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.

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.

Spectrum of health condition in methyl isocyanate (MIC)-exposed survivors measured after 30 years of disaster.

Health effects of methyl isocyanate (MIC) exposure were mostly reported on the one-time acute exposure in Bhopal population. Epidemiological survey conducted by the Indian apex body of health research has been reported as Technical Reports, which were lacking in peer review by the expert epidemiologic scientists. The present pilot survey was aimed to measure the health effects 30 years post disaster in MIC-exposed survivors. Questionnaire-based survey has captured every health complaint in 168 individuals and grouped as systemic functions for interpreting the long-term effects of MIC. Key health parameters, including reproductive outcome and respiratory/orthopedic/general morbidity, were prevalent among the severely exposed population compared to control and moderately exposed groups. The collective incidence of diabetes, hypertension, and cancer also was prevalent in the severely exposed group. Ophthalmic morbidity was almost similar in the three groups, rather with higher incidence in the control group, though not statistically significant. Among all health parameters, reproductive, ophthalmic, and respiratory effects were prevalent over others. Although the incidence of health problems has been declined among the survivors, long-term effect is apparent as scars of one-time acute exposure might trigger sequel of long-term effects. Additionally, acquisition of genetic rearrangements, survival of T cell sub-populations, variable latency of chemical effect on DNA nucleosides, nutritional status, occupational exposure, living environment, lifestyle, and overall gene-environment interaction might perturb individual immunity and favor onset of long-term illness in a scenario of background exposure to MIC. However, the exercise should be continued on a larger sample size for drawing a conclusive result on long-term MIC effect on survivors' health.

Isocyanates and human health: multistakeholder information needs and research priorities.

OBJECTIVES: To outline the knowledge gaps and research priorities identified by a broad base of stakeholders involved in the planning and participation of an international conference and research agenda workshop on isocyanates and human health held in Potomac, Maryland, in April 2013. METHODS: A multimodal iterative approach was used for data collection including preconference surveys, review of a 2001 consensus conference on isocyanates, oral and poster presentations, focused break-out sessions, panel discussions, and postconference research agenda workshop. RESULTS: Participants included representatives of consumer and worker health, health professionals, regulatory agencies, academic and industry scientists, labor, and trade associations. CONCLUSIONS: Recommendations were summarized regarding knowledge gaps and research priorities in the following areas: worker and consumer exposures; toxicology, animal models, and biomarkers; human cancer risk; environmental exposure and monitoring; and respiratory epidemiology and disease, and occupational health surveillance.

Reduction in diisocyanate and non-diisocyanate sensitizer-induced occupational asthma in Ontario.

OBJECTIVE: To investigate relative frequency and features of diisocyanate (ISO) and non-diisocyanate (N-ISO) allowed occupational asthma (OA) claims in Ontario, Canada, during a 5-year period (1998 to 2002). METHODS: Records were abstracted from the Ontario Workplace Safety and Insurance Board using methodology similar to our previous investigation that had identified 30 ISO and 30 N-ISO claims/yr during 1980 to 1993. RESULTS: There were 99 OA claims: 37 ISO (7.4 claims/yr) and 62 N-ISO (12.4 claims/yr). The ISO group had more males (86% vs 69%, p = 0.01), but there were no other significant differences. The commonest professions were spray painters (41%) and production workers (38%) in the ISO group and production workers (49%) and health care workers (8%) in the N-ISO group. CONCLUSIONS: ISO and N-ISO claims declined from the previous period, especially for ISO, perhaps because of effective surveillance programs.

Medical surveillance programme for diisocyanate exposure.

OBJECTIVES: Surveillance programmes for occupational asthma should reduce the severity of asthma both at the time of diagnosis and after removal from exposure as well as costs related to functional impairment. The aim of this study was to compare the severity and cost of diisocyanate-induced occupational asthma in workers participating in a surveillance programme and in twice the number of workers diagnosed after being referred by their physician. METHODS: Answers to a self-administered questionnaire led to possible referral for further assessment that included methacholine testing and specific inhalation challenges as the gold standard for confirming occupational asthma. Results Of the 2897 workers who participated, 182 (6.3%) had a positive questionnaire. 79/182 (43%) were referred for further medical assessment and 20 had confirmed occupational asthma by specific inhalation testing. At the time of diagnosis, the 20 screened subjects had a mean PC(20) of 3.35 mg/ml as compared to 1.50 mg/ml (p = 0.05) in the 66 controls. Two years after diagnosis and removal from exposure, the 20 subjects screened had a mean PC(20) of 4.81 mg/ml compared to 1.67 mg/ml (p = 0.03) in controls. Clinical remission occurred in 34% of the screened group compared to 16% of the control group (p = 0.02). The median costs for functional impairment were $C11,900 in screened subjects and $C19,600 in controls (p = 0.04). CONCLUSIONS: Subjects with occupational asthma screened by a medical surveillance programme have a better outcome both at the time of diagnosis and 2 years after removal from exposure, with lower compensation costs compared to controls.

Characterizing the health and safety needs of the collision repair industry.

Production workers in the collision repair industry are potentially exposed to many harmful chemicals, including isocyanates. Of particular concern is the burden of work-related asthma in this industry that likely reflects exposures to the isocyanates in two-part paints. The main objectives of this study were to gather information about: (i) the collision repair industry business model, (ii) the number of collision repair production workers potentially exposed to isocyanates, (iii) additional chemical and physical exposures of concern, (iv) current health and safety practices in the industry, (v) the health and safety perceptions and needs of business owners and managers, and (vi) strategies to reduce exposure and increase employer and worker awareness. Data were gathered using a combination of key informant interviews, field investigations, and a statewide needs assessment survey. Although a response bias cannot be excluded, the 69% response rate suggests that the survey results are likely representative of Washington State's collision repair industry. Collision repair was determined to be a male-dominated industry chiefly comprising small, nonunionized, family-run businesses. Many shops face numerous safety and health challenges resulting from a combination of misinformation within the industry, insufficient funds to address workplace health and safety concerns, and social barriers to enforcing best practices within the shops. Most notably, inappropriate selection and use of respirators and gloves likely contribute significantly to isocyanate exposures. Collision repair workers are potentially exposed to a variety of additional chemical and physical hazards that deserve attention. This industry requires health and safety intervention of both an educational campaign and technical assistance. Any such intervention must account for the financial, demographic, and social characteristics of this industry.

Ocular effects of exposure to industrial chemicals: clinical management and proteomic approaches to damage assessment.

Industrial chemicals in a variety of applications are often found in highly populated areas and their presence carries risks. The threat of serious consequences from inadvertent or intentional events involving hazardous chemicals is a possibility. Extremism and/or other illicit activities pose environmental threats from chemical exposures. We present here a review of the threat of ocular injury in small-and large-scale chemical releases and discuss mechanisms of damage and repair to the eyes. The emerging field of proteomics has been described in relation to its potential role in the assessment of ocular changes following chemical exposures and management of ocular trauma.

Skin exposure to aliphatic polyisocyanates in the auto body repair and refinishing industry: a qualitative assessment.

OBJECTIVES: Although respiratory exposures have been the primary concern with isocyanates, skin exposure also can occur and may contribute to sensitization and asthma. Methodologies to assess isocyanate skin exposure in the workplace are limited and skin exposure data scarce. The goals of this study were (i) to evaluate and validate the isocyanate colorimetric indicators against a quantitative assay, (ii) to evaluate the extent of isocyanate surface contamination and skin exposure among auto body shop workers and (iii) to evaluate isocyanate skin exposure determinants. METHODS: The colorimetric indicators were compared with a high-performance liquid chromatography (HPLC) quantitative assay based on the National Institute for Occupational Safety and Health Method 5525 using paired laboratory sampling. The colorimetric indicators were then used to assess surface contamination and skin exposure to aliphatic isocyanates in 35 auto body shops and 124 workers as part of an epidemiologic study. The positive sample rate was calculated for various surfaces, body parts and tasks. The color intensity of the colorimetric indicators was rated on a scale 0 (yellow color) to 5 (deep red). Side-by-side comparisons of the qualitative method with the quantitative HPLC assay were also performed in the field using paired samples. RESULTS: Laboratory and field evaluation validated use of the colorimetric indicators. The rate of positive surface samples for isocyanates was 46% (n = 145/313). Thirty-four percent (73/216) of samples were positive for unprotected skin and 20% (n = 22/111) for skin under latex gloves. The highest positive rate observed on skin samples was obtained after paint mixing and spraying tasks. The colorimetric indicators were highly specific for isocyanates, but false negatives occurred when compared with the more sensitive HPLC quantitative assay. The presence of surface contamination and the performance of paint-related tasks were the major determinants of isocyanate skin exposure. CONCLUSIONS: This study documents extensive surface contamination and skin exposure, including under gloves, to aliphatic polyisocyanates during painting and paint mixing tasks in auto body shop workers. Contaminated surfaces and aerosol deposition during spray painting may both contribute to skin exposure. The colorimetric indicator is a quick, practical and low-cost, but not highly sensitive, industrial hygiene tool to detect isocyanate surface contamination and skin exposures following the use of isocyanate-containing products.

Surveillance for isocyanate asthma: a model based cost effectiveness analysis.

AIMS: Because logistical and financial obstacles impede using large prospective cohort studies, surveillance decisions in occupational settings must often be made without evidence of relative benefits and costs. Using the example of isocyanate induced asthma, the most commonly reported immune mediated occupational asthma, the authors developed a model based approach to evaluate the costs and benefits of surveillance from both an employer and a societal perspective. METHODS: The authors used a mathematical simulation model of isocyanate asthma to compare annual surveillance to passive case finding. Outcome measures included symptom free days (SFD), quality adjusted life years (QALY), direct costs, productivity losses, and incremental cost effectiveness ratio (CER), measured from the employer and the societal perspectives. Input data were obtained from a variety of published sources. RESULTS: For 100,000 exposed workers, surveillance resulted in 683 fewer cases of disability over 10 years. Surveillance conferred benefits at an incremental cost of 24,000 dollars/QALY (employer perspective; 13.33 dollars/SFD) and was cost saving from the societal perspective. Results were sensitive to assumptions about sensitisation rate, removal rates, and time to diagnosis, but not to assumptions about therapy costs and disability rates. CONCLUSIONS: Baseline results placed the CER for surveillance for isocyanate asthma within the acceptable range. Costs from the societal and employer perspective differed substantially with a more attractive CER from the societal perspective, suggesting opportunities for employer/societal cost sharing. The analysis demonstrates the value of a model based approach to evaluate the cost effectiveness of surveillance programmes for isocyanate asthma, and to inform shared decision making among clinicians, patients, employers, and society. Such a modeling approach may be applicable to surveillance programmes for other work related conditions.

Host factors in occupational diisocyanate asthma: a Swiss longitudinal study.

OBJECTIVE: To investigate the usefulness of surrogates for individual susceptibility to organic diisocyanates in occupational asthma. SUBJECTS: All new cases declared to the Swiss National Accident Insurance Company (SUVA) for establishment of a case for compensable occupational disease during 1993. Sixty-nine persons, of whom three were women, were suspected of having occupational asthma due to isocyanates. Of these, 47 subjects fulfilled the criteria to be accepted as an occupational disease case. METHODS: All subjects were studied clinically and gave a blood sample for the phenotyping of their alpha-antitrypsin status and for immunological studies. The subjects were also given a peroral dose of caffeine for the determination of their N-acetylation capacity. Finally, those with an occupational disease were subjected to the methacholine provocation test. RESULTS: Forty-four persons with occupational disease, out of 47, were heterozygous antitrypsin carriers and/or slow acetylators of primary amines. In the bronchial provocation with methacholine, 12 of these subjects had an unaltered response and seven had a mild reaction, 13 a moderate one and 15 a severe reaction. INTERPRETATION: The study confirms the finding that slow N-acetylators are susceptible to asthma from exposure to common diisocyanate monomers at work. The same applies to heterozygous antitrypsin-phenotype carriers. Thus, the use of these markers may reinforce the diagnostic procedure, but they cannot completely replace the immunological tests.

Assessment of preferential Th1 or Th2 induction by low-molecular-weight compounds using a reverse transcription-polymerase chain reaction method: comparison of two mouse strains, C57BL/6 and BALB/c.

The aim of this study was to examine whether the RT-PCR method for various Th1/Th2 cytokines is appropriate for determination of response to allergens using C57BL/6 and Balb/c mice, which are known to preferentially demonstrate Th1 and Th2 responses, respectively. To this end, both strains of mice were sensitized by skin painting with the contact allergen dinitrochlorobenzene (DNCB) or the respiratory allergen trimellitic anhydride (TMA). We used the sensitizing protocol adopted by Kimber and coworkers (Toxicology 103, 63-73, 1995). At various time points after the last application, the levels of mRNA expression for Th1-type cytokines IFN-gamma, IL-18, and IL-12p40, as well as receptor IL-18R, and the Th2-type cytokine IL-4 and the receptor ST2L, in lymph nodes were measured. The results suggest that differential expression of IL-12p40 and IL-4 mRNA after 24 h allows clear discrimination between DNCB and TMA in C57BL/6 mice, more obviously than in Balb/c mice. Furthermore, to examine this method, C57BL/6 mice were exposed to OXA, DNFB, and TNCB (Th1-predominant allergens) or PA, TDI, and MDI (Th2-predominant allergens). Elevation of IL-12p40 expression was significant with the Th1 inducers, while the level of IL-4 was higher with Th2-predominant allergens. The results of the present study demonstrate, for the first time, that differential expression of IL-12p40 and IL-4 in C57BL/6 mice may be useful as a parameter for assessing influence of contact and respiratory allergens.

[Occupational activities and allergic respiratory manifestations]. / Activité professionnelle et manifestations respiratoires allergiques.

The authors review aspects of occupational asthma that are important for the practitioner. Insurance problems are dealt with in the light of local regulations. Detailed chapters are devoted to baker's asthma (various aeroallergens), to plastic workers and painters (isocyanates) and to respiratory diseases among farmers. Finally, a list of other occupations in which asthma typically occurs is given. The responsibility of practitioners in charge of young patients about to choose their future profession is underlined.

A two-centre study for the evaluation and validation of an animal model for the assessment of the potential of small molecular weight chemicals to cause respiratory allergy.

This study evaluated a single intradermal injection model in the guinea pig with subsequent inhalation challenge and serological analysis as a method to predict the potential of chemicals to induce respiratory allergy. Four known respiratory allergens (trimellitic anhydride, diphenyl methane diisocyanate, phthalic anhydride and toluene diisocyanate (TDI)) were screened by two industrial research laboratories using this protocol. Dinitrochlorobenzene, a potent contact allergen, was included as a negative control material. In both laboratories, the respiratory allergens, but not the contact allergen, induced high titre antigen-specific antibodies in treated animals. The inhalation challenge results were similar in both laboratories but were less conclusive in that exposure to free TDI failed to induce pulmonary responses, probably because it fails to penetrate to the deep lung in sufficient concentration. Although the assay shows promise as a means of identifying chemical respiratory sensitisers, its use as a routine screen for the prediction of the ability of materials to induce respiratory allergy in man is probably questionable.