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Many neurological disorders have complex etiologies that include noninheritable factors, collectively called the neural exposome. The National Institute of Neurological Disorders and Stroke is developing a new office with goals to advance our understanding of the multiple causes of neurological illness and to enable the development of more effective interventions.

Sarin is a highly toxic nerve agent that was developed for chemical warfare during World War II and is used in present conflicts. Immediate effects of acute sarin exposure are established; however, whether effects persist after initial signs have subsided is debated. The National Toxicology Program (NTP) conducted a systematic review to evaluate the evidence for long-term neurological effects following acute (

INTRODUCTION: Sarin (CASRN: 107-44-8) is a highly toxic organophosphorus nerve agent that was developed for chemical warfare during World War II and continues to be used in conflicts. Immediate effects of sarin exposure are well known, and although there are suggestions in the literature of neurological effects persisting after the initial signs have subsided, long-term neurological effects of acute exposure to sarin are not well characterized in humans. OBJECTIVE: The National Toxicology Program (NTP), on behalf of the National Institutes of Health (NIH) Countermeasures Against Chemical Threats program, conducted a systematic review to evaluate the evidence for long-term neurological effects in humans and nonhuman animals following acute exposure to sarin. (The terms "animal" and "animals" refer to nonhuman animals.). METHODS: A systematic review protocol was developed and utilized for this evaluation that followed the Office of Health Assessment and Translation approach for conducting literature-based health assessments. Any effect observed 24 hours after exposure (including days to years after exposure) was considered long term for this assessment. Because effects might vary based on time after exposure, the development of hazard conclusions was considered for three different time periods: initial (>24 hours-7 days after exposure), intermediate (8-364 days after exposure), and extended (greater than or equal to 1 year after exposure) periods. RESULTS AND EVIDENCE SYNTHESIS: The literature search and screening process identified 32 data sets within the 34 human studies and 47 data sets within the 51 animal studies (from 6,837 potentially relevant references) that met the objective and the inclusion criteria. Four main health effect categories of neurological response were identified as having sufficient data to reach hazard conclusions: (1) cholinesterase levels; (2) visual and ocular effects; (3) effects on learning, memory, and intelligence; and (4) morphology and histopathology in nervous system tissues. (This section of the abstract has been abridged.). DISCUSSION AND CONCLUSIONS: Hazard conclusions were considered for the four main health effect categories at all three time periods after exposure. The conclusions with the highest level of evidence for each time period were used to reach the overall conclusions. NTP concludes that acute sarin exposure is known to be a neurological hazard to humans in the initial time period of >24 hours-7 days after exposure based on suppression of cholinesterase. NTP concludes that acute sarin exposure is suspected to be a neurological hazard to humans in the intermediate time period of 8 days-1 year after exposure based on multiple effects, including suppression of cholinesterase, visual and ocular effects, and morphological and histological changes in nervous system tissues. NTP concludes that acute sarin exposure is suspected to be a neurological hazard to humans in the extended time period of greater than or equal to 1 year after exposure based on multiple effects, including effects on learning and memory and morphological and histopathological changes in nervous system tissues. DATA GAPS: This evaluation identified data gaps that contribute to lower confidence in the bodies of evidence for some endpoints and time periods after exposure. Future targeted research to assess the long-term neurological effects of sarin exposure should address areas with low confidence in the findings. Future research would benefit from the use of well-characterized human exposure data, use of exposed and appropriately matched control populations for neurological tests, and animal models that address the inconsistencies identified in this review using study design, conduct, and reporting practices to minimize bias. Given the hazard conclusions from this review, additional research on the four main health effect categories above may impact the confidence in the conclusions. Research may also be informative on a diverse range of neurological endpoints, identified in this report's appendices, for which there is inadequate evidence to determine whether there is an association with acute sarin exposure. Another area of research that the available data do not address is the effects of sarin on developing and aging brains. The current data are insufficient to assess if there are any susceptible populations.

Gray, WD, Jett, DM, Cocco, AR, Vanhoover, AC, Colborn, CE, Pantalos, GM, Stumbo, J, Quesada, PM, and Caruso, JF. Ergogenic and physiological outcomes derived from a novel skin cooling device. J Strength Cond Res 35(2): 391-403, 2021-Our study's purpose assessed a cooling headband's ergogenic and physiological impacts. Subjects (15 women and 13 men) completed six visits; the final 3 entailed rowing workouts with the following treatment conditions: no head cooling (NoHC), intermittent head cooling during exercise (HCex), and intermittent head cooling during exercise and post-exercise recovery (HCexpost). Data collection occurred at the following times (a) pre-exercise and post-warm-up, (b) between stages of up to eight 2-minute bouts, and (c) at 5, 10, 15, and 20 minutes post-exercise. In addition to distance rowed, thermal, cardiovascular, perceptual, and metabolic measurements were obtained. Results included a small yet significant intertreatment difference (HCex, HCexpostNoHC) for distance rowed. Our cardiovascular and metabolic indices exhibited sex and time differences but likely did not contribute to the ergogenic effect. Yet, left hand temperatures (LHT) exhibited significant 2-way and 3-way interactions that were the likely source of the ergogenic effect. Auditory canal temperature (AUDT) results suggest the head is sensitive to heat increases, yet LHT data show headband use evoked significantly greater temperature increases at the hand's palmar surface, indicative of heat transfer. We conclude, and our practical applications suggest, the headband's ergogenic effect was manifested by cold-induced vasodilation at the hand's palmar surface, rather than heat losses through the head.

Chemical warfare agents (CWAs) are highly toxic chemicals that have been used in military conflicts, beginning in World War I and continuing in more recent conflicts in Syria. Since the 2001 terrorist attacks in the United States, there has been an increased effort to better prepare for the potential deployment of chemical weapons and agents against civilian targets. Civilian threats include these CWAs, but also toxic industrial chemicals and other toxic chemicals deemed a threat to civilians. The National Institutes of Health (NIH) has developed and sustained a highly focused effort to support research on the development of safe and effective medical countermeasures for chemical threats. The NIH program has increased the knowledge base on chemical threats and had developed several new therapeutics that have transitioned to advanced development and some that have been approved for deployment. Research gaps and ongoing research at NIH are discussed in this chapter.

Phorate is a highly toxic agricultural pesticide currently in use throughout the world. Like many other organophosphorus (OP) pesticides, the primary mechanism of the acute toxicity of phorate is acetylcholinesterase (AChE) inhibition mediated by its bioactivated oxon metabolite. AChE reactivation is a critical aspect in the treatment of acute OP intoxication. Unfortunately, very little is currently known about the capacity of various oximes to rescue phorate oxon (PHO)-inhibited AChE. To help fill this knowledge gap, we evaluated the kinetics of inhibition, reactivation, and aging of PHO using recombinant AChE derived from three species (rat, guinea pig and human) commonly utilized to study the toxicity of OP compounds and five oximes that are currently fielded (or have been deemed extremely promising) as anti-OP therapies by various nations around the globe: 2-PAM Cl, HI-6 DMS, obidoxime Cl 2 , MMB4-DMS, and HLo7 DMS. The inhibition rate constants ( k i ) for PHO were calculated for AChE derived from each species and found to be low (i.e., 4.8 × 10 3 to 1.4 × 10 4 M − 1 min − 1 ) compared to many other OPs. Obidoxime Cl 2 was the most effective reactivator tested. The aging rate of PHO-inhibited AChE was very slow (limited aging was observed out to 48 h) for all three species. Conclusions: (1) Obidoxime Cl 2 was the most effective reactivator tested. (2) 2-PAM Cl, showed limited effectiveness in reactivating PHO-inhibited AChE, suggesting that it may have limited usefulness in the clinical management of acute PHO intoxication. (3) The therapeutic window for oxime administration following exposure to phorate (or PHO) is not limited by aging.

The increasing risk of a terrorist attack in the United States involving toxic chemicals has created new challenges for many programs across the federal government. Threat agents, including toxic i...

Intentional exposures to toxic chemicals can stem from terrorist attacks, such as the release of sarin in the Tokyo subway system in 1995, as well as from toxic industrial accidents that are much more common. Developing effective medical interventions is a critical component of the overall strategy to overcome the challenges of chemical emergencies. These challenges include the rapid and lethal mode of action of many toxic chemicals that require equally fast-acting therapies, the large number of chemicals that are considered threats, and the diverse demographics and vulnerabilities of those who may be affected. In addition, there may be long-term deleterious effects in survivors of a chemical exposure. Several U.S. federal agencies are invested in efforts to improve preparedness and response capabilities during and after chemical emergencies. For example, the National Institutes of Health (NIH) Countermeasures Against Chemical Threats (CounterACT) Program supports investigators who are developing therapeutics to reduce mortality and morbidity from chemical exposures. The program awards grants to individual laboratories and includes contract resource facilities and interagency agreements with Department of Defense laboratories. The range of high-quality research within the NIH CounterACT Program network is discussed.

Diisopropylfluorophosphate (DFP) is a potent acetylcholinesterase inhibitor commonly used in toxicological studies as an organophosphorus nerve agent surrogate. However, LD50values for DFP in the same species can differ widely even within the same laboratory, possibly due to the use of degraded DFP. The objectives here were to identify an efficient synthesis route for high purity DFP and assess the storage stability of both the in-house synthesized and commercial source of DFP at the manufacturer-recommended storage temperature of 4°C, as well as −10°C and −80°C. After 393 days, the commercial DFP stored at 4°C experienced significant degradation, while only minor degradation was observed at −10°C and none was observed at −80°C. DFP prepared using the newly identified synthesis route was significantly more stable, exhibiting only minor degradation at 4°C and none at −10°C or −80°C. The major degradation product was the monoacid derivative diisopropylphosphate, formed via hydrolysis of DFP. It was also found that storing DFP in glass containers may accelerate the degradation process by generating waterin situas hydrolytically generated hydrofluoric acid attacks the silica in the glass. Based on the results here, it is recommended that DFP be stored at or below −10°C, preferably in air-tight, nonglass containers.

Organophosphorus (OP) compounds, including pesticides and chemical warfare nerve agents (CWNA), are threats to the general population as possible weapons of terrorism or by accidental exposure whether through inadvertent release from manufacturing facilities or during transport. To mitigate the toxicities posed by these threats, a therapeutic regimen that is quick-acting and efficacious against a broad spectrum of OPs is highly desired. The work described herein sought to assess the protective ratio (PR), median effective doses (ED(50)), and therapeutic index (TI = oxime 24-h LD(50)/oxime ED(50)) of MMB4 DMS, HLö-7 DMS, and 2-PAM Cl against the OPs sarin (GB), VX, and phorate-oxon (PHO). All OPs are representative of the broader classes of G and V chemical warfare nerve agents and persistent pesticides. MMB4 DMS and HLö-7 DMS were previously identified as comparative efficacy leads warranting further evaluations. 2-PAM Cl is the U.S. FDA-approved standard-ofcare oxime therapy for OP intoxication. Briefly, PRs were determined in male guinea pigs by varying the subcutaneously (SC) delivered OP dose followed then by therapy with fixed levels of the oxime and atropine (0.4 mg/kg; administered intramuscularly [IM]). ED(50)s were determined using a similar approach except the OP dose was held constant at twice the median lethal dose (2 × LD(50)) while the oxime treatment levels were varied. The ED(50) information was then used to calculate the TI for each OP/oxime combination. Both MMB4 DMS and HLö-7 DMS provided significant protection, i.e., higher PR against GB, VX, and PHO when compared to atropine controls, but significance was not readily demonstrated across the board when compared against 2-PAM Cl. The ED(50) values of MMB4 DMS was consistently lower than that of the other oximes against all three OPs. Furthermore, based on those ED(50)s, the TI trend of the various oximes against both GB and VX was MMB4 DMS > HLö-7 DMS > 2-PAM Cl, while against PHO, MMB4 DMS > 2-PAM Cl > HLö-7 DMS.

Neurons that use acetylcholine in the developing brain make up an extremely important network that helps guide other systems during critical time periods of cell proliferation and differentiation. This important function and the variety of molecular targets for developmental neurotoxicants makes the central cholinergic system vulnerable to neurodevelopmental abnormalities that can result in functional deficits later in life. The alteration of brain development by toxicants, such as anticholinesterase pesticides, lead (Pb), nicotine, and others is supported by many studies that demonstrate concomitant changes in a variety of cholinergic targets, which provide clues to potential underlying mechanisms for their neurodevelopmental toxicity. This chapter discusses the role of the cholinergic system in brain development, special vulnerabilities related to brain region and developmental time periods, and human, animal, and mechanistic data on the neurodevelopmental toxicity of pesticides and other environmental chemicals.

The currently fielded pre-hospital therapeutic regimen for the treatment of organophosphorus (OP) poisoning in the United States (U.S.) is the administration of atropine in combination with an oxime antidote (2-PAM Cl) to reactivate inhibited acetylcholinesterase (AChE). Depending on clinical symptoms, an anticonvulsant, e.g., diazepam, may also be administered. Unfortunately, 2-PAM Cl does not offer sufficient protection across the range of OP threat agents, and there is some question as to whether it is the most effective oxime compound available. The objective of the present study is to identify an oxime antidote, under standardized and comparable conditions, that offers protection at the FDA approved human equivalent dose (HED) of 2-PAM Cl against tabun (GA), sarin (GB), soman (GD), cyclosarin (GF), and VX, and the pesticides paraoxon, chlorpyrifos oxon, and phorate oxon. Male Hartley guinea pigs were subcutaneously challenged with a lethal level of OP and treated at approximately 1min post challenge with atropine followed by equimolar oxime therapy (2-PAM Cl, HI-6 DMS, obidoxime Cl2, TMB-4, MMB4-DMS, HLö-7 DMS, MINA, and RS194B) or therapeutic-index (TI) level therapy (HI-6 DMS, MMB4-DMS, MINA, and RS194B). Clinical signs of toxicity were observed for 24h post challenge and blood cholinesterase [AChE and butyrylcholinesterase (BChE)] activity was analyzed utilizing a modified Ellman's method. When the oxime is standardized against the HED of 2-PAM Cl for guinea pigs, the evidence from clinical observations, lethality, quality of life (QOL) scores, and cholinesterase reactivation rates across all OPs indicated that MMB4 DMS and HLö-7 DMS were the two most consistently efficacious oximes.

The oral toxicity of phorate oxon (PHO), with emphasis on gender- and age-related effects, was characterized in the Sprague-Dawley rat. The oral LD50 (95% fiducial limits) for PHO in corn oil was 0.88 (0.79, 1.04) mg/kg in males and 0.55 (0.46, 0.63) mg/kg in females with a probit slope of 15. Females had higher baseline blood cholinesterase titers, but males were significantly more tolerant. Younger rats generally had lower absolute cholinesterase blood titers. However as PHO challenges increased, baseline-normalized cholinesterase inhibition was independent of age and gender. Butyrylcholinesterase (BChE) and especially acetylcholinesterase (AChE) in brains of younger females were affected more than that in either males or older females. In summary, while female rats, especially older females, had higher titers relative to males, female rats were more susceptible in terms of absolute cholinesterase inhibition and 24-hr lethality data, but the differences were not observed when titers were normalized to baseline levels.

Most hospital laboratories do not measure blood cyanide concentrations, and samples must be sent to reference laboratories. A simple method is needed for measuring cyanide in hospitals. The authors previously developed a method to quantify cyanide based on the high binding affinity of the vitamin B12 analog, cobinamide, for cyanide and a major spectral change observed for cyanide-bound cobinamide. This method is now validated in human blood, and the findings include a mean inter-assay accuracy of 99.1%, precision of 8.75% and a lower limit of quantification of 3.27 µM cyanide. The method was applied to blood samples from children treated with sodium nitroprusside and it yielded measurable results in 88 of 172 samples (51%), whereas the reference laboratory yielded results in only 19 samples (11%). In all 19 samples, the cobinamide-based method also yielded measurable results. The two methods showed reasonable agreement when analyzed by linear regression, but not when analyzed by a standard error of the estimate or paired t-test. Differences in results between the two methods may be because samples were assayed at different times on different sample types. The cobinamide-based method is applicable to human blood, and can be used in hospital laboratories and emergency rooms.

Anticholinesterases, such as organophosphorus pesticides and warfare nerve agents, present a significant health threat. Onset of symptoms after exposure can be rapid, requiring quick-acting, efficacious therapy to mitigate the effects. The goal of the current study was to identify the safest antidote with the highest therapeutic index (TI = oxime 24-hr LD50/oxime ED50) from a panel of four oximes deemed most efficacious in a previous study. The oximes tested were pralidoxime chloride (2-PAM Cl), MMB4 DMS, HLo-7 DMS, and obidoxime Cl2. The 24-hr median lethal dose (LD50) for the four by intramuscular (IM) injection and the median effective dose (ED50) were determined. In the ED50 study, male guinea pigs clipped of hair received 2x LD50 topical challenges of undiluted Russian VX (VR), VX, or phorate oxon (PHO) and, at the onset of cholinergic signs, IM therapy of atropine (0.4 mg/kg) and varying levels of oxime. Survival was assessed at 3 hr after onset clinical signs. The 3-hr 90th percentile dose (ED90) for each oxime was compared to the guinea pig pre-hospital human-equivalent dose of 2-PAM Cl, 149 µmol/kg. The TI was calculated for each OP/oxime combination. Against VR, MMB4 DMS had a higher TI than HLo-7 DMS, whereas 2-PAM Cl and obidoxime Cl2 were ineffective. Against VX, MMB4 DMS > HLo-7 DMS > 2-PAM Cl > obidoxime Cl2. Against PHO, all performed better than 2-PAM Cl. MMB4 DMS was the most effective oxime as it was the only oxime with ED90 < 149 µmol/kg against all three topical OPs tested.

Purpose: Aldicarb and methomyl are carbamate pesticides commonly implicated in human poisonings. The primary toxic mechanism of action for carbamate poisoning is cholinesterase (ChE) inhibition. As such, it is logical to assume that the currently accepted therapies for organophosphate poisoning (muscarinic antagonist atropine and the oxime acetylcholinesterase reactivator pralidoxime chloride [2-PAM Cl]) could afford therapeutic protection. However, oximes have been shown to be contraindicated for poisoning by some carbamates. Methods: A protective ratio study was conducted in guinea pigs to evaluate the efficacy of atropine and 2-PAM Cl. The ChE activity was determined in both the blood and the cerebral cortex. Results: Coadministration of atropine free base (0.4 mg/kg) and 2-PAM Cl (25.7 mg/kg) demonstrated protective ratios of 2 and 3 against aldicarb and methomyl, respectively, relative to saline. The data reported here show that this protection was primarily mediated by the action of atropine. The reactivator 2-PAM Cl had neither positive nor negative effects on survival. Both blood acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) activities were significantly reduced at 15 minutes postchallenge but gradually returned to normal within 24 hours. Analysis of cerebral cortex showed that BChE, but not AChE, activity was reduced in animals that succumbed prior to 24 hours after challenge. Conclusion: The results suggest that coadministration of atropine and 2-PAM Cl at the currently recommended human equivalent doses for use in the prehospital setting to treat organophosphorus nerve agent and pesticide poisoning would likely also be effective against aldicarb or methomyl poisoning.

Pesticides represent one of the largest classes of toxic chemicals produced, stored, and used in the United States and abroad. These chemicals are designed to be toxic and many, besides being toxic to the pests they are intended to control, are also toxic to nontarget species including humans. The article gives a brief review of their toxicity to humans with emphasis on their effects on the nervous system. Examples of case studies are included to illustrate their toxicity. A discussion of the possible contribution of occupational and other pesticide exposures to neurologic diseases and disorders is also included.

Publisher Summary The use of chemical warfare agents (CWAs) has historically been a primary concern of military personnel due to their devastating physical and psychological effects when employed on the battlefield. As a result of these new threats, agencies within the Department of Health and Human Services (DHHS) and other federal agencies have sustained a highly focused effort to assess and, if necessary, improve upon current emergency response capabilities in the event of an actual terrorist event. These efforts include both nonmedical countermeasures such as personal protective equipment for first responders who must enter a contaminated site, and medical capabilities such as safe and effective antidotes and diagnostic tools to reduce mortality and morbidity after a chemical attack or accident. In 2006, the US Congress appropriated funds to the National Institutes of Health (NIH) to implement the National Strategic Plan and Research Agenda for Medical Chemical Countermeasures. The goal of the CounterACT program is to develop safer and more effective therapeutics to treat victims exposed during a chemical incident. Complementing this goal, the CounterACT program also supports the research and development of novel diagnostic technologies, which could be utilized after a chemical incident to determine the proper course of medical intervention.

It has been suggested that pesticide exposure may be a contributing factor underlying the increased incidence of asthma in the United States and other industrialized nations. To test this hypothesis, airway hyperreactivity was measured in guinea pigs exposed to chlorpyrifos, a widely used organophosphate pesticide. Electrical stimulation of the vagus nerves caused frequency-dependent bronchoconstriction that was significantly potentiated in animals 24 h or 7 days after a single subcutaneous injection of either 390 mg/kg or 70 mg/kg of chlorpyrifos, respectively. Mechanisms by which chlorpyrifos may cause airway hyperreactivity include inhibition of acetylcholinesterase (AChE) or dysfunction of M3 muscarinic receptors on airway smooth muscle or of autoinhibitory M2 muscarinic receptors on parasympathetic nerves in the lung. AChE activity in the lung was significantly inhibited 24 h after treatment with 390 mg/kg of chlorpyrifos, but not 7 days after injection of 70 mg/kg of chlorpyrifos. Acute exposure to eserine (250 μg/ml) also significantly inhibited lung AChE but did not potentiate vagally induced bronchoconstriction. Neuronal M2 receptor function was tested using the M2 agonist pilocarpine, which inhibits vagally induced bronchoconstriction in control animals. In chlorpyrifos-treated animals, pilocarpine dose-response curves were shifted significantly to the right, demonstrating decreased responsiveness of neuronal M2 receptors. In contrast, chlorpyrifos treatment did not alter methacholine-induced bronchoconstriction, suggesting that chlorpyrifos does not alter M3 muscarinic receptor function on airway smooth muscle. These data demonstrate that organophosphate insecticides can cause airway hyperreactivity in the absence of AChE inhibition by decreasing neuronal M2 receptor function.

The National Institutes of Health has developed a comprehensive research program that includes research centers of excellence, individual research projects, small business projects, contracts, and interagency agreements to conduct basic, translational, and clinical research aimed at the discovery and/or identification of better medical countermeasures against chemical threat agents. Chemical threats include chemical warfare agents, toxic industrial and agricultural chemicals, and toxins and other chemicals that could be used intentionally as an act of terror or by large-scale accidents or natural disasters. The overarching goal of this research program is to enhance our medical response capabilities during an emergency. The program is named Countermeasures Against Chemical Threats (CounterACT). It supports translational research, applying ideas, insights, and discoveries generated through basic scientific inquiry to the treatment or prevention of mortality and morbidity caused by chemical threat agents. The categories of research supported under this program include creation and development of screening assays and animal models for therapy development, identification of candidate therapeutics, obtaining preliminary proof-of-principle data on the efficacy of candidate therapeutics, advanced efficacy and preclinical safety studies with appropriate animal models using Good Laboratory Practices (GLP), and clinical studies, including clinical trials with new drugs. Special consideration is given to research relevant to people who are particularly vulnerable, including the young, the elderly, and individuals with pre-existing medical conditions.

This study tested the hypothesis that the inhibition of acetylcholinesterase is greater when the insecticide chlorpyrifos (CPF) is in the presence of several polycyclic aromatic hydrocarbons (PAHs) found in house dust. CPF-oxon (CPFO) inhibition curves of purified AChE (electric eel) were generated in the presence or absence of different concentrations of the PAHs pyrene, benzo(a)pyrene, anthracene, and fluoranthene. Without CPF-oxon, all four PAHs themselves inhibited AChE activity with IC50 values in the range 8.2-17 microM. The IC50 for benzo(a)pyrene with human recombinant AChE was 1.5 microM. When AChE was incubated with CPF-oxon together with the PAHs, the inhibitory effect on AChE was additive. This was exemplified by large (60-80%) and significant (P

We examined spatial learning in the Morris water maze after daily acute bilateral micro-injection of 13.9 ng sodium acetate (NaAc) or 37.9 ng lead acetate (PbAc) in 1 microliter volumes into the dorsal hippocampus of normal adult rats. After six days of injections and water maze training, rats injected with NaAc were able to find a hidden platform in 8.3 s, and those injected with PbAc were significantly slower (15.2 s; p0.02). In a second experiment, rats were trained to find a hidden platform before injections began and then tested in order to determine if intrahippocampal injections of Pb affected the recall of a previously learned task. The escape latency on the first day after injections began was increased slightly when compared to the last day of training before injections, however the NaAc and PbAc groups were not significantly different over three days of injections. Both treatment groups performed as well as they did before injections began by the second day of injections. These results suggest that the direct injection of Pb into the hippocampus impairs the acquisition but not the recall of the spatial learning task in the Morris water maze.

The neurobehavioral toxicity of developmental exposure to lead (Pb) was investigated by conducting tests of spatial learning in the Morris water maze. Female Long–Evans rats were exposed to 0 or 250 ppm Pb acetate in the diet beginning 10 days prior to breeding and continued throughout gestation and lactation. Pups were weaned onto the same diets as the dams at postnatal day 20 (PN20). Increased levels of Pb were detected in the hippocampus of the 250 ppm Pb acetate group relative to controls. The highest concentration of Pb measured in the hippocampus was at PN21 with decreasing levels at older ages. In the Morris Water Maze, a statistically significant ( p < 0.03; female rats) or near significant ( p < 0.07; male rats) increase in the time required to find the hidden platform (escape latency) was observed when Pb-treated rats were tested in a reference memory paradigm. This effect was only observed when rats were tested at PN21 and not at older ages. No signif- icant effects of developmental Pb exposure were measured when rats were tested in a working memory paradigm of the Morris water maze at any age. These initial studies indicate an impairment of performance in the swim task in PN21 rats exposed to Pb during development. The age-dependent effect of Pb in this learning paradigm is consistant with previous studies in experimental animals and with the observation that children are more susceptible to Pb-induced cognitive deficits than adults. The Morris water maze may be useful for studying the effects of Pb on learning and memory, and their neurochemical basis.

The Chemical Events Working Group of the Global Health Security Initiative has developed a flexible screening tool for chemicals that present a risk when accidentally or deliberately released into the atmosphere. The tool is generic, semi-quantitative, independent of site, situation and scenario, encompasses all chemical hazards (toxicity, flammability and reactivity), and can be easily and quickly implemented by non-subject matter experts using freely available, authoritative information. Public health practitioners and planners can use the screening tool to assist them in directing their activities in each of the five stages of the disaster management cycle.

The National Institutes of Health (NIH) supports research about and the development of better therapies for treating exposure to toxic chemicals that could be used in a terrorist attack or released during an industrial accident. A review of recent research published by NIH investigators working in this field indicates that scientific advances in this area also have implications for reducing the burden of other neurological diseases and disorders. Some key examples discussed include studies on the development of therapeutic drugs to treat seizures and the neuropathology caused by chemical nerve agents, which may help find better cures for epilepsy, stroke, and neurodegenerative diseases.

The effects of the organophosphorus anticholinesterase paraoxon on the binding of radioactive ligands to the M3 subtype of the muscarinic receptor and receptor-coupled synthesis of second messengers in intact rat submaxillary gland (SMG) cells were investigated. The binding of [3H]quinuclidinyl benzilate ([3H]QNB) was most sensitive to atropine and the M3-specific antagonist 4-DAMP followed by pirenzepine and least sensitive to the cardioselective M2 antagonist AFDX116. This, and the binding characteristics of [3H]4-DAMP, confirmed that the muscarinic receptors in this preparation are of the M3 subtype. Activation of these muscarinic receptors by carbamylcholine (CBC) produced both stimulation of phosphoinositide (PI) hydrolysis and inhibition of cAMP synthesis, suggesting that this receptor subtype couples to both effector systems. Paraoxon (100 microM) reduced Bmax of [3H]4-DAMP binding from 27 +/- 4 to 13 +/- 3 fmol/mg protein with nonsignificant change in affinity, suggesting noncompetitive inhibition of binding by paraoxon. Like the agonist CBC, paraoxon inhibited the forskolin-induced cAMP formation in SMG cells with an EC50 of 200 nM, but paraoxon was greater than 500 fold more potent than CBC. However, while the inhibition by CBC was counteracted by 2 microM atropine, that by paraoxon was unaffected by up to 100 microM atropine. It suggested that this effect of paraoxon was not via binding to the muscarinic receptor. Paraoxon did not affect beta-adrenoreceptor function in the preparation, since it did not affect the 10 microM isoproterenol-induced cAMP synthesis, which was inhibited totally by 10 microM propranolol and partially by CBC. Paraoxon had a small but significant effect on CBC-stimulated PI metabolism in the SMG cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Chemicals pose a threat to the United States military and civilian populations because they are relatively inexpensive and easy to obtain, and have the potential to cause mass casualties if released by accident, natural disaster, or by a deliberate act. Chemical threat agents include nerve agents, vesicating or blister agents, pulmonary agents, and blood agents or metabolic poisons. The traditional chemical warfare agents include the organophosphorus nerve agents such as sarin and VX, and the mustard vesicating agents. These remain in several stockpiles around the world. Toxic industrial chemicals (TICs) manufactured, stored, and transported across the nation are not regulated as strictly as traditional warfare agents, and many are stored in large amounts and are readily available. Unfortunately and unlike biological and radiation threats, there are many examples of human suffering and death after deliberate releases of chemical welfare agents (CWAs) (sarin, sulfur mustard) and TICs (cyanide, chlorine). Requirements for an effective response to a civilian chemical event include (i) post exposure treatments that are effective within an often short therapeutic time window, (ii) drugs and devices that can be used by medical personnel at the scene of the event or in a pre hospital setting to treat many victims, (iii) drugs and devices that are appropriate for a diverse population including pediatric and elderly victims, and individuals with preexisting medical conditions, and (iv) rapid and effective diagnostic technologies to determine the chemical agent and pathophysiology. The National Institutes of Health (NIH) has developed a research program called Countermeasures Against Chemical Threats (CounterACT) to develop new and improved diagnostic technologies and therapies. This program has identified several new opportunities to develop better medical intervention strategies that treat the most severe symptoms, and broad spectrum drugs that target common physiological mechanisms of injury that could be used to treat victims exposed to many different kinds of chemicals. CounterACT and other programs at Department of Health and Humman Services (DHHS) charged with this important goal are essential to national security and will require strong support to meet the challenges ahead. It will require a commitment that is consistent with the cost and time required for drug discovery and development, and should support research at the basic, translational, and clinical levels. Keywords: chemical threats; medical countermeasures; research; Counter ACT; NIH; sarin; VX; cyanide; sulfur mustard; chlorine; phosgene

Presented below is a brief description of research supported by the National Institutes of Health (NIH) on cholinesterases that was discussed at the IXth International Meeting on Cholinesterases in Suzhou, China. It is a partial description of the research conducted by researchers at academic and other institutions supported by the NIH, and by some of the researchers in NIH intramural laboratories. It does not represent a comprehensive survey of all research supported by the NIH related to cholinesterases, but rather a brief discussion of some of the studies discussed at the IXth International Meeting on Cholinesterases. The article describes exciting basic, translational and clinical research on therapies for neurological and other diseases. In addition, cholinesterases that may treat substance abuse are discussed, and pesticide and chemical warfare agents that inhibit cholinesterases are highlighted as part of the NIH portfolio. It is the intent of this article to share with the international community some of the research being supported by the NIH on cholinesterases that complements many of the studies being conducted elsewhere. The information was obtained only from published articles or from abstracts available to the public within the NIH CRISP database (http://crisp.cit.nih.gov/).

Publisher Summary The chapter discusses the role of acetylcholinesterase (AChE) inhibition in the toxicity of organophosphorus (OP) and carbamate (CM) pesticides, with particular emphasis on lower doses that are sublethal, but induce changes in cholinergic receptors, either by indirect modulation of ACh levels via AChE inhibition or by direct interaction with the receptor. Case studies are presented that suggest that some of the toxicities associated with OP insecticides may not involve AChE or cholinergic receptors. Inhibition of AChE and subsequent cholinergic dysfunction represent a common mechanism of toxicity after acute exposure to high levels of OP and CM pesticides. However, the range of toxicities among different compounds is broad, and the potency of a compound for the inhibition of AChE is not a good predictor of its toxicity. Furthermore, toxicities associated with repeated exposures to lower levels of OPs do not correlate well with AChE inhibition. These observations suggest that significant interaction of OPs and CMs with other critical molecules within the cell may be important in explaining some of the toxicity of these compounds not attributable to AChE inhibition. Examples are presented with AChE null mice, direct interactions with muscarinic receptors and other molecules, impairment of cognitive development in weaning rats, OP-induced airway hyperreactivity mediated by M2 receptors, and direct effects on axonal growth in cell culture. The case studies provide strong evidence that noncholinesterase mechanisms are involved in the toxicity of OPs, and they have identified important gaps in the understanding of the mechanism of action of these important classes of insecticides.

Evidence that children are widely exposed to organophosphorus pesticides (OPs) and that OPs cause developmental neurotoxicity in animal models raises significant concerns about the risks these compounds pose to the developing human nervous system. Critical to assessing this risk is identifying specific neurodevelopmental events targeted by OPs. Observations that OPs alter brain morphometry in developing rodents and inhibit neurite outgrowth in neural cell lines suggest that OPs perturb neuronal morphogenesis. However, an important question yet to be answered is whether the dysmorphogenic effect of OPs reflects perturbation of axonal or dendritic growth. We addressed this question by quantifying axonal and dendritic growth in primary cultures of embryonic rat sympathetic neurons derived from superior cervical ganglia (SCG) following in vitro exposure to chlorpyrifos (CPF) or its metabolites CPF-oxon (CPFO) and trichloropyridinol (TCP). Axon outgrowth was significantly inhibited by CPF or CPFO, but not TCP, at concentrations > or =0.001 microM or 0.001 nM, respectively. In contrast, all three compounds enhanced BMP-induced dendritic growth. Acetylcholinesterase was inhibited only by the highest concentrations of CPF (> or =1 microM) and CPFO (> or =1 nM); TCP had no effect on this parameter. In summary, these compounds perturb neuronal morphogenesis via opposing effects on axonal and dendritic growth, and both effects are independent of acetylcholinesterase inhibition. These findings have important implications for current risk assessment practices of using acetylcholinesterase inhibition as a biomarker of OP neurotoxicity and suggest that OPs may disrupt normal patterns of neuronal connectivity in the developing nervous system.