Identifying emerging novel psychoactive substances (NPS), using liquid chromatography high-resolution tandem mass spectrometry (LC-HR-MS/MS), is often hindered by their frequent absence from mass spectral reference libraries. We developed a graph neural network (GNN) classification model to detect amphetamine derivatives, a subclass of NPS, based on their fragmentation patterns. The data set was generated by transforming high-resolution mass spectra from mass/charge (m/z) values into the chemical compositions of the fragments, from which fully connected graphs were constructed. Spectra were extracted from the NIST23 database, using only protonated HCD spectra to ensure consistency, and collision energy was included as a parameter to enhance spectral informativeness. The network architecture and hyperparameters were systematically optimized, including network layout and pooling layer type. To address the class imbalance in the data set, where amphetamine derivatives were outnumbered 100:1 by nonamphetamines, the loss function was adjusted to penalize false negatives 50 times more than false positives. Evaluation of model performance on a randomly selected 10% test subset yielded a recall of 0.86 and a precision of 0.11. The model successfully detected the spiked amphetamine derivative in a human plasma sample, achieving a 1% false positive rate and demonstrating its applicability to complex biological matrices.

Lipids, either as food components or as drug delivery formulations, are known to enhance the absorption of poorly water-soluble drugs in the gastrointestinal tract. Most research on the effects of different lipids and their lipolysis on dissolution and solubilization of poorly water-soluble drugs is conducted using liquid lipids, including liquid meals. In the present study, we examine the effects of a pure solid lipid, Tristearin (TS), compared to a liquid lipid, Triolein (TO), on the in vitro dissolution and the in vivo bioavailability of the BCS class II drug griseofulvin. Solid hydrophobic drugs may adsorb to solid lipids, and potentially decrease their bioavailability, however, we found that griseofulvin adsorbs negligibly to TS, which undergoes only partial lipolysis and therefore does not seem to contribute to griseofulvin dissolution nor to delay it and therefore TS is not expected to aid or impede griseofulvin bioavailability. Using the swine model to validate this prediction, we observed faster gastric emptying in the fasted state but similar overall bioavailabilities in the fasted and fed states, regardless of meal composition. This prompted an examination of the swine gastrointestinal contents for fed and fasted animals. Similar and high bile acids concentrations were found in fasted and fed states, suggesting a lack of difference in bile levels between the two states which may explain the observed lack of food effect. The use of the swine model to investigate lipid-enhanced bioavailability should be further explored, paying attention to the differences between different swine breeds. The results from in vitro experiments suggest that solid lipids are not expected to affect the bioavailability of BCS class II drugs.

Botulinum neurotoxin A (BoNT/A) is one of the most toxic substances known. The bacterium Clostridium botulinum produces it as a 150-kDa polypeptide with relatively low potency. Surprisingly, the endogenous protease that transforms this polypeptide into an activated, full-potency toxin, consisting of a 100-kDa heavy chain connected to a 50-kDa light chain (LC) by a disulfide bond, is still unknown. This study aimed to identify the BoNT/A-activating protease. We screened cation-exchange chromatography fractions of C. botulinum A culture supernatant for activity using a toxin-simulating substrate comprising the LC and the translocation domain (HN). Proteomic analysis of the active fraction identified α-clostripain as a candidate BoNT/A-activating protease. Recombinant α-clostripain cleaved the simulating substrate between the toxin LC and HN. However, incubation of recombinant α-clostripain with recombinant inactivated BoNT/A (rBoNT/Ai) resulted in non-specific digestion of the toxin. Since similar non-specific digestion was observed also by C. botulinum A culture supernatant, we hypothesized that the toxin should be protected by an accessory protein to prevent non-specific cleavage. Indeed, incubation of rBoNT/Ai with α-clostripain or culture supernatant in the presence of recombinant NTNH (non-toxic non-hemagglutinin) resulted in specific cleavage of the toxin into 100- and 50-kDa fragments. Subsequently, we evaluated the activation of rBoNT/A by α-clostripain in a mouse model. Cleavage of rBoNT/A by α-clostripain in the presence of NTNH resulted in a 77-fold increase in toxicity, corresponding to toxin activation. To the best of our knowledge, these results elucidate the mechanism of BoNT/A activation for the first time.

Air pollution interpolation is crucial for civil management. Various methods, including deterministic, geostatistical, and Machine Learning (ML)-based techniques have been utilized. Each method has limitations: deterministic methods do not adequately model environmental complexity, geostatistical methods struggle with small-scale areas, and ML depends heavily on data availability. On the other hand, air pollution simulators, which are driven by physicochemical dispersion models, capture intricate pollution dispersion patterns but are unsuitable for real-time interpolation.This study introduces an air pollution interpolation approach combining simulated air-dispersion patterns through an educated machine that includes machine learning and environmental modeling that considers boundaries, pollution sources, obstacles, wind dynamics, and topography. Specifically, we present a combined ML-based linear regression model designed to infer dense concentration maps from a sensor array using state-of-the-art simulation methods. We dub this the Ridiculously Simple data-driven air pollution Interpolation Method (RSIM). This method was evaluated on both synthetic and real-life-based simulation models.

Background

We investigated the safety and immunogenicity of the IIBR-100 (rVSV-SARS-CoV-2-S) vaccine, a recombinant VSV-ΔG-spike vaccine against SARS-CoV-2 virus.

Methods

In a phase 1/2, randomized, observer-blind, placebo-controlled study (ClinicalTrials.gov : NCT 04608305), healthy younger (18–55 years) and older (56–85 years) adults were recruited from eight clinical sites in Israel. In the phase 1 study, low (1 × 10⁵ PFU), mid (1 × 10⁶ PFU), and high doses (1 × 10⁷ PFU) of IIBR-100 were tested in a single-dose treatment regimen, with an additional booster dose for the low-dose group only. Based on the phase 1 study results, only mid- and high-dose prime-boost regimens alongside an additional top dose (1 × 10⁸ PFU) were tested in the phase 2 study. Participants, randomly assigned to either IIBR-100 or placebo, were followed for 12 months from the last vaccination for safety and immunogenicity outcomes.

Findings

No safety concerns were observed in the phase 1 study (N = 82); therefore, the study moved on to phase 2. In phase 2 (N = 762), for both age groups, the most common AEs included injection-site pain (20–64 %), fatigue (21–33 %), and headache (15–22 %). In the top-dose group, neutralizing and binding antibody titers peaked on Days 35 and 42, respectively; seroconversion rates reached maximal levels by Day 56 for neutralizing antibody and binding antibody (spike and RBD).

Interpretation

The IIBR-100 vaccine against SARS-CoV-2 in prime-boost regimens at 1 × 10⁷ PFU/mL and 1 × 10⁸ PFU/mL doses is safe, well tolerated, and immunogenic in healthy adults.

Clinical trial registry

This trial is registered at ClinicalTrials.gov (NCT 04608305) and in the trial registry of the Israeli Ministry of Health website.

Encapsulation of mRNA in lipid nanoparticles (LNPs) has established the LNP–mRNA platform as the strategy of choice for the rapid development of vaccines against both existing and emerging pathogens. However, despite its widespread global implementation during the COVID-19 pandemic, the immunological mechanisms underlying its efficacy remain incompletely understood. In this study, we investigated in a murine model, the early and robust innate immunity events elicited following immunization with an LNP–mRNA vaccine. Using mRNAs encoding two different proteins as well as a non-coding sequence, it is demonstrated that the mRNA component—rather than the LNP or the encoded antigen —is essential for inducing a potent innate immune response. This response is characterized by rapid activation of dendritic cells, recruitment of monocytes to draining lymph nodes, and systemic cytokine responses involving activation of various innate immune cell populations. Notably, these effects are all dependent on signaling through the type I interferon receptor (IFNAR). Importantly, we show that even a brief and transient inhibition of IFNAR signaling significantly enhances the ability of the LNP–mRNA vaccine to elicit adaptive immune responses, as evidenced by increased frequencies of antigen-specific CD8+ T cells and elevated titers of antigen-specific antibodies. Together, our findings reveal that the strong IFNAR-dependent innate response induced by mRNA can attenuate subsequent adaptive immunity. These insights should be considered in the future design and optimization of LNP–mRNA vaccine platforms.

Novichoks are highly toxic chemical warfare agents (CWAs), which gained significant attention after being used in the assassination attempt of Sergei Skripal (2018, Salisbury). Since then, only a limited number of experimental studies on Novichoks were conducted with focus on their physicochemical properties, toxicology and pharmacology. Their stability on various matrices has received even minor consideration. Herein, we present a comprehensive study on the fate of A232, a prominent Novichok agent, on a variety of previously unexamined environmental and urban matrices, using solid and solution state ³¹P NMR techniques. The matrices studied included limestone soil, desert soil, silicate sand, asphalt, fresh and old concrete and sawdust, all in dry and wet forms. A232 was found to be highly persistent on most matrices, with degradation being very slow or nonexistent. Once degradation occurred, its products stemmed from either a PN bond or a PF bond cleavage or double hydrolysis of both leaving groups, depending on the matrix. The results indicate that A232 poses significant, long-term threat to the environment across a diverse range of realistic scenarios. Importantly, A232 adsorbed on solid support was rapidly and completely hydrolyzed to a nontoxic product once treated with only a few equivalents of NaOH.

Sulfur mustard (SM) is a chemical warfare agent that causes severe cellular damage by alkylating DNA and proteins, with the skin, eyes, and respiratory system being the most affected. Acute respiratory injury is the primary cause of long-term lung damage following exposure. Despite its long history of use, SM's mechanism of action remains poorly understood, and no effective treatments exist. Proteomics enables systematic identification and characterization of proteins in specific tissues, yet no comprehensive proteomic studies have examined lungs exposed to SM. This study presents the first in vivo, time-resolved proteomic characterization of rat lungs at multiple time points after whole-body exposure to SM vapor (155 μg/L/10 min). Using mass spectrometry, differentially expressed proteins were categorized into four groups reflecting different phases of injury: naive (unexposed), 4 days post-exposure (early acute injury), 1 week post-exposure (advanced acute injury), and 4 weeks post-exposure (recovery). Bioinformatics integration of these data revealed stage-specific molecular pathways involved in inflammation, tissue remodeling, oxidative stress, and immune regulation. Importantly, several identified proteins overlapped with known markers of SM-induced damage in human samples, reinforcing their translational relevance. By linking protein groups to disease associations and drug databases, we proposed a dynamic therapeutic framework tailored to the evolving phases of injury. This included specific regimens and identified FDA-approved drugs that target key proteins. By focusing on proteomic profiling changes in response to SM exposure, this study provides novel insights into SM's mechanism of action and supports the development of targeted treatments customized to each stage of injury.

Background: Botulism is a rare but potentially fatal neuroparalytic illness caused by Clostridium botulinum neurotoxins (BoNTs). While adult cases usually result from foodborne exposure or wound infection, intestinal colonization is exceedingly uncommon. Diagnosis can be delayed by overlap with other neuromuscular syndromes, and confirmation requires specialized assays. Anti-GQ1b antibodies, classically associated with Miller–Fisher syndrome, have rarely been reported in confirmed botulism, raising questions about shared pathophysiology. Case Presentation: We describe an adult patient with acute dyspnea, xerostomia, and cranial neuropathies. No foodborne source was identified, but intestinal colonization of BoNT/B toxigenic Clostridium botulinum was confirmed by stool enrichment and mouse lethality bioassay. The patient improved promptly following heptavalent antitoxin. Unexpectedly, anti-GQ1b antibodies were detected during recovery, a finding typically linked to MFS rather than botulism. Discussion: This case illustrates the diagnostic challenges of sporadic cases of botulism, especially when respiratory compromise and autonomic dysfunction dominate the initial presentation. The autoantibodies finding raises the possibility of molecular mimicry, whereby toxin–ganglioside interactions expose neuronal epitopes and trigger an immune response. While causality cannot be proven, the overlap between botulism and GQ1bpositive neuropathies merits further investigation. Conclusions: Clinicians should maintain high suspicion for botulism in adults with acute dyspnea especially when associated with cranial neuropathies, even in the absence of foodborne exposure. Anti-ganglioside antibodies in this context should be interpreted with caution, as they do not exclude botulism but may highlight immunological overlap with autoimmune neuropathies.

Background

Neutrophils are critical first responders of the innate immune system, rapidly recruited to sites of infection or sterile injury. Upon activation by pathogen- or damage-associated molecular patterns, neutrophils initiate antimicrobial responses, including cytokine release, phagocytosis, and the formation of neutrophil extracellular traps (NETs). While NETosis plays a protective role, excessive NET formation can exacerbate inflammation and tissue damage. Pulmonary exposure to ricin, a potent toxin derived from Ricinus communis, results in acute lung injury characterized by neutrophil infiltration, cytokine production, vascular leakage, and pulmonary edema. This study investigated the contribution of NETosis to ricin-induced lung pathology and explored the therapeutic potential of targeting NETosis with a long acting recombinant DNase I (PRX-119) to attenuate lung injury.

Methods

CD1 outbreed mice were pulmonary exposed to ricin, and bronchoalveolar lavage fluid (BALF) and lung tissues were collected at various time points post-exposure. NETosis was assessed by immunofluorescence and Western blot analysis of markers, including peptidyl arginine deiminase 4 (PAD4), citrullinated histone H3 (citH3) and myeloperoxidase (MPO). Therapeutic intervention included administration of a NET-degrading DNase agent in combination with an anti-ricin antibody. Cell-free DNA levels, NETosis markers, neutrophil infiltration, lung histopathology, vascular permeability and the expression of pro- and anti-inflammatory mediators were evaluated. Weight loss and survival were also monitored and compared between anti-ricin monotherapy and combined anti-ricin and plant-produced human recombinant long acting (LA) DNase I (PRX-119), a novel NET degradation therapy.

Results

Ricin exposure led to elevated pulmonary levels of PAD4, citH3 and MPO, accompanied by extensive NET formation in both BALF and lung tissue. Mice receiving combined therapy with a newly developed DNase I - based agent (PRX-119) and an anti-ricin antibody treatment exhibited significantly improved survival and reduced weight loss compared to antibody monotherapy. The combined treatment not only significantly reduced NETosis markers, but also improved lung histopathology, reduced vascular leakage and pulmonary edema and altered the levels of proteins involved with pro- or anti-inflammatory effects, Dkk-1, CD93 and Periostin.

Conclusions

These findings demonstrate for the first time that NETosis plays a significant pathological role in ricin-induced lung injury. Moreover, they underscore the therapeutic potential of combining advanced NET-degrading agents, specifically PRX-119, an advanced DNAse I under development, with toxin-neutralizing antibodies as a promising strategy to reduce acute lung damage and enhance clinical outcomes.