1. Pentobarbital (PTB) is metabolised by cytochrome P450 (CYP) enzymes, being a model compound for evaluating CYP-mediated drug-drug interactions, has become difficult to obtain for animal studies in Japan. The metabolic pathway of thiopental (TP), a barbiturate with similar pharmacological properties, remains insufficiently characterised. This study investigated TP metabolic profile, with emphasis on CYP modulator-induced drug-drug interactions.2. TP was administered intraperitoneally to either phenobarbital (PB), chloramphenicol (CP)-pretreated, or untreated rats. Plasma TP concentrations were quantified using HPLC-DAD and a plate reader. Hypnotic and sedative properties were assessed by time to loss of righting reflex and recovery from hindlimb paralysis. TP metabolism was further evaluated in vitro using rat liver microsomes (RLMs) and recombinant CYP enzymes, and hepatic CYP mRNA levels were quantified by real-time RT-PCR.3. Compared with TP alone, PB pretreatment shortened TP-induced anaesthesia and decreased plasma TP concentrations, whereas CP pretreatment prolonged the duration of action and increased plasma TP levels. TP metabolism was mediated by CYP2B1 and CYP2C11. PB pretreatment induced CYP2B1 and enhanced TP metabolism, whereas CP pretreatment suppressed CYP2C11, inhibited CYP2B1, and reduced TP metabolism.4. These results suggest that TP is a useful alternative model compound for investigating pharmacokinetics/pharmacodynamics and CYP-mediated drug-drug interactions.

Chactas exsul (Werner, 1939) is a specialized, saproxylic scorpion belonging to the diverse family Chactidae, inhabiting the humid tropical forests of Costa Rica and Panama. Characterized by robust pedipalps and a relatively small telson, C. exsul primarily relies on mechanical immobilization for prey capture, though it may utilize venom during challenging encounters. Despite its evolutionary importance, the venom of most of its members remains largely uncharacterized. In this study, we provide a comprehensive proteomic and biochemical analysis of the venom of C. exsul from the Caribbean lowlands of Costa Rica. Our results reveal a venom peptidome dominated by putative scorpine-like antimicrobial peptides, calcins, and orphan components such as La1-like peptides. Sodium channel-modulating toxins (NaTxs) and common non-buthid potassium toxins (α- and κ-KTx) were not identified. The protein fraction exhibited significant complexity, featuring metalloproteinases, serine proteinases, phospholipases A2, and two putative hyaluronidase isoforms, among others. CAP-domain secretory proteins show several variants in the venom. These findings suggest a dual-functional trophic and, probably, defensive strategy: with a peptide fraction likely targeting insect-specific K+ and Ca2+ channels, which could induce paralysis, while the enzymatic cocktail could facilitate toxin maturation and spreading, as well as tissue predigestion. These activities and the role of putative scorpines as antimicrobials against soilborne pathogens inhabiting humid microhabitats remain to be determined.

Intraoperative neuromonitoring (IONM) is commonly used during thyroid surgery to facilitate recurrent laryngeal nerve (RLN) identification and potentially reduce related postoperative complications. However, the efficacy of routine IONM in decreasing complication rates remains debated. In this study, we investigated the impact of IONM on postoperative complications after thyroidectomy. This retrospective single-center study included 214 thyroidectomy operations performed between 2021 and 2024. Patients were divided into 2 groups: Group 1 (n = 107) underwent surgery with routine IONM, and Group 2 (n = 107) without IONM, using only conventional direct visualization. Postoperative complications (RLN palsy, hypocalcemia, hypoparathyroidism, and incidental parathyroidectomy) were compared between groups. The primary outcome was any RLN palsy, analyzed at the nerve level. Prespecified propensity score matching and sensitivity analyses were also performed. Preoperative calcium levels were higher in Group 1 than in Group 2 (P = .002). Postoperative calcium and parathyroid hormone levels and the rates of hypocalcemia and hypoparathyroidism did not differ between the groups; the smallest P value across these outcomes was.04 before and.40 after adjustment for multiplicity. There were 198 nerves at risk in Group 1 and 204 in Group 2. Any RLN palsy occurred in 3/198 nerves (1.5%) in Group 1 and 2/204 (1.0%) in Group 2 (P = .68). Transient RLN palsy occurred in 3/198 (1.5%) and 1/204 (0.5%), respectively (P = .37), whereas permanent RLN palsy occurred in 0/198 (0%) and 1/204 (0.5%), respectively (P > .99). No difference in any RLN palsy was observed in the propensity score-matched cohort (P > .99), and comparable findings were observed in the pre specified sub cohort and sensitivity analyses. Routine IONM was not associated with a significant reduction in RLN palsy, hypocalcemia, or hypoparathyroidism compared with conventional direct visualization. The very small number of RLN events and the wide confidence intervals indicate limited statistical precision, and the study was not sufficiently powered to exclude clinically meaningful differences in the risk of RLN palsy.

BackgroundGuillain-Barré syndrome (GBS) can be difficult to recognise in advanced untreated HIV, particularly where cerebrospinal fluid analysis and electrophysiology are unavailable.MethodsWe retrospectively reviewed adults with advanced HIV who were not receiving antiretroviral therapy and presented with acute symmetrical weakness at an Indian HIV specialty centre during 2019-2020. Cases were classified as clinically probable GBS with Brighton Collaboration level 3 diagnostic certainty.ResultsEight cases were identified among 139 admissions (5.8%). Mean CD4 count 78 cells/mm3. All eight had multiple concurrent infectious conditions. Herpesvirus-associated syndromes included ulcerative oesophagitis compatible with a herpesvirus-associated process (n=5), antecedent herpes zoster (n=2), or diarrhoeal illness with supportive CMV IgG serology (n=1). Seven recovered after multimodal treatment; one died.ConclusionsThis repeated clinical pattern of coexistence is hypothesis-generating but does not establish active viral reactivation, directionality, or causality. Pragmatic syndromic recognition and treatment remain necessary when definitive diagnostics are unavailable.

Chronic kidney disease (CKD) is associated with reflecting innate immune impairment. Chronic inflammation, oxidative stress, metabolic dysregulation, and prolonged uraemia are the primary causes of this condition. Monocytes, macrophages, and their progenitors may undergo drastic epigenetic changes in response to these systemic stresses. This has lowered the activation of histone marks, such as H3K4me3, and destabilised repressive marks, like H3K9me3. Critical immune response genes are silenced by chromatin closure. This epigenetic change exhausts the immune system and impairs its ability to clear pathogens. This causes 'uraemic immunodeficiency', a poor vaccination response, and increased susceptibility to infection. On the other hand, the Bacillus Calmette-Guérin (BCG) vaccination may produce trained immunity, a favourable innate immunological memory. BCG stimulates the NOD2 and Akt/mTOR pathways. This increases H3K4me3 near cytokine gene promoters and stabilises H3K9me3 in non-essential genomic areas. This progressive remodelling creates transcriptionally permissive chromatin that increases inflammatory reactivity. This increases the phagocytosis and antimicrobial activity of monocytes, macrophages, and neutrophils. BCG also reprogrammes hematopoietic stem and progenitor cells to create sustained myeloid cells. Comparing CKD-induced immunological paralysis to BCG-induced immune extension indicates that metabolic and environmental factors strongly influence the innate immune epigenome. BCG-induced trained immunity may represent a low-cost, host-directed strategy to reduce infection susceptibility and improve vaccine responsiveness in patients with CKD. This review shows that the BCG restores chromatin accessibility and transcriptional ability to treat CKD-induced immunological deficiencies. To restore immunological function in CKD patients, BCG-based therapies and epigenetic modulators should be studied.

Sepsis and septic shock result from a dysregulated host response to infection, encompassing both hyperinflammatory and immunosuppressed phenotypes. Although immunomodulatory adjunctive therapies have been extensively investigated over the past decades, none have demonstrated consistent clinical efficacy. In contrast, low circulating immunoglobulin levels are common in septic patients and are associated with greater disease severity and poorer outcomes, providing a biological rationale for the use of polyclonal intravenous immunoglobulins (IVIg) as phenotype-targeted adjunctive therapies. This expert consensus sought to systematically evaluate the appropriateness of IVIg therapy across predefined and clinically relevant scenarios in critically ill patients and deliver pragmatic, phenotype-oriented guidance. A multidisciplinary Italian panel comprising experts in intensive care, infectious diseases, transplant medicine, and oncology applied a modified RAND/UCLA Appropriateness Method. The process integrated a structured literature review (PubMed, EMBASE, and the Cochrane Library) with anonymous 9-point Likert scale ratings. Indications were classified according to the median score as appropriate (7-9), uncertain (4-6), or inappropriate (1-3), and interpanel disagreement was assessed using the Interpercentile Range Adjusted for Symmetry. The expert panel evaluated seven phenotypes that were identified as the most clinically relevant and extensively addressed in the available literature. IVIg use was judged appropriate without formal disagreement in severe community-acquired pneumonia with hyperinflammation plus shock (median 8), low IgM < 60 mg/dL (median 8), or prior immunosuppression (median 8), whereas hyperinflammation without shock was uncertain (median 6). Appropriateness was also supported for abdominal infection with endotoxaemic shock, immune suppression/paralysis, or low IgM (all median 8); sepsis in solid-organ transplant recipients (median 7), particularly with low IgM (median 8), while prophylaxis in high-risk transplant recipients was uncertain (median 5). IVIg was rated appropriate for overwhelming septic shock with severe hyperinflammation (median 8), especially with low IgM (median 8) and toxin-mediated syndromes (median 9); for post-cardiac surgery sepsis with immune suppression (median 7) but not clearly for hyperinflammation (uncertain, median 6); for multidrug-resistant pathogen sepsis with immune suppression (median 7-8); and in haematologic malignancies, as prophylaxis or treatment primarily when immunoglobulins are low (medians 7-8), with prophylaxis otherwise uncertain. Across all phenotypes, IgM-enriched preparations were consistently preferred (median 8-9). Available evidence suggests no major safety signals, although adverse events are likely to be underreported. Overall, the consensus supports personalised, phenotype-oriented consideration of IgM-enriched IVIg as an adjunctive strategy to optimise standard care, while underscoring persistent evidence gaps and the need for adequately powered trials to define timing, dosing, and responsive subgroups.

The prolonged neuroparalytic effect of botulinum neurotoxin type A (BoNT/A) is largely attributable to the high intracellular stability of its light chain (LC), highlighting the need for strategies to promote LC clearance. Although deubiquitinating enzymes (DUBs) regulate LC turnover through both proteasomal and non-proteasomal pathways, the latter remains poorly understood. Here, we investigated the role of the DUB USP9X in regulating LC stability. Using a stable LC-overexpressing cell model, we found that pharmacological inhibition of USP9X with Degrasyn markedly promoted LC clearance. Transcriptomic analysis following lentivirus-mediated USP9X knockdown revealed enrichment of autophagy-related pathways, accompanied by suppression of PI3K-AKT signaling. Mechanistically, USP9X knockdown did not affect total mTOR protein levels but suppressed AKT-mTOR signaling, as shown by reduced phosphorylation of AKT and mTOR. This was associated with decreased ULK1 phosphorylation at the inhibitory site Ser757, increased phosphorylation at the active site Ser555, elevated p-AMPK levels, increased LC3-II/LC3-I ratio, and reduced p62 levels. All of those pointed to enhanced autophagic activity in response to USP9X inhibition. In BoNT/A-intoxicated rats, chloroquine-mediated autophagy inhibition delayed recovery from paralysis, whereas rapamycin-induced autophagy activation accelerated muscle strength recovery. Consistently, systemic administration of Degrasyn promoted functional recovery. Collectively, these findings indicate that USP9X inhibition promotes intracellular LC clearance, likely through an autophagy-lysosome-dependent mechanism involving suppression of AKT-mTOR signaling and activation of AMPK-ULK1-related pathways. Our study identifies USP9X as a host negative regulator of LC clearance and a potential therapeutic target for accelerating recovery following botulinum toxin intoxication.

Echovirus 19 (E19) is a rarely reported non-polio enterovirus (NPEV), although it has been associated with a spectrum of severe syndromes, including encephalitis, meningitis, and multisystem hemorrhagic disease in infants. Moreover, Echovirus 19 has been detected in patients with acute flaccid paralysis (AFP), suggesting a potential association between viral infection and CNS damage. Here we describe molecular epidemiology of Echovirus 19 in Russia and Central Asia from 1999 to 2024 using data on poliomyelitis surveillance. Echovirus 19 accounted for no more than 4% of annual NPEV isolates, however, in 2022 abundance rate dramatically increased making this virus the most frequently isolated NPEV from AFP cases (14.3% of NPEV-positive AFP cases). Phylogenetic analysis revealed that contemporary (2020-2025) regional isolates formed a distinct, well-supported clade sharing a common ancestor with Echovirus 19 variants isolated from AFP cases in Iran and Pakistan in 2013. Notably, these recent isolates lack sustained country-specific sublineages indicating ongoing cross-border circulation. A novel A83S mutation in the BC-loop of the VP1 antigenic region was identified in 75% isolates collected in 2022-2025. The emergence of this variant highlights the need for continued surveillance and further investigation into the Echovirus 19 possible role in neurological diseases.