"Isocyanates and isocyanides - life-threatening toxins or essential compounds?"

Isocyanides and isocyanates are some of the most reactive compounds in organic chemistry, making them perceived as compounds with high potential for use in both the laboratory and industry. With their high reactivity also comes several disadvantages, most notably their potentially high toxicity. The following article is a collection of information on the toxic effects of the isocyanide group on the human body and the environment. Information on the mechanism of how these harmful substances affect living tissues and the environment, worldwide information on how to protect against these chemicals, current regulations, and exposure limits for specific countries is compiled. The latest research on the application uses of isocyanates and isocyanides is also outlined, as well as the latest safer and greener methods and techniques to work with these compounds. Additionally, the presented article can serve as a brief guide to the organic toxicity of a group of isocyanates and isocyanates.

4,4'-Methylene diphenyl diisocyanate exposure induces expression of alternatively activated macrophage-associated markers and chemokines partially through Krüppel-like factor 4 mediated signaling in macrophages.

Occupational exposure to the most widely used monomeric diisocyanate (dNCO), 4,4'-methylene diphenyl diisocyanate (MDI), may lead to the development of occupational asthma (OA). Alveolar macrophages with alternatively activated (M2) phenotype have been implicated in allergic airway responses and the pathogenesis of asthma. Recent in vivo studies demonstrate that M2 macrophage-associated markers and chemokines are induced by MDI-exposure, however, the underlying molecular mechanism(s) by which this proceeds is unclear.Following MDI exposure (in vivo and in vitro) M2 macrophage-associated transcription factors (TFs), markers, and chemokines were determined by RT-qPCR, western blots, and ELISA.Expression of M2 macrophage-associated TFs and markers including Klf4/KLF4, Cd206/CD206, Tgm2/TGM2, Ccl17/CCL17, Ccl22/CCL22, and CCL24 were induced by MDI/MDI-GSH exposure in bronchoalveolar lavage cells (BALCs)/THP-1 macrophages. The expression of CD206, TGM2, CCL17, CCL22, and CCL24 are upregulated by 3.83-, 7.69-, 6.22-, 6.08-, and 1.90-fold in KLF4-overexpressed macrophages, respectively. Endogenous CD206 and TGM2 were downregulated by 1.65-5.17-fold, and 1.15-1.78-fold, whereas CCL17, CCL22, and CCL24 remain unchanged in KLF4-knockdown macrophages. Finally, MDI-glutathione (GSH) conjugate-treated macrophages show increased chemotactic ability to T-cells and eosinophils, which may be attenuated by KLF4 knockdown.Our data suggest that MDI exposure may induce M2 macrophage-associated markers partially through induction of KLF4.

Reconstruction of exposure to methylene diphenyl-4,4'-diisocyanate (MDI) aerosol using computational fluid dynamics, physiologically based toxicokinetics and statistical modeling.

OBJECTIVES: This study employed computational fluid dynamics (CFD), physiologically based toxicokinetics (PBTK), and statistical modeling to reconstruct exposure to methylene diphenyl-4,4'-diisocyanate (MDI) aerosol. By utilizing a validated CFD model, human respiratory deposition of MDI aerosol in different workload conditions was investigated, while a PBTK model was calibrated using experimental rat data. Biomonitoring data and Markov Chain Monte Carlo (MCMC) simulation were utilized for exposure assessment. RESULTS: Deposition fraction of MDI in the respiratory tract at the light, moderate, and heavy activity were 0.038, 0.079, and 0.153, respectively. Converged MCMC results as the posterior means and prior values were obtained for several PBTK model parameters. In our study, we calibrated a rat model to investigate the transport, absorption, and elimination of 4,4'-MDI via inhalation exposure. The calibration process successfully captured experimental data in the lungs, liver, blood, and kidneys, allowing for a reasonable representation of MDI distribution within the rat model. Our calibrated model also represents MDI dynamics in the bloodstream, facilitating the assessment of bioavailability. For human exposure, we validated the model for recent and long-term MDI exposure using data from relevant studies. CONCLUSION: Our computational models provide reasonable insights into MDI exposure, contributing to informed risk assessment and the development of effective exposure reduction strategies.

Isocyanate induces cytotoxicity via activation of phosphorylated alpha synuclein protein, nitrosative stress, and apoptotic pathway in Parkinson's Disease model-SHSY-5Y cells.

BACKGROUND: Neurotoxic disorders account for a significant portion of the diseases that influence the worldwide disease burden. Parkinson's disease is one such disease that is linked with environmental toxin exposure. Isocyanates are a highly reactive industrial intermediate used widely in manufacturing plastic products, paints, etc. This study aims to delineate the neurotoxic potential of isocyanate in Parkinson's cell model-SHSY-5Y cells. METHODOLOGY: SHSY-5Y cells were treated with isocyanate analogue (N succinimidyl N methyl carbamate) in time and dose dependant manner. Different parameters were assessed like protein expression, nitrosative stress level, antioxidant enzymes level and apoptosis. RESULTS: Our findings demonstrate that dose- and time-dependent isocyanate exposure increases reactive nitrogen species and decreases the glutathione, SOD, and catalase levels. Further, increased phosphorylated alpha-synuclein protein and activation of caspase 3 exert cytotoxicity in SHSY-5Y cells. CONCLUSION: Our research reveals that widely used isocyanate induces cytotoxicity, apoptosis, nitrosative stress, and protein dysfunction, which might constitute a potential mechanism of neurodegeneration in Parkinsonism.

New Method to Biomonitor Workers Exposed to 1,6-Hexamethylene Diisocyanate.

Isocyanates such as 1,6-hexamethylene diisocyanate (HDI), 4,4'-methylenediphenyl diisocyanate, and toluene diisocyanate are highly reactive compounds that have a variety of commercial applications, including manufacturing polyurethane foam, elastomers, paints, adhesives, coatings, insecticides, and many other products. Their primary route of occupational exposure is through inhalation. Due to their high chemical reactivity, they are toxic and have adverse effects at the cellular and subcellular levels, leading to irritative and immunological reactions associated with lung disease. High concentrations of isocyanates are strong respiratory irritants. Bronchial sensitization and asthma are among the major adverse clinical reactions associated with low-level chronic exposure to isocyanates. Albumin adducts have been linked to the mechanism of occupational asthma caused by isocyanates. Isocyanates react in vivo with albumin, which is recognized by the immune system. Albumin adducts of isocyanates trigger immune responses and are probably the antigenic basis for isocyanate asthma. Sensitization to isocyanates is the main pathway for adverse health effects. Therefore, markers for the biologically effective dose such as albumin adducts of HDI are needed. A new isocyanate adduct of HDI with lysine─Nε-[(6-amino-hexyl-amino)carbonyl]-lysine (HDI-Lys)─was synthesized and characterized by 1H-NMR, 13C-NMR, and mass spectrometry (MS). Appropriate internal standards─HDI-Lys-4,4'-5,5'-d4 (HDI-d4-Lys) and Nε-[(7-amino-heptyl-amino)carbonyl]-lysine (Hep-Lys)─were synthesized to establish a LC-MS/MS method for the analysis of HDI adducts in in vitro modified albumin and in workers. The presence of HDI-Lys was found after pronase digestion of albumin and confirmed by two independent chromatographic approaches: with a C8 reversed-phase column and with a hydrophilic interaction liquid chromatography column. Quantification was performed with positive electrospray ionization (ESI)-MS. The adduct peak found in vivo was confirmed with the less sensitive negative ESI-MS. In summary, these are new compounds and methods to determine isocyanate-specific adducts with albumin in workers exposed to HDI.

An in vitro alveolar model allows for the rapid assessment of chemical respiratory sensitization with modifiable biomarker endpoints.

Diisocyanates are commonly used in polyurethanes where use includes industrial, commercial, and residential applications and can exist as respirable contaminants. These respirable contaminants exist in the air we breathe. Yet, there is no rapid assay available to test for potential respiratory sensitizers. To assess these hazards, as well as to decrease animal numbers used in testing, investigations that lead to verifiable in vitro methods are needed. We describe an easy, reliable, verified cell culture model that can be adopted by any lab capable of performing molecular toxicology. The architecturally relevant alveolar model consists of epithelial cells, macrophage cells, and dendritic cells in a simply maintained submerged system ideal for high-throughput testing. Exposures to contaminants that verify biomarker identification include a known pulmonary sensitizer (isophorone diisocyanate) and a positive control for cellular activation (phorbol 12-myristate 13-acetate/ionomycin). The mitochondrial reactive oxygen species generation and cytostructural changes were assessed with confocal laser scanning microscopy; cell morphology was assessed with scanning electron microscopy; biochemical reactions were assessed via protein arrays; genetic alterations were assessed via gene arrays; and cell surface activation markers were assessed via flow cytometry. Results showed that compared to untreated cultures, isophorone diisocyanate increased markers for dendritic cell activation, trafficking, and antigen presentation; number and length of dendritic protrusions; oxidative stress; and genetic and cytokine expression of neutrophil chemoattractants. The chemokines and cytokines CCL7, CXCL5, IL-6, and IL-8 were identified as biomarkers indicative of respiratory sensitization. By including multiple methods to assess endpoints, the in vitro model described can serve as a high-throughput assay to identify substances which may lead to respiratory sensitization.

Methylenediphenyl diisocyanate lysine conjugates in the urine of workers exposed to methylenediphenyl diisocyanate.

Diisocyanates have long been a leading cause of occupational asthma. As control often relies on personal protective equipment and there is the potential for skin uptake, biological monitoring is often used to assess worker exposure. Current routine biological monitoring methods do not distinguish between a diisocyanate and the corresponding diamine exposure in urine samples; therefore, a specific urinary biomarker is desirable. Urine samples were obtained from a group of workers exposed to methylenediphenyl diisocyanate (MDI) where aerosol generation was unlikely. Lysine conjugates of MDI were extracted from urine by solid phase extraction; analysis was performed by liquid chromatography tandem mass spectrometry. Acetylated MDI-lysine (acMDI-Lys) conjugates were detected in 73% of samples tested from persons with exposure to MDI compared to 93% of samples that were positive for methylene dianiline (MDA) in hydrolysed urine. There was a weak but significant positive correlation between the two biomarkers (r2 = 0.377). This is the first report detecting and quantifying acMDI-Lys in the urine of workers exposed to MDI, and acMDI-Lys may be a useful non-invasive biomarker in discriminating between MDI and MDA exposures.

HBM4EU Diisocyanates Study-Research Protocol for a Collaborative European Human Biological Monitoring Study on Occupational Exposure.

Diisocyanates have long been a leading cause of occupational asthma in Europe, and recently, they have been subjected to a restriction under the REACH regulations. As part of the European Human Biomonitoring project (HBM4EU), we present a study protocol designed to assess occupational exposure to diisocyanates in five European countries. The objectives of the study are to assess exposure in a number of sectors that have not been widely reported on in the past (for example, the manufacturing of large vehicles, such as in aerospace; the construction sector, where there are potentially several sources of exposure (e.g., sprayed insulation, floor screeds); the use of MDI-based glues, and the manufacture of spray adhesives or coatings) to test the usability of different biomarkers in the assessment of exposure to diisocyanates and to provide background data for regulatory purposes. The study will collect urine samples (analysed for diisocyanate-derived diamines and acetyl-MDI-lysine), blood samples (analysed for diisocyanate-specific IgE and IgG antibodies, inflammatory markers, and diisocyanate-specific Hb adducts for MDI), and buccal cells (micronucleus analysis) and measure fractional exhaled nitric oxide. In addition, occupational hygiene measurements (air monitoring and skin wipe samples) and questionnaire data will be collected. The protocol is harmonised across the participating countries to enable pooling of data, leading to better and more robust insights and recommendations.

Assessment of personal inhalation and skin exposures to polymeric methylene diphenyl diisocyanate during polyurethane fabric coating.

Methylene diphenyl diisocyanate (MDI) monomers and polymeric MDI (pMDI) are aromatic isocyanates widely used in the production of polyurethanes. These isocyanates can cause occupational asthma, hypersensitivity pneumonitis, as well as contact dermatitis. Skin exposure likely contributes toward initial sensitization but is challenging to monitor and quantitate. In this work, we characterized workers' personal inhalation and skin exposures to pMDI in a polyurethane fabric coating factory for subsequent health effect studies. Full-shift personal and area air samples were collected from eleven workers in representative job areas daily for 1-2 weeks. Skin exposure to hands was evaluated concomitantly with a newly developed reagent-impregnated cotton glove dosimeter. Samples were analyzed for pMDI by liquid chromatography-tandem mass spectrometry. In personal airborne samples, the concentration of 4,4'-MDI isomer, expressed as total NCO, had a geometric mean (GM) and geometric standard deviation (GSD) of 5.1 and 3.3 ng NCO/m3, respectively (range: 0.5-1862 ng NCO/m3). Other MDI isomers were found at much lower concentrations. Analysis of 4,4'-MDI in the glove dosimeters exhibited much greater exposures (GM: 10 ng/cm2) and substantial variability (GSD: 20 ng NCO/cm2; range: 0-295 ng NCO/cm2). MDI inhalation exposure was well below occupational limits for MDI for all the job areas. However, MDI skin exposure to hands was substantial. These findings demonstrated the potential for substantial isocyanate skin exposure in work settings with very low airborne levels. This exposure characterization should inform future studies that aim to assess the health effects of work exposures to MDI and the effectiveness of protective measures.

Exploring structure/property relationships to health and environmental hazards of polymeric polyisocyanate prepolymer substances-2. Dermal sensitization potential in the mouse local lymph node assay.

The sensitization potencies of twenty custom-designed monomer-depleted polymeric polyisocyanate prepolymer substances and their associated toluene diisocyanate (TDI), methylene diphenyl diisocyanate (MDI), hexamethylene diisocyanate (HDI), and isophorone diisocyanate (IPDI) monomer precursors were investigated by means of the mouse Local Lymph Node Assay (LLNA). These polymeric prepolymers were designed to represent the structural features and physical-chemical properties exhibited by a broad range of commercial polymeric polyisocyanate prepolymers that are produced from the reaction of aromatic and aliphatic diisocyanate monomers with aliphatic polyether and polyester polyols. The normalization of LLNA responses to the applied (15-45-135 mM) concentrations showed that the skin sensitization potency of polymeric polyisocyanate prepolymers is at least 300 times less than that of the diisocyanate monomers from which they are derived. The sensitization potency of the prepolymers was shown to be mainly governed by their hydrophobicity (as expressed by the calculated octanol-water partition coefficient, log Kow) and surfactant properties. Neither hydrophilic (log Kow <0) nor very hydrophobic (log Kow >25) prepolymers stimulated lymphocyte proliferation beyond that of the dosing vehicle control. The findings of this investigation challenge the generally held assumption that all isocyanate (-N=C=O) bearing substances are potential skin (and respiratory) sensitizers. Further, these findings can guide the future development of isocyanate chemistries and associated polyurethane applications toward reduced exposure and health hazard potentials.

A brief overview of properties and reactions of diisocyanates.

By way of introduction to the special issue on diisocyanates and their corresponding diamines, this brief overview presents, for the most commonly used diisocyanate monomers, a selection of physical-chemical properties that are relevant to exposure in the workplace and in the general environment, as well as a concise overview of diisocyanate reactions and some of their toxicological implications.

[Isocyanate-induced asthma].

Isocyanates are often found in workplaces in e.g., glue, paint, plastics and foam products. Asthma caused by isocyanates is one of the most common forms of occupational asthma, though it is difficult to diagnose, as described in this review. It is not possible to demonstrate sensitisation with available allergy tests. A certain diagnosis can only be made with the help of specific bronchial provocation, which is performed at three centres in Denmark. A correct diagnosis is important, as it helps provide optimal treatment as well as alerting the employer, that improvements are needed at the workplace.

Absorption, distribution, metabolism, and excretion of methylene diphenyl diisocyanate and toluene diisocyanate: Many similarities and few differences.

Methylene diphenyl diisocyanate (MDI) and toluene diisocyanate (TDI) are high production volume chemicals used for the manufacture of polyurethanes. For both substances, the most relevant adverse health effects after overexposure in the workplace are isocyanate-induced asthma, lung function decrement and, to a much lesser extent, skin effects. Over the last two decades many articles have addressed the reactivity of MDI and TDI in biological media and the associated biochemistry, which increased the understanding of their biochemical and physiological behavior. In this review, these new insights with respect to similarities and differences concerning the adsorption, distribution, metabolism, and excretion (ADME) of these two diisocyanates and the implications on their toxicities are summarized. Both TDI and MDI show very similar behavior in reactivity to biological macromolecules, distribution, metabolism, and excretion. Evidence suggests that the isocyanate (NCO) group is scavenged at the portal-of-entry and is not systemically available in unbound reactive form. This explains the lack of other than portal-of-entry toxicity observed in repeated-dose inhalation tests.

Evaluation of animal toxicity studies with diisocyanates regarding presence of thresholds for induction and elicitation of respiratory allergy by quantitative weight of evidence.

Animal toxicity studies on diisocyanates were evaluated using quantitative weight of evidence (QWoE) to test the hypothesis that the dose-response curve shows a threshold for the induction and/or elicitation of respiratory sensitization. A literature search identified 59 references that included at least two concentration groups of the diisocyanate and a vehicle-exposed concurrent control in the study design. These studies were subjected to a QWoE-assessment applying scoring criteria for quality and relevance/strength of effects relevant to the selected endpoint of respiratory sensitization. Overall, the studies assessing dose/concentration-response for diisocyanates with the endpoint, respiratory sensitization, were heterogenous regarding study design, animal models used, endpoints assessed, and quality. Only a limited number of the studies subjected to the QWoE-assessment allowed drawing conclusions about possible thresholds for respiratory sensitization. Highest quality and relevance/strength of effects scores were obtained by a series of studies specifically designed to investigate a potential threshold for elicitation of respiratory sensitization in the Brown Norway (BN) rat. These studies applied an elaborate study design to optimize induction of respiratory sensitization and reduce interference by respiratory tract irritation. In summary, the available studies provided moderate to good support for the existence of a threshold for elicitation and limited to moderate support for a threshold regarding induction of respiratory allergy by diisocyanates in experimental animals. However, a quantitative extrapolation of threshold values established in rodents to humans remains complex.

Severe asthma and death in a worker using methylene diphenyl diisocyanate MDI asthma death.

Diisocyanates are well-recognized to cause occupational asthma, yet diisocyanate asthma can be challenging to diagnose and differentiate from asthma induced by other allergens. The present study assesses the potential contribution of methylene diphenyl diisocyanate (MDI) to a workplace fatality. Examination of medical records, tissue, and blood from the deceased worker were undertaken. Formalin-fixed paraffin-embedded lung tissue sections were assessed through histologic and immunochemical stains. Serum MDI-specific IgE and IgG, and total IgE, were measured by enzyme-linked immunosorbent assays and/or Western blot. Information about potential chemical exposures and industrial processes in the workplace were provided by the employer and through interviews with co-workers. Review of the worker's medical records, occupational history, and autopsy findings were consistent with severe asthma as the cause of death, and ruled out cardiac disease, pulmonary embolism, or stroke. Lung pathology revealed hallmarks of asthma including smooth muscle hypertrophy, eosinophilia, basement membrane thickening, and mucus plugging of bronchioles. Immunochemical staining for MDI was positive in the thickened basement membrane of inflamed airways. MDI-specific serum IgE and IgG were significantly elevated and demonstrated specificity for MDI versus other diisocyanates, however, total serum IgE was normal (24 IU/ml). The workplace had recently introduced MDI into the foundry as part of a new process, but MDI air levels had not been measured. Respirators were not required. In summary, post-mortem findings support the diagnosis of diisocyanate asthma and a severe asthma attack at work as the cause of death in a foundry worker.

Inhibitory regulation of purple sweet potato polysaccharide on the hepatotoxicity of tri-(2,3-dibromopropyl) isocyanate.

Tri-(2,3-dibromopropyl) isocyanate (TBC), a new emerged persistent organic pollutant, is widely used in fields of flame retardant, textile, rubber and plastic with strong hepatotoxicity. Purple Sweet Potato Polysaccharide (PSPP) has antioxidant and hepatoprotective effects. This study aims to answer the scientific question whether PSPP has a protective effect on TBC induced liver injury. The effect of PSPP on the apoptosis of HepG2 cells was detected by MTT assay, the morphological changes were observed by morphological observation, and the apoptosis rate was determined by flow cytometry. The apoptotic genes were detected by qPCR assay, the relevant protein express was detected by western blot. The correlation between proteins and genes in the apoptosis pathway of HepG2 cells was calculated. To further reveal the apoptosis mechanism of TBC hepatotoxicity in vivo, 19 target genes and 14 apoptotic related proteins of inhibiting apoptosis via death receptor and mitochondria were discussed, all the above results proved that PSPP had protective effect on liver injury induced by TBC. This study not only provided a scientific basis for clarifying the mechanism of TBC hepatotoxicity and the protective effect of PSPP, but also generated the new point and method in terms of the prevention in advance and early intervention of diseases caused by environmental pollution.

Is Isocyanate Exposure and Occupational Asthma Still a Major Occupational Health Concern? Systematic Literature Review.

Isocyanate, whose disease-inducing mechanism is poorly understood, with poor prognosis, is widely used. Asthma is the most frequent manifestation of prolonged exposure. We assessed the evolution of the incidence of isocyanate-induced occupational asthma over time. PubMed and Cochrane databases were systematically searched for studies published since 1990 that assessed the relationship between occupational exposure to isocyanates and asthma. We identified 39 studies: five retrospective cohort studies, seven prospective cohort studies, three of which were inception cohorts), seven observational cross-sectional studies, five literature reviews, two case series, and 13 registry studies. The incidence of occupational asthma secondary to isocyanate exposure has decreased from more than 5% in the early 1990s to 0.9% in 2017 in the United States. Despite the wide use of optimal collective and individual protection measures, the risk of occupational asthma has stabilized. Occupational asthma risk can be assessed with good sensitivity using self-questionnaires and pulmonary function tests. Occupational avoidance should be implemented as soon as possible after the first symptoms appear because the prognosis becomes increasingly poor with the persistence of exposure. It is now necessary to study specifically cutaneous sensitization to isocyanates and to define what protective equipment is effective against this mode of exposure.

Susceptibility of the Bhopal-methyl isocyanate (MIC)-gas-tragedy survivors and their offspring to COVID-19: What we know, what we don't and what we should?

There is credible evidence that the 1984-Bhopal-methyl isocyanate (MIC)-gas-exposed long-term survivors and their offspring born post-exposure are susceptible to infectious/communicable and non-communicable diseases. Bhopal's COVID-19 fatality rate suggests that the MIC-gas tragedy survivors are at higher risk, owing to a weakened immune system and co-morbidities. This situation emboldened us to ponder over what we know, what we don't, and what we should know about their susceptibility to COVID-19. This article aims at answering these three questions that emerge in the minds of public health officials concerning prevention strategies against COVID-19 and health promotion in the Bhopal MIC-affected population (BMAP). Our views and opinions presented in this article will draw attention to prevent and reduce the consequences of COVID-19 in BMAP. From the perspective of COVID-19 prophylaxis, the high-risk individuals from BMAP with co-morbidities need to be identified through a door-to-door visit to the severely gas-affected regions and advised to maintain good respiratory hygiene, regular intake of immune-boosting diet, and follow healthy lifestyle practices.

Isocyanate induced allergic contact dermatitis in a university undergraduate student: An occupational dermatitis case report, review of laboratory safety regulations, and implications for campus research.

A 19-year-old female college undergraduate developed an intensely swollen, erythematous and pruritic rash on the face and hands while working in an optical fabrication lab producing photosensitive polymers. She had no respiratory symptoms. The rash was consistent with contact dermatitis and there was no clinical evidence of respiratory involvement with normal spirometry. A review of the safety data sheets of chemicals used in the laboratory revealed several known sensitizers, including 6-hexamethylene diisocyanate (HDI), dibutyl phthalate, and 2,4,6-tribromophenyl acrylate. Patch testing confirmed the patient's sensitization to HDI. A subsequent worksite visit identified several hazardous chemicals that were used without appropriate hazard communication, training, standard operating procedures, or personal protective equipment. Simple exposure controls were recommended and instituted, and the patient was able to return to work in the laboratory without the recurrence of symptoms. This case demonstrates the importance of hazard identification, communication, and safety training in academic laboratories, for students and workers. A medical evaluation can identify hazards as well as lead to improvements in exposure controls and safe return to research.

Alleviation of methyl isocyanate-induced airway obstruction and mortality by tissue plasminogen activator.

Methyl isocyanate (MIC, "Bhopal agent") is a highly reactive, toxic industrial chemical. Inhalation of high levels (500-1000 ppm) of MIC vapor is almost uniformly fatal. No therapeutic interventions other than supportive care have been described that can delay the onset of illness or death due to MIC. Recently, we found that inhalation of MIC caused the appearance of activated tissue factor in circulation with subsequent activation of the coagulation cascade. Herein, we report that MIC exposure (500 ppm for 30 min, nose-only) caused deposition of fibrin-rich casts in the conducting airways resulting in respiratory failure and death within 24 h in a rat model (LC90-100 ). We thus investigated the effect of airway delivery of the fibrinolytic agent tissue plasminogen activator (tPA) on mortality and morbidity in this model. Intratracheal administration of tPA was initiated 11 h post MIC exposure and repeated every 4 h for the duration of the study. Treatment with tPA afforded nearly 60% survival at 24 h post MIC exposure and was associated with decreased airway fibrin casts, stabilization of hypoxemia and respiratory distress, and improved acidosis. This work supports the potential of airway-delivered tPA therapy as a useful countermeasure in stabilizing victims of high-level MIC exposure.