Diazepam (DZP) is a widely prescribed benzodiazepine that is increasingly detected in aquatic systems and known to impair neural function in aquatic organisms. However, its potential to elicit intergenerational neurobehavioral disruption, along with the underpinning mechanism in fish, remains uncharacterized. In this study, zebrafish (Danio rerio) embryos were exposed to DZP (0.1-1000 μg/L) during early development, and behavioral endpoints were assessed from larval to adult stages in the parental (F0) generation and in F1 offspring not directly exposed via water. Embryonic DZP exposure reduced larval locomotor activity and altered anxiety-related thigmotaxis and social interaction in F0 adults; similar behavioral alterations were also observed in F1 larvae not directly exposed via water. Transcriptomic profiling identified that the top enriched pathways altered in DZP-exposed fish were GABA receptor signaling. Targeted neurochemistry and fluorescence imaging revealed sustained activation of the GABAergic system in the F0 and F1 fish brain, as evidenced by elevated GABA levels and increased gad1b-associated signals. Collectively, these results suggest that exposure to DZP can cause long-lasting and intergenerational behavioral changes in zebrafish. This highlights the importance of considering the developmental stage at which exposure occurs when assessing the ecological risk of benzodiazepines.

Aims: Morin is a flavonoid with potential neuroprotective and anti-inflammatory properties. This study evaluated the anticonvulsant and anxiolytic effects of morin-loaded PLGA-chitosan nanoparticles (Morin-PLGA-CS NPs) in male Wistar rats. Methods: Morin-PLGA-CS NPs were synthesized using a modified single emulsion-solvent evaporation method followed by CS coating. NPs were characterized by dynamic light scattering (DLS), scanning electron microscopy (SEM), and in vitro drug release analysis. Adult male Wistar rats received intraperitoneal injections of free morin (25 mg/kg), Morin-PLGA-CS NPs, diazepam (1 mg/kg), blank NPs, or vehicle. Behavioral assessments included the open-field test (OFT), elevated-plus maze (EPM), novel object recognition (NOR) test, and pentobarbital-induced sleep test. Anticonvulsant activity was evaluated using PTZ-induced seizure latency. Cytokine concentrations in cortical and hippocampal tissue lysates were quantified by ELISA at 12 h post-PTZ. Hippocampal relative mRNA expression of Nrf2, HO-1, GFAP, and Iba1 was quantified by quantitative real-time PCR (qRT-PCR). Results: Morin-PLGA-CS NPs demonstrated a hydrodynamic diameter of 221.6 nm, a zeta potential of +23.3 mV, and an encapsulation efficiency of 81%. FTIR spectroscopy showed spectral changes compatible with morin incorporation and possible hydrogen-bonding interactions. The NPs exhibited approximately 81.4% morin release over 72 h in vitro. Compared to free morin, Morin-PLGA-CS NPs increased center-zone exploration in the OFT and open-arm behavior in the EPM, but also reduced total distance traveled in the OFT, indicating that motor suppression or sedation may have contributed to the behavioral profile (p < 0.001). It also improved the discrimination index (DI) in the NOR test and elevated sleep duration in the pentobarbital test (p < 0.001). The NPs also prolonged seizure latency (137.2 s vs. 114.5 s for free morin; p < 0.001) and markedly reduced IL-1β, IL-6, and TNF-α levels in both the cortex and hippocampus. At the molecular level, Morin-PLGA-CS NPs were associated with significantly increased hippocampal Nrf2 and HO-1 mRNA transcript levels and decreased GFAP and Iba1 transcript levels relative to free morin and the PTZ-challenged vehicle control group. Conclusions: Morin-PLGA-CS NPs produced greater behavioral, anticonvulsant, inflammatory, and redox effects than free morin in male rats. Molecular data revealed changes in hippocampal mRNA expression, including increased Nrf2 and HO-1 transcripts and decreased GFAP and Iba1 transcripts. However, these transcript-level findings are preliminary and require protein-level validation.

Ovine coenurosis, caused by the larval stage of Taenia multiceps (Coenurus cerebralis), is a parasitic disease that affects the central nervous system of livestock. While diagnosis traditionally relies on clinical signs, imaging, and postmortem examination, electroencephalography (EEG) remains underutilized despite its potential to detect brain abnormalities. This case report describes EEG and quantitative EEG (qEEG) findings in a 2-year-old sheep presenting with clinical signs suggestive of a chronic focal encephalopathy (ataxia, head tilt and circling) associated with lethargy and progressive weight loss. Computed tomography (CT) confirmed a single, discretely bilobate intra-axial cystic lesion within the left cerebral hemisphere, with a severe mass effect, marked deformation and displacement of the ventricular system, compression of the thalamic structures, and contralateral deviation of the median line. A solitary parasitic cyst with no lesion in the right hemisphere was subsequently confirmed at necropsy and on histopathology. Under ketamine-diazepam sedation, both EEG recordings showed a continuous, delta-dominant background consistent with the anesthetic protocol. Epoch-wise asymmetry indices with bootstrap confidence intervals distinguished an unstable component (which reversed in hemispheric sign between recordings) from a reproducible one confined to the occipital derivations, where the asymmetry lay over the right hemisphere in both recordings in every band except delta. This reproducible preponderance was contralateral to the cyst, over a hemisphere in which no structural lesion was demonstrable either on imaging or at necropsy, and absolute powers indicated that it arose principally from attenuation of the signal recorded over the diseased hemisphere. These findings provide the first electrophysiological characterization of ovine coenurosis and show that in the presence of a severe mass effect, the side of the surface qEEG abnormality may not indicate the side of the lesion (an electrophysiological false localizing sign described in human patients but not previously in a ruminant). EEG therefore reflects the functional brain disturbance caused by Coenurus cerebralis but should be regarded as a complement to, rather than a substitute for, cross-sectional imaging in neurolocalization.

The objective of this prospective alternating crossover study was to compare quality of induction and recovery and cardiopulmonary function in horses administered guaifenesin or diazepam as co-induction drugs. Six healthy horses were anesthetized twice to undergo an MRI and surgical biopsies, with 5 wk between treatments. Horses were sedated intravenously (IV) with xylazine, 0.6 mg/kg BW, and IV induction started 4 min later. Group G received guaifenesin, 100 mg/kg BW, over 2 min and ketamine, 2 mg/kg BW, within 1 to 2 min of starting the guaifenesin. Group D received diazepam, 0.1 mg/kg BW, within 5 s and ketamine, 2 mg/kg BW, 0.5 to 1 min after and the quality of induction assessed. Horses were positioned on an MRI-compatible table, the trachea intubated, allowed to breathe spontaneously for the initial 10 min, then connected to a rebreathing circuit and anesthetic machine and mechanically ventilated under anesthesia [end-tidal isoflurane (FE'Isoflurane): 1.3 to 1.5%] during the MRI. Direct arterial blood pressure, heart rate, end-tidal CO2 (PE'CO2) and FE'Isoflurane were recorded at 10, 20, 30, and 40 min post-induction. Requirements for dobutamine and/or phenylephrine for arterial blood pressure support were recorded. A 2-way analysis of variance (ANOVA) was used to compare arterial blood pressure and a Wilcoxon matched-pairs signed rank test for comparing sedation, induction, and recovery scores (significance was set at P < 0.05). Arterial pressures were not significantly different between groups; however, systolic (P = 0.004), diastolic (P < 0.0001), and mean (P = 0.004) arterial pressures decreased significantly at 20, 30, and 40 min in Group D. Dose of dobutamine and/or phenylephrine was similar between groups. Sedation, induction, and recovery scores were similar. Induction scores were good to excellent [both groups: 0.5 (0, 2); median (range)]. In conclusion, guaifenesin and diazepam at the doses used in this study produced similar effects on arterial blood pressure and on the quality of induction and recovery.

Acyl CoA binding protein (ACBP, encoded by the gene diazepam binding inhibitor, DBI) is a tissue stress hormone involved in pathological aging that mediates immunosuppression. The antibody-mediated neutralization of ACBP/DBI enhances anticancer immune responses in the context of chemoimmunotherapy. ACBP/DBI inhibition also upregulates the expression of the anti-aging gene Lmna coding for Lamin A in tumor-infiltrating T lymphocytes. This study was designed to explore the role of Lmna in the anticancer effects of ACBP/DBI neutralization.

Amyotrophic lateral sclerosis (ALS) can cause autonomic storms characterized by blood pressure fluctuations, particularly in the advanced stages of the disease and among ventilator-dependent patients. Although benzodiazepines are used to manage these symptoms, the optimal agent is unclear. We present the case of a 78-year-old home-care patient with ventilator-dependent ALS who developed autonomic storms. Clotiazepam did not adequately control the episodes; however, diazepam was associated with resolution of the episodes, followed by improved sleep and nutritional intake. This report highlights the importance of recognizing an autonomic storm in advanced ALS patients and the potential benefit of diazepam in home-care settings.

Diazepam is a widely prescribed benzodiazepine that undergoes extensive hepatic metabolism. Patients with liver cirrhosis may exhibit altered diazepam pharmacokinetics (PK), resulting in increased systemic exposure and a higher risk of adverse effects. This study aimed to develop a physiologically-based-pharmacokinetic (PBPK) model of diazepam and evaluate the impact of hepatic impairment on drug exposure and dose requirements. A PBPK model of diazepam was developed using PK-Sim® based on published clinical PK data from healthy subjects. Drug-specific physicochemical and metabolic parameters were incorporated into the model. The model was externally qualified using independent intravenous and oral clinical datasets. Subsequently, disease-specific physiological and biochemical changes associated with Child-Pugh A (CP-A) and Child-Pugh B (CP-B) liver cirrhosis were incorporated to establish hepatic impairment PBPK models. Model-based simulations were performed to assess PK alterations and optimize dosing strategies. The developed PBPK model adequately described diazepam plasma concentration-time profiles across multiple dosing regimens and administration routes. All predicted-to-observed ratios for AUC and Cmax were within the predefined twofold error range. Simulations demonstrated progressive increases in diazepam exposure with worsening hepatic impairment. Following repeated oral administration of 5 mg twice daily, predicted steady-state AUC increased by 43.6% and 72.6% in CP-A and CP-B patients, respectively, compared with healthy subjects. Model-based extrapolation and dose optimization indicated that dose reductions of approximately 30% for CP-A and 42% for CP-B patients were required to achieve exposure comparable to that observed in healthy individuals. Sensitivity analysis identified CYP3A4-mediated metabolism, liver volume, plasma protein binding, and fraction unbound as key determinants of diazepam PKs. The developed PBPK model characterized diazepam PKs in healthy subjects and provided a disease-extrapolation framework for patients with liver cirrhosis. The model provides a quantitative framework for predicting hepatic impairment-associated PK changes and supports rational dose adjustment of diazepam according to the severity of liver dysfunction.

Benzodiazepines are the recommended first-line treatment for canine seizure emergencies, and non-parenteral routes are essential when intravenous access is unavailable. Clinical trials show that intranasal midazolam controls emergency seizures more reliably than rectal diazepam, yet the cortical electrophysiological correlates of these routes remain uncharacterised in dogs. In a two-session, fixed-order (non-randomised) study, the quantitative electroencephalographic (qEEG) profiles of intranasal midazolam (IN-MDZ; 0.2 mg/kg), volume-matched intranasal saline (IN-SAL) and intrarectal diazepam (IR-DZP; 0.5 mg/kg) were compared in six healthy adult dogs. Propofol (1%, 0.5 mL/kg IV) was given in both sessions for electrode placement, and recording followed clinical recovery. Metrics were computed at baseline and 1, 3 and 5 min and analysed with the dog as the unit of replication (n = 6), using Friedman and exact Wilcoxon tests with false-discovery-rate (FDR) control. IN-MDZ was associated with a coherent, rapidly emerging cortical-slowing signature: at 1-3 min it separated from both controls across relative delta, beta, delta/beta, theta/alpha, alpha and entropy, unanimously and drug-specifically, the correlated indices reflecting a single coherent spectral shift reproduced at two time points. This slowing, in contrast to the beta activation reported in seizing human patients, is consistent with a genuine, rapidly acting GABAergic effect, and is concordant with the established clinical efficacy of intranasal midazolam. No contrast survived false-discovery-rate correction; the core indices nonetheless persisted on baseline-corrected change scores (r = 0.7-0.9). The study defines the qEEG endpoints, effect sizes and sample size required for an adequately powered confirmatory trial.

Posterior reversible encephalopathy syndrome (PRES) is a reversible neurological disorder most commonly triggered by eclampsia and characterized on imaging by vasogenic edema, typically involving the parieto-occipital regions. We describe a 26-year-old primigravida at 35 weeks of gestation who presented with headache, dyspnea, and a generalized tonic-clonic seizure, with severe hypertension (200/110 mmHg), 3+ proteinuria, serum creatinine (1.1 mg/dL), and platelet count (100,000/mm[3]) consistent with severe preeclampsia with superimposed eclampsia. She underwent emergency lower-segment cesarean section (LSCS) for delivery of a live neonate, with a second intraoperative seizure controlled with intravenous diazepam and magnesium sulfate. Postoperatively, she developed sudden bilateral visual loss without focal neurological deficit. Brain MRI showed bilateral, symmetric T2- and FLAIR-hyperintense signal in the parieto-occipital white matter without evidence of restricted diffusion on DWI/ADC a pattern of vasogenic edema diagnostic of PRES. She was treated with intravenous magnesium sulfate, antihypertensive therapy, and levetiracetam, with complete clinical recovery and radiological resolution on follow-up imaging. This case highlights eclamptic PRES as a cause of postoperative cortical visual loss and illustrates how the combination of T2/FLAIR signal change with preserved diffusion on MRI distinguishes reversible vasogenic edema from ischemic injury, supporting prompt, targeted management even in resource-limited settings.