Philip K. Hopke, Andrea R. Ferro, Peter Jaques Clarkson University Resuspension of Aeroallergens and Exacerbation of Inflammation in Asthmatic Children Philip K. Hopke, Andrea R. Ferro, Peter Jaques Clarkson University

Background to this project TAD project with Air Innovations The origin of this project is a Technical Application and Demonstration (TAD) contract for the Center of Excellence to Air Innovations, Inc. to test the effectiveness of their HEPAIRx to reduce symptoms in asthmatic children. Clarkson is the subcontractor to Air Innovations and will be performing the field study in 45 homes.

Air Innovations HEPAIRx This system includes a HEPA filter with an air-to-air heat exchanger to provide approximately 12 air changes per hour.

Background to this Project [1] TAD project with Air Innovations The 45 subjects will be split into two groups approximately equal size. During the first part, group A has the cleaners and group B does not. During part 2, both groups have the cleaners, and in part C, Group A turns the cleaners off and Group B keeps theirs running. Each experimental period would run for ~ 6 weeks. Each room in which the HEPAIRx is deployed will be equipped with an Air Advice monitor for at least 1 week during the period when the unit is operating and 1 week when the unit is not operating. In addition, measurements will be made in as many homes as practical using portable condensation particle and optical particle counters.

Background to this Project [2] TAD project with Air Innovations Exhaled breath condensate samples will be collected using the Respiratory Research, Inc. RTube system. This system is specifically designed to be simple and easy to use without the supervision of a trained technician. The RTube captures the biomarker data on acute airway disease with the ability to non-invasively monitor a subject’s airway pH and other biomarkers such as nitrate, nitrite and RNOS (nitrosothiols integral in the mechanism of pulmonary inflammation experienced by asthmatics). Subjects exhale normally into the RTube while the moisture from their breath is condensed in the collection chamber. Each subject will collect a pair of tubes every 6th day for the 18 weeks of the study with the first tube being collected just before going to bed and the second collected on rising in the morning.

RTube EBC Collector This system includes a HEPA filter with an air-to-air heat exchanger to provide approximately 12 air changes per hour.

Resuspended Aeroallergen EPA03 Project [T6.A.4] Human inhalation of asthmagens in settled indoor dust can occur when the dust resuspends into the indoor air via human activity or air currents. Several studies have reported high indoor and personal airborne particle concentrations as a result of human activities indoors that resuspend indoor dust including the work being conducted by Prof. Ferro under EPA03 support. Airborne concentrations and size distributions of dust particles carrying cat allergen due to common household activities such as vacuuming, for example, have also been studied in the laboratory. However, similar studies have not been conducted in actual residences where environmental conditions can vary substantially from the laboratory setting. Paired indoor dust and airborne samples are necessary to understand the relationship between the settled dust and human exposure.

Resuspended Aeroallergen EPA ’03 Project The existing project offers the opportunity to study the influence of resuspension as a source of airborne allergens and compare the effects of different microenvironments on the expression of the asthma in terms of markers of inflammation. The specific aims of this project are as follows: To collect paired measurements of settled indoor dust and airborne particles to better understand inhalation exposure to pollutants of concern in the dust arising from resuspension. To examine the use of microenvironmental exposure models to estimate the personal exposure of the subjects to airborne allergens To examine the use of exhaled breadth condensate analysis as a quantitative measure of the effect of resuspended allergens to increase inflammation in asthmatic children with the relationships depending on the extent of the disease

Resuspended Aeroallergen EPA ’03 Project Sampling: Within the homes that we will be studying for the effect of the IERV, we propose an expanded monitoring program to determine the quantities of aeroallergens and endotoxins in settled dusts and in the indoor aerosol in selected rooms. The allergen/endotoxin sampling will involve personal PM10 sampling and paired settled dust and airborne PM10 sampling in the child’s three main microenvironments: In the child’s bedroom In the main living area of the child’s home In the child’s principal school classroom Sampling will occur for two 24-hr periods for 25 selected participants. The details of the sample collection are presented in Table 1.

Resuspended Aeroallergen Location 24-h PM10 Settled Dust Personal 2   Bedroom Living Room Classroom Total samples per participant 8 6 Total – selected participants (x 25) 200 150 Duplicates (add 10%) 20 15 Blanks (add 5%) 10 5 Study Total 230 170

Resuspended Aeroallergen EPA ’03 Project Aeroallergens using monoclonal antibody enzyme-linked immunosorbent assays Cat (Fel d 1), dust mite (Der p 1, Der f 1), cockroach (Bla g 1), and dog (Can f 1) Endotoxin using a recombinant DNA form of Factor C (PyroGene Recombinant Factor C Endotoxin Detection System and fluorescence For the subjects 10 and older, the severity of asthma will be ascertained using Methacholine challenge. The tests will be administered by Canton Potsdam Hospital and will permit the classification of the subjects’ asthma to mild, medium and severe. Exhaled breath condensate will be collected after school before going home, before bed and upon arising in the morning.

Resuspended Aeroallergen EPA ’03 Project It is expected that exposures to inhaled resuspended allergens (including dust mites and pet dander) will stimulate the inflammatory metabolic pathway and increase the production of nitric oxide and its metabolites, including nitrite and nitrate, along the lung lining, which will be measured in the exhaled breath as gas phase NO (measured from eNO) and NO2-, NO3-, and RNOS dissolved in the liquid phase measured from EBC condensate. It is expected that these exhaled biomarkers of pulmonary inflammation will be positively correlated with exposures to aeroallergens, and that a greater correlation will exist as a function of asthma severity.

Resuspended Aeroallergen EPA ’03 Project Current Status: QAPP and IRB Submitted and Approved Pilot has been conducted using 4 subjects on the TAD project Revised questionnaire for TAD developed and revised IRB submitted 9 new asthmatic kids scheduled for training About 10 more homes having 11-12 asthmatic kids have been identified Letters have been sent by Canton Potsdam Hospital to all of the childhood asthma cases handled by CPH physicians. Plate Reader has been ordered and we will develop the lab analyses in August PM10 PEMS have been acquired. Thus, the project is moving rapidly into operation.