Characterization of Fungal and Indoor Air Exposures

真菌和室内空气暴露的特征

• 批准号: 8338006
• 金额: $ 29.68万
• 依托单位: NATIONAL INSTITUTE OF ENVIRONMENTAL HEALTH SCIENCES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8338006
• 关键词: A549; Address; Adult; Adverse effects; Affect; Agreement; Agriculture; Air; Allergens; Allergic; Amines; Anabolism; Animals; Antibodies; Antigens; Area; Asthma; Benzoquinones; Binding; Biological Markers; Cell Line; Cell surface; Chaetomium; Chemical Structure; Chemicals; Chemistry; Child; Chronic; Chronic berylliosis; Cities; Clinical; Collaborations; DNA; Data; Delayed Hypersensitivity; Dermal; Detection; Development; Disease; Dust; Ear; Environment; Environmental Exposure; Environmental Health; Epithelial Cells; Evaluation; Experimental Animal Model; Exposure to; Gases; Gene Targeting; General Population; Genes; Genetic; Genetic Polymorphism; Genetic Variation; Goals; Hazard Assessment; Hazard Identification; Health; Home environment; Human; Hypersensitivity; IgE; Immune; Immune Response Genes; Immune system; Immunoglobulin G; Immunologic Techniques; Immunologics; In Vitro; Individual; Infiltration; Inflammatory; Interferons; Investigation; Iowa; Irritants; Kansas; Kinetics; Libraries; Light; Local Lymph Node Assay; Lung; Lymphocyte; Mass Fragmentography; Mediating; Methodology; Minority; Mitogens; Modeling; Molds; Mus; Mycoses; Mycotoxins; National Institute of Environmental Health Sciences; National Toxicology Program; North Carolina; Occupational; Occupational Asthma; Occupational Diseases; Occupational Exposure; Outcome; Ozone; Participant; Patient Recruitments; Patients; Penicillium; Performance; Peripheral Blood Mononuclear Cell; Pesticides; Phase; Play; Polymerase Chain Reaction; Population; Prevalence; Principal Investigator; Probability; Process; Production; Public Health; Publishing; Reaction; Reproduction spores; Respiratory System; Respiratory tract structure; Rhinitis; Risk; Risk Assessment; Rodent; Role; Sampling; Scientist; Series; Serological; Serum; Silicosis; Solvents; Spectrophotometry; Sulfhydryl Compounds; Swelling; System; Techniques; Testing; Time; Toxicology; Universities; Upper respiratory tract; Woman; Work; Workplace; Xenobiotics; age group; air contaminant; airborne allergen; airway hyperresponsiveness; animal data; atopy; base; chemical addition; chronic rhinosinusitis; cohort; cooperative study; cytokine; disorder prevention; farmer; field study; fungus; immune function; improved; in vivo Model; insight; interest; limonene; lymph nodes; male; neutrophil; research study; respiratory; response

The goal of this interagency agreement is to provide support of National Toxicology Program (NTP) hazard identification (or hazard assessment) activities targeted toward the prevention of diseases or adverse effects caused by environmental exposure to chemical or physical agents. These cooperative studies continue to improve the risk assessment process by determining quantitatively what constitutes an adverse health effect on the immune system in humans. These studies evaluate unique cohorts of individuals from professions associated with immune-mediated occupational diseases including asthma, respiratory and contact allergy, chronic beryllium disease, rhinitis, and silicosis. Occupational cohorts are being studied for a number of endpoints including, impact of genetic polymorphisms on inflammatory disease development and clinical outcomes, the role that genetic variations play in environmental and workplace related diseases and identification of unique immunological biomarkers for disease. Occupational rhinitis is a chronic inflammatory disease of the upper respiratory tract that is observed in greater than 90% of workers with occupational asthma. A cohort of chronic rhinosinusitis patients is being evaluated for subjects with allergies to molds. Patient recruitment and testing has been completed. Data has been collected and statistically analyzed. Analysis of allergic sensitization has shown that patients with chronic rhinosinusitis are more likely to be sensitized to a variety of environmental aeroallergens. The results derived from this study have provided further insight into the role of allergic sensitization in chronic rhinosinusitis. To investigate the diversity of fungi in the indoor built environment, air and dust samples have been collected from 30 homes of asthmatic children in the Kansas City area, and are currently in storage. Efforts have focused on developing an effective DNA isolation technique, both in terms of reducing bias against smaller-spore species, and in terms of sensitivity. Isolation of DNA from the Kansas City air and dust samples, amplifying and sequencing the fungal ribosomal internal transcribed spacer (ITS) regions, and assessing the diversity of fungi within these samples has begun. The results will then be incorporated into the health and environmental data of the homes to examine a possible correlation between the fungi present and the health of the inhabitants. In addition, a polymerase chain reaction (PCR) primer/probe set for the detection of toxigenic fungi within the Chaetomium genus is being developed. Species within the Chaetomium genus are often associated with moisture infiltration, and their presence can be an indication of mold problems. A Chaetomium globosum gene that shares strong similarity to the Che a chaetoglobosin (mycotoxin) biosynthesis gene in Penicillium expansum has been identified and a primer/probe set was developed to target this gene and tested against 13 other common environmental fungi. To date, the data indicate that this primer/probe set is specific to C. globosum, and it currently being evaluated against C. strumarium and C. atrobrunneum (two Chaetomium species implicated in clinical mycoses). Patient recruitment and serological sampling has been completed for an evaluation of mold allergy in 677 adult male farmers from the Agricultural Health Study, a cohort of pesticide applicators from North Carolina and Iowa. All sera have been analyzed for both total immunoglobulin E and specific IgE to molds using a mixture of mold antigens. Preliminary results suggest that agricultural practices may be associated with mold sensitization and may differ from those associated with atopy. Work is progressing in assessing an amine probe (pyridoxylamine, PYDA) for both identification and potential potency assessment of electrophilic contact allergens by kinetic spectrophotometric analyses. To date, 24 known sensitizers and 4 non-sensitizers have been assessed. This methodology has been set up at Portland State University Chemistry Department where stopped flow spectrophotometry for assessment of initial kinetic and very rapidly reacting allergens is being assessed. A reactivity constant (Ka) is determined for each allergen and compared to published values for sensitization. A series of benzoquinones is initially being assessed to identify structural activity relationships to binding, and mechanism(s) rates of binding. Work is planned on evaluating the potential incorporating less polar solvents to allow assessment of poorly soluble allergens and to expand the library of allergen Ka¿s for binding to both the amine and thiol spectrophotometric probes. Finally, ongoing studies are identifying and quantifying the volatile reaction products of gas-phase compounds present in the indoor environment, especially dicarbonyls, and investigating immunotoxic and hypersensitivity effects of these reaction products in both in vitro and in vivo models. Studies to characterize the effects of dicarbonyls on the respiratory tract have been conducted using an in vitro indoor air exposure system. Changes in inflammatory cytokine expression were evaluated in a pulmonary epithelial cell line (A549) after exposure to (i) a mixture of limonene and ozone, (ii) air contaminated only with limonene, and (iii) clean air. Preliminary data indicate that exposure to limonene was more inflammatory than exposure to clean air and a mixture of limonene and ozone was more inflammatory than exposure to limonene alone. Additional experiments are underway using different chemical concentrations and time points to confirm these findings. The indoor air contaminant 4-oxopentanal (4-OPA) was examined for irritancy and hypersensitivity following both dermal and pulmonary exposure using a murine model. 4-OPA was tested in a combined local lymph node assay and identified to be an irritant and strong sensitizer (EC3=0.08%) with a delayed hypersensitivity response supported by a positive ear-swelling test. Pulmonary exposure to 4-OPA caused a significant elevation in nonspecific airway hyperreactivity, with increased numbers of lung associated lymphocytes and neutrophils, and increased interferon-g production by lung associated lymph nodes. Further investigation into the role of structurally similar indoor air chemicals present in mixtures in the indoor environment and identification of conserved chemical structures such as di-oxygenated organic compounds among indoor air reaction products that may affect immunotoxic potential is ongoing. Improved chemical derivatization and gas chromatography/mass spectrometry techniques will be used for sampling of indoor air immunotoxic compounds.

该互动协议的目的是提供国家毒理学计划（NTP）危害识别（或危害评估）活动，该活动针对预防疾病或因环境暴露于化学或物理剂而引起的不良影响。这些合作研究继续通过定量确定对人类免疫系统构成不利健康影响的方法来继续改善风险评估过程。这些研究评估了与免疫介导的职业疾病有关的专业人员的独特人群，包括哮喘，呼吸和接触过敏，慢性腹部疾病，鼻炎和硅氧化疾病。正在研究许多终点的职业队列，包括遗传多态性对炎症性疾病发展和临床结果的影响，遗传变异在环境和工作场所相关疾病中的作用以及对疾病的独特免疫生物标志物的鉴定。职业鼻塞炎是上呼吸道的一种慢性炎性疾病，在90％以上的职业哮喘工人中观察到。正在评估对霉菌过敏的受试者的一组慢性鼻鼻涕患者。患者招募和测试已经完成。数据已收集并统计分析。对过敏感应化的分析表明，患有慢性鼻孔炎的患者更有可能对各种环境空气过敏原敏感。从这项研究中得出的结果为过敏性感觉在慢性鼻 - 鼻窦炎中的作用提供了进一步的见解。为了调查室内建筑环境中真菌的多样性，已经从堪萨斯城地区的30个哮喘儿童家庭中收集了空气和灰尘样品，目前正在存储。努力集中在开发有效的DNA隔离技术方面，无论是在减少针对较小物种的偏见和灵敏度方面。从堪萨斯城的空气和灰尘样品中分离DNA，扩大和测序真菌核糖体内部转录间隔（ITS）区域，并开始评估这些样品中真菌的多样性。然后，结果将纳入房屋的健康和环境数据中，以检查现场真菌与居民健康之间的可能相关性。此外，正在开发用于检测Chaetomium属内毒真菌的聚合酶链反应（PCR）引物/探针。 Chaetomium属内的物种通常与水分浸润有关，它们的存在可能表明霉菌问题。已经确定了与CHE的CHAETOGLOBOSIN（霉菌毒素）生物合成基因具有很强相似之处的球体基因，在青霉膨胀中的生物合成基因已被鉴定出来，并开发了一个引物/探针集来靶向该基因并针对其他13种其他13个常见环境真菌进行测试。迄今为止，该数据表明该引物/探针集特定于Globosum，并且目前正在针对Strumurium和C. actrobrunneum（临床真菌中实现的两个Chaetomium strem）进行评估。患者招募和血清学采样已完成，以评估来自北卡罗来纳州和爱荷华州的一批农药施用者的农业健康研究的677名成年男性农民的霉菌过敏。使用霉菌混合物分析了所有血清的总免疫球蛋白E和特异性IgE的霉菌。初步结果表明，农业实践可能与霉菌敏感性有关，并且可能与与特应性相关的实践有所不同。 通过动力学分光光度法分析评估胺探针（吡ido胺，PYDA）的工作正在进行中。迄今为止，已经评估了24个已知传感器和4个非传感器。该方法已在波特兰州立大学化学系建立，在该系中，正在评估最初动力学和非常快速反应的过敏原的停止流量分光光度法。确定每种过敏原的反应性常数（KA），并将其与已发表的敏化值进行比较。最初正在评估一系列苯喹酮，以识别结构活动与结合的结构性关系和结合速率。计划评估较少的极性求解器的潜在的工作，以评估固体过敏原的不良过敏原，并扩展过敏原ka。与胺分光光度计学问题的结合。 最后，正在进行的研究正在识别和量化室内环境中存在的气相化合物的挥发性反应产物，尤其是二核酸，并研究了这些反应产物在体外和体内模型中这些反应产物的免疫毒性和超敏反应作用。已经使用体外室内空气暴露系统进行了表征二骨对呼吸道影响的研究的研究。在暴露于（i）（i）柠檬烯和臭氧的混合物后，（ii）仅被柠檬烯污染的空气和（iii）清洁空气后，评估了炎症性细胞因子表达的变化（A549）（a549）。初步数据表明，暴露于柠檬烯比暴露于清洁空气更具炎症性，而柠檬烯和臭氧的混合物比单独暴露于柠檬烯更炎症。使用不同的化学浓度和时间点正在进行其他实验，以确认这些发现。使用鼠模型检查了皮肤和肺部暴露后，检查了室内空气污染物4-氧化剂（4-OPA）的刺激性和超敏反应。在合并的局部淋巴结测定中测试了4-OPA，并确定为刺激性且强的传感器（EC3 = 0.08％），其延迟的超敏反应由正阳性耳塞测试支持。肺暴露于4-OPA导致非特异性气道高反应性的显着升高，并提高了化学衍生化和气相色谱/质量的提高。改善的化学衍生化和气相色谱/质谱技术将用于室内免疫毒性化合物的采样。