Nerve Agents Research Articles

Efficacy of the oxime HI-6 dimethanesulphonate in the treatment of guinea-pigs exposed to the nerve agents GB and GD

The bispyridinium oxime HI-6 DMS is in development as an improved therapy for the treatment of patients exposed to organophosphorus nerve agents. The aim of the work described in this paper was to provide non-clinical data to support regulatory approval of HI-6 DMS, by demonstrating efficacy against an oxime-sensitive agent, GB and an oxime-resistant agent, GD. We investigated the dose-dependent protection afforded by therapy including atropine, avizafone and HI-6 DMS in guinea-pigs challenged with GB or GD. We also compared the efficacy of 30 mg.kg−1 of HI-6 DMS to an equimolar dose of the current in-service oxime P2S and the dichloride salt of HI-6 (HI-6 Cl2). In the treatment of GB or GD poisoning there was no significant difference between the salt forms. The most effective dose of HI-6 DMS in preventing lethality following challenge with GB was 100 mg.kg−1; though protection ratios of at least 25 were obtained at 10 mg.kg−1. Protection against GD was lower, and there was no significant increase in effectiveness of HI-6 DMS doses of 30 or 100 mg.kg−1. For GD, the outcome was improved by the addition of pyridostigmine pre-treatment. These data demonstrate the benefits of HI-6 DMS as a component of nerve agent therapy. © Crown copyright (2023), Dstl.

Rheologically improved microemulsion for deactivation of simulants of blister and nerve agents

The efficiency of decontamination of blister and nerve agents was studied using the example of nucleophilic decomposition of paraoxone (O,O-diethyl-O-4-nitrophenyl phosphate) and oxidation of methylphenyl sulfide. Hydrogen peroxide solutions in an oil-in-water microemulsion containing synthetic nanoclay Laponite EP and polyvinylpyrrolidone polymer were studied as reactive decontamination systems. The base of the microemulsion consisted of an aqueous phase, a codetergent (isopropanol), oil (hexane), with a variation of detergent (cetylpyridinium chloride, sodium dodecyl sulfate, and Triton X-100). It was shown that the solubility of paraoxone and methylphenyl sulfide in the studied microemulsions increases by an average of 100 times or more compared to the solubility in water, and the substrate binding constants are 2–3 times higher than the binding constants in similar microemulsion media. It was found that the presence of nanoclay in the microemulsion provides a catalytic effect, i.e. an increase in the rate of decomposition of paraoxone and methylphenyl sulfide by at least 2 times. In addition, nanoclay thickens the microemulsion and, together with the polymer, increases the viscosity of the reaction medium. The determined kinetic parameters of decontamination and solubility of substrates allow us to conclude that the use of the investigated microemulsion system provides an acceleration of nucleophilic substitution and oxidation reactions by 150–350 times compared to the reaction rate in water.

Chemistry, Metabolism and Neurotoxicity of Organophosphorus Insecticides: A Review

Organophosphorus compounds (OPs) are phosphoric acid derivatives represented by the formula (R2XP=O/S), R as organic groups; however, they need not contain a direct carbon-phosphorus bond. The organophosphorus compounds can be categorized into three classes, viz., organophosphates, carbamates nerve agents. The OPs having application as insecticides are generally phosphorothioates (i.e., containing P=S bond). These sulfur analogs are first bioactivated (in vivo) and converted to oxygen analogs responsible for exerting toxic action. These organophosphorus compounds are esters, fluorides, anhydrides, and amides of phosphoric, phosphorothioate, and phosphorodithioic acids. The toxicity of OPs is related to their molecular structure, metabolism in the targeted organisms, concentration, mode of decomposition, application, ingestion in organisms, etc. Exposure to OPs leads to the appearance of neurological symptoms followed by acute poisoning by targeting the target primarily, acetylcholine (AChE). However, secondary targets and other harmful effects besides nerve system problems are also reported. Organophosphates poison insects and other animals, including birds, amphibians, and mammals. These chemicals can have neural effects (Neurotoxicity), non-neuronal effects, or acute toxicity, which may also result in fatality. Their uncontrollable widespread became a significant threat to the environment; thus, corrective measures have been essential to save living beings and the environment from further damage.

Current Perspectives onthe Management ofPatients Poisoned With Novichok: A Scoping Review.

Nerve agents have emerged as a global threat since their discovery in the 1930s, posing severe risks due to their inhibition of acetylcholinesterase and the subsequent accumulation of acetylcholine in nerve synapses. Despite the enforcement of the Chemical Weapon Convention to control chemical weapons, including nerve agents, recent events, such as the Novichok attacks on Sergei Skripal and Alexei Navalny, have highlighted the persistent threat. Novichok, a distinct class of nerve agents, raises specific concerns regarding its management due to limited understanding. This article aims to comprehensively analyze existing literature. A scoping review was employed to comprehensively assess the current state of knowledge on managing patients poisoned with Novichok. Following the PRISMA-ScR guidelines, relevant literature was identified in peer-reviewed journals covering symptoms, diagnosis, treatment, decontamination, and long-term effects. Searches were conducted on February 1, 2023, across four electronic databases (PubMed, EMBASE, MEDLINE, and Web of Science) using "Novichok" as a keyword. No restrictions were applied, and additional studies were sought from the references of identified papers. Eligible papers included discussions on Novichok or its specific properties impacting management, regardless of study type, language, or publication date, while those unrelated to the study's conceptual framework were excluded. A total of 170 records were identified from the initial database search, with 86 studies screened after removing duplicates. Among these, 28 publications met the eligibility criteria and were included in the analysis. An additional relevant study was identified from the citation lists of included studies, bringing the total to 29. The review encompasses studies published from 2018 onwards, indicating the growing interest in this topic. While most studies are reviews on Novichok or nerve agents in general, several theoretical and experimental investigations were also found. This review highlights the significant uncertainties and knowledge gaps surrounding the management of patients poisoned with Novichok. While some aspects align with other nerve agents, limited research likely due to safety and ethical challenges leads to assumptions and uncertainties in patient care. The review identifies areas with ongoing research, such as decontamination and biomarker recognition, while other aspects remain understudied. The possible inefficacy of current treatment options and the need for further research on oximes, bioscavengers, and long-term effects emphasize the necessity for increased research to optimize patient outcomes. More studies are essential to clarify the actual threat and toxicity of Novichok. Moreover, raising awareness among medical staff is crucial for early diagnosis, prompt treatment, and safety. This review offers valuable insights into managing Novichok-poisoned patients and calls for increased research and awareness in this critical area.

Protective Activity of Novel Hydrophilic Synthetic Neurosteroids on Organophosphate Status Epilepticus-induced Chronic Epileptic Seizures, Non-Convulsive Discharges, High-Frequency Oscillations, and Electrographic Ictal Biomarkers.

Nerve agents and organophosphates (OP) are neurotoxic chemicals that induce acute seizures, status epilepticus (SE), and mortality. Long-term neurologic and neurodegenerative effects manifest months to years after OP exposure. Current benzodiazepine anticonvulsants are ineffective in preventing such long-term neurobehavioral and neuropathological changes. New and effective anticonvulsants are needed for OP intoxication, especially for mitigating the long-term sequelae after acute exposure. We developed neurosteroids as novel anticonvulsants and neuroprotectants in OP exposure models. In this study, we evaluated the long-term efficacy of novel synthetic neurosteroids in preventing the development of chronic epilepsy and hyperexcitable ictal events in a rat OP model of SE. Rats were exposed to the OP nerve agent surrogate diisopropylfluorophosphate (DFP), and the experimental groups were treated with the synthetic neurosteroid valaxanolone (VX) or lysaxanolone (LX) 40 minutes post-exposure in conjunction with midazolam. Video-electroencephalography was monitored for two months to assess spontaneous recurrent seizures (SRS), epileptiform discharges, interictal spikes, and high-frequency oscillations (HFOs). Within 60 days of DFP exposure, rats developed chronic epilepsy characterized by frequent SRS, epileptiform discharges, and HFOs. LX treatment was associated with a dose-dependent reduction of epilepsy occurrence and overall seizure burden with a significant decrease in SRS and epileptiform discharges. It also significantly reduced the occurrence of epileptic biomarkers of HFOs and interictal spikes, indicating potential disease-modifying activity. Similarly, the neurosteroid analog VX also significantly attenuated SRS, discharges, HFOs, and ictal events. These results demonstrate the long-term protective effects of synthetic neurosteroids in the OP-exposed post-SE model, indicating their disease-modifying potential to prevent epilepsy and ictal abnormalities. SIGNIFICANCE STATEMENT: The effects of nerve agents and organophosphate (OP) exposure are persistent, and survivors suffer from a number of devastating, chronic neurological dysfunctions. Currently, there is no specific therapy for preventing this disastrous impact of OP exposure. We propose synthetic neurosteroids that activate tonic inhibition provide viable options for preventing the long-term neurological effects of OP intoxication. The results from this study reveal the disease-modifying potential of two novel synthetic neurosteroids in preventing epileptogenesis and chronic epileptic seizures after OP-induced SE.

Adsorption behavior of VX nerve agent on X12Y12 nanocages: a density functional theory study

Adsorption behavior of VX nerve agent on X12Y12 nanocages: a density functional theory study

Thin colorimetric film array for rapid and selective detection of v-type nerve agent mimic in potentially contaminated areas

The expeditious detection and quantification of V-series nerve agents (VX) on potentially contaminated surfaces are crucial for the prevention of regional conflict incidents, acts of terrorism, or illicit activities. However, the low volatility and high toxicity of VX make these tasks challenging. Herein, we designed two novel colorimetric thin polymeric films to rapidly and sensitively detect demeton-S, a VX mimic, in contaminated areas. The polymeric films were specifically engineered to include a coordination site for Au (III) ions. Initially, these films were coordinated with Au (III), causing a discernible alteration in color due to enhancement in intramolecular charge transfer process. In the presence of demeton-S, the Au (III) ligands in the films are displaced with demeton-S, resulting in the restoration of the original color of the film, as the enhanced intramolecular charge transfer process is inhibited and thereby serving as an indicator of the presence of demeton-S. The polymeric films exhibit remarkable selectivity toward demeton-S compared to G-type nerve agents and other interference. The reusability of the polymeric films for demeton-S detection was achieved owing to the reversibility of the films during the alternative exposure of Au (III) and demeton-S. The polymeric films demonstrated their applicability for demeton-S detection and quantification in several contaminated areas, including different water, soil, and skin, rendering them highly suitable for on-site measurements.

Simple Chemical and Cholinesterase Methods for the Detection of Nerve Agents Using Optical Evaluation.

The extreme toxicity of nerve agents and the broad spectrum of their physical and chemical properties, enabling the use of these agents in a variety of tactical situations, is a continuing challenge in maintaining the knowledge and capability to detect them, as well as in finding new effective methods. Despite significant advances in the instrumentation of the analysis of nerve agents, relatively simple methods based on the evaluation of colour signals (absorption and fluorescence), in particular those using the cholinesterase reaction, continue to be of importance. This review provides a brief presentation of the current status of these simple methods, with an emphasis on military applications, and illustrates the high interest of the professional community in their further development. At the same time, it also contains some peculiarities (high reliability and durability, resistance to extreme climatic conditions, work in deployed means of protection, low purchase prices, economic availability especially in a state of war, etc.) that the authors believe research and development of simple methods and means for the detection of nerve agents should respect.

Dual-chromophore-functionalized silica mesoporous nanoparticles for fast visual detection of nerve agent simulant DCP

In the present work, a mesoporous silica nanoparticle (MSN@RB-NA)-based fluorescent nanomaterial was developed for fast and selective detection of a nerve agent simulant, diethylchlorophosphate (DCP), where two chromophores, naphthalimide and rhodamine B, were co-functionalized on the surface of MSN through their silane derivatives. The successful immobilization of the two fluorescent derivatives on MSN was demonstrated by a variety of characterization methods including FT-IR, BET-BJH, XPS, TEM, TGA, and solid-state 13C NMR, etc. A filter-paper sensor based on the dual-chromophore-functionalized MSN was tested for DCP vapor detection. Fluorescence measurements revealed that DCP can induce fluorescence reduction of naphthalimide unit at 476 nm and enhanced emission of rhodamine B at 575 nm of the paper sensor. Control experiments reveal that the piperazine group connected with the naphthalimide unit is responsible for DCP recognition, and a FRET process from naphthalimide to rhodamine B is believed the reason for the observed enhanced rhodamine emission. The paper sensor exhibits high sensitivity to DCP vapor with detection limit at 4.58 ppb, excellent selectivity over other 17 interferents, and fast response speed (< 1 s). Moreover, the present work provides an easy way for fast visual detection of DCP vapor.

High Capacity Adsorption and Degradation of a Nerve Agent Simulant and a Pesticide by a Nickel Pyrazolate Metal-Organic Framework.

The development of materials that enable the efficient removal of toxic compounds is important for the improvement of current protective materials or decontamination technologies. Current materials rely either on agent removal by adsorption or by effecting (catalytic) degradation. Ideally, both of these mechanisms are combined in a single material in order to target a more broad spectrum of toxic agents and to improve the performance of the materials. Recent attempts to combine materials with either adsorptive or catalytic properties into a composite material are promising, although the overall performance often suffers from competition for the agent between the adsorptive and catalytic domains in the composites. In this work, we propose that metal-organic frameworks (MOFs) could feature both adsorptive properties as well as catalytic properties in a single structural domain, thereby avoiding a reduction in the overall performance originating from competitive agent interactions. We showcase this concept using the MOF Ni3(BTP)2, which exhibits strong affinity and high capacity for the storage of a nerve agent simulant and a pesticide. Moreover, it is demonstrated that the adsorbed agents are efficiently degraded and that the nontoxic degradation products are rapidly expelled from the MOF pores. Its ability to catalyze the hydrolytic degradation of both organophosphate and organophosphorothioate compounds highlights another unique feature of this material. The presented concept illustrates the feasibility for developing materials that target a broader spectrum of agents via adsorption, catalysis, or both and by their broader reactivity toward different types of agents.

The effect of disease misclassification on the ability to detect a gene-environment interaction: implications of the specificity of case definitions for research on Gulf War illness

BackgroundSince 1997, research on Gulf War illness (GWI) has predominantly used 3 case definitions—the original Research definition, the CDC definition, and modifications of the Kansas definition—but they have not been compared against an objective standard.MethodsAll 3 case definitions were measured in the U.S. Military Health Survey by a computer-assisted telephone interview in a random sample (n = 6,497) of the 1991 deployed U.S. military force. The interview asked whether participants had heard nerve agent alarms during the conflict. A random subsample (n = 1,698) provided DNA for genotyping the PON1 Q192R polymorphism.ResultsThe CDC and the Modified Kansas definition without exclusions were satisfied by 41.7% and 39.0% of the deployed force, respectively, and were highly overlapping. The Research definition, a subset of the others, was satisfied by 13.6%. The majority of veterans meeting CDC and Modified Kansas endorsed fewer and milder symptoms; whereas, those meeting Research endorsed more symptoms of greater severity. The group meeting Research was more highly enriched with the PON1 192R risk allele than those meeting CDC and Modified Kansas, and Research had twice the power to detect the previously described gene-environment interaction between hearing alarms and RR homozygosity (adjusted relative excess risk due to interaction [aRERI] = 7.69; 95% CI 2.71–19.13) than CDC (aRERI = 2.92; 95% CI 0.96–6.38) or Modified Kansas without exclusions (aRERI = 3.84; 95% CI 1.30–8.52) or with exclusions (aRERI = 3.42; 95% CI 1.20–7.56). The lower power of CDC and Modified Kansas relative to Research was due to greater false-positive disease misclassification from lower diagnostic specificity.ConclusionsThe original Research case definition had greater statistical power to detect a genetic predisposition to GWI. Its greater specificity favors its use in hypothesis-driven research; whereas, the greater sensitivity of the others favor their use in clinical screening for application of future diagnostic biomarkers and clinical care.

Neurosteroids as Novel Anticonvulsants for Refractory Status Epilepticus and Medical Countermeasures for Nerve Agents: A 15-Year Journey to Bring Ganaxolone from Bench to Clinic.

This article describes recent advances in the use of neurosteroids as novel anticonvulsants for refractory status epilepticus (RSE) and as medical countermeasures (MCs) for organophosphates and chemical nerve agents (OPNAs). We highlight a comprehensive 15-year journey to bring the synthetic neurosteroid ganaxolone (GX) from bench to clinic. RSE, including when caused by nerve agents, is associated with devastating morbidity and permanent long-term neurologic dysfunction. Although recent approval of benzodiazepines such as intranasal midazolam and intranasal midazolam offers improved control of acute seizures, novel anticonvulsants are needed to suppress RSE and improve neurologic function outcomes. Currently, few anticonvulsant MCs exist for victims of OPNA exposure and RSE. Standard-of-care MCs for postexposure treatment include benzodiazepines, which do not effectively prevent or mitigate seizures resulting from nerve agent intoxication, leaving an urgent unmet medical need for new anticonvulsants for RSE. Recently, we pioneered neurosteroids as next-generation anticonvulsants that are superior to benzodiazepines for treatment of OPNA intoxication and RSE. Because GX and related neurosteroids that activate extrasynaptic GABA-A receptors rapidly control seizures and offer robust neuroprotection by reducing neuronal damage and neuroinflammation, they effectively improve neurologic outcomes after acute OPNA exposure and RSE. GX has been selected for advanced, Biomedical Advanced Research and Development Authority-supported phase 3 trials of RSE and nerve agent seizures. In addition, in mechanistic studies of neurosteroids at extrasynaptic receptors, we identified novel synthetic analogs with features that are superior to GX for current medical needs. Development of new MCs for RSE is complex, tedious, and uncertain due to scientific and regulatory challenges. Thus, further research will be critical to fill key gaps in evaluating RSE and anticonvulsants in vulnerable (pediatric and geriatric) populations and military persons. SIGNIFICANCE STATEMENT: Following organophosphate and nerve agent intoxication, refractory status epilepticus (RSE) occurs despite benzodiazepine treatment. RSE occurs in 40% of status epilepticus patients, with a 35% mortality rate and significant neurological morbidity in survivors. To treat RSE, neurosteroids are better anticonvulsants than benzodiazepines. Our pioneering use of neurosteroids for RSE and nerve agents led us to develop ganaxolone as a novel anticonvulsant and neuroprotectant with significantly improved neurological outcomes. This article describes the bench-to-bedside journey of bringing neurosteroid therapy to patients, with ganaxolone leading the way.

Graphene based Mo6 cluster hybrid for detecting simulant nerve agent DMMP

Nerve agents are highly toxic chemical compounds that pose significant threats to human health and safety. Detecting and identifying these agents quickly is crucial for preventing their exposure and providing timely treatment. This study proposes the use of nanohybrids comprising crystalline octahedral molybdenum iodide cluster material supported on graphene (Mo6@Graphene) as a novel sensing material for the detection of dimethyl methylphosphonate (DMMP), a simulant of sarin nerve agent. The electrical sensing performance towards different concentrations of DMMP and at room temperature demonstrated fast and sensitive responses to DMMP, with almost full recovery of the sensor baseline in a few minutes. The calibration curve obtained showed a linear relationship between the sensor response and DMMP concentration, enabling the estimation of the limit of detection (LOD) of 270 ppb. The robustness of the Mo6@Graphene nanomaterial towards DMMP was assessed using various techniques, including Raman spectroscopy, X-ray diffraction and photoelectron spectroscopy. The binding energies (BEs) between DMMP and the supported Mo6 clusters determined by DFT calculations revealed H-bonding interactions involved in the sensing mechanism. The excellent sensing performance of the Mo6@Graphene film, combined with its low power consumption and high stability, makes it a promising candidate for the detection of nerve agents and their simulants.

Discriminative detection of soman or VX exposure using europium chelated microparticle-based immunofluorescence microfluidic chip

The retrospective detection of organophosphorus nerve agents (OPNAs) exposure has been achieved by the off-site analysis of OPNA-human serum albumin (HSA) adducts using mass spectrometry-based detection approaches. However, few specific methods are accessible for on-site detection. To address this, a novel immunofluorescence microfluidic chip (IFMC) testing system combining europium chelated microparticle (EuCM) with self-driven microfluidic chip assay has been established to unambiguously determine soman (GD) and VX exposure within 20 min, respectively. The detection system was based on the principle of indirect competitive enzyme-linked immunosorbent assay. The specific monoclonal antibodies that respectively recognized the phosphonylated tyrosine 411 of GD-HSA and VX-HSA adducts were labeled by EuCM to capture corresponding adducts in the exposed samples. The phosphonylated peptides in the test line and goat-anti-rabbit antibody in the control line were utilized to bind the EuCM-labeled antibodies for signal exhibition. The developed IFMC chip could discriminatively detect exposed HSA adducts with high specificity, demonstrating a low limit of detection at exposure concentrations of 0.5 × 10−6 mol/L VX and 1.0 × 10−6 mol/L GD. The exposed serum samples can be qualitatively detected following an additional pretreatment procedure. This is a novel rapid detection system capable of discriminating GD and VX exposure, providing an alternative method for rapidly identifying OPNA exposure.

Differentiating Between V- and G-Series Nerve Agent and Simulant Vapours Using Fluorescent Film Responses.

In-field rapid and reliable identification of nerve agents is critical for the protection of Defence and National Security personnel as well as communities. Fluorescence-based detectors can be portable and provide rapid detection of chemical threats. However, most current approaches cannot differentiate between dilute vapors of nerve agent classes and are susceptible to false positives due to the presence of common acids. Here a fluorescence-based method is shown for rapid differentiation between the V-series and phosphonofluoridate G-series nerve agents and avoids false positives due to common acids. Differentiation is achieved through harnessing two different mechanisms. Detection of the V-series is achieved using photoinduced hole transfer whereby the fluorescence of the sensing material is quenched in the presence of the V-series agent. The G-series is detected using a turn-on mechanism in which a silylated excited state intramolecular proton transfer sensing molecule is selectively deprotected by hydrogen fluoride, which is typically found as a contaminant and/or breakdown product in G-series agents such as sarin. The strategy provided discrimination between classes, as the sensor for the G-series agent class is insensitive to the V-series agent, and vice versa, and neither responded to common acids.

Inhibition with simultaneous spontaneous reactivation and aging of acetylcholinesterase by organophosphorus compounds: Demeton-S-methyl as a model

The kinetic analysis of esterase inhibition by acylating compounds (organophosphorus, carbamates and sulfonylfluorides) sometimes cannot yield consistent results by fitting simple inhibition kinetic models to experimental data of complex systems. In this work kinetic data were obtained for demeton-S-methyl (DSM) with human acetylcholinesterase in two kinds of experiments: (a) time progressive inhibition with a range of concentrations, (b) progressive spontaneous reactivation starting with pre-inhibited enzyme. DSM is an organophosphorus compound used as pesticide and considered a model for studying the dermal exposure of nerve agents such as VX gas. A kinetic model equation was deduced with four different molecular phenomena occurring simultaneously: (1) inhibition; (2) spontaneous reactivation; (3) aging; and (4) ongoing inhibition (inhibition during the substrate reaction). A 3D fit of the model was applied to analyze the inhibition experimental data. The best-fitting model is compatible with a sensitive enzymatic entity. The second-order rate constant of inhibition (ki = 0.0422 μM−1 min−1), the spontaneous reactivation constant (ks = 0.0202 min−1) and the aging constant (kg = 0.0043 min−1) were simultaneously estimated. As an example for testing the model and approach, it was tested also in the presence of 5 % ethanol (conditions as previously used in the literature), the best fitting model is compatible with two apparent sensitive enzymatic entities (17 % and 83 %) and only one spontaneously reactivates and ages. The corresponding second-order rate constants of inhibition (ki = 0.0354 and 0.0119 μM−1 min−1) and the spontaneous reactivation and aging constants for the less sensitive component (kr = 0.0203 min−1 and kg = 0.0088 min−1) were estimated. The results were also consistent with a significant ongoing inhibition. These parameters were similar to those deduced in spontaneous reactivation experiments of the pre-inhibited samples with DSM in the absence or presence of ethanol. The two apparent components fit was interpreted by an equilibrium between ethanol-free and ethanol-bound enzyme. The consistency of results in inhibition and in spontaneous reactivation experiments was considered an internal validation of the methodology and the conclusions.

Sarin-Induced Neuroinflammation in Mouse Brain Is Attenuated by the Caspase Inhibitor Q-VD-OPh.

Organophosphates cause hyperstimulation of the central nervous system, leading to extended seizures, convulsions, and brain damage. Sarin is a highly toxic organophosphate nerve agent that has been employed in several terrorist attacks. The prolonged toxicity of sarin may be enhanced by the neuroinflammatory response initiated by the inflammasome, caspase involvement, and generation/release of proinflammatory cytokines. Since neurodegeneration and neuroinflammation are prevalent in sarin-exposed animals, we were interested in evaluating the capacity of quinolyl-valyl-O-methylaspartyl-[-2,6-difluorophenoxy]-methyl ketone (Q-VD-OPh), a pan caspase inhibitor to attenuate neuroinflammation following sarin exposure. To test this hypothesis, sarin-exposed C57BL/6 mice were treated with Q-VD-OPh or negative control quinolyl-valyl-O-methylglutamyl-[-2,6-difluorophenoxy]-methyl ketone, sacrificed at 2- and 14-day time points, followed by removal of the amygdala and hippocampus. A Bio-Rad 23-Plex cytokine analysis was completed on each tissue. The results suggest that exposure to sarin induced a dramatic increase in interleukin-1β and 6 other cytokines and a decrease in 2 of the 23 cytokines at 2 days in the amygdala compared with controls. Q-VD-OPh attenuated these changes at the 2-day time point. At 14 days, six of these cytokines were still significantly different from controls. Hippocampus was less affected at both time points. Diazepam, a neuroprotective drug against nerve agents, caused an increase in several cytokines but did not have a synergistic effect with Q-VD-OPh. Treatment of sarin exposure with apoptosis inhibitors appears to be a worthwhile approach for further testing as a comprehensive counteragent against organophosphate exposure. SIGNIFICANCE STATEMENT: A pan inhibitor of caspases (Q-VD-OPh) was proposed as a potential antidote for sarin-induced neuroinflammation by reducing the level of inflammation via inflammasome caspase inhibition. Q-VD-OPh added at 30 minutes post-sarin exposure attenuated the inflammatory response of a number of cytokines and chemokines in the amygdala and hippocampus, two brain regions sensitive to organophosphate exposure. Apoptotic marker reduction at 2 and 14 days further supports further testing of inhibitors of apoptosis as a means to lessen extended organophosphate toxicity in the brain.

24 Longitudinal Neuropsychological Functioning in Gulf War Veterans Exposed to Neurotoxicants and War-Related Trauma

Objective:Gulf War (GW) veterans were exposed to many neurotoxicants during the 1990-1991 Gulf War. Neurotoxicants included: chemical warfare such as sarin nerve gas, combustion byproducts from oil well fires and diesel fuels from tent heaters, pesticides, and prophylactic anti- nerve gas pyridostigmine bromide pills (PB); all of which have been associated with both cognitive and mood concerns. There are few longitudinal studies that have examined cognitive functioning regarding these toxicant exposures. In our longitudinal Fort Devens cohort, we found decrements over time in the area of verbal learning and memory but no differences in measures of nonverbal memory and executive function. To describe changes more accurately over time in this GW veteran cohort, we examined cognitive functioning in those with probable Post-Traumatic Stress Disorder (PTSD) versus those without.Participants and Methods:The FDC is the longest running cohort of GW veterans with initial baseline cognitive, mood, exposure and trauma assessments in 1997-1998 and follow-up evaluations in 2019-2022. FDC veterans (N=48) who completed both time points were the participants for this study. Veterans were categorized into dichotomous (yes/no) groups of PTSD classification. The PTSD checklist (PCL) was used to determine PTSD case status. Symptom ratings on the PCL were summed (range:17-85) and a cutoff score of 36 or higher was utilized to indicate probable PTSD. Neuropsychological measures of mood (POMS) and memory (Visual Reproductions from the Weschler Memory Scale-R; California Verbal Learning Test Second Edition; CVLT2) and executive function and language; (Delis-Kaplan Executive Function System- Color Word and Verbal fluency- Animals) were compared overtime using Paired T-tests.Results:The study sample (N=48) was 92% male and 96% reported active-duty status at the time of the GW. Mean current age was 58 years. All veterans reported exposure to at least one war-related toxicant. 48% met criteria for probable PTSD (N = 23) while 52% did not (n=25). No differences between groups were found in any of the POMS subscales, nor were differences seen in verbal memory, executive function, or language tasks. There were, however, significant differences in nonverbal memory in those with probable PTSD showing fewer details recalled during delay on the WMS-R Visual Reproductions (p&lt;0.05).Conclusions:In this longitudinal analysis, GW veterans with PTSD showed declines in nonverbal memory and consistent levels of function in all other tasks. Basic mood scales did not show decline; therefore, these results are not due to generalized changes in mood. All participants reported at least one neurotoxicant exposure and we did not have the power to examine the impact of the individual exposures, thus we cannot rule any contributing factors other than PTSD. This study highlights the importance of longitudinal follow up and continual documentation of GW veterans’ memory performance and their endorsement of mood symptoms overtime. Specifically, these findings reveal that future studies should examine the prolonged course of memory and mood symptomatology in GW veterans who have endorsed a traumatic experience.

Liquid-phase surface enhanced Raman spectroscopic detection of nerve agent motifs using gold nanostars

Organophosphates and their derivatives are deadly compounds owing to their ability to disrupt key regulators of biological activity and neuronal transmission. As such, typical uses for synthetic and natural organophosphates include nerve agents and toxic pesticides. Organophosphate pesticides alone are estimated to make up around 3–10 % of the international pesticide poisoning deaths, or approximately 5–15,000 deaths annually. Mechanistically, they irreversibly bind to acetylcholinesterase, a crucial enzyme dealing with motor control that has minimal structural differences across all walks of life explaining the universal toxicity of organophosphates. In this study, we present the fabrication of a nanoparticle-based detection method to detect two commonly used organophosphates: dimethyl methylphosphonate (DMMP) which is used as the starting material for many chemical warfare agent syntheses, and dimethyl chlorophosphate (DMCP) which is a potent acetylcholinesterase inhibitor and readily harmful to the body through dermal absorption or ingestion. We further demonstrate accurate and precise micromolar detection of these organophosphates with a simple portable Raman spectrometer, and additionally demonstrate the ability to easily detect and distinguish between both analytes in a heterogenous solution using this methodology. The nanoparticle-based detection relies on a method of Surface-enhanced Raman spectroscopy (SERS) in liquid phase. Throughout the duration of the experiment, liquid solutions and a portable Raman spectrometer were utilized to demonstrate efficacy of the detection method even with instruments and conditions that would be typically seen in the environment of organophosphate use, such as in a less technology developed warfare or rural setting. Through this detection method, we found a reliable fingerprint peak for DMMP (710.63 cm−1) with a limit of detection (LOD) of 9.11 mM and a limit of quantitation (LOQ) of 27.6 mM, with 1.5 ± 0.11 SERS. Similarly, for DMCP (755.97 cm−1), a LOD of 17.79 mM, a LOQ of 53.90 mM, and 1.3 ± 0.01 SERS enhancement were observed. Our theoretical studies predict possible SERS enhancement of >104, with significantly stronger signal compared with spherical nanoparticles. Time and pH studies show that our detection method is both fast and facile, without any required field modifications for baseline detection.

COF-supported zirconium oxyhydroxide as a versatile heterogeneous catalyst for Knoevenagel condensation and nerve agent hydrolysis

A composite of catalytic Lewis acidic zirconium oxyhydroxides (8 wt%) and a covalent organic framework (COF) was synthesized. X-ray diffraction and infrared (IR) spectroscopy reveal that COF's structure is preserved after loading with zirconium oxyhydroxides. Electron microscopy confirms a homogeneous distribution of nano- to sub-micron-sized zirconium clusters in the COF. 3D X-ray tomography captures the micron-sized channels connecting the well-dispersed zirconium clusters on the COF. The crystalline ZrOx(OH)y@COF's nanostructure was model-optimized via simulated annealing methods. Using 0.8 mol% of the catalyst yielded a turnover number of 100-120 and a turnover frequency of 160-360 h-1 for Knoevenagel condensation in aqueous medium. Additionally, 2.2 mol% of catalyst catalyzes the hydrolysis of dimethyl nitrophenyl phosphate, a simulant of nerve agent Soman, with a conversion rate of 37% in 180min. The hydrolytic detoxification of the live agent Soman is also achieved. Our study unveils COF-stabilized ZrOx(OH)y as a new class of zirconium-based Lewis+ Bronsted-acid catalysts.