The accumulation of acetaminophen (APAP) in agricultural soils and water systems poses risks to ecosystems and public health. This study characterizes Acinetobacter sp. DL27, an APAP-degrading strain with broad temperature and pH adaptability, demonstrating its bioremediation potential in soil and wastewater. Through high-resolution mass spectrometry, we identified three novel metabolic intermediates, thereby refining the bacterial APAP degradation pathway. Multiomics analysis elucidated metabolic coordination and stress-tolerance mechanisms, leading to the identification of a novel amidase (AdA). Recombinant AdA exhibited activity over 10-60 °C and pH 4.0-10.0, with a Km of 8.96 ± 1.03 μM and a kcat/Km of 9.04 μM-1s-1, indicating higher catalytic efficiency than previously reported APAP amidases. Molecular dynamics simulations and site-directed mutagenesis confirmed that a Ser161-Ser185-Lys82 triad constitutes the catalytic center driving amide cleavage. These findings provide mechanistic insights into bacterial APAP biodegradation and highlight the practical application potential of strain DL27 and AdA.

Acetaminophen (APAP) overdose is a leading cause of severe liver injury and acute liver failure (ALF). Increased hepatic macrophages promote inflammation and clear necrotic cell debris, thereby promoting a regenerative state for liver repair during APAP-induced liver injury. Vacuole membrane protein 1 (VMP1) is an ER transmembrane protein essential for autophagy. Deletion of VMP1 in the liver impairs autophagy. The role of VMP1 in APAP-induced liver injury is unknown. In this study, human APAP ALF RNA sequencing database was analyzed. Liver-specific Vmp1 knockout (Vmp1LKO) and matched wild-type (Vmp1LWT) mice were administered with 500 mg/kg APAP. Liver injury, necrosis and other biochemistry analyses were assessed. Increased mRNA levels of VMP1 and autophagy-related genes were found in human APAP ALF livers. Serum ALT and hepatic necrosis were increased in WT mice following APAP treatment. LC3B-II and p62 were increased, but VMP1 protein decreased in WT mouse livers following APAP treatment. Liver injury was markedly decreased in Vmp1LKO mice compared with their WT littermates. Basal hepatic CYP2E1 and glutathione (GSH) levels were similar between genotypes, but Increased lipid-laden macrophages (foam cells) were present in Vmp1LKO livers. Interestingly, APAP protein adduct formation decreased in Vmp1LKO mouse livers but mitochondrial damage was similar in both phenotypes. Peroxynitrite formation was observed in APAP-treated Vmp1LWT hepatocytes but was limited in foam cells of Vmp1LKO mouse livers. Furthermore, GSH replenishment and liver regeneration were enhanced in Vmp1LKO mouse livers. In conclusion, loss of hepatic VMP1 protects APAP-induced liver injury by replenishing GSH, modulating inflammation and enhancing liver regeneration.

This overview of systematic reviews aimed to identify effective pain management strategies for neonatal blood sampling. Searches were conducted in four databases from 2014 to April 2026. Eligible studies were systematic reviews including randomized or quasi-randomized controlled trials assessing pain management in neonates during blood sampling. Reviews were required to have searched at least two databases within the past 10 years, reported their search strategy, and conducted a risk-of-bias assessment. The search identified 2463 records; 22 reviews (13 Cochrane and two non-Cochrane pairwise reviews, and seven network meta-analyses) met inclusion criteria. Interventions evaluated included opioids, paracetamol, topical anesthetics, breastfeeding or breastmilk, sucrose, music medicine, acupuncture, and other non-pharmacological methods. Sucrose with or without non-nutritive sucking was effective in reducing pain scores during heel pricks in preterm and term neonates (mean difference -1.74, 95% confidence interval [-2.11, -1.37], moderate-certainty). Skin-to-skin contact also reduced pain scores compared with control (mean difference -3.47 [-5.55, -1.38], moderate-certainty). Recent network meta-analyses generally supported parent-led interventions and oral sweet solutions as the most effective strategies. Breastfeeding, supplemental breastmilk, acupuncture, and light reduction were promising but require further evidence. For other interventions, evidence was insufficient to draw conclusions. Evidence from systematic reviews and network meta-analyses indicates that sucrose and skin-to-skin contact effectively reduce procedural pain in neonates. Parent-led and other non-pharmacological interventions, including breastfeeding, also appear beneficial and should increasingly be considered part of family-centred neonatal pain care where feasible.

The issue of drug-alcohol interactions is a major concern in drug therapy and can endanger patient safety. Paracetamol is one of the most widely used analgetic drugs, while ethanol is commonly consumed addictive substance. This study investigated the toxic effects of paracetamol, ethanol, and their mixture at different concentrations on the embryonic development of zebrafish (Danio rerio, D. rerio).

Currently, the manufacture of injectable drug products relies on a quality-by-test approach, where product quality is evaluated at fixed manufacturing steps. The introduction of the quality-by-design approach, supported on process analytical technologies (PAT) is changing this paradigm. The US Food and Drug Administration established the foundations for the application of PAT in real-time control of the drug manufacturing process. One of the most versatile PAT tools is the Raman spectroscopy. Nevertheless, the application of this technique to injectables is still underexplored. This work intends to expand the use of Raman spectroscopy as a PAT tool for injectable drug products. A Raman spectroscopy method was developed for the assay of paracetamol in paracetamol infusion solutions, using a PLS (partial least squares) algorithm. The method is specific, with a root mean square error of calibration (RMSEC), cross-validation (RMSECV) and prediction (RMSEP) of 1.067%, 2.591%, and 2.601%, respectively, with R[2] of 0.999 and Q[2] of 0.992. The method was developed with a hand-held Raman spectrometer and was tested as an at-line PAT tool, rendering similar error levels to the ones found for the validation, with results readily available in ca. 1 min. This work presents a further extension of PAT applied to injectable drug products, specifically with the development of an at-line paracetamol assay method. This approach enables flexible sampling in several manufacturing steps with a single instrument and without adaptation of the manufacturing line. This work marks a significant step towards a comprehensive monitorization and control of the injectable manufacturing process.