Acetaminophen (APAP) overdose induces systemic oxidative and inflammatory responses, yet its effects on splenic homeostasis remain poorly characterized. We investigated whether prophylactic Viburnum opulus L. protects against APAP-induced splenic injury and the molecular pathways potentially underlying this protection. Forty female Wistar rats were allocated to control, APAP, N-acetylcysteine (NAC), V. opulus 100 (VO-100), and 300 mg/kg (VO-300) groups. NAC or V. opulus was administered orally for seven days, followed by a single oral APAP dose of 3 g/kg. Splenic histomorphology and immunohistochemical expression of cleaved Caspase-3, Ki-67, NF-κB, iNOS, SOD1, NRF2, and GPX4 were evaluated; network pharmacology and independent splenic transcriptomic analyses contextualized the experimental findings. APAP disrupted white- and red-pulp organization, induced lymphoid depletion, follicular disorganization, congestion, and sinusoidal dilatation, and significantly altered all seven markers, increasing cleaved Caspase-3, NF-κB, and iNOS immunoreactivity while reducing Ki-67, SOD1, and NRF2. GPX4 also decreased, but the control-APAP comparison lost significance after multiple-testing correction. VO-300 markedly attenuated the histomorphological alterations, reduced inflammatory and apoptosis-associated marker expression, and increased proliferative and antioxidant markers toward control levels. Network pharmacology identified 44 shared targets between major V. opulus constituents and APAP-toxicity-associated genes, with AKT1, NFKB1, PTGS2, and STAT3 as key hub proteins linked to oxidative-stress, apoptotic, and NF-κB-related pathways; 28 of these targets were confirmed in independent spleen transcriptomic data. Prophylactic V. opulus, particularly at 300 mg/kg, therefore, limits APAP-induced splenic injury by preserving tissue architecture, proliferative activity, and antioxidant defenses while attenuating inflammatory and apoptosis-associated responses.

Acetaminophen (APAP) is the most widely used analgesic during pregnancy, yet its effects on prenatal human brain development remain incompletely understood. Epidemiological studies have reported inconsistent associations between prenatal APAP exposure and later neurodevelopmental outcomes, underscoring the need for mechanistic evaluation in human-relevant developmental models. Here, we examined how APAP influences early cortical development using induced pluripotent stem cell-derived cortical organoids (COs) generated from six independent lines. Organoids were exposed to physiologically relevant APAP concentrations (25, 50, and 100 μM) for 5 days beginning at day 21 of differentiation, corresponding to late first-trimester cortical development. We assessed organoid growth, apoptosis, differentiation, synaptic maturation, transcriptomic profiles using bulk and single-nucleus RNA sequencing (snRNA-seq), and functional network activity using multielectrode array recordings up to 4 months. APAP exposure did not affect organoid size, cytoarchitecture, or viability. Neuronal and progenitor cell proportions, as well as synaptic puncta density were unchanged. Bulk RNA-seq revealed subtle transcriptional changes only at the highest dose (16 differentially expressed genes at 100 μM), enriched for neurodevelopmental pathways. In contrast, snRNA-seq at 3 months revealed no changes in cell type composition or gene expression. Consistent with the limited molecular changes observed, electrophysiological measures including firing rate, burst frequency, and network synchrony, showed no significant differences from controls. Together, these results indicate that direct exposure to parent APAP at the concentrations tested produces minimal molecular perturbations. These findings reflect responses to parent acetaminophen in vitro and do not exclude metabolite-mediated or systemic effects in vivo.

In this work, a liver-targeted NIR fluorescent probe CA-Cys for Cys was developed by integrating benzothiazole hemicyanine, an acrylate moiety responsive to Cys, and a cholic acid (CA) ligand. The CA group of CA-Cys recognizes the sodium-taurocholate cotransporting polypeptide (NTCP) specifically expressed on hepatocytes, enabling specific liver targeting and excellent resistance to disease interference. CA-Cys exhibits an excellent linear response to Cys with a detection limit of 0.54 μM, rapid response within 30 s, and favorable selectivity, without interference by other reactive sulfur species such as homocysteine, glutathione, hydrogen sulfide, etc. CA-Cys exhibits liver-targeted ability both in vitro and in vivo. Moreover, CA-Cys distinguishes liver cancer cells from normal liver cells by sensing intracellular Cys levels. CA-Cys can track the Cys changes in drug-induced liver injury (DILI) cell models and evaluate the therapeutic effect of N-acetylcysteine (NAC). In vivo and ex vivo imaging of mice showed that Cys accumulation induced by acetaminophen (APAP) mainly occurred in the liver, and NAC treatment effectively restored hepatic Cys homeostasis. This liver-targeted CA-Cys enables non-invasive real-time visualization of hepatic Cys at both cellular and animal levels, offering a practical tool for pathological research, diagnosis, and therapeutic monitoring of DILI.

Powder segregation is a critical quality risk in solid dosage manufacturing, yet current risk assessment is resource-intensive hence poorly suited to early formulation screening. This work addresses that by introducing a mechanistic predictive framework linking constituent particle properties and blend flowability to segregation risk. Binary blends of two ibuprofen grades (IBU25, IBU70) and micronized acetaminophen (mAPAP) with eight pharmaceutical excipients (n = 24 total) were characterized for particle size, shape, density, and flow function coefficient (FFC). Segregation was quantified by NIR spectroscopy (SPECTester) using Segregation Intensity (SI) and Cumulative Segregation Area (CSA), a composition-integrated metric. Particle size-density disparity between API and excipient establishes the mechanical driving force for segregation, while blend flowability determines how fully that potential is expressed; neither factor alone is sufficient, but together they could be predictive. Consequently, a dimensionless Particle Mechanical Property Factor (PMPF), combining D3,2 ratio and bulk density ratio, was introduced as a pre-formulation disparity descriptor, leading to a novel parameter called the Segregation Propensity Number (SPN) as the product PMPF × FFC. It was used in a power-law model y = k(SPN)n, trained on IBU25 and mAPAP blends (n = 16) and validated on held-out IBU70 blends (n = 8) achieved R2_ext = 0.95 for CSA and R2_ext = 0.84 for SI, with CSA showing superior generalizability and minimal systematic bias. This framework is expected to enable a priori segregation risk assessment from routinely collected particle data paired with a single blend flowability measurement, supporting QbD-driven excipient selection and early formulation development.

Functional dispersion within microbial communities is a major constraint on lignin humification during composting. Lignin degradation relies on extracellular enzymes whose activities can be modulated by phenolic units released during lignin depolymerization. Accordingly, acetaminophen (APAP) and mature composting products (MC) were combined in the MCA treatment to evaluate their synergistic effects on microbial functional specialization and humification. The MC treatment modestly enhanced humification, increasing HD and final HA content by 30.3 % and 16.4 % relative to CK, respectively. APAP produced larger increases of 119.4 % and 24.2 %, respectively, but reduced community stability. By contrast, the combined MCA treatment produced the greatest response, with HD and final HA content increasing by 172.7 % and 40.9 %, respectively, at the end of composting. Activities of laccase, lignin peroxidase, and manganese peroxidase also increased, while pathways associated with aromatic compound degradation and energy metabolism were coordinately enriched. These responses suggest broader metabolic activation accompanied by greater functional focus. Together, the findings suggest that combining phenolic small molecules with mature composting products promoted microbial functional specialization. They also provide complementary support for a conceptual model linking substrate induction, microbial community assembly, and enzyme responses. Overall, the combined treatment enhanced humification, providing a useful basis for optimizing straw composting and resource utilization.

Ferroptosis, often characterized by glutathione depletion-associated oxidative stress and lipid peroxidation, contributes importantly to many pathological conditions, including drug-induced liver injury. Protein disulfide isomerase (PDI or PDIA1) has emerged as a pivotal upstream regulator of chemically-triggered ferroptosis and also an important therapeutic target. In this work, we identified raloxifene (RAL), a clinically-used selective estrogen receptor modulator, as a suppressor of chemically-induced oxidative ferroptosis and liver toxicity both in vitro and in vivo. Mechanistically, RAL can interact directly with PDI in both purified systems and cultured hepatocytes, leading to inhibition of its enzymatic activity. This inhibition decreases the dimerization (i.e., catalytic activation) of the inducible nitric oxide synthase (iNOS) and the subsequent buildup of cellular nitric oxide (NO), and these changes are associated with reduced accumulation of cellular reactive oxygen species, including lipid peroxides. RAL treatment markedly mitigates acetaminophen-evoked liver injury in vivo, as reflected by normalized serum enzyme levels, decreased lipid oxidative damage, and improved tissue morphology. Together, these results reveal that RAL can effectively limit oxidative ferroptotic injury in hepatocytes in both cellular and animal settings via its direct inhibition of PDI-mediated oxidative signaling cascade. The findings reveal a novel estrogen receptor-independent, PDI-targeted mechanism for the hepatoprotective actions of RAL, and also illustrate its promise for treating ferroptosis-associated liver injuries.

Drug-induced liver injury (DILI) is a serious adverse reaction with limited therapeutic options. Taraxasterol (TAS), a natural triterpenoid with hepatoprotective potential, suffers from poor aqueous solubility and low oral bioavailability, hindering its clinical application. Here, we develop an integrated oral delivery platform comprising TAS nanocrystals (TAS NCs) stabilized by D-α-tocopherol polyethylene glycol succinate (TPGS), embedded within chitosan-carbopol polyelectrolyte complex (EPC) micropatches, and encapsulated in enteric-coated capsules (TAS NCs@EPC/CAP). This design sequentially overcomes multiple absorption barriers through a multi-stage process: nanocrystallization enhances dissolution and permits TPGS-mediated P-glycoprotein inhibition and hepatocyte uptake; the mucoadhesive EPC micropatches prolong intestinal residence and transiently open tight junctions; and enteric coating ensures site-specific release in the small intestine. In mice, TAS NCs@EPC/CAP dramatically improved oral bioavailability (AUC increased 3.1-fold vs. free TAS) and achieved sustained liver enrichment. In both acetaminophen-induced intrinsic and isoniazid/rifampicin-induced idiosyncratic DILI models, TAS NCs@EPC/CAP exerted superior hepatoprotection, as evidenced by normalized liver function markers (ALT, AST, LDH), attenuated oxidative stress (ROS, MDA, GSH, SOD), suppressed inflammation (IL-1β, IL-6, TNF-α), and reduced hepatocyte apoptosis. Integrated network pharmacology and experimental validation revealed that TAS NCs@EPC/CAP potentiates the effects of TAS, leading to enhanced activation of the Nrf2/HO-1 antioxidant signaling, more pronounced inhibition of JNK-mediated stress pathways, and greater modulation of the Bax/Bcl-2/caspase-3 apoptotic cascade. This study establishes a clinically translatable oral delivery strategy for TAS and provides a versatile platform for enhancing the bioavailability of poorly soluble natural products, with potent efficacy against diverse forms of DILI.

Supraparticles (SPs) are secondary particles formed through nanoparticle self-assembly with porous architectures and bioinspired structural features suitable for drug delivery applications. Although binder-mediated structural control influences SP assembly, how different loaded drugs systematically alter amyloid nanofibril (ANF) and SP structures remains unclear. In this study, neutral, basic, and acidic model drugs-acetaminophen (ACP), fluconazole (FCZ), and proglumide (PGM), respectively-were incorporated into spray-dried SPs composed of silica nanoparticles (SiNPs) and ANFs to investigate the effects of drug properties on SP structure and release behavior. All drugs exhibited nearly 100% loading efficiency, and powder X-ray diffraction analysis revealed that they were in an amorphous state within the SPs. ACP- and FCZ-loaded SPs maintained their characteristic ANF-induced wrinkled morphology, whereas PGM-loaded SPs exhibited a pronounced loss of wrinkling at pH 2, indicating that acidic drugs can disrupt ANF-mediated assembly during SP formulation. Drug release studies further showed that ANFs suppressed the initial burst release and slowed the overall release by restricting diffusion pathways within the SP structures. This suppression was the most pronounced in PGM-loaded SPs, likely owing to drug retention within the assembled ANF domains. These findings demonstrate that drug acid-base properties critically modulate SP morphology and release behavior, highlighting the importance of drug-biomaterial interactions in designing SP-based delivery platforms.

Poisoning by acetaminophen is a serious public health concern because of its accessibility and the ease with which overdoses can occur accidentally or on purpose. The liver metabolizes acetaminophen, and when taken in excess, it is transformed into a poisonous metabolite that seriously damages the liver and may result in hepatic failure and death. The cornerstone of treatment is the early administration of N-acetylcysteine, which is most effective within 8 hours of ingestion. Co-ingestants, therapy timing, and pre-existing liver diseases are some of the factors that affect toxicity. Both unintentional overdoses and deliberate poisonings have legal and forensic ramifications, especially where minors or self-harm are involved. In fatal cases, a toxicological investigation is essential to identify the cause of death and differentiate acetaminophen poisoning from other possible causes. Regulations such as age restrictions, pack-size constraints, and improved labeling have demonstrated the potential to reduce the prevalence of acetaminophen toxicity. Intentional and accidental overdoses can be avoided in part by raising public awareness and educating healthcare professionals. To lessen the impact of acetaminophen poisoning on public health systems, more research is required to enhance early identification, improve treatment techniques, and guide policy decisions. This review aims to familiarize clinicians and forensic professionals with the pharmacology, clinical presentation, management, and medicolegal aspects of acetaminophen toxicity..

Objective: To describe the timing, frequency, and accessibility of Instagram posts from pediatric health institutions during the 2025 acetaminophen-autism controversy.Methods: Cross-sectional audit of public Instagram accounts of 62 pediatric health institutions from September 22 to November 15, 2025. The a priori sampling frame comprised the top 50 US children's hospitals for pediatric behavioral health, the top 10 Honor Roll children's hospitals (US News and World Report), 5 national medical societies, and health departments of the 10 states with the largest pediatric populations. Posts were extracted with a web-scraping tool and manually verified. Two reviewers independently coded relevant posts for prespecified content and accessibility features. Readability was assessed with two validated measures.Results: Twelve of 62 institutions (19%) posted relevant content. The American Academy of Pediatrics and the American College of Obstetricians and Gynecologists posted on day 0, while hospital time to first post ranged from 3 to 51 days. No state health department posted. Nonresponding institutions remained active: 1 hospital published 108 unrelated posts. The 26 relevant posts had a mean Flesch-Kincaid Grade Level of 11.4 and SMOG Index of 10.2. Among coded features, 88% included resource links, 73% directed audiences to a health care provider, 46% featured a named clinician, 42% discussed risks and benefits, and none were multilingual.Conclusions: Most sampled institutions published no relevant content during an active health controversy despite remaining active. Readability of relevant posts exceeded recommended targets, and none were multilingual. These gaps in institutional communication infrastructure could be addressed.

Recent studies indicate that injury-induced dedifferentiation generates transient regenerative progenitors (TRPs), which support tissue repair across multiple organs, including the liver. In vivo partial reprogramming using Yamanaka factors (Oct4, Sox2, Klf4, and c-Myc; OSKM) produces intestinal TRPs closely resembling those arising from injury-induced dedifferentiation, but transgenic or viral approaches raise safety concerns because of prolonged expression. Here, we show that transient OSKM induction generates two distinct liver progenitor-like cell (LPLC) populations without injury and with limited activation of injury-associated immune responses. Single-nucleus RNA sequencing shows that one population resembles injury-induced LPLCs observed in acetaminophen-injured mouse and human livers. To develop a temporally controlled regenerative strategy, we establish a liver-targeted lipid nanoparticle system for OSKM mRNA delivery. OSKM mRNA-LNP treatment induces hepatocyte dedifferentiation and Sox9[+] LPLC-like cell formation and is associated with enhanced regeneration after acetaminophen-induced injury. Together, these findings identify transient hepatic reprogramming as a strategy for inducing regenerative plasticity and support liver-targeted OSKM mRNA delivery as a potential approach for acute liver injury.

Real-world evidence on systemic diclofenac patch use in cancer pain is limited. We described prescribing patterns in oncology practice. In this single-center retrospective descriptive study, we reviewed consecutive patients with cancer who received a systemic diclofenac patch (75 mg per patch) for cancer-related pain at a Japanese university hospital (n = 48). The usual regimen was two 75-mg patches once daily (150 mg/day); one patient started one 75-mg patch/day. Among 48 patients, 35 (73%) retained oral medication intake and 32 (67%) were receiving ongoing anticancer therapy at patch initiation. Gastroprotective agents were co-prescribed in 31 patients (65%), any acetaminophen in 29 (60%; scheduled acetaminophen in 28 [58%]), and opioids in 28 (58%). Transdermal fentanyl-containing regimens were observed in 9 patients (19%), whereas strict all-transdermal scheduled baseline analgesia was identified in 3 (6%). No recipient had documented clinically apparent gastrointestinal bleeding or symptomatic peptic ulcer during available follow-up. The systemic diclofenac patch was prescribed to selected patients with cancer-related pain in routine Japanese practice and was commonly incorporated into multimodal analgesic regimens. These findings describe prescribing patterns and chart-documented observations only; prospective studies with standardized outcomes are needed to clarify effectiveness, safety, and patient-centered impact.