Acetaminophen (APAP) also known as paracetamol, is the only antipyretic and analgesic agent that can be administered to pregnant women. In addition to hepatotoxicity in cases of an overdose, the developmental neurotoxicity of APAP has been reported in model organisms. In this study, developing zebrafish were used to assess the neurotoxic effects of waterborne APAP exposure (0.039-2.5 mM). At 120 hpf, the spontaneous swimming distance was significantly reduced at all tested APAP concentrations under alternating light-dark conditions. Touch-evoked escape velocity, which assesses the motor function, was not significantly affected at any concentration and it increased at moderate APAP doses. The optokinetic response was only reduced at 2.5 mM APAP concentrations, suggesting that the decrease in spontaneous swimming at low APAP doses is unlikely to be due to motor or visual impairments. Vibration-evoked responses, indicative of the sensory function, were slightly but significantly diminished at 0.625 and 2.5 mM APAP concentrations. At 72 h post-fertilization (hpf), acridine orange and terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) staining showed that exposure to 2.5 mM APAP resulted in regulated cell death in the cerebellum, retina, and dorsal trunk. Some APAP-induced cell death in the dorsal trunk was confirmed to be of neuronal origin using Tg(eno2:Cerulean) transgenic zebrafish. N-acetylcysteine, a well-known antioxidant and antidote for APAP toxicosis in humans, partially attenuated APAP-induced cell death. These results suggest that APAP induces developmental neurobehavioral and sensory organ toxicity in the absence of detectable regulated cell death under the present experimental conditions.

An electrochemical sensor based on a novel composite of electrochemically reduced graphene oxide, polypyrrole-coated magnetic nanoparticles (MNPs@PPy), and Nafion was developed for dopamine (DA) detection. The structural, thermal, and electrochemical properties of the composite were validated through a combination of advanced spectroscopic techniques, thermal profiling, electron microscopy, and impedance analyses. Under optimized conditions, differential pulse voltammetry (DPV) revealed a high sensitivity (1.573 A·M[-1]·cm[-2], R[2] = 0.9874) and a limit of detection (LOD) of 5.4 × 10[-9] M for DA. The sensor displayed excellent selectivity, showing minimal interference from ascorbic acid, uric acid, and acetaminophen. Repeatability and reproducibility were confirmed (coefficient of variation ~8%). Real-sample analysis of urine and blood demonstrated recovery rates between 75 and 116%.

Natural Killer (NK) cell-mediated immunosurveillance is a cornerstone of anti-tumor defense, yet its efficacy can be compromised by common clinical medications. Paracetamol (APAP) and metformin (MET) are widely used for pain and metabolic management in cancer patients, but their unintended effects on the immune-tumor interface remain poorly understood. MCF-7 breast cancer cells (Luminal A subtype) were treated with APAP or MET and co-cultured with primary expanded NK cells (CD3[-]CD56[+]CD16[+]). We evaluated cell proliferation, cell cycle distribution, and the enrichment of the CD44[+]CD24[-] cancer stem-like cell (CSC-like) subpopulation. Transcriptional changes in immune checkpoints (PD-L1/L2), stress ligands (MICA/B), and costimulatory molecules CD80/86 were quantified via RT-qPCR. Despite inhibiting MCF-7 growth (IC50 at 48 h: 11.86 mM for APAP; 21.11 mM for MET), both drugs induced a "therapeutic paradox" by promoting an immune-evasive phenotype. APAP and MET significantly enriched the CD44[+]CD24[-] CSC-like subpopulation to 75.31% and 68.31%, respectively, compared to 11.00% in controls. Molecular analysis revealed a robust upregulation of PD-L1 (19.9-fold by APAP) and PD-L2 (16.4-fold by MET), alongside increased CD80/86 and MICA/B transcription. Consequently, drug-treated cells exhibited marked resistance to NK-mediated apoptosis and necrosis. NK cells preferentially eliminated non-stem cells (non-CSCs), inadvertently further concentrating the highly resistant CSC-like subpopulation. Additional experiments revealed that APAP directly impaired NK-cell survival and reduced the proportion of CD3[-]CD56[+] cells, whereas MET exerted minimal effects on NK cells, suggesting distinct mechanisms underlying the observed reduction in NK-mediated cytotoxicity. Under the experimental conditions employed in this study, APAP and MET were associated with reduced susceptibility of MCF-7 cells to NK-mediated killing through distinct but partially overlapping mechanisms, including CSC-like enrichment and transcriptional activation of immune-evasion pathways. Although these findings were obtained in a mechanistic in vitro model using supra-physiological drug concentrations, they identify potential mechanisms that warrant further validation in physiologically relevant experimental systems and in vivo models.

Objective: Our objective was to compare clinical safety and postoperative efficacy between transvesicoscopic Politano-Leadbetter (TPL) and laparoscopic Lich-Gregoir (LLG) ureteral reimplantation in infants and toddlers under 36 months diagnosed with primary obstructive megaureter (POM). Methods: This was a single-center retrospective cohort study of pediatric patients with POM who underwent ureteral reimplantation between January 2018 and April 2025. Patients were stratified into TPL and LLG groups by surgical approach. Baseline characteristics, perioperative outcomes, perioperative complications, imaging findings, renal functional outcomes, and long-term complications were collected and analyzed. Primary group comparisons were unadjusted, with additional multivariable linear regression performed for operative duration. Results: We enrolled 65 children: 30 in the TPL group and 35 in the LLG group. Baseline characteristics showed no statistically significant differences across the two study cohorts. There were no open conversions in either group. Total operative duration did not differ significantly between the TPL and LLG groups (145.8 ± 30.6 min vs. 136.9 ± 25.2 min; mean difference, 8.9 min; 95% CI, -4.9 to 22.7; p = 0.210). In multivariable linear regression, TPL showed a non-significant trend toward longer operative time after adjustment for age, preoperative ureteral diameter, and surgery year (B = 10.4 min; 95% CI, -3.7 to 24.4; p = 0.141). Time to postoperative oral intake was slightly shorter in the TPL group (6.1 ± 2.3 h vs. 8.2 ± 2.9 h; mean difference, -2.1 h; 95% CI, -3.4 to -0.8; p = 0.002). No statistically significant differences were detected in estimated blood loss, ureteral tapering rate, double-J stent placement rate, postoperative pain score, acetaminophen use, urinary catheterization duration, or length of hospital stay. Perioperative complications occurred in 3 patients in the TPL group and 5 patients in the LLG group (10.0% vs. 14.3%; risk difference, -4.3%; 95% CI, -20.1% to 11.5%; p = 0.716). No Clavien-Dindo grade III or higher complications occurred in either group. During follow-up, both groups showed reductions in anteroposterior pelvic diameter (APD) and ureteral diameter (UD), with preserved differential renal function (DRF). Low-grade postoperative vesicoureteral reflux (VUR) was detected in two LLG patients, including one grade I case at 4 months and one grade II case at 6 months after surgery; both were managed conservatively. Neither group experienced recurrent ureterovesical junction obstruction (UVJO) nor required reoperation during follow-up. Conclusions: In this single-center retrospective cohort of infants and toddlers with POM, no statistically significant differences were detected in the main perioperative and follow-up outcomes between TPL and LLG. TPL enables intravesical reconstruction and better preserves the natural anatomical course of the ureter and may serve as an alternative minimally invasive option for this patient population.

Background: Although genetic-environmental interactions are established in autism spectrum disorder (ASD), how environmental toxicants confer susceptibility remains unclear. This study aimed to investigate potential relationship between genetically predicted cerebrospinal fluid (CSF), metabolite levels and ASD liability, and to prioritize regulatory genes, key pathways, and candidate environmental toxicants. Methods: Using two ASD GWAS datasets (exploration data: 18,381 ASD cases/27,969 controls; validation data: 18,235 ASD cases/36,741 controls), we applied multi-omics approaches to prioritize ASD-associated CSF metabolites, regulatory SNPs, and genes. Enrichment analysis and protein-protein interaction (PPI) network analysis were performed on these metabolite-related genes to explore the potential mechanisms linking CSF metabolic disturbances to ASD. Finally, candidate environmental neurotoxicants were screened through protein-chemical interaction analysis, with binding relationships assessed via molecular docking prediction. Results: Two-sample Mendelian randomization (MR) analysis prioritized adenine and proline as candidate CSF metabolites with potential risk associations with ASD. Summary-data-based MR (SMR) prioritized 39 brain-specific quantitative trait loci (QTL) involving 35 candidate regulatory genes, including dual-metabolite modulator GRM8. Functional enrichment analyses suggested potential associations with mitochondrial dysfunction, Hippo signaling pathway, and microtubule dynamics impairment, with protein-protein interaction networks highlighting KATNA1/KATNAL2 as hubs. Protein-chemical interaction screening nominated 14 candidate environmental toxicants, including established chemicals (acetaminophen, valproic acid, estradiol) and novel candidates (SB-431542, K 7174, benzo[a]pyrene), with docking affinity assessed computationally. Conclusions: Our study provides suggestive evidence that elevated adenine and proline may be potential risk factors for ASD and suggests possible involvement of the mitochondrial-Hippo-microtubule pathway. We also propose benzo[a]pyrene as a candidate environmental toxicant that may perturb CSF metabolism. However, given the limited statistical significance, these findings require further validation.

Baicalin (BAI) is a natural flavonoid with hepatoprotective potential; however, its poor aqueous solubility and low oral bioavailability limit its therapeutic efficacy. Here, a carrier-free, self-assembled mangiferin (MAN) hydrogel was developed for oral BAI delivery without additional polymeric carriers or chemical crosslinkers. The resulting BAI@MAN HG formed a porous network with favorable hydration and viscoelastic properties while retaining antioxidant activity. FTIR, Raman spectroscopy, XRD, and DSC collectively revealed altered molecular organization, reduced crystalline order, and changes in the solid-state environment of BAI following hydrogel formation, consistent with a possible noncovalent association between BAI and MAN. BAI@MAN HG enhanced the apparent solubility, gastrointestinal release, and oral absorption of BAI, resulting in greater systemic exposure than free BAI. In a preventive mouse model of acetaminophen-induced acute liver injury, oral administration of BAI@MAN HG markedly improved liver morphology, reduced hepatocellular damage, normalized serum transaminase levels, and alleviated oxidative stress and inflammation. These protective effects were accompanied by restoration of redox homeostasis and reduced hepatic levels of HMGB1, NF-κB and other inflammatory mediators. Overall, MAN functioned as both a structural gelator and a bioactive matrix, providing a promising carrier-free platform for enhancing the oral delivery and in vivo efficacy of poorly water-soluble bioactives.

Background/Objectives: Opiates comprise naturally occurring opium alkaloids and their semisynthetic derivatives. Routine urine drug screening relies on enzyme immunoassays (EIAs) to rapidly detect opiate exposure; however, EIAs provide limited insight into opiate-associated metabolic patterns. Methods: We retrospectively analyzed liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-qToF-MS) datasets from comprehensive urine drug screening of 363 patients at the University of Pittsburgh Medical Center Clinical Toxicology Laboratory. Multiple statistical analyses were applied to identify the features associated with opiate (OPIA)-EIA-positive, oxycodone (OXY)-EIA-positive, and 6-monoacetylmorphine (6MAM)-EIA-positive specimens (42, 34, and seven specimens, respectively) designated as EIA-associated discovery feature sets. The feature sets selected by ≥2 statistical analyses were defined as EIA-associated consensus feature set and further evaluated using MS-FINDER for feature annotation. Results: Among 14,883 features, 138, 121, and 104 features were assigned to the OPIA-, OXY-, and 6MAM-EIA discovery feature sets, respectively. Consensus feature sets included oxycodone/opiate metabolites, acetaminophen metabolites, and norfentanyl for OPIA-EIA; oxycodone metabolites, α-phenylalanylaspartic acid, and 4-pyridoxic acid for OXY-EIA; and norfentanyl, 6-monoacetylmorphine, and 3-hydroxycotinine artifact for 6MAM-EIA. Conclusions: These metabolomic patterns indicate a dominant exposure gradient model, in which OXY-EIA-positive specimens primarily reflect prescribed oxycodone exposure, 6-MAM-EIA-positive specimens reflect illicit heroin/fentanyl exposure with polysubstance/recreational use signature, and OPIA-EIA-positive specimens occupy an intermediate, mixed profile shaped by immunoassay cross-reactivity and real-world co-exposures. Associations involving α-phenylalanylaspartic acid and 4-pyridoxic acid are hypothesis-generating and require further validation. These findings illustrate the value of archived clinical toxicology datasets for metabolomic discovery and as a foundation for sentinel laboratory-based surveillance of evolving drug and chemical exposures.

Acetaminophen (APAP) overdose frequently leads to acute liver injury (ALI), driven by oxidative stress and inflammation from toxic metabolites. Current treatments like N-acetylcysteine have a narrow therapeutic window, highlighting the need for novel herbal alternatives. This study investigated the hepatoprotective mechanisms of Lonicera confusa (SYH) against APAP-induced ALI using an integrated network pharmacology and molecular docking approach. We identified 33 active compounds and 55 potential targets. Protein-protein interaction analysis revealed five core targets: IL6, TNF, EGFR, CASP3, and TLR4. Enrichment analyses indicated SYH modulates key pathways related to oxidative stress and inflammation, including PI3K-AKT, NF-κB, and TNF pathways. Through molecular docking, strong binding affinities were confirmed, and apigetrin exhibited high affinity for the core targets. Subsequent in vivo validation demonstrated that apigetrin effectively alleviated APAP-induced ALI and inflammatory responses in mice. Collectively, these results suggest SYH mitigates ALI by inhibiting the pro-inflammatory TLR4/MYD88/NF-κB pathway and activating the antioxidant Keap-1/Nrf2/HO-1/NQO1 pathway.