Clinical trial NCT03444298
Conditions: Airway Inflammation Asthma
Sponsor: University of North Carolina, Chapel Hill
Start date: 2018-06-08; Completion date: 2023-03-06
Enrollment: 16 participants; Allocation: RANDOMIZED; Intervention model: CROSSOVER

Brief summary:
Purpose: To determine the efficacy of 1400 mg gamma tocopherol-enriched supplement for mitigating inhaled wood smoke particle-induced airway inflammation in healthy adults with no more than mild asthma.

Detailed description:
Particulate matter (PM) is a leading cause of respiratory tract and cardiovascular disease in the United States and world-wide. Wood smoke particles (WSP) derived from wildland and other fires account for a significant fraction of ambient air PM. Health effects associated with WSP include acute bronchitis, asthma exacerbation, pneumonia, cough and systemic inflammation. While these effects are seen in both healthy and asthmatic individuals, many studies indicate that asthmatics have increased susceptibility to the effects of WSP. The investigators have developed a 500 μg/m3 WSP exposure protocol (levels similar to those encountered by firefighters and residents in close proximity to wildland burn sites) that induces airway and systemic inflammation in healthy volunteers. As with other pollutants, these inflammatory responses modulate non-specific bronchial reactivity (NSBR), inflammatory cell recruitment to the airways (primarily neutrophils), and potentially cardiovascular function.

The investigators have focused on gamma tocopherol (γT) as a nutritional intervention to prevent inflammatory responses to air pollutants such as WSP. Building on animal and in vitro preclinical studies, the investigators have established that 1400 mg/day of oral γT-enriched supplement for 7 and 14 days in healthy volunteers and mild asthmatics, respectively, inhibited neutrophil influx into the airways, reduced production of sputum mucins, and improved mucociliary clearance following challenge with inhaled endotoxin, another common component of PM. The findings occurred in the context of significantly increased plasma concentrations of γT and its active metabolite 2,7,8-trimethyl-2-(β-Carboxy-Ethyl)-6-Hydroxychroman (γ-CEHC). Given the findings in these early phase clinical trials, γT supplementation is an attractive approach to prevent WSP-induced adverse health effects. The investigators propose to use γT supplementation in a human model of WSP inhalation to mitigate key features of airway inflammation: inflammatory cell recruitment, production of inflammatory cytokines and mucous, and changes in airway physiology.

Gamma tocopherol will be administered in softgel form, with each softgel containing 700 mg of tocopherols, 89.5% of which is d-gamma tocopherol. Subjects will consume two softgels by mouth once daily for 7 days. This dosing regimen was chosen based on the results of the investigators' previous early phase clinical trials examining the impact of gamma tocopherol on lipopolysaccharide (LPS) -induced airway inflammation in healthy adults and adults with asthma. These studies tested a 7 and 14 day course of treatment, respectively, and found similar plasma concentrations of γT and active metabolites in both studies. Furthermore, the investigators showed in both studies that γT significantly reduced LPS-induced sputum neutrophilia compared to placebo. Based on the previous findings, the investigators will now study the efficacy of γT for mitigating WSP-induced airway inflammation.

January 2022 update: The existing, custom source of gamma tocopherol expired during the protocol paused period during Covid. The Gamma t is replaced by Gamma E Gems, manufactured by Carlson Labs, each capsule with 577mg of gamma tocopherol, based on Certificate of Analysis. Subjects will continue to ingest 2 capsules at each dosing, for a total dose of 1154mg. The safflower oil placebo is replaced with a neutral oil capsule.

Arms: arm 1: Participants that are randomized to placebo treatment will take a short treatment course of Neutral Oil followed by chamber exposure with wood smoke particulate. After a 4-week washout period, participants will cross over to the gamma Tocopherol (active) treatment group. arm 2: Participants that are randomized γT treatment will take a short treatment course of gamma Tocopherol followed by chamber exposure with WSP. After a 4-week washout period, participants will cross over to the placebo treatment group.
Interventions: intervention 1: Gamma Tocopherol intervention 2: Placebo
Intervention descriptions: intervention 1: Each dose consists of two (700 mg) capsules by mouth once daily for a total of 7 days. intervention 2: Each dose consists of two capsules by mouth once daily for a total of 7 days.

Protocol documents:
Prot_SAP_000.pdf: 
 
 
A Study of Gamma Tocopherol-enriched Supplement on Lower Airway 
Responses to Inhaled Wood Smoke in Healthy Adults (SmokeyT) 
 
 
 
NCT number NCT03444298 
Document Date 01/24/2023 
 


Gamma Tocopherol (Vitamin E) and airway inflammatory responses, IND 136873 
Sponsor:  David Peden, MD, MS 
 
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6.0. PROTOCOL 
 
 
 
A PHASE II RANDOMIZED, DOUBLE BLINDED, PLACEBO 
CONTROLLED STUDY OF GAMMA TOCOPHEROL-ENRICHED 
SUPPLEMENT ON LOWER AIRWAY RESPONSE TO INHALED 
WOOD SMOKE 
 
 
Regulatory Sponsor/Primary  
Investigator:  
 
 
 
David Peden, MD, MS 
 
 
 
 
 
 
CEMALB 
 
 
 
 
 
 
104 Mason Farm Road 
 
 
 
 
 
 
CB# 7310 
 
 
 
 
 
 
Chapel Hill, NC 27599-7310 
 
 
 
 
 
 
(919) 843-2714 
 
Study Product: 
Gamma E Gems (J.R. Carlson Laboratories, Inc., 600 W University Drive 
Arlington Heights, IL 60004) 
 
 
 
 
 
 
 
 
 
 
 
Version Date: January 2023 
 


Gamma Tocopherol (Vitamin E) and airway inflammatory responses, IND 136873 
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Table of Contents 
STUDY SUMMARY ...................................................................................................................................... 5 
1 
INTRODUCTION .................................................................................................................................. 6 
1.1 
BACKGROUND ................................................................................................................................ 6 
1.2 
INVESTIGATIONAL AGENT ................................................................................................................ 7 
1.3 
DOSE RATIONALE AND RISK/BENEFITS ............................................................................................ 7 
2 
STUDY OBJECTIVES ......................................................................................................................... 8 
3 
STUDY DESIGN ................................................................................................................................... 9 
3.1 
GENERAL DESIGN .......................................................................................................................... 9 
3.2 
PRIMARY STUDY ENDPOINTS .......................................................................................................... 9 
3.3 
SECONDARY STUDY ENDPOINTS ..................................................................................................... 9 
3.4 
PRIMARY SAFETY ENDPOINTS ....................................................................................................... 10 
4 
SUBJECT SELECTION AND WITHDRAWAL .................................................................................. 11 
4.1 
INCLUSION CRITERIA .................................................................................................................... 11 
4.2 
EXCLUSION CRITERIA ................................................................................................................... 11 
4.3 
WITHDRAWAL OR TERMINATION ..................................................................................................... 13 
5 
STUDY PROCEDURES ..................................................................................................................... 13 
5.1  
SCHEDULE OF EVENTS ................................................................................................................. 13 
5.1.1 
Screening Visits ..................................................................................................................... 14 
5.1.2 
Visit 0: Baseline ..................................................................................................................... 14 
5.1.3 
Visit 1: Training/Dispense Study Drug .................................................................................. 14 
5.1.4 
Visits 2 and 5: WSP ............................................................................................................... 14 
5.1.5 
Visit 4: Dispense Study Drug ................................................................................................. 15 
5.1.6 
Visits 3 and 6: 24 Hours Post-Exposure ............................................................................... 15 
5.1.7 
Visit 7: Study Completion ...................................................................................................... 15 
5.1.8 
Table of Study Procedures .................................................................................................... 16 
5.2 
PROTOCOLS FOR MEASUREMENTS AND OBSERVATIONS .................................................................. 17 
5.2.1 
WSP Chamber Exposure ...................................................................................................... 17 
5.2.2 
Induced Sputum .................................................................................................................... 17 
5.2.3 
Spirometry ............................................................................................................................. 17 
5.2.4 
Flow Mediated Dilation/Brachial Artery Ultrasound ............................................................... 18 
5.2.5 
Left Ventricular Strain ............................................................................................................ 18 
5.2.6 
WSP Exposure Symptom Questionnaire .............................................................................. 19 
5.2.7 
GI Symptom Diary ................................................................................................................. 20 
6 
STATISTICAL PLAN ......................................................................................................................... 21 
6.1 
SAMPLE SIZE DETERMINATION ...................................................................................................... 21 
6.2 
STATISTICAL METHODS ................................................................................................................ 21 
6.3 
DATA MANAGEMENT ................................................................................................... 22 
7 
MONITORING AND RISK MINIMIZATION........................................................................................ 22 
7.1 
DEFINITION OF ADVERSE EVENT AND SERIOUS ADVERSE EVENT ...................................................... 22 
7.1.1 
Grading criteria ...................................................................................................................... 23 
7.1.2 
Protocol specific potential AE’s ............................................................................................. 23 
7.2 
RISKS TO SUBJECTS ..................................................................................................................... 24 
7.2.1 
Risks associated with Gamma Tocopherol ........................................................................... 24 
7.2.2 
Risks associated with WSP Exposure .................................................................................. 24 
7.2.3 
Risks associated with Induced Sputum Procedure ............................................................... 25 
7.2.4 
Risks associated with Spirometry.......................................................................................... 25 
7.2.5 
Risks associated with Measuring Flow Mediated Dilation (FMD) ......................................... 25 
7.2.6 
Risks associated with Measuring Left Ventricular Strain (LVS) ............................................ 26 


Gamma Tocopherol (Vitamin E) and airway inflammatory responses, IND 136873 
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7.2.7 
Risks associated with Other Study Procedures .................................................................... 26 
7.3 
MEASURES TO MINIMIZE RISKS ...................................................................................................... 26 
7.3.1 
Protections to Minimize Risks with Gamma Tocopherol ....................................................... 26 
7.3.2 
Protections to Minimize Risks with WSP Exposure .............................................................. 26 
7.3.3 
Protections to Minimize Risks with Induced Sputum Procedure ........................................... 27 
7.3.4 
Protections to Minimize Risks with Spirometry ..................................................................... 27 
7.3.5 
Protections to Minimize Risks with Measuring FMD ............................................................. 28 
7.3.6 
Protections to Minimize Risks with Measuring LVS .............................................................. 28 
7.3.7 
Protections to Minimize Risks with Other Study Procedures ................................................ 28 
7.3.8 
Protections to Minimize Risks with Breach of Confidentiality ................................................ 28 
7.4 
REPORTING OF AES AND SAES .................................................................................................... 29 
7.5 
INFORMED CONSENT .................................................................................................................... 29 
7.6 
CONFIDENTIALITY ......................................................................................................................... 29 
7.7 
INVESTIGATORS, FACILITIES, AND INSTITUTIONAL REVIEW BOARD .................................................... 29 
8 
REFERENCES ................................................................................................................................... 30 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
List of Abbreviations 
5-NO2-γT: 5-nitro-gamma tocopherol 


Gamma Tocopherol (Vitamin E) and airway inflammatory responses, IND 136873 
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γT: gamma tocopherol 
γ-CEHC: 2,7,8-trimethyl-2-(B-carboxy-ethyl)-6-hydroxychroman 
αT: alpha tocopherol 
α-TTP: alpha tocopherol transfer protein 
ANC: absolute neutrophil count 
AUC: area under the curve 
BAR: brachial artery reactivity 
BSA: body surface area 
COX-2: cyclooxygenase-2 
CYP3A: cytochrome P450 3A 
FA: filtered air 
FMD: flow mediated dilation 
FDR: fractional disappearance rate 
FEF25-75: forced expiratory flow during mid portion (25%-75%) of forced vital capacity 
FEV1: forced expiratory volume in 1 second 
FRC: functional residual capacity 
FVC: forced vital capacity 
GSTM1:  glutathione-S-transferase mu 1 
HF: high frequency 
HRV: heart rate variability 
IDS: investigational drug service 
LDMS: lab data management system 
LF: low frequency 
LPS: lipopolysaccharide 
LVS: left ventricular strain 
mT-γT: mixed tocopherol-γT 
NSBR: nonspecific bronchial reactivity 
O3: ozone 
PBMC: peripheral blood mononuclear cell 


Gamma Tocopherol (Vitamin E) and airway inflammatory responses, IND 136873 
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PC20: provocative concentration resulting in 20% reduction in FEV1 
PF: peak flow 
PGD2/PGE2: prostaglandin D2/prostaglandin E2 
PM: particulate matter 
PPM: parts per million 
PMN: polymorphonuclear leukocytes 
PNN50: mean number of times per hour in which the change in consecutive normal sinus (NN) 
intervals exceeds 50 milliseconds 
QTVI: QT variability index 
SDNN: standard deviation of normal to normal R-R intervals 
TEOM: tapered element oscillating microbalance 
TH1: T helper type 1 
VAPS: versatile air pollution sampler 
VLF: very low frequency 
WSP: wood smoke particles 
 
Study Summary 
Title 
A phase II randomized, double blinded, placebo-controlled study 
of gamma tocopherol-enriched supplement on lower airway 
responses to inhaled wood smoke in healthy adults 
Short Title 
Gamma tocopherol for wood smoke-induced airway inflammation 
IRB Number 
17-2303 
Phase 
II 
Study Duration 
3 years 
Study Center(s) 
University of North Carolina Hospitals in Chapel Hill, North 
Carolina, 27599-7310 
Objectives 
To determine the efficacy of 1100 mg gamma tocopherol-enriched 
supplement for mitigating inhaled wood smoke particle-induced 
airway inflammation in healthy adults with no more than mild 
asthma  
Number of Subjects 
35 subjects 


Gamma Tocopherol (Vitamin E) and airway inflammatory responses, IND 136873 
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Diagnosis and Main 
Inclusion Criteria 
Healthy adult volunteers with no more than mild asthma 
Study Product, 
Dose, Route, 
Regimen 
Gamma E Gems  (J.R. Carlson Laboratories, Inc., 600 W 
University Drive Arlington Heights, IL 60004).) 
 
Statistical 
Methodology 
Change in sputum % neutrophils (%PMNs) [Δ postWSP-preWSP 
(γT ) vs Δ postWSP-preWSP (placebo)]. Statistically significant 
differences will be determined using two-sample tests, either 
parametric (t-test) or non-parametric (Wilcoxon signed-rank test) 
depending on whether the normality assumption is met.  
 
1. Introduction 
This document is a protocol of a human research study. This study is to be conducted according 
to US and international standards for Good Clinical Practice (FDA Title 21 part 312 and 
International Conference on Harmonization guidelines), applicable government regulations and 
Institutional research policies and procedures. 
 
1.1 Background 
Asthma is among the most prevalent chronic diseases in the U.S. (1) and represents a source of 
significant burden to patients and healthcare systems. Acute exacerbation of asthma is 
characterized by airway inflammation, bronchoconstriction, increased production of airway 
mucous, and decreased mucociliary clearance with formation of mucous plugs (2, 3). While 
corticosteroids are often the treatment of choice, their use is associated with a number of adverse 
effects, particularly when administered systemically. Currently there is an unmet need for non-
steroidal treatment options for asthma, and Vitamin E has been proposed as one such treatment 
to combat inflammatory features of asthma.   
Among the isoforms of vitamin E that can be tested in asthma are α-tocopherol (αT), the 
predominant isoform found in over the counter Vitamin E supplements, and gamma tocopherol 
(γT), the predominant isoform found in foods. Intervention trials of αT in humans with asthma 
have been generally disappointing (4, 5). However, γT and its primary metabolite 2,7,8-
trimethyl-2-(B-carboxy-ethyl)-6-hydroxychroman (γ-CEHC) have been shown to have a number 
of anti-inflammatory activities not observed with αT and its metabolites (6-10). These actions 
include scavenging reactive nitrogen species to form 5-nitro-y-tocopherol (5-NO2-γT) (6) and 
inhibition of cyclooxygenase-2 (COX-2) and 5-lipooxygenase, reducing inflammatory 
eicosanoid production (7-9).  
Particulate matter (PM) is a leading cause of respiratory tract and cardiovascular disease in the 
United States and world-wide. Wood smoke particles (WSP) derived from wildland and other 
fires account for a significant fraction of ambient air PM. Health effects associated with WSP 
include acute bronchitis, asthma exacerbation, pneumonia, cough and systemic inflammation. 
While these effects are seen in both healthy and asthmatic individuals, many studies indicate that 
asthmatics have increased susceptibility to the effects of WSP. Our center has developed a 500 
μg/m3 WSP exposure protocol (levels similar to those encountered by firefighters and residents 


Gamma Tocopherol (Vitamin E) and airway inflammatory responses, IND 136873 
Sponsor:  David Peden, MD, MS 
 
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in close proximity to wildland burn sites) that induces airway and systemic inflammation in 
healthy volunteers. As with other pollutants, these inflammatory responses modulate non-
specific bronchial reactivity (NSBR), inflammatory cell recruitment to the airways (primarily 
neutrophils), and potentially cardiovascular function. 
Our team has focused on gamma tocopherol as a nutritional intervention to prevent inflammatory 
responses to air pollutants such as WSP. Building on animal and in vitro preclinical studies, we 
have established that 1400 mg/day of oral γT-enriched supplement for 7 and 14 days in healthy 
volunteers and mild asthmatics, respectively, inhibited neutrophil influx into the airways, 
reduced production of sputum mucins, and improved mucociliary clearance following challenge 
with inhaled endotoxin, another common component of PM (10, 11). The findings occurred in 
the context of significantly increased plasma concentrations of γT and its active metabolite γ-
CEHC. Given our findings in these early phase clinical trials, γT supplementation is an attractive 
approach to prevent WSP-induced adverse health effects. We propose to use γT supplementation 
in a human model of WSP inhalation to mitigate key features of airway inflammation: 
inflammatory cell recruitment, production of inflammatory cytokines and mucous, and changes 
in airway physiology. 
 
1.2. Investigational Agent 
 
Gamma E Gems 548 mg softgels,  (J.R. Carlson Laboratories, Inc., 600 W University Drive 
Arlington Heights, IL 60004.) 
 
1.3. Dose Rationale and Risk/Benefits 
 
Gamma tocopherol will be administered in softgel form, with each softgel containing 874.1 mg 
of tocopherols, , including: 
 
Alpha Tocopherol: 
141.0 mg/sg (softgel) 
Beta Tocopherol: 
  18.1 mg/sg 
Gamma Tocopherol:  548.0 mg/sg  
Delta Tocopherol:  
167.0 mg/sg 
 
Subjects will consume two softgels by mouth once daily for 7 days. This dosing regimen was 
chosen based on the results of our previous early phase clinical trials examining the impact of 
gamma tocopherol on LPS-induced airway inflammation in healthy adults (11) and adults with 
asthma (26). These studies tested a 7 and 14 day course of treatment, respectively, and found 
similar plasma concentrations of γT and active metabolites in both studies. Furthermore, we 
showed in both studies that γT significantly reduced LPS-induced sputum neutrophilia compared 
to placebo. Based on our previous findings, we will now study the efficacy of γT for mitigating 
WSP-induced airway inflammation.  
 
We have discussed the potential benefits of γT supplementation for protection against ambient 
air pollutant-induced airway inflammation in healthy volunteers and adults with mild asthma in 
section 1.1. In terms of risk, the most significant safety concern for Vitamin E supplements in 
general is increased bleeding risk, and most trials have used supplements containing 
predominantly αT as opposed to γT. The Alpha Tocopherol Beta Carotene cancer prevention 


Gamma Tocopherol (Vitamin E) and airway inflammatory responses, IND 136873 
Sponsor:  David Peden, MD, MS 
 
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study, a randomized, double blind, placebo controlled trial of αT and b-carotene supplementation 
in middle aged male smokers, found that alpha tocopherol supplementation was associated with 
increased risk of fatal subarachnoid hemorrhage (13). To our knowledge, there are no published 
studies of γT supplementation that show an increased risk of bleeding. In our own previous 
studies, we have not observed any significant bleeding events nor any changes in PT, aPTT, or 
INR with γT supplementation at doses similar to the proposed study. 
An additional area of concern was raised by findings from the Coronary Artery Risk 
Development in Young Adults (CARDIA) study, a population-based observational study. In this 
study, higher plasma γT levels in young adults (not taking Vitamin E supplements) were 
associated with lower lung function measurements (FEV1 and FVC) (14). However, others have 
shown that increased dietary vitamin E intake had protective effects on lung function in adults 
(15). Plasma γT levels have not been significantly associated with risk of asthma or asthma 
outcomes. However, increased dietary vitamin E, which is predominantly gamma tocopherol 
(γT), has been associated with decreased occurrence of asthma in both children (16-18) and 
adults (19). 
Based on our own previous studies, the most common adverse effects of γT are diarrhea, 
flatulence, abdominal bloating, and nausea. In our most recent randomized, placebo controlled 
crossover study evaluating 14 days of γT-enriched supplementation (1400 mg daily) in 23 adult 
volunteers with asthma (26), diarrhea was reported in 6 participants (26%) on active treatment, 
compared to 1 participant (4.3%) on placebo. Nausea was reported in 6 participants (26%) on 
active treatment, compared to 2 participants (8.7%) on placebo. Bloating and flatulence were 
reported in 2 participants (8.7%) and 1 participant (4.3%) on active treatment, respectively, with 
no reports of these symptoms in subjects receiving placebo. One participant withdrew from the 
study due to intolerable gastrointestinal symptoms. In all other participants, gastrointestinal 
symptoms were transient in nature, beginning after the first or second dose of study drug and 
self-resolving despite continued treatment for the 14-day period.  
 
2. Study Objective 
The primary objective of this study is to examine the effectiveness of gamma tocopherol for 
mitigating inhaled WSP-induced airway inflammation in healthy adults and mild asthmatics, 
specifically airway inflammatory cell recruitment and cytokine production, mucous production, 
and non-specific bronchial reactivity. 
 
 
 
 
 
3. Study Design 
 
3.1. General Design 
This study is designed to evaluate the effectiveness of gamma tocopherol for mitigating features 
of airway inflammation induced by inhaled WSP by examining airway inflammatory cell 
recruitment and cytokine production, and mucous production,. We anticipate 3 years will be 
required for completion of study visits for all participants. 
 


Gamma Tocopherol (Vitamin E) and airway inflammatory responses, IND 136873 
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Upon successful completion of two screening protocols (one as a general screen to determine 
eligibility for airway challenge studies and a second to identify those individuals who are 
deemed “responders” to WSP inhalation, defined as ≥10% increase in sputum PMNs following 
exposure), participants will be enrolled to participate in a randomized, double blinded, placebo-
controlled, crossover study consisting of 8 visits. Participants will be randomized to γT or 
placebo treatment followed by chamber exposure with WSP. After a 4-week washout period, 
participants will cross over to the alternate treatment group.  
Following randomization to placebo or gamma tocopherol treatment, subjects will return for 
baseline (V0) and training visits (V1). Participants will consume 1400 mg of γT-enriched 
supplement or placebo once daily for 7 days. Participants will undergo WSP exposure (V2), then 
return the following day for a 24 hour post-exposure visit (V3). After a 4 week wash out period, 
subjects will present for a review of medical history and adverse events and cross over to the 
alternate treatment (V4). They will undergo a second WSP exposure (V5) followed by a 24 hour 
post-exposure visit (V6). Five to 10 days after the exposure, participants will return for a study 
completion visit (V7).  
 
3.2. Primary Study Endpoint 
The primary endpoint will be change in sputum %PMNs [Δ (postWSP (4 hr) – preWSP)[γT] vs 
Δ (postWSP (4 hr)– preWSP)[placebo]] and change in sputum %PMNs [Δ (postWSP (24 hr) – 
preWSP)[γT] vs Δ (postWSP (24 hr)– preWSP)[placebo]]. 
 
3.3. Secondary Study Endpoints 
The secondary endpoints of the study will be change in peripheral blood absolute neutrophil 
count (ANC) in cells x109/L [Δ (postWSP (4 hr) – preWSP)[γT] vs Δ (postWSP (4 hr)– 
preWSP)[placebo]] and [Δ (postWSP (24 hr) – preWSP)[γT] vs Δ (postWSP (24 hr)– 
preWSP)[placebo]].   
 
Exploratory endpoints of interest include: 
1. Role of asthma as a modifier of γT treatment effect on WSP-induced airway 
inflammation 
2. Spirometry measurements – forced vital capacity (FVC), forced expiratory volume in 1 
second (FEV1), FEV1/FVC ratio 
3. Secretion of sputum mucins (total sputum mucins, MUC5AC, MUC5B) will be 
determined by Western blot  
4. Serum cytokine concentrations via Mesoscale: 
 
a. TH1 cytokines: IL-1b, IL-6, IL-8, TNFa 
5. Sputum cytokine concentrations via Mesoscale: 
 
a. TH1 cytokines: IL-1b, IL-6, IL-8, TNFa 
6. Flow mediated dilation (FMD) using brachial artery ultrasound 
7. Left ventricular strain (LVS) 
8. Serum concentrations of gT aT gCEHC and aCEHC. 
 
3.4. Primary Safety Endpoints 


Gamma Tocopherol (Vitamin E) and airway inflammatory responses, IND 136873 
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Safety Endpoints: Safety endpoints include respiratory symptoms such as wheezing or shortness 
of breath requiring albuterol, significant bleeding events, or significant change in blood 
coagulation markers (PT, aPTT) such that these measurements are outside of normal range, and 
adverse events (AEs) and severe adverse events (SAEs). Exploratory safety endpoints include GI 
symptoms with γT vs Placebo. Adverse event data will be collected through 7 days post study 
drug treatment for both treatment periods. UNC Chapel Hill Human Research Protection 
Program Standard Operating Procedures will be followed for reporting adverse 
events/unanticipated problems to the Biomedical IRB. All SAEs will be reported to the CDER of 
the FDA as well as to the UNC Biomedical IRB within 24 hours of recognition of the event. 
Adverse events will be reported to both the FDA and UNC IRB on no less than an annual basis, 
or when the protocol is completed.   
 
3.4.1. Criteria for safety prior to initiation of wood smoke inhalation challenge 
(Subjects not meeting these criteria the morning of challenge will not proceed with wood smoke 
inhalation challenge): 
1. FEV1 of at least 75% of predicted (without use of bronchodilating medications for 12 
hours) consistent with lung function of persons with no more than mild intermittent or 
mild persistent asthma.   
2. Baseline oxygen saturation of at least 93%. 
3. No history of viral respiratory tract symptoms within 4 weeks of challenge. 
4. No current symptoms of rhinorrhea, sneezing, nasal or ocular pruritus consistent with 
allergic rhinoconjunctivitis. 
3.4.2. Criteria for safety of a given individual following challenge which would suspend the 
individual from further participation in the study will include: 
1. No greater than 25% relative decrease in FEV1 or FVC from WSP pre-challenge values, 
which does not improve within the first 2 hours of challenge.  
2. FEV1 must recover without treatment to within 90 % of baseline within 24 hours. 
3. A symptom score no greater than 45 (representing a ranking of “moderate” on each of 15 
criteria, with a maximum score of 60, see section 5.2.8. below) with improvement to a 
score of 15 or less within 6 hours without therapy. 
4. Specifically, no symptom score greater than “moderate” (3 on a scale of 0-4) for 
shortness of breath or cough at 24 hours post-challenge. 
5. Oxygen saturation of ≥93% throughout the exposure period. 
6. If albuterol therapy is needed more than once during an observation period. 
7. Any significant abnormalities in PT and aPTT during the treatment periods will be 
assessed and reported to CDER, IRB, and the funding agency. 
 
Criteria for safety within the entire protocol (failure of which would result in suspension of 
further study until consultation with the CDER of the FDA and the UNC IRB) will include the 
following: 
 


Gamma Tocopherol (Vitamin E) and airway inflammatory responses, IND 136873 
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1. The study will be suspended if 3 of the first 10 participants fail the individual safety 
criteria outlined above. If this occurs, the study will remain suspended until consultation 
with the CDER of the FDA. 
2. Once 10 subjects have been enrolled, study suspension will occur if more than 25% of all 
participants fail the individual safety criteria outlined above 
3. No occurrence of any SAE 
 
Subjects will be under direct supervision for 6 hours from the start of WSP challenge and will 
have contact information for after-hours access to a study physician.  In the event of an 
unexpected event, such as wheezing or bronchospasm, the physician will follow the usual 
medical standard of care to treat the subject.   
 
4. Subject Selection and Withdrawal 
Preliminary results from our screening WSP exposure study have shown that approximately 57% 
of healthy volunteers were responsive to the inflammatory effects of WSP. Based on this 
experience, we estimate we will need to screen 50 volunteers to identify 28 who are responsive 
to inhaled WSP. Anticipating at attrition rate of 20%, we will need to identify 35 volunteers for 
the intervention study, requiring an overall screen of a minimum of 64 volunteers. There will be 
no gender or ethnic restrictions. The target completion goal is 28 healthy volunteers and mild 
asthmatics. Individuals who meet all of the following criteria are eligible for enrollment as study 
participants. 
 
4.1. Inclusion Criteria  
1. Age 18-45 years, inclusive, of both genders 
2. Negative pregnancy test for females who are not s/p hysterectomy with oopherectomy 
3. FEV1 of at least 75% of predicted (without use of bronchodilating medications for 12 hours), 
consistent with lung function of persons with no more than mild intermittent or mild persistent 
asthma. 
4. Oxygen saturation of ≥93% and blood pressure within the following limits: (Systolic between 
150-85 mmHg, Diastolic between 90-50 mmHg). 
5. Ability to provide an induced sputum sample. 
6. Subject must demonstrate a ≥ 10% increase in sputum neutrophils following inhaled WSP 
exposure, when compared to baseline sputum (to be completed in a separate protocol). 
7. Ability/willingness to discontinue inhaled corticosteroids, montelukast, and cromolyn for 2 
weeks without increased symptoms or increased need for beta agonist rescue medication prior to 
screening and through the course of the study. 
8.  Documented vaccine to Covid 19  
 
4.2. Exclusion Criteria 
Patients who meet any of these criteria are not eligible for enrollment as study participants: 
1. Clinical contraindications: 
a. Any chronic medical condition considered by the PI as a contraindication to the 
exposure study including significant cardiovascular disease, diabetes, chronic 
renal disease, chronic thyroid disease, history of chronic 
infections/immunodeficiency. 
b. Viral upper respiratory tract infection within 4 weeks of challenge. 


Gamma Tocopherol (Vitamin E) and airway inflammatory responses, IND 136873 
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c. Any acute infection requiring antibiotics within 4 weeks of exposure or fever of 
unknown origin within 4 weeks of challenge. 
d. Abnormal physical findings at the baseline visit, including but not limited to 
abnormalities on auscultation, temperature of 37.8° C, Systolic BP > 150mm Hg 
or < 85 mm Hg; or Diastolic BP > 90 mm Hg or < 50 mm Hg, or pulse oximetry 
saturation reading less than 93%. 
e. Physician directed treatment for an asthma exacerbation, including requirement 
for oral corticosteroids, within the preceding 12 months. 
f. Moderate or severe asthma 
g. Asthma symptoms more than 2 times a week, which would be characteristic of a 
person with moderate or severe persistent asthma.  
h. Daily requirement for albuterol due to asthma symptoms (cough, wheeze, chest 
tightness) which would be characteristic of a person with moderate or severe 
persistent asthma (not to include prophylactic use of albuterol prior to exercise) 
i. Requirement for asthma controller therapy greater than low dose ICS or 
leukotriene receptor antagonist. Persons requiring medium or high-dose ICS, 
ICS/LABA, ICS plus leukotriene receptor antagonists, omalizumab or other 
biologic therapies for asthma control will not be enrolled. 
a. Nighttime symptoms of cough or wheeze greater than 1x/week at baseline (not 
during a clearly recognized viral induced asthma exacerbation) which would be 
characteristic of a person of moderate or severe persistent asthma. 
b. History of intubation for asthma 
c. If there is a history of allergic rhinitis, subjects must be asymptomatic of allergic 
rhinitis at the time of study enrollment. 
d. Mental illness or history of drug or alcohol abuse that, in the opinion of the 
investigator, would interfere with the participant’s ability to comply with study 
requirements. 
e. Cigarette smoking > 1 pack per month 
f. Unwillingness to use if sexually active (IUD, implant, sterilization, or when used 
in combination: birth control pills/patch and barrier methods). (i.e., those that, 
alone or in combination, result in a failure rate of <1% per year when used 
consistently and correctly). Note that male condom use, hormonal contraceptive 
pills, or hormonal contraceptive patches used alone have estimated failure rates 
that do not meet these criteria. 
g. Abnormal PT or aPTT values at screening or during the treatment period. Normal 
values will be those published by the clinical lab (Labcorp, INC). 
h. Use of immunosuppressive or anticoagulant medications including routine use of 
NSAIDS. Oral contraceptives are acceptable, as are Antidepressants and other 
medications may be permitted if, in the opinion of the investigator, the medication 
will not interfere with the study procedures or compromise safety and if the 
dosage has been stable for 1 month. 
i. Orthopedic injuries or impediments that would preclude bicycle or treadmill 
exercise. 
j. Inability to avoid NSAIDS, Multivitamins, Vitamin C or E or herbal supplements. 
k. Allergy/sensitivity to study drugs or their formulations 
l. Known hypersensitivity to methacholine or to other parasympathomimetic agents 


Gamma Tocopherol (Vitamin E) and airway inflammatory responses, IND 136873 
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m. Unwillingness to avoid coffee, tea, cola drinks, chocolate, or other foods 
containing caffeine after midnight on the days that methacholine challenge testing 
is to be performed. 
n. Positive test for Covid 19 in the past 90 days 
2. Pregnant/nursing women and children (< 18 years as this is age of majority in North 
Carolina) will also be excluded since the risks associated with woodsmoke exposure to 
the fetus or child, respectively, are unknown and cannot be justified for this non-
therapeutic protocol. Individuals over 45 years of age will not be included due to the 
increased possibility of co-morbidities and need for prohibited medications. 
3. Inability or unwillingness of a participant to give written informed consent. 
 
4.3. Withdrawal or Termination 
Participants will be terminated early from the study for the following reasons: 
1. The participant elects to withdraw consent from all future study activities including 
follow up. 
2. The participant is “lost to follow up” (i.e., no further follow up is possible because 
attempts to reestablish contact with the participant have failed). 
3. The participant dies. 
4. The participant develops a medical condition or is started on a new medication(s) not 
previously mentioned in the list of prohibited medications that, in the opinion of the 
investigator, may pose additional risks from participation in the study, may interfere with 
the participant’s ability to comply with study requirements or that may impact the quality 
of the data obtained from the study. 
5. The participant meets any of the individual stopping rules as delineated. 
6. The study is terminated for any reason. 
Participants with early termination from this study will not be replaced.  
5. Study Procedures 
 
5.1. Schedule of Events 
Each subject will have 8 study visits (excluding the general screening visit and wood smoke 
exposure screening visit to identify responders, which are under separate protocols): 
 
5.1.1. Screening Visits 
General Screening: To identify non-asthmatic and asthmatic subjects suitable for WSP exposure, 
subjects will complete a general screening protocol (UNC IRB approved study #98-0799, 
Database and Screening Protocol for Research Studies of the Center for Environmental 
Medicine & Lung Biology). 
 
Screening WSP exposure: This protocol will identify subjects who experience a ≥ 10% increase 
in sputum neutrophilia after WSP exposure, compared to baseline sputum (UNC IRB approved 
study #15-1775, To identify persons who are susceptible to WSP-induced inflammation and 
examine the role of GSTM1 and other factors in this susceptibility). This protocol involves a 
detailed review of the subject’s medical history and medication use, lung function (spirometry), 
collection of an induced sputum sample, and WSP exposure.  


Gamma Tocopherol (Vitamin E) and airway inflammatory responses, IND 136873 
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14 
 
5.1.2. Visit 0: Baseline, determine eligibility (at least 1 month after screening WSP exposure): 
1. Obtain informed consent. 
2. Review any change in medical status since the last visit. 
3. Vital signs, oxygen saturation, resting blood pressure 
4. Urine pregnancy test for females 
5. Physical exam 
6. Obtain 12-lead EKG 
7. Spirometry 
8. Sputum induction  
9. Venipuncture (CBC with differential, serum cytokines, serum C-reactive peptide, lipid 
panel, LDH, fibrinogen, PT, aPTT, INR, gT and metabolite levels). 
10. HRV will be collected throughout the entire visit with a Spacelabs monitor. A 30 minute 
controlled rest will be performed at the end of visit.  
5.1.3. Visit 1: Training/Dispense Study Drug (Period 1) (≤ 1 month from baseline visit and at 
least 1 week prior to V2 Exposure Visit and up to 3 weeks) (we request participants to be fasting 
for this visit, therefore it is scheduled in the morning) 
1. Review any changes in medical status since the last visit. 
2. Vital signs, oxygen saturation, and GI symptom questionnaire. 
3. Urine pregnancy test for females. 
4. Spirometry 
5. Baseline flow mediated dilation (FMD) measurement 
6. Baseline left ventricular strain (LVS) measurement 
7. Volunteers will be trained on the cycle ergometer to reach an exercise level producing a 
minute ventilation of 20 L/min/m2 body surface area (BSA). Subjects will be monitored 
with a standard 3 lead ECG during exercise. 
8. Dispense study drug and symptom diary 
5.1.4. Visits 2 (Period 1) and 5 (Period 2): WSP exposure 
1. Vital signs, oxygen saturation, resting blood pressure 
2. GI and WSP Exposure symptom questionnaire 
3. Collect symptom diary 
4. Urine pregnancy test for females 
5. HRV will be collected throughout the entire visit with a Spacelabs monitor. The monitor 
will be worn until the 24 hours post exposure visit. 
6. Spirometry 
7. Venipuncture (CBC with differential, serum cytokines, serum C-reactive peptide, lipid 
panel, LDH, fibrinogen, gT and metabolite levels) 
8. 500 μg/m3 WSP chamber exposure for 2 hours, with intermittent exercise and rest. 
Subjects will alternate 15 minutes of exercise on a cycle ergometer (intensity needed to 
achieve 20 L/min/m2 BSA determined at baseline visit) with 15 minutes of rest. 
9. Symptom questionnaire (immediately prior to exit from the chamber) 
10. Spirometry and resting blood pressure (immediately following exit from the chamber) 
11. Lunch - low fat meal is provided. The meal must be the same during each exposure. 
12. Obtain 30 minute controlled rest HRV, FMD, and LVS (90 minutes post-WSP exposure) 


Gamma Tocopherol (Vitamin E) and airway inflammatory responses, IND 136873 
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15 
 
13. Venipuncture (CBC) with differential, serum cytokines, serum C-reactive peptide, lipid 
panel, LDH, fibrinogen) (4 hours post-WSP exposure) 
14. GI and WSP Exposure Symptom questionnaire (4 hours post-WSP exposure) 
15. Sputum induction (4 hours post-WSP exposure) 
 
5.1.5. Visit 4: Dispense Study Drug (Period 2) (following 4 week washout and at least 1 week 
prior to V5 Exposure Visit and up to 3 weeks) 
1. Review medical history and adverse events 
2. Urine pregnancy 
3. Vital signs, oxygen saturation, GI symptom questionnaire 
4. Dispense study drug 
5. Lung function testing for participants who have asthma 
 
5.1.6. Visits 3 (Period 1) and 6 (Period 2): 24 hours post-exposure 
1. Review any adverse events 
2. Vital signs, resting blood pressure, oxygen saturation 
3. WSP Exposure symptom questionnaire 
4. Venipuncture (CBC with differential, serum cytokines, serum C-reactive peptide, lipid 
panel, LDH, fibrinogen, PT, aPTT, INR, gT and metabolite levels) 
5. Spirometry 
6. Sputum induction 
7. .  
At least a 4 week, but no more than an 8 week washout is required between Period 1 and Period 
2.  
5.1.7. Visit 7: Study completion, 5-10 days post-exposure 
1. Review any adverse events 
2. Vital signs, oxygen saturation, symptom questionnaire 
3. If the participant has any health symptoms or concerns, they will receive a physical 
examination and evaluation by a study physician. 
We will contact participants 1 month after participation to follow up and ask about health status.  
If the participant develops an upper or lower respiratory tract illness between V0 and V1 or 
between periods, we will postpone the next study visit for a minimum of 4 weeks after resolution 
of symptoms. In this case, if V1 occurs >1 month from V0, or if V4 occurs >2 months from V3, 
to prioritize participant safety we will bring the subject in for an additional visit to collect safety 
labs, perform spirometry, and review interval history. 
If the participant has an illness mid-dosing of the gamma tocopherol/placebo or fails to adhere to 
the dosing regimen, participants will return 4 weeks after they have taken their last dose or 4 
weeks after resolution of symptoms.  At that visit, gamma tocopherol/placebo will be re-
dispensed.  To prioritize participant safety we will review interval history.  Subjects with an 
illness during dosing, we will additionally collect safety labs, and perform spirometry.   


Gamma Tocopherol (Vitamin E) and airway inflammatory responses, IND 136873 
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16 
 
All subjects with a symptomatic Covid 19 infection will be deferred for 90 days.  Baseline 
spirometry and safety labs will be assessed prior to any gamma tocopherol/placebo dosing and 
WSP exposure. 
 
5.1.8. Table of study procedures 
 
 
General 
Screen 
Screening 
WSP 
Exposure 
V0 
(Baseline) 
V1 
(Training) 
V2 (period 1) 
V5 (period 2) 
V4 
(Dispense 
study drug) 
V3 (period 1)  
V6 (period 2) 
V7 
(Completion 
visit) 
Consent 
 
X 
X 
X 
 
 
 
 
 
Review 
history/AE’s 
 
X 
X 
X 
X 
X 
X 
X 
X 
Vital Signs 
 
X 
X 
X 
X 
X 
X 
X 
X 
Urine hCG 
(females only) 
 
X 
X 
X 
X 
X 
X 
 
 
Dispense 
study drug 
 
 
 
X 
 
X 
 
 
WSP 
Symptom 
Questionnaire 
 
X 
 
 
X 
 
X 
 
GI symptom 
diary 
 
 
 
X 
X 
X 
 
 
Spirometry 
(FEV1) 
 
X 
X 
X 
X 
X 
X (asthmatic 
participants 
only) 
X 
 
Allergy Skin 
Prick Test 
 
X 
 
 
 
 
 
 
 
Physical Exam 
 
 
X 
X 
 
 
 
 
 
Venipuncture 
 
X 
X 
 
X 
 
X 
 
CV measure-
ments: FMD, 
LVS 
 
 
 
X (FMD, 
LVS only) 
X 
 
) 
 
Sputum 
Induction 
 
X 
X 
X 
 
X 
 
X 
 
2 hour WSP 
chamber 
exposure 
 
 
X 
 
 
X 
 
 
 
 
 
5.2. Protocols for Measurements and Observations 
 
5.2.1 Woodsmoke particle chamber exposure 
We will employ the exposure protocol described by (20). We will heat whiteoak wood on an 
electric heating element (Brinkmann, Dallas, Texas, USA) in a Quadrafile 3100 woodstove 
(Colville, Washington, USA) to generate wood smoke. The smoke will be extracted from the 
chimney and injected into the chamber air stream, and wood smoke particle concentration will be 


Gamma Tocopherol (Vitamin E) and airway inflammatory responses, IND 136873 
Sponsor:  David Peden, MD, MS 
 
17 
 
controlled using a tapered element oscillating microbalance (TEOM; Thermo Fisher Scientific, 
Franklin, Massachusetts, USA) to measure the chamber concentration. A DustTrak 8530 aerosol 
monitor (TSI, Shoreview, Minnesota, USA) will also be used as an alternative monitor for 
independent verification of proper TEOM operation. The particle number concentration will be 
measured with a Model 3022A CPC (TSI, Shoreview, Minnesota, USA) and the number and size 
distribution will be measured with a Model 3080L SMPS (TSI). The particle concentration will 
be verified by weighing filters obtained using a versatile air pollution sampler (VAPS; URG, 
Chapel Hill, North Carolina, USA). The chamber temperature and humidity will be controlled to 
approximately 22°C and 40%, respectively. The target average concentration of WSP we will 
use for this study will be 500 μg/m3, with mild exercise to increase minute ventilation (20 
L/min/m2 BSA) for 15 minute periods, alternating with 15 minute rest periods across an overall 2 
hour total exposure period.  
 
5.2.2. Induced sputum 
Sputum samples will be collected using a specific sputum induction protocol in which FEV1 and 
FVC will be measured to determine the baseline FEV1 and FVC values. Asthmatic individuals 
will be pretreated with 2 to 4 puffs of albuterol prior to the procedure, and post-albuterol 
spirometry will be collected. The FEV1 values that match a 10% and 20% fall from baseline will 
be calculated. An ultrasonic nebulizer will be filled with 15 mL of 3% hypertonic saline 
(inhalation grade for respiratory use only) to begin the test. The nebulizer will be set to the 
maximum output setting and turned on. The subject will be instructed to breathe normally (i.e. 
tidal breaths) through the nebulizer mouthpiece for 7 minutes. The saline will be nebulized 
through the mouthpiece in a jet stream and inhaled. The nose will not be occluded for this 
procedure. Prior to expectoration, subjects will blow their nose, rinse their mouth with water, and 
clear their throat to avoid the inclusion of non-airway fluid samples. The subject will also be 
encouraged to come off the mouthpiece at any time to cough if a sputum sample from the lower 
airways (i.e. not from the back of the throat) is ready for expectoration. The sample will be 
coughed into a sterile specimen jar and capped. Following repeat measurement of FEV1 after the 
first 7 minute inhalation period, the concentration of saline will be increased from 3% to 4%, 
provided the FEV1 falls by <10% from the baseline value. The same procedure will be followed 
for the final 7 minute inhalation period using 5% hypertonic saline. The nebulization will be 
stopped at that point, or earlier, if a sputum sample of good quality is obtained (i.e. visible 
sputum plugs). The 4% and 5% saline solutions will be mixed for each induction by study staff 
just prior to induction and a volume of 15-20 mL will be used on each occasion.  
5.2.3. Spirometry 
Spirometry will be performed according to American Thoracic Society guidelines. Subjects will 
inhale as deeply as possible, then exhale as rapidly and completely as possible into the 
spirometer. Measurements obtained from each maneuver include the FVC, the FEV1, the 
maximal mid-expiratory flow rate (FEF 25-75%) and the peak flow (PF). The largest FVC and 
FEV1, from at least 3 acceptable trials will be selected for analysis and subject must meet the 
appropriate inclusion/exclusion criteria based on comparison of the subject’s measurements to 
the predicted value for an individual of his/her gender, age, race and height with NHANES III as 
the predicted set. 
5.2.4. Flow mediated dilation measured by brachial artery ultrasound: 


Gamma Tocopherol (Vitamin E) and airway inflammatory responses, IND 136873 
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18 
 
Vascular endothelial function will be assessed by measuring flow-mediated dilation of the 
brachial artery. Ultrasound images of the right brachial artery proximal to the antecubital fossa 
will be acquired using a Philips EPIQ 7 ultrasound machine equipped with a high-frequency (L9-
3) broadband linear-array transducer. The baseline images will be obtained after 10 minutes of 
supine rest. Flow-mediated (endothelium-dependent) dilation will be assessed by determining the 
change in arterial diameter in response to reactive hyperemia. Reactive hyperemia will be 
induced by inflating a pneumatic occlusion cuff placed around the upper forearm to a 
suprasystolic pressure (approximately 200 mmHg) for five minutes. Images of the artery will be 
recorded for two minutes after cuff deflation. Gated end-diastolic images will be stored in a 
digital format on a personal computer for subsequent offline quantification. Measurements will 
be performed using customized software (Brachial Tools, Medical Imaging Applications, LLC). 
Arterial diameter will be measured from the lumen-intimal interfaces of the proximal and distal 
arterial walls. Data from at least ten end-diastolic frames will be averaged for the baseline 
measurement and from at least three frames at maximum dilation during reactive hyperemia 
FMD will be calculated as the percent change in diameter from baseline. 
 
5.2.5. Left ventricular strain 
LV strain will be assessed by measuring global longitudinal strain using 2-dimensional speckle 
tracking echocardiography.  Speckle tracking imaging is a relatively new technique with which 
myocardial mechanics can be quantified by tracking groups of intramyocardial speckles.  Global 
longitudinal strain, the average fractional change in myocardial segment length, is a more 
sensitive index of LV contractility than ejection fraction or other standard measures.  Apical 4-
chamber, 2-chamber, and long axis 2-dimensional ultrasound images of the heart will be 
acquired using a Philips EPIQ 7 ultrasound machine and stored for subsequent off-line analysis.  
Global longitudinal strain will be quantified using Philips QLAB advanced quantification 
software. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


Gamma Tocopherol (Vitamin E) and airway inflammatory responses, IND 136873 
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19 
 
5.2.7. WSP Exposure Symptom Questionnaire 
Please indicate if you are experiencing any of the symptoms or restrictions listed below, using 
the following scale to indicate the severity. Circle the number. 
 
0 = NONE 
 
 
 
(not present) 
1 = TRACE/NOTICED 
 
(barely detectable) 
2 = MILD/LIGHT 
 
 
(present, but not annoying) 
3 = MODERATE 
 
 
(present, but somewhat annoying) 
4 = SEVERE/HEAVY 
 
(present and very annoying and painful) 
______________________________________________________________________________ 
SYMPTOMS  
 
 
 
NONE     TRACE 
MILD   MODERATE SEVERE 
______________________________________________________________________________
_ 
1. HEADACHE 
0 
1 
2 
3 
4 
 
2. IRRITATION OF THE NOSE 
0 
1 
2 
3 
4 
 
3. STUFFY NOSE/SINUS CONGESTION 0 
1 
2 
3 
4 
 
4. RUNNY NOSE 
0 
1 
2 
3 
4 
 
5. DRY/SORE THROAT 
0 
1 
2 
3 
4 
 
6. PAIN on DEEP INSPIRATION 
0 
1 
2 
3 
4 
 
7. UNUSUAL FATIGUE OR TIREDNESS 0 
1 
2 
3 
4 
 
8. EYE IRRITATION 
0 
1 
2 
3 
4 
 
9. SHORTNESS OF BREATH 
0 
1 
2 
3 
4 
 
10. SNEEZING 
0 
1 
2 
3 
4 
 
11. COUGH 
0 
1 
2 
3 
4 
 
12. WHEEZING/WHISTLING in CHEST 
0 
1 
2 
3 
4 
 
13. CHEST TIGHTNESS 
0 
1 
2 
3 
4 
 
14. SWEATING 
0 
1 
2 
3 
4 
 
15. Other___________________________ 
0 
1 
2 
3 
4 
 
 
 
 


Gamma Tocopherol (Vitamin E) and airway inflammatory responses, IND 136873 
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5.2.8. GI Symptom Questionnaire 
Please indicate if you are experiencing any of the symptoms or restrictions listed below, using 
the following scale to indicate the severity. Circle the number. 
 
0 = NONE 
 
 
 
(not present) 
1 = MILD 
(minimally noticeable but would not cause you to stop 
normal activities such as going to work or school or 
physical exercise) 
2 = MODERATE 
(symptom level would not ordinarily cause you stop normal 
activities such as going to work or school although it might 
stop you from doing physical exercise) 
3 = SEVERE 
(symptom would cause you to consider not going to work 
or school or to seek medical attention such as calling a 
healthcare provider, going to student health, or going to the 
emergency room) 
______________________________________________________________________________ 
 
SYMPTOMS  
 
 
 
NONE     MILD 
MODERATE 
  SEVERE 
_____________________________________________________________________________ 
1. INCREASE IN STOOLS 
0 
1 
2 
3 
 
2. DIARRHEA 
0 
1 
2 
3 
 
3. BLOATING/FULLNESS 
0 
1 
2 
3 
 
4. STOMACH CRAMPING/PAIN 
0 
1 
2 
3 
 
5. INCREASE IN PASSING GAS 
0 
1 
2 
3 
 
6. VOMITING 
0 
1 
2 
3 
 
7. NAUSEA 
0 
1 
2 
3 
 
8. HEADACHE 
0 
1 
2 
3 
 
9. EASY BRUISING OR BLEEDING 
0 
1 
2 
3 
 
10. Other___________________________ 
0 
1 
2 
3 
 
 
 
 
 
 
 
 


Gamma Tocopherol (Vitamin E) and airway inflammatory responses, IND 136873 
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21 
 
6. Statistical Plan 
 
6.1 Sample Size Determination 
The primary endpoint of this study is Δ sputum %PMNs (post WSP – pre WSP [γT]) at 4 hour 
vs (post WSP – pre WSP [placebo]) at 4 hours,  and (post WSP – pre WSP [γT]) at 24 hours vs 
(post WSP – pre WSP [placebo]) at 24 hours. Preliminary analyses from our screening study of 
the effects of 500 µg/m3 WSP exposure on sputum neutrophilia in healthy volunteers and mild 
asthmatics demonstrated that, among “responders” (those participants with a ≥ 10% increase in 
sputum neutrophils following WSP exposure), the mean increase in sputum %PMNs was 
28.5±13%. In our studies of γT vs placebo for mitigating LPS-induced change in sputum 
%PMNs in healthy volunteers (11) and mild asthmatics (26), we found a γT treatment effect size 
of 14.5% ±25.9. Assuming β=0.8 and α=0.05, a sample size of 28 healthy volunteers and mild 
asthmatics would be required to detect the same γT treatment effect size. To account for a 25% 
attrition rate with human studies, we will inflate the sample size to 35 healthy volunteers and 
mild asthmatics. Importantly, we anticipate that we will decrease subject variability by only 
recruiting persons with ≥ 10% increase in WSP-induced %PMNs in sputum during the screeing 
protocol (the so-called “responders”). 
 
6.2. Statistical Methods 
Our analytic plan was developed in collaboration with Dr. Zhou, the biostatistician for the 
CEMALB, who will oversee all statistical analysis. For analysis of sputum and circulating PMN 
responses as well as cytokine changes within each treatment group, we will use (i) descriptive 
analysis, (ii) two-sample comparison test, and (iii) random effects regression analysis (Equation 
1) designed to take advantage of the crossover design (22). Specifically, we will use the tabular 
and graphic methods to describe/explore the basic relationship between the groups. We will then 
follow up by comparing the Δ sputum %PMNs (post WSP – pre WSP [γT]) vs (post WSP – 
pre WSP [placebo]) using two-sample tests, either parametric (T-Test) or nonparametric 
(Wilxocon signed-rank test), depending on whether the normality assumption is met. We 
will transform the data to achieve normality if needed. Finally, we will employ a random 
effect regression modeling technique (Equation 1) that allows us to assess treatment effect while 
accounting for potential period effect and carry-over effects. This approach is similar to those we 
have previously described to assess genotype effects (including the GSTM1 null genotype) on 
response to pollutants (11, 23-25) and is shown below. In all analyses, criterion for significance 
will be p≤0.05. To address potential missing data, we will employ multiple imputation methods 
to address this issue when appropriate; using techniques we have previously described (24).  
 
Linear mixed model approach for global unified analysis of data for secondary and exploratory outcomes: 
 
 
 
 
 
 
 
 
Y_11k=μ+π_1+τ_1+βX_k+s_1k+ϵ_11k; Y_12k=μ+π_2+τ_2+λ_1+βX_k+s_1k+ϵ_12k 
 
Y_21k=μ+π_1+τ_2+βX_k+s_2k+ϵ_21k; Y_22k=μ+π_2+τ_1+λ_2+βX_k+s_2k+ϵ_22k     
μ is the grand mean, π1 and π2 are period effects, τ1 and τ2 are treatment effects, λ1 and λ2 
are carryover effects for different sequences, ς1k and ς2k are individual random effects 
within each sequence, and ε's are random errors. Subscript k is for subjects. 


Gamma Tocopherol (Vitamin E) and airway inflammatory responses, IND 136873 
Sponsor:  David Peden, MD, MS 
 
22 
 
 
6.3. 
Data Management 
Study coordinators will be responsible for efforts and computations for database management. 
Data will be initially collected by study coordinators on paper documents. Clinical data such as 
health history will be collected by study staff as interview or directly measured, such as vital 
signs, spirometry, etc. All paper forms (source documentation) are maintained by a study 
coordinator or PI in a binder, 1 binder per subject. All data will be entered into REDCap by a 
study coordinator within 2 weeks of collection, and will be verified by a 2nd person within 4 
weeks to ensure accuracy and completeness. REDCap is a secure web application developed at 
Vanderbilt University and can be used to collect virtually any type of data (including 21 CFR 
Part 11, FISMA, and HIPAA-compliant environments). It is specifically geared to support online 
or offline data capture for research studies and operations. Missing data will be noted and a 
comment made in the database to explain the missing data. Time stamps will be used when 
appropriate. At minimum, the visit number will be indicated.  
 
Biological samples collected by coordinators will be delivered to the CEMALB lab by the 
coordinators in properly labeled containers. On receipt in the lab, samples are initially entered 
into a software program called LDMS (Lab Data Management System) which tracks samples. 
Processing of these samples is done per the SOP maintained in the CEMALB quality assurance 
plan, as some samples are processed immediately and others are processed in batches at later 
time points. This data is entered into REDCap at appropriate intervals, based on sample analysis. 
 
We will use the data dictionary or REDCap codebook as our course for data codes. All data will 
be collected by qualified staff using CEMALB SOPs and will be entered into REDCap. Once the 
data is entered and confirmed by a 2nd person, it will be considered “locked” and will not be 
changed without documentation with the reason for the change. REDCap limits and monitors 
who can change data, a feature developed to ensure data integrity. 
 
6.4. Randomization plan 
The randomization schedule will be generated by Dr. Haibo Zhou (biostatistician) and provided 
to the investigational pharmacy. Only Dr. Zhou and the pharmacist will have access to the 
randomization schedule. Subjects will be allocated to begin the first period of the crossover study 
with placebo or γT treatment using permuted block randomization with a block size of 4 (2 
placebo, 2 γT for the first treatment period of the protocol).  
 
7. Monitoring and Risk Minimization 
 
7.1. Definition of Adverse Event (AE) and Serious Adverse Event (SAE) 
Adverse events and serious adverse events will be recorded and reported according to 21 CFR 
312.32. An adverse event for a given volunteer will be defined as any untoward medical 
occurrence associated with the use of a drug whether or not considered drug related and will also 
include failure of any of the safety criteria outlined in section “3.4.”. Additionally, minor upper 
respiratory tract infections occurring within 96 hours of the exposure will be considered adverse 
events.  Other non-specified clinical illnesses or symptoms, which occur during the treatment 
period or within 96 hours of each challenge, will also be reported as an AE.  Any decrease in 
lung function or increase in symptom score, as outlined in section “3.4,” will be considered an 


Gamma Tocopherol (Vitamin E) and airway inflammatory responses, IND 136873 
Sponsor:  David Peden, MD, MS 
 
23 
 
adverse event. Any symptoms that induce a volunteer to seek medical attention from any 
provider during study drug treatment or within 7 days of completing study drug treatment or 
within 96 hours of challenge will be considered an adverse event.  A serious adverse event will 
also be defined as any event that requires hospitalization or results in life threatening illness or 
injury, permanent (or likely to be permanent) illness or injury, or death if these events occur 
during study drug treatment or within 7 days of completing study drug treatment or within 96 
hours of challenge (or if the clinical scenario leading up to hospitalization, illness, injury or death 
begins during study drug treatment, within 7 days of completing study drug treatment or within 
96 hours of a challenge).   
7.1.1. Grading criteria 
In addition to determining whether an adverse event fulfills criteria for a serious adverse event or 
not, the severity of adverse events experienced by study participants will be graded according to 
the criteria set forth in the Toxicity Grading Scale for Healthy Adult and Adolescent Volunteers 
Enrolled in Preventive Vaccine Clinical Trials except for those items listed in 7.1.2 below.   This 
document provides a common language to describe levels of severity, to analyze and interpret 
data, and to articulate the clinical significance of all adverse events.   
All adverse events whether or not listed in the Toxicity Grading Scale will be graded on a 
scale from 1 to 4 according to the following standards (A semi-colon indicates ‘or’ within the 
description of the grade.): 
Grade 1 = Mild; asymptomatic or mild symptoms; clinical or diagnostic observations only; 
intervention not indicated. 
Grade 2 = Moderate; minimal, local or noninvasive intervention indicated; limiting age-
appropriate instrumental ADL (preparing meals, shopping for groceries or clothes, using 
the telephone, managing money, etc). 
Grade 3 = Severe or medically significant but not immediately life-threatening; 
hospitalization or prolongation of hospitalization indicated; disabling; limiting self care 
ADL (bathing, dressing and undressing, feeding self, using he toilet, taking medications, 
and not bedridden).  
Grade 4 = Life-threatening consequences; or urgent intervention indicated. 
 
7.1.2. Protocol specific potential AE’s 
Adverse events which have relative specificity for this protocol will be recorded and graded 1 to 
4 according to the grade definition provided below:  
 
 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
Potentially life-
threatening 
(Grade 4) 
Gastrointestinal 
distress  
(i.e. diarrhea, 
nausea, 
vomiting, 
flatulence) 
Asymptomatic 
or mild 
symptoms; 
clinical or 
diagnostic 
observations 
only; 
Moderate; 
minimal, local 
or noninvasive 
intervention 
indicated; 
limiting age 
appropriate 
Severe or 
medically 
significant but not 
immediately life-
threatening; 
hospitalization or 
prolongation of 
Life-threatening 
consequences; 
urgent 
intervention 
indicated 


Gamma Tocopherol (Vitamin E) and airway inflammatory responses, IND 136873 
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24 
 
 
intervention 
not indicated 
instrumental 
ADL 
existing 
hospitalization; 
disabling; 
limiting self care 
ADL 
Coagulopathy 
with or without 
bleeding 
Asymptomatic 
or mild 
symptoms; 
clinical or 
diagnostic 
observations 
only; 
intervention 
not indicated 
Moderate; 
minimal, local 
or noninvasive 
intervention 
indicated; 
limiting age 
appropriate 
instrumental 
ADL 
Severe or 
medically 
significant but not 
immediately life-
threatening; 
hospitalization or 
prolongation of 
existing 
hospitalization; 
disabling; 
limiting self care 
ADL 
Life-threatening 
consequences; 
urgent 
intervention 
indicated 
 
7.2. Risks to Subjects 
 
7.2.1. Potential risks of gamma tocopherol 
There is a rare chance that higher doses of αT may effect coagulation (PT and aPTT) and 
predispose to bleeding. It is possible that γT carries the same risk, though this has not been 
demonstrated in prior studies of γT. An observational study of un-supplemented individuals 
suggested that higher plasma γT levels were associated with reduced lung function, specifically 
FVC and FEV1 (14). Finally, our prior studies with γT have demonstrated risk for mild 
gastrointestinal symptoms, including diarrhea, nausea, bloating, and flatulence that tended to 
occur early on during treatment and to self-resolve within a few days despite continued 
treatment. In our most recent randomized, placebo controlled crossover study evaluating 14 days 
of γT supplementation in adults with asthma, diarrhea was reported in 26% of subjects on active 
treatment, compared to 4.3% of subjects on placebo.  
 
7.2.2. Potential risks of WSP 
Concentrations of wood smoke exhaust proposed for this study (500 μg/m3) are below those 
reported for individuals using biomass burning for heating and food preparation and those living 
in the area of a forest fire or agricultural burning. No major adverse health effects were observed 
in a recently completed study which exposed healthy volunteers to concentrated Chapel Hill air 
particles at the EPA Human Studies facility (Physiological and Biochemical Changes Associated 
with Exposure to Air Pollution Particles; Andrew Ghio, MD, PI; IRB# 95-EPA-310)1. The mean 
particulate matter (PM) was 485 + 84 μg/m3 during the WSP exposure (vs. “below the detectable 
limit” for filtered air (FA)), and analyses of chamber concentrations of total hydrocarbon and 
carbon monoxide during WSP exposure were also increased compared to FA (4.8 ± 0.5 vs. 2.0 ± 
0.1 ppm and 2.7 ± 0.7 vs. 0.0 ± 0.0 ppm, respectively). However, these healthy subjects had no 
significant change in pulmonary function and no nasal irritation, rhinitis, inspiratory pain, 


Gamma Tocopherol (Vitamin E) and airway inflammatory responses, IND 136873 
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25 
 
shortness of breath, cough, wheezing or chest tightness after exposure to WSP or filtered air. 
There were complaints of fatigue, headache and eye irritation, but these were noted by volunteers 
as frequently before as after both the filtered air and particle exposures. Pulmonary function 
testing demonstrated no significant changes from pre-exposure values following exposures to 
either filtered air or WSP. 
There is potential for increased airway inflammation in asthmatics. Thus, we are limiting 
inclusion to mild asthmatics only. Seven participants with asthma have completed an ongoing 
screening protocol (UNC IRB approved study #15-1775, To identify persons who are susceptible 
to WSP-induced inflammation and examine the role of GSTM1 and other factors in this 
susceptibility) to identify mild asthmatics who experience a ≥ 10% increase in sputum 
neutrophilia after WSP exposure for inclusion into this study. In addition to the mild, transient 
symptoms experienced by healthy volunteers as noted above, these seven asthmatic participants 
have experienced transient chest tightness following WSP exposure. This symptom has not been 
accompanied by any significant changes in spirometry, and chest tightness has resolved 
spontaneously within a few minutes following exposure and without any intervention. We will 
assess for any symptoms following WSP exposure with the WSP exposure questionnaire 
immediately after exit from the chamber and again 4 hours later. Spirometry will also be 
performed immediately following exit from the chamber with additional spirometry to be 
performed for any persistent symptoms and/or physician discretion. If FEV1 drops by 20% or 
more immediately after wood smoke exposure, or if the subject has intolerable symptoms, 
albuterol will be given for rescue. Finally, our Center has previously performed inhaled 
challenges with ozone and LPS in the allergic asthmatic population. With ozone and LPS we 
have not experienced any serious adverse events, and we expect the response to the wood smoke 
challenge to be similar.  
 
7.2.3. Potential Risks of the Induced Sputum Procedure 
The saline solutions (3%, 4% and 5%) nebulized during sputum may cause throat irritation, but it 
is uncommon for the duration of discomfort to last more than a few minutes post procedure. The 
procedure also has a minimal risk of inducing bronchospasm, which is a potential concern for 
individuals with asthma. Thus, all volunteers with a history of asthma will be pre-treated with 2-
4 puffs of albuterol prior to beginning sputum induction. Additional albuterol is employed as a 
rescue agent if needed, and a physician is available for all sputum induction procedures.  
 
7.2.4. Potential Risks of Spirometry 
Potential risks include possible lightheadedness or wheezing. 
 
The adhesive patches used during monitoring may cause itching or skin irritation. 
 
7.2.5. Potential Risks of Measuring Flow Mediated Dilation with Brachial Artery 
Ultrasound 
There are no significant risks associated with ultrasound imaging of the brachial artery, or with 
the 5 minutes of reduced blood flow that is part of the test. Cutting off blood flow to the arm may 
result in mild discomfort or temporary sensations of tingling or numbness in the hand until the 
blood pressure cuff is released. About 1 in 200 patients develop a painless rash on the arm where 
the blood pressure cuff is placed; this disappears over several days. 


Gamma Tocopherol (Vitamin E) and airway inflammatory responses, IND 136873 
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26 
 
 
7.2.6. Potential Risks of Measuring Left Ventricular Strain 
There are no significant risks associated with ultrasound imaging to measure left ventricular 
strain. 
7.2.7. Potential Risks associated with other study procedures 
Bruising during the venipuncture and vasovagal response to phlebotomy are the anticipated risks 
of venipuncture. There are no anticipated risks for completion of questionnaires or collection of a 
urine sample from female subjects. Light bicycle exercise (used to increase minute ventilation) 
presents a potential, but minimal risk. Side effects of exercise include occasional muscle 
soreness, cramps or general fatigue. These effects are temporary and not typically harmful to 
volunteers. It is also possible that exercise might uncover a previously unidentified pre-existing 
cardiac condition that could present a health risk to a volunteer. 
  
Additionally there is a risk of Breach of Confidentiality and Privacy. 
 
7.3. Measures to Minimize Risk 
 
7.3.1. Protections to Minimize Risk of Gamma Tocopherol 
Tocopherols may affect coagulation. We will query volunteers for bruising and bleeding prior to 
enrollment and during the dosing period, and will monitor PT and aPTT values prior to the first 
dose of study drug and at the end of the dosing period. They may be checked at any time the 
subjects experiences abnormal bruising. Any participant with an abnormal baseline PT, aPTT or 
INR will be excluded from receipt of study treatment. Spirometry is performed at regular 
intervals throughout the γT dosing period to monitor for any changes in lung function. As to the 
incidence of gastrointestinal symptoms, we will recommend that γT supplements be taken with a 
high-fat meal to promote intestinal absorption of γT and mitigate many associated diarrhea or 
bloating. Participants will receive a GI symptom diary to be completed daily while taking the 
study drug and returned to study coordinators on the morning of the challenge visits (Visit 2 or 
5). All female subjects, except those who have had a hysterectomy with oophorectomy, will 
undergo urine pregnancy testing prior to beginning treatment and prior to WSP exposure. A 
positive pregnancy test will exclude the subject, and they will be withdrawn if they become 
pregnant during the course of the study. Study medication, including placebo, will be dispensed 
by the Investigational Drug Service of UNC Hospitals. 
 
7.3.2. Protections to Minimize Risk of Wood smoke exposure 
Prior to enrollment, subjects will have completed either the CEMALB screening protocol at 
which time a detailed medical history will be obtained. Only otherwise healthy volunteers who 
meet study inclusion/exclusion criteria will be considered for this study, and the maximum age 
of participants will be 45 years in order to limit the likelihood of occult disease. Subjects will be 
deferred if they have had a viral upper respiratory tract infection within 4 weeks of exposure 
challenge. Also, unspecified illnesses, which in the judgment of the investigator increase the 
risks associated with woodsmoke, will be a basis for exclusion. All subjects will need to fulfill 
objective lung function and symptom criteria outlined above in section 3.4.1. prior to chamber 
entry. During wood smoke exposure, subjects will be monitored by direct observation or via 
closed-circuit television. Subjects will have ECG telemetry leads attached to monitor cardiac rate 
and rhythm during exercise throughout the exposure. Subjects will be aware that they can 


Gamma Tocopherol (Vitamin E) and airway inflammatory responses, IND 136873 
Sponsor:  David Peden, MD, MS 
 
27 
 
terminate their exposure for any reason and still receive compensation for their participation up 
to that point. A physician familiar with the protocol will be available for all challenge 
procedures. The investigator or duty physician will end the exposure if the subject is found to be 
suffering from any major adverse effect. An emergency “crash cart” with standard emergency 
medications, IV fluids and a defibrillator are is also readily available at the CEMALB in the 
unlikely event of a medical emergency during any challenge or study visit.  
 
For safety reasons, pulse oximetry will be performed during the exposures to wood smoke and 
filtered air, and the subject will be withdrawn from the chamber if the value is <90%. The risk of 
discomfort associated with exercise will be limited because ability and response to exercise will 
be assessed at the training visit, and subjects with orthopedic issues that limit ability to pedal a 
bicycle will be excluded. All female subjects, except those who have had a hysterectomy with 
oophorectomy, will undergo urine pregnancy testing on the morning (prior to) of each exposure. 
A positive pregnancy test will exclude the subject. Various engineering system controls will be 
used during woodsmoke exposures. A Tapered Element Oscillating Microbalance (TEOM®) 
will be used control and monitor wood smoke particle concentration in the chamber. If the 
concentration measured by the TEOM® exceeds a predetermined level, the exposure will be 
terminated and the subject removed from the chamber. A DataRAMTM particulate measurement 
device will also be used during exposure as an alternative monitor for independent verification of 
proper TEOM® operation. Additionally, gas analyzers continually monitor the chamber 
environment during exposures and will alarm if the carbon monoxide level exceeds 35 ppm. 
 
7.3.3. Protections to Minimize Risks of the Induced Sputum Procedure 
Throat irritation will be minimized by providing the subject with a drink (water, juice etc) and a 
snack after induction. All subjects with a history of asthma will be pretreated with 2-4 puffs of 
albuterol prior to procedure initiation. An additional albuterol multidose inhaler will be 
immediately available in the event the subject experiences symptoms of bronchoconstriction. A 
physician on duty in the facility will be available during sputum inductions. If the covering 
physician feels that the subject’s respiratory status is such that providing an induced sputum 
sample would place them at increased risk for significant bronchospasm, sputum sampling will 
be deferred. Baseline FEV1 and force vital capacity (FVC) will be measured before the start of 
the induction and at the end of each level (concentration) of saline inhalation. The FEV1 values 
that match a 10% and 20% reduction from baseline will be calculated and recorded. Subjects 
who have a 10-20% decrease in FEV1 after the 3% or 4% level of saline will not be advanced to 
a higher concentration of saline and will continue at the same level. If the FEV1 drops by > 20%, 
the induction will be stopped and the subject will be treated with 2 puffs of albuterol. Additional 
albuterol may be given if needed. 
 
7.3.4. Protections to Minimize Risks of Spirometry 
Subjects will be seated in a non-rolling chair when spirometry is performed and standard 
methodology conforming to the American Thoracic Society guidelines for measurement of 
spirometry will be used. Subjects will be instructed to notify the study staff if they feel 
lightheaded, and albuterol will be available in the event the subject experiences any 
unanticipated bronchoconstriction. 
 
7.3.5. Protection to Minimize Risks Associated with FMD 


Gamma Tocopherol (Vitamin E) and airway inflammatory responses, IND 136873 
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28 
 
If any subject finds this procedure intolerable, he or she will be withdrawn. In the rare event of a 
rash in the area where circulation is restricted, the subject will be monitored until it resolves. 
 
7.3.6. Protections to Minimize Risks Associated with Measuring Left Ventricular Strain 
If any subject finds this procedure intolerable, he or she will be withdrawn. 
 
7.3.7. Protections to Minimize Risks Associated with Other Study Procedures 
In order to minimize harm from the procedure, venipuncture will only be performed by qualified 
personnel. Subjects will have access to juice and a small snack after blood draws and will have 
the opportunity to lie recumbent after the procedure. Subjects with a known serious vagal 
response to phlebotomy will not be enrolled. A fully stocked medical “crash” cart will be present 
and readily available in the unlikely event that a major vagal episode occurs. Subjects will be 
screened for orthopedic injuries or conditions which would increase the likelihood of injury. 
Additionally, subjects will be trained on the proper use of the bicycle, and will be monitored 
during bicycle use. 
 
7.3.8. Protections for Minimizing Risks Associated with Breach of Confidentiality and 
Privacy 
All phone conversations/interviews as well as collection of personal medical history will be 
carried out in private offices or private examination rooms in the CEMALB research facility. All 
physical examinations will be carried out in a private examination room. Access to subject’s 
identity will be limited to the investigators and staff directly involved in the protocol. 
CRF’s/worksheets generated as part of these studies will be kept in a locked room, and the 
database will not contain personal identifiers and will also be password protected and accessible 
only to the appropriate study coordinators and investigators. Subjects’ names will not be used in 
any publication. Risks to Confidentiality will be also be minimized by labeling samples for in-
house analysis with coded identifiers. The ID label for any samples sent outside of the UNC 
Health Care System will be also be coded and without any personal identifiers (i.e., no DOB, 
gender etc will be indicated on the tube or lab request form). Clinical blood samples (PT, aPTT, 
WBC) are typically sent to Labcorp INC, Burlington, NC, without identifiers. All subjects are 
discharged to home with 24 hour contact information for a physician who is familiar with the 
study. 
 
 
7.4. Reporting of AEs and SAEs 
All SAEs will be reported to the CDER of the FDA as well as to the UNC Biomedical IRB 
within 24 hours of recognition of the event. Adverse events will be reported to both the FDA and 
UNC IRB on no less than a quarterly basis, or when the protocol is completed.  If criteria for 
suspension of the protocol are met, then the FDA and UNC IRB will be notified within 24 hours.  
All subjects with a non-fatal SAE will be evaluated medically by a study physician, in concert 
with their own physician as appropriate. Likewise all subjects with an adverse event will be 
examined and evaluated by a study physician. All assessments will include the same lung 
function and vital sign assessments outlined for challenge observation. Other assessments will be 
undertaken as needed. Any unspecified event, which in the judgment of the PI of the study, 
constitutes an unusual, unexpected or prolonged event (greater than 96 hours) will be reported to 
both the FDA and the UNC IRB.  


Gamma Tocopherol (Vitamin E) and airway inflammatory responses, IND 136873 
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29 
 
 
7.5. 
Informed Consent  
Investigators and study staff will explain all study procedures and the benefits and risks of the 
study to potential participants as part of obtaining informed consent. Subjects may withdraw 
their consent at any time during the study.  If they withdraw from the study it will not impact the 
care they receive at UNC or its affiliated hospitals and clinics.  
 
7.6. 
Confidentiality  
Risks to subject confidentiality will be minimized by storing records with personal identifiers in 
an office in CEMALB which is locked when unattended by the study coordinators. All samples 
will be stored with codes only (no personal identifiers). The CEMALB is located in the US 
Environmental Protection Agency’s Human Studies Facility on the UNC campus, which has a 
security guard and limited access 24 hours/day, 7 days/week.  
 
All data and specimens will be coded with unique identifiers assigned to the subject at the time 
of enrollment, and actual identity cannot be ascertained exclusively from these data. 
Conversations and interviews with the subject, as well as collection of personal medical history, 
will be carried out in private examination rooms or private offices in the CEMALB research 
facility. All physical examinations will be carried out in a private examination room. Access to 
study records containing identifiable private information will be provided only to those 
individuals associated with the study who require access to the data to perform their duties. 
These individuals will have completed ethics training as required by the UNC Biomedical IRB 
and will be listed on the approved IRB protocol. Records containing PHI will be kept in a locked 
office. 
 
7.7. Investigators, Facilities, and Institutional Review Board 
 
Principal 
Investigator 
David B. Peden, MD, MS 
CEMALB, 104 Mason Farm Road 
The University of North Carolina 
CB#7310  
Chapel Hill, NC 27599-7310 
Professor of Pediatrics & 
Director, Center for 
Environmental Medicine, 
Asthma and Lung Biology  
Co-Investigator 
Neil Alexis, PhD 
CEMALB, 104 Mason Farm Road 
The University of North Carolina 
CB#7310 
Chapel Hill, NC 27599-7310 
Assistant Professor of Pediatrics 
& Investigator, 
Center for Environmental 
Medicine, Asthma, and Lung 
Biology 
Co-Investigator 
Haibo Zhou, PhD 
3104C McGavran-Greenberg Hall 
The University of North Carolina 
CB #7420 
Chapel Hill, NC 27599 
Professor of Biostatistics 
Director of the Biostatistics 
Core, CEMALB 


Gamma Tocopherol (Vitamin E) and airway inflammatory responses, IND 136873 
Sponsor:  David Peden, MD, MS 
 
30 
 
 
Curriculum Vitae for investigators are appended to this application. 
 
Facilities: Volunteers for these studies will be recruited, screened and undergo challenge 
procedures at the Center for Environmental Medicine, Asthma and Lung Biology, CB#7310, 104 
Mason Farm Road, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 
27599-7310. All necessary clinical research equipment, medical equipment, and laboratory 
equipment is located within the Center. 
 
Institutional Review Board: This study will be reviewed and approved by the  
Biomedical IRB for the UNC School of Medicine, The University of North Carolina at Chapel 
Hill CB# 7097, Medical School Building 52 Chapel Hill, NC 27599-7097 prior to subject 
enrollment. 
 
 
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Gamma Tocopherol (Vitamin E) and airway inflammatory responses, IND 136873 
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Gamma Tocopherol (Vitamin E) and airway inflammatory responses, IND 136873 
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tocopherol-enriched supplement reduces sputum eosinophilia and endotoxin-induced 
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