SGLT1 mRNA and protein expression, including BBM localization, were unchanged. Basolateral Na⁺/K⁺-ATPase activity was decreased, excluding enhanced Na⁺ gradient generation as the mechanism for SGLT1 stimulation. In contrast, DRA/PAT1 activity was significantly increased in HFD-fed mice, and kinetic studies revealed elevated Vmax without a change in Km, indicating increased transport capacity. DRA/PAT1 mRNA, total protein, and BBM expression were all significantly elevated. NHE3 activity and expression remained unchanged. These findings demonstrate that DIO enhances intestinal glucose absorption by increasing SGLT1 affinity and chloride absorption by upregulating DRA/PAT1 transcription. These transporter-specific alterations may amplify nutrient absorption and contribute to metabolic dysregulation in obesity. Full article

Background and Objectives: The objective of this study is to evaluate whether deferoxamine modulates cell biological properties, such as proliferation and wound closure of porcine corneal endothelial cells (CECs) in vitro, and whether the treatment of CECs with deferoxamine results in an enhanced expression of vascular endothelial growth factor (VEGF). Materials and Methods: Corneal endothelial cells were extracted from porcine globes within 24 h postmortem. Immunohistochemistry for the endothelial Na⁺/K⁺-ATPase was performed to confirm the cells’ endothelial origin. To assess CEC viability and proliferation, a water-soluble tetrazolium salt (WST-1) and 5-bromo-2′-deoxyuridine (BrdU) assay were performed. Corneal endothelial wound closure was evaluated using a wound closure assay. VEGF mRNA expression was evaluated using real-time polymerase chain reaction (rt-PCR). Results: The extracted corneal endothelial cells showed a typical hexagonal morphology with Na⁺/K⁺-ATPase staining of the cell membrane. The treatment with 200 µM deferoxamine significantly increased CEC viability to 121 ± 24% compared to the control group (p = 0.0024). Corneal endothelial cell proliferation did not show any significant changes under the treatment with deferoxamine (p > 0.05). Both 100 µM and 200 µM deferoxamine led to a significantly smaller remaining wound area of 82.4 ± 6.7% and 78.7 ± 6.2% (p < 0.0001) in comparison to the control group after 24 h of treatment in the wound closure assay. Treatment with 200 µM deferoxamine significantly induced VEGF mRNA expression to 1.67- ± 0.57-fold from 1.00- ± 0.03-fold in the control group (p = 0.0006). Conclusions: Deferoxamine effectively enhances corneal endothelial cell viability and wound healing associated with an overexpression of VEGF. Thus, deferoxamine is a potent modulator of cell biological properties of corneal endothelial cells and maintains their integrity in vitro. Full article