Clinical trial NCT02885025
Conditions: Rhinitis,Allergic
Keywords: hypersensitivity antioxidants
Sponsor: VA Office of Research and Development
Start date: 2016-10-01; Completion date: 2019-03-12
Enrollment: 47 participants; Allocation: RANDOMIZED; Intervention model: PARALLEL

Brief summary:
Allergic rhinitis is a common illness suffered among US Veterans. There are medications that help relieve allergy symptoms, including nasal steroid sprays and antihistamines. Some patients have increase symptoms with exposure to their trigger, such as a grasses when combined with pollution due to oxidative stress from pollution. In this study, patients with allergic rhinitis to grass will be given broccoli sprout extract that contains an antioxidant sulforaphane do see if there is beneficial effect in these patients.

Detailed description:
Allergic rhinitis is a common illness suffered among US Veterans. Despite the availability of medications, many patients fail to get adequate control of symptoms especially in high pollutant areas. Southern California air pollutants, like diesel exhaust particles may act as adjuvants to allergens leading to an exaggerated allergic response in certain individuals. These individuals may lack adequate Glutathione transferase activity; necessary to protect cells against pollutant induced oxidative stress. Sulforaphane has been shown to protect against oxidative stress. The investigators hypothesize consumption of foods containing high levels of sulforaphane are beneficial to US Veterans with GST deficiencies suffering from allergic rhinitis. In the investigators' preliminary data, the investigators discovered administration of broccoli sprout extract rich in sulforaphane leads to diminished nasal inflammatory reaction brought on by diesel exhaust particles though did not have sufficient patient numbers to prove an association with GST deficiency. The investigators propose a 3 week randomized clinical trial comparing broccoli sprout extract consumption with nasal corticosteroid administration after subjects with allergic rhinitis undergo a Timothy, Bermuda or Johnson grass nasal challenge. In Aims 1 and 2, the investigators will compare clinical (nasal symptom scores and peak nasal inspiratory flows) and laboratory (inflammatory cytokines, eosinophil cationic protein, tryptase) measurements between four groups: nasal corticosteroid, broccoli sprout extract, nasal corticosteroid plus broccoli sprout extract, and placebo. Aim 3 focuses on exploratory genetic analysis of the 3 glutathione S-transferase (GST) genes to determine if correlations exist between the inability to produce GST enzyme and the response to broccoli sprout extract. At the conclusion of this study, the investigators will gain further knowledge of which patients benefit from anti-oxidant nutritional supplementation in treating allergic rhinitis.

Arms: arm 1: subjects will be randomized into 1 of 4 arms:

1. Broccoli Sprout Extract + Nasal Fluticasone
2. Broccoli Sprout Extract + normal saline nasal spray
3. Placebo Pill + Nasal Fluticasone
4. Placebo Pill + normal saline nasal spray arm 2: subjects will be randomized into 1 of 4 arms:

1. Broccoli Sprout Extract + Nasal Fluticasone
2. Broccoli Sprout Extract + normal saline nasal spray
3. Placebo Pill + Nasal Fluticasone
4. Placebo Pill + normal saline nasal spray arm 3: subjects will be randomized into 1 of 4 arms:

1\. Broccoli Sprout Extract + Nasal Fluticasone 2 Broccoli Sprout Extract + normal saline nasal spray 3. Placebo Pill + Nasal Fluticasone 4. Placebo Pill + normal saline nasal spray arm 4: subjects will be randomized into 1 of 4 arms:

1. Broccoli Sprout Extract + Nasal Fluticasone
2. Broccoli Sprout Extract + normal saline nasal spray
3. Placebo Pill + Nasal Fluticasone
4. Placebo Pill + normal saline nasal spray
Interventions: intervention 1: Broccoli Sprout Extract intervention 2: fluticasone nasal intervention 3: broccoli sprout extract placebo intervention 4: normal saline nasal spray
Intervention descriptions: intervention 1: Broccoli Sprout Extract contains the antioxidant Sulforaphane which is the active ingredient being studies. intervention 2: fluticasone is a nasal steroid that is currently approved for treatment of allergic rhinitis. intervention 3: a tablet similar to the actual broccoli sprout extract though without BSE. intervention 4: normal saline to replace nasal fluticasone in specific arms of the study

Protocol documents:
Prot_SAP_000.pdf: 
 
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Effects of Broccoli Sprout Extract on Allergic Rhinitis     
 
NCT02885025  
 
 
 
Study Protocol and Statistical Analysis Plan  
 
 
November 2, 2018  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


 
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RESEARCH PLAN 
 
 
BACKGROUND AND SIGNIFICANCE 
 
Safe alternative nutritional supplementation is needed for atopic disorders 
suffered by US veterans:  
Allergic Rhinitis a major condition impairing nasal health, effects 10-30% worldwide and 8% 
nationwide (1). A 10 year retrospective review revealed the majority of patients receiving care at 
the VA Greater Los Angeles Healthcare System (VAGLAHS) suffered from Allergic Rhinitis (2). 
From suffering comes cost, both indirect and direct. This can manifest as lost days of work and 
medication expense. Among atopic disorders, completed surveys determined allergic rhinitis as 
the highest cost in regards to both presenteeism and absenteeism, with the average total 
productivity loss per employee surveyed at 593 dollars per year (3). Over 3.4 billion dollars is 
spent directly treating allergic rhinitis, with 46.6% attributed to prescription medications. Despite 
no cost effective treatment for allergic rhinitis, nasal corticosteroids appear to be the most 
beneficial.   Patients suffering from allergic rhinitis seek alternative therapies since 2/3rds report 
daily impairment from rhinitis symptoms despite the use of available medications (4). 
 
The Association between Pollutants and Allergy: 
The increase prevalence in allergic disorders may be attributable to pollutant factors (5). Bench 
Research led to a valid explanation of how pollutants can act as an adjuvant to allergens in 
susceptible patients. Original work discovered pollutant diesel exhaust particles (DEP) increase 
reactive oxygen species (ROS) in respiratory epithelium cells.  This overabundance of oxidative 
stress leads increases intracellular nuclear factor kappa beta (NfKB) and Mitogen Activated 
Protein Kinase (MAP-kinase) cascades causing atopic cytokine production (6-9).    
 
Animal Models investigating the association between Pollutants and Allergy: 
Mice models validated the association between pollutant exposure and allergic response 
increase. Intranasal administration of diesel exhaust particles (DEP) caused an enhanced IgE 
response (10). More specifically administering DEP to dust mite sensitized mice enhanced the 


 
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production of dust mite specific IgE (11). Further experiments on mice solidified the concept that 
pollutants cause an increase allergic response when mice given DEP plus allergen were later 
found to have increased levels of atopic cytokines IL4 and IL5 within lungs and lymph nodes 
(12, 13). 
 
Human Models investigating the association between Pollutants and Allergy: 
These findings were later replicated in human studies. Purified human B cells cultured with TH2 
cytokines and DEP significantly increased IgE production when compared to B cells cultured 
with cytokines alone (14).  Human studies demonstrated instillation of DEP through nasal 
lavage triggered an increase in nasal mucosa IgE production.  (15). Nasal instillation of both 
DEP and ragweed produced increased atopic cytokines IL4, 5,6,10 and 13 and ragweed IgE 50 
fold (16). Further review of these studies noted this increase in allergic biomarkers varied 
among individuals, with some having a drastic response and others no response (17).      
 
Phase II enzymes and Allergic Rhinitis: 
The exaggerated atopic response from DEP when present with allergen exposure can be 
prevented by antioxidant production. Genetic polymorphisms in phase 2 antioxidant enzymes 
among individuals can determine how pollutants influence atopy.  Endogenous anti-oxidant 
phase 2 enzymes GSTT1, GSTM1 and GSTP1 play a vital role in preventing the production of 
cytokines through restoration of redox balance between reduced and oxidized glutathione. Null 
genotypes for GSST1 and GSTM1 are more prevalent in atopic subjects (18), which could 
explain their diminished ability to produce phase II antioxidants leading to unchecked balance of 
oxidation. When exposed to pollutants, patients having null mutations in GST increase 
production of IgE and histamine levels respectively (19, 20).  Patients with GST null mutations 
suffering from allergic rhinitis may respond best to antioxidant therapy.   
  
The use of antioxidants in treating Allergic Conditions: 
Success rates in treating atopic conditions with antioxidants have been mixed (21-23). The 
importance of evaluating the patient for GST polymorphisms is supported by studies showing 
patients with GSTM1 null genotypes were more likely to have a greater detrimental loss of 
pulmonary function with exposure to ozone and a more positive response to consumption of 
Vitamin C/Vitamin E antioxidants supplementation (24, 25). 
 
Sulforaphane for treating allergic conditions: 
The antioxidant Sulforaphane increases phase II enzyme production through activation of the 
Nrf2 transcription factor and the anti-oxidant response element (ARE) leading to an increase 
production of phase II enzymes (26-30). Sulforaphane also interferes with the pro-inflammatory 
pathways NkKB and AP1 responsible for the production of TH2 cytokines and IgE.    
 
Sulforaphane has been shown to block B cell IgE production (31) and to inhibit IL8 production in 
human bronchial airway epithelial cell lines after exposure to DEP via inducing nuclear factor 
like 2, leading to enhanced Phase II enzyme production (32). A recent study showed that ova 
sensitized mice treated with SFN had a diminished inflammatory response after ova challenge 
leading to diminished hyper responsive airways (33). 
 
Broccoli Sprout for improving nasal health: 
Broccoli sprouts (BSE), members of the cruciferous vegetable family, have extremely high 
concentrations of sulforaphane (26). The first placebo controlled BSE human study performed at 
our partnering institution involved administering escalating doses of broccoli sprout homogenate 
(BSH) to subjects. Phase II enzymes were shown to increase proportionally to the dose of BSE 


 
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given. Furthermore the induction of individual Phase II enzymes was inversely correlated with 
the expression of phase II enzymes (glutathione-s-transferase M1, glutathione-s-transferase P1, 
NADPH quinone oxidoreductase, and hemoxygenase-1) as revealed by genetic testing. 
Patients with GSTM1 null or GSTP1 Ile/Ile105 polymorphisms known to endogenously produce 
the lowest Phase II enzyme levels responded best to BSE (34). 
 
PRELIMINARY STUDIES: 
 
Administering BSE to allergic subjects was first performed in a recent NIH sponsored study by 
Dr. Li and colleagues at the University of California, Los Angeles, a partnering institution to the 
VAGLAHS. Subjects with cat allergy through skin testing diagnosed allergic rhinitis were 
challenged with 300µg  DEP (equivalent to daily PM exposure levels on LA freeway) then 
underwent 4 weeks of washout, followed by daily BSE administered as a single oral morning 
dose (11umol sulforaphane equivalent to that formed from consuming 100 to 200 grams of 
broccoli) for four days. The diesel exhaust particle challenge was repeated after BSE 
consumption. The average nasal white blood cell counts increased by at least 4 fold (p<0.0005) 
within 24 hours post-DEP exposure and there was significant attenuation of cell count post-DEP 
exposure when patients were given daily BSE for four days prior to DEP challenge (p<0.01 at 6 
hours and p<0.0001 at 24 hours following DEP challenge). There was no correlation between 
the inflammatory response and the presence of GSTM1 null orGSTP1 Ile/Ile105 polymorphisms 
(35). Failure to determine a statistically significant difference in response among the genetic 
polymorphisms may have been due to the low subject number. 
 
We are proposing a double blinded randomized placebo controlled study to investigate the 
effectiveness of BSE on nasal health among US veterans with a diagnoses seasonal grass 
allergic rhinitis in an urban environment where DEP levels are high. 
 
During the study, GST genotype data will be collected in order to obtain data for future 
exploratory analysis.          
 
RESEARCH DESIGN AND METHODS 
 
Objectives: 
Primary Objective:   
To conduct a double-blind, randomized, placebo-controlled trial to compare clinical 
measurements for allergic rhinitis following Timothy, Bermuda or Johnson grass allergen 
(phleum pretense, cynodon dactylon, or songhum halepense) challenge before and after 
initiation of BSE supplementation. Clinical measurements will be derived from the total nasal 
symptoms score (TNSS) and peak nasal inspiratory flow (PNIF).   
 
Secondary Objectives: 
To conduct a randomized controlled trial to compare biomarker measurements for allergic 
rhinitis following grass allergen challenge before and after initiation of BSE supplementation. 
Biomarkers will include the following inflammatory cytokines: Interleukin 4, Interleukin 5, 
Interleukin 13 along with Tryptase and Eosinophil cationic protein (ECP) obtained through 
polyurethane sponges placed into both nostrils.   
 
To conduct an exploratory genetic analysis of the 3 glutathione S-transferase (GST) 
genes (GSTM1, GSTT1, and GSTP1) for future investigation into how genetic polymorphisms 
effect the response to BSE supplementation in atopic disorders.  


 
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Overview of Study Design 
The proposed study is a double-blind, randomized, placebo-controlled trial consisting of four 
arms. A total of 190 subjects diagnosed with grass (p. pretense, c. cynodon, and/or s songhum) 
specific allergic rhinitis will be randomly assigned to one of four groups: 
1. nasal corticosteroid + Placebo tablet  
2. placebo nasal spray + Placebo tablet 
3. nasal corticosteroid + Avmacol®  
4. placebo nasal spray + Avmacol®  
 
Avmacol is a supplement that supports sulforaphane production by providing glucoraphanin and 
an active myrosinase enzyme via our Sulforaphane Production System.  Subjects will be given 
4 tablets to take with their evening meal.   
 
Following consent, a list of antioxidant foods (including sulforaphane containing foods) and  
prohibited medications to avoid will be given to all subjects then subjects will be scheduled for 
visit 1 within <20 days.   
 
At visit 1, Subjects will undergo epicutaneous hypersensitivity skin testing to grass allergens 
prevalent in southern California: Johnson Grass (Sorghum Halepense), Bermuda Grass 
(Cynodon Dactylon), and Timothy Grass (Phleum Pretense), along with dog, cat and perennial 
allergens: dust mite (dermatophagoides pteronyssinus, dermatophagoides farina), and common 
mold allergens (alternaria, aspergillus, cladosporium, and penicillium) (36). Subjects with 
positive skin test results to one or more grass allergens and either negative skin test to 
perennial allergens (dust mite or mold) or positive tests to perennial allergens (dust mite or 
mold) without a correlating clinical history consistent with perennial allergic rhinitis  will undergo 
grass allergen nasal provocation testing. Subjects with a positive skin test to cat or dog and 
clinical symptoms consistent with cat or dog triggered allergic rhinitis will be asked to avoid cats 
and or dogs for the entire study. For subjects who have an epicutaneous allergen skin or blood 
test on file completed by the VA Allergy Clinic within <1 year that confirms a positive to grass 
pollens the results will be used to proceed onto the up-dosing nasal challenge. Subjects 
experiencing a positive nasal provocation test defined by a total nasal symptom score (TNSS) 
over 7, or PNIF is reduced by > 50% from baseline redaing will be tested for serum GSTM1 
polymorphism and baseline sulforaphane levels and will proceed to visit 2. Subjects failing to 
mount a positive response with the highest concentration (10,000 BU/cc for p. phleum and c. 
cynodon, or 1:100 w/v for s. sorghum) will be withdrawn from the study..     
 
Nasal challenge will repeat at visit 2, two weeks after visit 1, and visit 3, 3 weeks after beginning 
intervention at visit 2.  This challenge was developed and perfected by Guy W. Scadding PhD 
and colleagues and showed reproducibility in a 2012 study (36).  Prior to and following each 
challenge, subjects will undergo spirometry.  Challenges will not be performed in patients with 
forced expiratory volume in 1 second (FEV1) values less than 70%. The visit 1 nasal challenge 
will be considered a graded up-dosing challenge needed to determine the dose used at visits 2 
and 3.   
 
At visit 2 (2 weeks following visit 1) and visit 3 (5 weeks following visit 1), subjects will undergo a 
single nasal allergen challenge with the effective dose determined during the up-dosing 
challenge at visit 1. Total nasal symptom score and peak nasal inspiratory flow will be recorded 
throughout all nasal challenge procedures.   
  


 
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Biomarkers collected through sponges placed in both nasal passages during the challenge will 
include IL4, IL5, IL13, eosinophil cationic protein and tryptase. At the end of visit 2, patients will 
be randomized and will be instructed to begin the following evening, to return in 3 weeks for 
another nasal challenge.     
 
Throughout the study, serum sulforaphane levels for each subject will be checked at visits 1, 2 
and 3 to monitor adherence.   
 
All study procedures will be conducted at the VAGLAHS. Subjects will be assigned an 
appointment day and appointment time for each scheduled visit. All appointments will be at 
approximately the same time of the day for all subjects to avoid diurnal variation. 
 
Figure 1. Overview of Study Procedures 
 
Study Population 
General Considerations: 
Subjects participating in the study are followed at two VA medical centers and one academic 
medical center.  All subjects are non-smoking male or female US veterans aged 18 years and 
older with symptoms consistent with seasonal summer allergic rhinitis (rhinorrhea, nasal 
congestion, postnasal drainage sneezing, and/or nasal or eye pruritus). There will be no 
exclusion based on gender or ethnic/racial background.   
 
Inclusion Criteria 
1. 
Females and males 18 years or older.  
2. 
History consistent with seasonal allergic rhinitis consistent with grass allergy (symptoms               
during summer months, June through August, for at least two consecutive seasons).   
3. 
Not currently taking any medications for allergic rhinitis. 
4.         Provide written informed consent. 
5.  
Willing and able to comply with all aspects of the protocol.     
 
Exclusion Criteria 
1. 
The subject has any uncontrolled or serious disease, or any medical or surgical or 
condition that in the opinion of the investigator(s) could affect the subject’s safety and/or 
interfere with the study assessments. 
2. 
History of anaphylaxis to environmental allergens.or an unknown trigger.   
3. 
History of broccoli allergy. 
4. 
Recent upper respiratory infection (less than 4 weeks prior to study) or other active 
Infection. 


 
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5. 
Active smoker.   
6. 
Currently receiving allergy immunotherapy.  
7. 
History of rhinitis exacerbation within the past 2 weeks. 
8. 
Use of non-selective Beta-Blocker. 
9. 
Inability to give written informed consent. 
10. 
History or evidence of non-stable cognitive capacity within less than 1 year (i.e. 
Alzheimer’s disease, dementia, bipolar disorder) that in the opinion of the investigator(s) 
could affect the subject’s safety and/or interfere with the study assessments.    
11. 
Pregnancy. 
12.      Perennial rhinitis. 
13.     Uncontrolled asthma.   
14.     FEV1 (forced expiratory volume in 1 s) <70% predicted at screening.    
 
Recruitment: 
In order for 152 subjects to complete the study, 475 subjects will be recruited for enrollment 
under the assumption 50% (237) of subjects will have seasonal allergic rhinitis to grass 
confirmed on skin testing, 80% (190) of subjects will have a positive nasal challenge and 80% 
(152)  of subjects will complete the study. 
 
Subject recruitment will occur at three facilities:  VA Greater Los Angeles Health Care System 
(VA GLA) , VA Long Beach Health Care System (VA LB) and the University of California Los 
Angeles (UCLA) Medical Center.  Within each facility, recruitment will occur at primary and 
specialty (allergy) clinics.   
 
Recruitment at VA Greater Los Angeles and VA Long Beach Health Care Centers:   
Primary Care Clinic Recruitment:    
Investigators will work together with providers assigned to Patient Aligned Care Teams (PACT).  
On average, PACTs follow 44,000 patients at VA Long Beach and 63,000 at VA Greater Los 
Angeles.  Assuming involvement with primary care providers following 10,000 subjects at VA 
GLA and 10,000 subjects at VA LB, At each facility, 2,000 (20%) will should have a diagnoses 
of allergic rhinitis.  600 (30%) will experience allergic rhinitis symptoms during the summer 
months and 300 (50%) of these subjects will have symptoms limited to summer months.  We 
estimate at total of 600 potential recruitment subjects from primary care clinics at both VA GLA 
and VA LB.       
 
Allergy Clinic Recruitment: 
Within the past year, 40% (320) of new consultations at VA Greater Los Angeles Healthcare 
System and 68% (208) of new consultations at VA Long Beach Healthcare System were 
requested to evaluate allergic rhinitis. Based on these figures, we would anticipate a total of 
30% having allergic rhinitis associated with summer months and then 50% of these patients 
having allergic rhinitis limited to summer months or 79 (48 + 31) potential subjects at VA GLA 
and VA LB combined.  
 
Recruitment at the University of California Los Angeles Medical Center:   
In Southern California, patients are seen at 16 satellite primary care clinics affiliated with UCLA 
Medical Center.  Adult Allergists practice at four of these locations (Porter Ranch CA, Santa 
Monica CA, Thousand Oaks CA, Torrance CA.)  Investigators will work with providers at UCLA 
affiliated clinics in identifying potential subjects.  (see Contingency Plan)  
 
Use of Flyers at Institutions:  


 
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In addition to the above recruitment strategy, flyers will be placed in primary care, ear nose and 
throat, and dermatology clinics at VAGLAHS and VALBHCS with contact information for 
enrollment. Potential subjects who call will be preliminary screened via phone call screening 
script to determine possible eligibility. Those who pass the preliminary screening process will 
have an appointment scheduled. 
 
Recruitment from VA GLA Pharmacy Database: 
In addition, we will work with pharmacy to retrieve patients prescribed nasal corticosteroids 
limited to summer months.  These patients have a higher likelihood of suffering from grass 
pollen allergic rhinitis.      
 
Contingency Plan: 
Participation recruitment will be followed closely.  If needed, we will submit a request for 
expansion to involve non veterans.  The VA GLA is close to UCLA.  Students/staff/potential 
subjects attending UCLA may qualify for inclusion into the study.  
 
Medical History: 
A medical history will be obtained to confirm whether patient has clinical symptoms consistent 
with summer seasonal rhinitis (during grass season) allergic rhinitis. 
 
Procedures at Each Visit: 
 
Consenting (<20 days to Visit 1)  
-Affirm eligibility, check medication use. 
-Review study with patient and if subject agrees to enroll, obtain subjects written 
informedconsent. 
 -Instructions for avoidance of sulforaphane containing food products in diet. 
- Review concomitant medication use. 
-Instructions for avoidance of the following medications: oral antihistamines, nasal 
  corticosteroids, nasal mast cell stabilizers, nasal antihistamines, leukotriene antagonists. 
-Proceed onto visit 1, or schedule subject for visit 1 <20 days from providing written consent.   
 
Visit 1 (Day 1) 
 
-Spirometry performed (Note: subjects’ have up to 3 attempts to FEV1 of >70% to continue on 
with this study).  
-Collect blood pressure, heart rate, height and weight. 
- Check concomitant medication use since last visit.  
-General physical examination. 
-Nasal examination. 
-Epicutaneous Allergen Skin Testing. Patients who qualify based on skin test results will 
proceed  with up-dosing nasal challenge to the positive skin test grass allergen. Subjects that 
have completed epicutaneous allergen skin or blood test at the VA Allergy Service within < 1 
year with positive results to grass pollens will to proceed onto the up-dosing nasal challenge 
using the results on file.  
-Updosing nasal challenge. Patients with positive nasal challenge will continue as follows:   
-Obtain blood work to determine sulforaphane baseline level and genotype for  GSTM1 
polymorphisms. 
- Schedule subject for visit 2. 
 


 
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Visit 2 (Day 14) 
-Check concomitant medication use since last visit. 
-Collect blood pressure, heart rate and weight. 
-Spirometry performed 
-General physical examination. 
-Nasal  examination. 
-nasal lavage  
-Obtain blood work for sulforaphane level check. 
-Grass Nasal Challenge. 
-Randomize patients into one of four groups. 
-Medication Dispensing (to begin evening of Day 15).      
 
Visit 3 (Day 35) 
-Check concomitant medication use since last visit. 
- Collect medication dispensed at last visit (research team complete medication compliance).  
-Collect blood pressure, heart rate and weight. 
-Spirometry performed 
-General physical examination. 
-Nasal   examination. 
-Nasal lavage  
-Obtain blood work for sulforaphane level check.  
-Grass nasal Challenge.  
 
Study Schedule: 
 
The typical schedule is shown as an example for consenting.  
 
Consenting (<20 days prior to visit 1) Activitity (in sequence): 
 
Affirm eligibility, check medication use. 
 
Review study with patient and if subject agrees to enroll, obtain subjects written 
informedconsent. 
 
Instructions for avoidance of sulforaphane containing food products in diet prior to visit 1 and 
throughout the study. 
 
Instructions for avoidance of the following medications: oral antihistamines, nasal 
corticosteroids, nasal mast cell stabilizers, nasal antihistamines, leukotriene antagonists prior to 
visit 1 and throughout the study.  
 
Proceed onto visit 1, or schedule subject for visit 1 within < 20 days from providing written 
consent.   
 
The typical schedule is shown as an example for the enrollment visit (Visit 1).  
 
Visit 1 (Day 1) Activity (in sequence):  
 
Subject arrives at the Allergy/Immunology Clinic.  


 
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Spirometry will be performed before skin test. (Note: subjects’ have up to 3 attempts to FEV1 of 
>70% to continue on with this study.) 
 
Vital signs will be collected (heart rate, blood pressure, weight, height) and general physical 
examination along with nasal examination. 
 
Subjects undergo epicutaneous skin testing. (Subjects who qualify based on skin test will 
continue with nasal challenge). For subjects who have an epicutaneous allergen skin or blood 
test on file completed by the VA Allergy Clinic within < 1 year that confirms a positive to grass 
pollens the results will be used to proceed onto the up-dosing nasal challenge.   
 
Up-dosing nasal allergen challenge will be performed  
 
Subjects testing positive to nasal challenge will proceed with blood work to check sulforaphane 
Baseline levels and genotype for GSTM1 polymorphisms 
 
 
The typical schedule is shown as an example for a follow up evaluation day for visit 2.  
 
Visit 2 Activity (in sequence): 
 
Subject arrives at the Allergy/Immunology Clinic. 
                                                                 
Vital signs (heart rate, blood pressure and weight) , nasal examination , concomitant medication 
and diet checked since last visit.    
  
Spirometry will be performed. (Note: subjects’ have up to 3 attempts to FEV1 of >70% to 
continue on with this study.) 
 
general physical examination along with nasal examination. 
 
Nasal lavage performed.  
 
Blood work obtained for sulforaphane level    
 
Nasal allergen Challenge followed by monitoring explained below  
 
Randomization occurs.  Instructions for avoidance of  sulforaphane containing food products in 
diet and for avoidance of medications reiterated.   
  
The typical schedule is shown as an example for a follow up evaluation day for visit 3.  
 
Visit 3 Activity (in sequence):         
                                       
Subject arrives at the Allergy/Immunology Clinic. 
 
Study medication disepensed at visit 2 are returned by the subject and a compliance check is 
performed the study team.  
 


 
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Vital signs (heart rate, blood pressure and weight), nasal examination, concomitant medication 
and diet checked since last visit.   
 
Spirometry will be performed. (Note: subjects’ have up to 3 attempts to FEV1 of >70% to 
continue on with this study.) 
 
general physical examination along with nasal examination. 
 
Nasal lavage performed. 
 
Blood work obtained for sulforaphane level.   
 
Nasal Allergen Challenge followed by monitoring explained below      
 
Study completed.     
 
Study Outcome Measures: 
1. The Total Nasal Symptoms Score (TNSS), performed to determine adequate dosing for nasal 
challenge (visit 1) and during the nasal challenges (visit 2, 3).  During the challenge the TNSS 
will be taken 30 minutes after nasal lavage, and then repeated at 5, 15, and 30 minutes then 
hourly to 4 hours post-challenge.     
2. The peak nasal inspiratory flow (PNIF) performed at the up-dosing nasal challenge visit (visit 
1) and during nasal challenges (visit 2 and 3).  During the challenge, PNIF will be taken  30 
minutes after the nasal lavage, then repeated at 5, 15 and 30 minutes after the challenge, then 
hourly to 4 hours post-challenge.    
3. Biomarker measurements taken from nasal fluid collected by nasal sponge at visit 2 and visit 
3 will include:   
a) IL-4: 0 mintue, 5 minutes, 15 minutes, 30 minutes, then hourly to 4 hours post challenge  
b) IL-5: 0 minute, 5 minutes, 15 minutes, 30 minutes, then hourly to 4 hours post challenge 
c) IL-13: 0 minute, 5 minutes, 15 minutes, 30 minutes, then hourly to 4 hours post challenge   
d) Eosinophil Cationic Protein (ECP): 0 minute, 5 minutes, 15 minutes, 30 minutes, then hourly 
to 4 hours post challenge  
e) Tryptase: 0 minute, 5 minutes, 15 minutes, 30 minutes, then hourly to 4 hours post challenge   
 
Study Regimen: 
Study Medications and Placebo 
Nasal Corticosteroids: Fluticasone propionate nasal spray prescribed as 2 sprays in each nostril 
daily will be the nasal corticosteroid of choice for the current study. Placebo will be normal 
saline based. Both fluticasone and placebo will be ordered through the VAGLAHS Research 
Pharmacy.  For optimal control of nasal symptoms a treatment period of 1 week is required. 
Fluticasone will be given for 3 weeks in order to maximize effect.    
 
Active Avmacol Enteric Coated Tablets:Water, HPMC, Sodium Alginate, Purified Stearic Acid, 
Ethylcellulose, Medium Chain Triglycerides, Oleic Acid, Triactin, Mineral Oil, Sodium Lauryl 
Sulfate, Titanium Dioxide 
 
Avmacol, a blend of broccoli seed and sprout extracts contains the 2 essential ingredients 
needed for sulforaphane production: glucoraphanin and the myrosinase enzyme.   Avmacol 
contains ~ 30 - 35 umoles sulforaphane per 2/tablet recommended serving.  


 
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Placebo Avmacol Coated Tablets:Water, HPMC, Triactin, Mineral Oil, Sodium Lauryl Sulfate, 
Titanium Dioxide 
 
Concomitant Medications 
Concomitant medication is defined as any medication, which is taken during the study. All 
prescription, herbal supplements, and over-the-counter drugs which are being taken by subjects 
upon entry to the study or which are taken during the study are regarded as concomitant 
treatments and will be documented. Basic medication use information will be confirmed at the 
time of screening, in order to determine if subject meets exclusion criteria regarding 
medications. 
 
Rescue Medications 
Subjects in the study are atopic. During the course of the study, subjects may develop nasal 
symptoms. They will be allowed to treat their symptoms with saline nasal spray. If their 
symptoms persist and they require additional medications, they will be discontinued from the 
study. 
 
Prohibited Medications  
The days below indicate last dose received prior to visit 1 and throughout the subjects’ total 
participation time:  
 
1. Topical Nasal Steroid use in the past 14 days, 
2. Systemic steroid use in the past 14 days, 
3. Oral antihistamine use in the past 5 days,  
4. Intranasal antihistamine or cromolyn use in week prior to nasal challenge. 
6. currently receiving allergy immunotherapy, or use anytime during the protocol. 
7. short acting beta agonists in the past 6 hours. 
8. nasal cromolyn in the past 7 days, 
9. oral decongestants in the past 3 days,  
10. antihistamine-decongestant tablets/liquids in the past 3 days,  
 
Nasal Sprays: 
Both the active fluticasone proprionate (50mcg) and the placebo will be obtained from the VA 
Research pharmacy.     
 
Criteria for Premature Termination of the Study 
 
Participant Withdrawal Criteria 
In the best interest of the subject, participants may be prematurely terminated 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” 
3. The participant takes any of the prohibited medications during the study. 
4. The participant develops an adverse event that, in the opinion of the Protocol Chair 
(Principal Investigator) or the Safety Monitoring Board of the study, increases his/her 
risks if study participation continues. 
5. The participant dies 
 


 
Version  date: 11/2/2018 
Page 13 of 18 
 
Study Stopping Rules 
The Data Safety Monitoring Board (DSMB) will be provided summaries of all adverse events on 
a regular basis.  The DSMB will have the authority to terminate the trial at any time based on 
reported adverse events.   
 
Study enrollment and ongoing study treatments will be suspended pending expedited review of 
all pertinent data by the VAGLAHS Institutional Review Board (IRB), the Data Safety Monitoring 
Board (DSMB) if a significant adverse event as deemed by DSMB occurs  that is possibly 
related to the study procedures. The study may be terminated by the DSMB upon review of data 
with observations or findings that such action is necessary. (National Cancer Institute’s 
Common Terminology Criteria for Adverse EventsVersion 4 (published May 28, 2009)).   
 
Grade 1=Mild adverse event 
Grade 2=Moderate adverse event. 
Grade 3=Severe and undesirable adverse event 
Grade 4=Life-threatening or disabling adverse event 
Grade 5=Death 
 
Screening Visit 
At the screening visit, potential subjects are given a copy of the IRB-approved informed consent 
form for review and are invited to ask any questions or express concerns after reading the 
consent. The research study is clearly explained to the subject, including the rationale, risks, 
and benefits. The subject is asked to explain what he/she understands to be required in the 
study. The subject is informed that all participation is voluntary and that he/she is free to 
discuss the study with family or friends when considering enrollment. Subjects are told that only 
if the study protocol is fully understood should they sign the consent form. After the subject 
provides written consent, potential subjects’ eligibility will be confirmed based on the determined 
inclusion/exclusion criteria. Once the subject has signed the informed consent form, a study 
investigator will also sign it, and the subject will receive a copy. 
 
Procedures performed 
Skin Testing:  Epicutaneous skin testing is considered highly reliable, safe and widely used in 
diagnosing allergic rhinitis (37).   After application of the specific allergen to the sterile tip of the 
applicator, the subject’s skin will be gently lifted and the tip containing allergen will be applied.  
Along with application of the allergens mentioned, histamine and a saline control will be applied 
to subject’s skin. Results will be interpreted 20 minutes following and will be considered positive 
if the wheal and erythema are at least 3mm larger than the saline control. For subjects who 
have an epicutaneous allergen skin or blood test on file completed by the VA Allergy Clinic 
within < 1 year that confirms a positive to grass pollens the results will be used to proceed onto 
the up-dosing nasal challenge.   
 
Nasal Challenge: Nasal Challenge is considered a safe and useful diagnostic tool in diagnosing 
causes of both allergic and nonallergic rhinitis (38). The nasal challenge chosen was based on 
repeated reliability using the TNSS and PNIF scores along with the biomarkers chosen for this 
study. (36)       
 
Subjects will undergo an up-dosing nasal challenge performed at visit 1 in order to determine 
the threshold dose to be given at the subsequent nasal challenges (visit 2 and visit 3).  During 
the up-dosing nasal challenge, the TNSS will be recorded immediately before each subsequent 
dose.  Grass allergen extract (c. cynodon, p. pretense or s. sorghum,) will be used in the 


 
Version  date: 11/2/2018 
Page 14 of 18 
 
challenge.  For c. cynodon (Bermuda grass), dosing will start at 30 BU/ml and will continue with 
the following concentrations until a TNSS score reaches 7 or greater or PNIF is reduced by 
>50% from baseline reading: 300, 1000, 3000 and 10,000 BU/ml. For p. pretense(Timothy 
grass), dosing will start at 300 and will continue with the following concentrations until a TNSS 
score reaches 7 or greater or baseline PNIF is reduced by > 50% from baseline reading: 
1,000,3,000,10,000 and 100,000 BU/ml.  For s. sorghum (Johnson grass) dosing will start at 
1:200,000 and will continue with the following concentrations until a TNSS score reaches 7 or 
greater or baseline PNIF is reduced by > 50% from baseline reading: 1:20,000, 1:2000, 1:200, 
and 1:20 w/v. Subjects will receive three sprays equivalent to 100 µl for each concentration into 
each nostril using an atomizer device, part of the standardized equipment at VAGLAHS for 
patients requiring nasal sprays during procedures applied every 10 minutes until maximal 
concentration or a TNSS score of 7 or PNIF reduction by > 50% from baseline reading is 
achieved.  Prior to subsequent challenges at visit 2 and 3, nasal lavage (SinusRinse) will be 
performed.  A baseline PNIF and TNSS will be recorded 30 minutes following nasal lavage for 
visits 2 and 3. Nasal challenge with 100 µl of the final dose achieved at visit 1 will then be 
applied into each nostril.  The PNIF and TNSS will be recorded following challenge at 5, 15, 30 
minutes following challenge and then at 1 hour, continuing to record each hour for 4 hours.  
During these intervals, nasal sponges (Plastocell & Co.) will be applied into each nostril onto the 
nasal mucosa, beyond the nasal vestibule, and alongside the inferior turbinate with the use of a 
crock  forceps (Phoenix surgical instruments, Harlow, UK)  and a Thuddicum’s nasal speculum 
(Phoenix surgical instruments, Harlow, UK).  After two minutes of placement, the sponges will 
be removed and then added to 2 ml centrifuge tubes with indwelling .22µm cellulose acetate 
filters (Costar Spin-X, Corning, NY, USA) and placed immediately into ice before centrifuging 
within the hour.  The liberated fluid will then be freezed at -80º until analysis occurs.    
  
General physical examination: A physical examination to determine general health to include 
measurement of vitals and a cardiopulmonary examination will be performed in the VA GLA 
Allergy/Immunology Clinic by the attending physician. A nasal examination will be performed at 
every visit before any nasal challenge or lavage is performed. On Day 1, a urine pregnancy test 
will be performed for women of child-bearing potential. 
  
The Immediate Total Nasal Symptom Score (TNSS): The TNSS has been used in prior 
interventional studies to evaluate different treatment modes for allergic rhinitis (39, 40, and 41). 
The score will consist of subjects rating 4 symptoms (nasal congestion/postnasal drainage, 
nasal pruritus, rhinorrhea, or sneezing) on a scale of 0 to 3, where 0=no symptoms present, 
1=mild symptoms that do not interfere with activity, 2=moderate symptoms that are slightly 
bothersome symptoms with slight interference with activity and/or nighttime sleep, or 3=severe 
symptoms with interference with activity and nighttime sleep. 
 
The Peak Nasal Inspiratory Flow (PNIF):  the PNIF (Alliance Tech Medical) has been used in 
prior interventional studies involving allergic rhinitis (42, 43, and 44).  Patients exhales fully and 
quickly after checking that an air tight seal covers the nose and seals the mouth.      
 
Nasal Examination:  Nasal examination will be performed at each visit prior to nasal challenge.   
Each nasal examination will include use of an otoscope with specula inserted into the nasal 
passage.  Nasal passages and nasal turbinates will be inspected for color, swelling, exudate or 
bleeding.    
  
Precautions: 


 
Version  date: 11/2/2018 
Page 15 of 18 
 
Since allergen will be administered through nasal challenge, it is imperative that the study take 
place in a controlled environment that is prepared to treat hypersensitivity reactions. The 
VAGLAHS Allergy/Immunology clinic has the appropriate equipment and trained personnel to 
treat allergic reactions including anaphylaxis. A crash cart is available inside the clinic and 
epinephrine is readily available.  The clinic is located within the VAGLAHS hospital where there 
are physicians trained in advanced cardiac life support (ACLS). 
 
Visit Target Windows: 
Subject visit times during the treatment phase of the protocol should be consistent, within -+10 
days of targeted scheduled visit days. 
 
Randomization: 
This is a double blind randomized study. The subject randomization will be carried out based on 
a computer generated randomization list with a random blocked design of block sizes of 4 and 
8. Only the study statistician and pharmacist will maintain the randomization list, the treatment 
administrators and outcome evaluators are blind to the randomization. The investigator 
performing and recording the results of the challenge will be blinded to the subjects assigned 
treatment arm.    
 
PAYMENT OFFERED FOR PARTICIPATION 
 
Twenty dollars ($20.00) will be paid to participants who complete up to the skin test 
procedure at visit 1, and do not meet study qualifications (skin test results determine 
that you not allergic to grass) to continue participation in the study. 
Forty-five dollars ($45.00) will be paid to participants who meet study qualifications and  
complete visit 1.  
Ninety dollars ($90.00) will be paid to participants who complete visit 2. 
Ninety dollars ($90.00) will be paid to participants who complete visit 3. 
Additionally, those participants who will comply with study protocol and complete all three study 
visits will receive the completion of study payment in amount of $75 at the end of the study. 
 
Study Material and analysis: 
Choosing cytokines mediators:    
TH2 cytokines (IL4, IL15, IL13):  These cytokines direct production of IgE antibodies which latter 
attach to mast cells.   
 
Tryptase, Eosinophilic Cationic Protein (ECP):  Tryptase is immediately released from mast 
cells upon activation.  ECP marks presence of eosinophils which are involved in the delayed 
reaction of allergic rhinitis.   
 
Both the above cytokines, tryptase and ecp have been found in nasal mucus and levels have 
been replicated in prior nasal challenge (36)      
 
Analysis of biomarkers:  
All the following analysis will be performed at the biomarker lab, UCLA Center for Human 
Nutrition.  Measurements of IL-4, IL-5, and IL-13 will be  performed using MSD Human TH1/TH2 
7-Plex, Ultra-Sensitive Kit according to the manufacturer's instructions (MS6000 7 spot, Meso 
Scale Discovery, Maryland, USA). Plates will read using anMSD SECTOR® 6000 instrument. 
With this method, validation procedures demonstrated that the lower quantification limit for all 
cytokines was approximately 5 pg/ml, with an assay range of 5–5000 pg/ml in nasal secretions. 


 
Version  date: 11/2/2018 
Page 16 of 18 
 
All measurements will be performed on undiluted nasal fluid samples in duplicate and reported 
as mean values, though cytokine levels will be standardized with ng/ml protein concentration.   
Tryptase and Eosinophilic Cationic Protein (ECP) will be measured through ImmunoCAP at the 
minimum level of detection of 2mg/ml.  Each plate/Immunocap carousel is run with standards 
and curve controls.   
 
Genomic DNA Isolation: 
Peripheral blood will be drawn into a BD Vacutainer K2 EDTA tube (BD Biosciences; San Jose, 
CA). Genomic DNA will be extracted from blood using the GenElute Mammalian Genomic DNA 
Miniprep Kit (Sigma-Aldrich; St. Louis, MO) according to manufacturer’s instructions. Briefly, 
200 μl blood will be treated with Proteinase K followed by RNase for 2 min before lysing cells in 
a 55 deg C water for 10 min. DNA will be precipitated with 100% ethanol and bound to a spin 
column. After two washes, DNA will be eluted from column with TE buffer. TaqMan Polymerase 
Chain Reaction Genotyping of GSTP1 Ile105Val Genotyping will be performed using SNP 
Genotyping Assays for GSTP1 (rs1695) (Applied Biosystems; Foster City, CA) on a 
StepOnePlus Real-Time PCR Machine (Applied Biosystems).  Real-time PCR will be performed 
with 25 ng DNA in a 20 μl reaction containing TaqMan Genotyping Master Mix and SNP 
genotyping assay. Reactions will be heated for 95 oC for 10 minutes following by 40 cycles of 
92 oC for 15 sec with 60 oC for 1 min. Alleles will be determined using the allelic discrimination 
plot in the StepOne software genotyping program.  Each sample will be performed in 
quadruplicate. 
 
GSTM1 Copy Number-GSTM1 copy number will be determined using Pre-designed TaqMan 
Copy Number Assays (Applied Biosystems). GSTM1 will run in a duplex assay with RNase P as 
a reference gene. Reactions will be heated for 95 oC for 10 minutes following by 40 cycles of 92 
oC for 15 sec with 60 oC for 1 min. Results will be analyzed using Copy Caller version 1.0 
software (Applied Biosytems). Each sample is performed in quadruplicate. 
 
Measurement of sulforaphane: 
HPLC-MS/MS identification of SFN metabolites in plasma R, S-sulforaphane will be purchased 
from LKT Laboratories, Inc. (Minnesota, USA). All solvents and other chemical used will be 
HPLC grade and were purchased from Fisher Scientific (USA). Water will be ultrafiltered using a 
Milli-Q pure water system. To the 4° C plasma sample (200 μL) in a 1.5mL micro centrifuge tube 
(Fisher Scientific, USA), pre-cooled (4° C) trifluoroacetic acid (20 μL) (TFA) will be added to 
precipitate plasma proteins (30 seconds) in the sample followed by brief mixing and refrigerated 
centrifugation at 15900×g for 10 min. The resulting supernatant will be filtered (0.2 μm pore 
size) and 20 μL injected for analysis. 
  
A LCQ Advance mass (Thermo Finnegan) and a Survey HPLC equipped with binary pump, 
degasser, cooled auto-sampler and a PDA detector (USA) were used for the LC–MS analysis. 
The HPLC column will be a Zorbax SB- will be 0.25 mL/min and gradient elution will be used: 
solution A (1% acetic acid solution): solution B (acetonitrile). The gradient started at 2% solution 
B increasing over 20 min to 30% B and 50% in 30 minutes, finally re-equilibrating to 2% B for 10 
min. The mobile phase solutions were filtered through 0.22 μm pore size cellulose nitrate or 
nylon membrane filters using a vacuum filtration apparatus. The LC eluent flow of 0.25 mL/min 
will be sprayed into the mass spectrometer interface without splitting. Electrospray ionization in 
the positive mode (ESI+) will be applied. The analytes are identified by molecular weight and 
their MS/MS, respectively. The identification of the MS/MS of SFN-Cys, SFN-Cys-Gly; SFN-
NAC; and SFN-GSH at m/z of 178; 179; 178 and 179, respectively were used to confirm 
consumption of the BSE supplement by subjects in the study. 


 
Version  date: 11/2/2018 
Page 17 of 18 
 
 
Peak Nasal Inspiratory Flow (PNIF): 
PNIF will be performed on each subjectthroughout nasal allergy challenge testing.  
Measurements will be performed using the In-Check Inspiratory Flow Measurement Device 
(Alliance Tech Medical), a portable inspiratory flow meter that can monitor nasal patency.  Nasal 
peak inspiratory flow will be measured in L/min.  The subject will place the device over the nose 
and mouth.  The subject will be asked to take a deep, quick forced inspiration through the nose.  
The test will be repeated three times and the highest value will be recorded.      
 
Statistical Considerations and Analytical Plan: 
Summary statistics (mean, standard deviation, median, interquartile range and frequency 
distribution) will be generated for baseline demographic and clinical information to characterize 
the study population for subjects of 4 treatment arms, respectively. All outcomes of interest 
(TNSS, PNIF and biomarkers) will also be summarized by treatment arms and repeated 
measurement time points. Box plots and histograms will be generated to visualize the variable 
distributions, and scatter plot will be used to explore the relationship among variables. Primary 
analysis: outcomes of interest will be collected at different time points in visits 2 (baseline) and 3 
(after 3 weeks of treatment).  TNSS will be recorded before 30 minutes after nasal lavage, 5, 15, 
30, mintues then hourly to 4 hours after challenge initiation.  According to Bareille (figure 2), 
TNSS is stable after 3 hours following challenge, therefore we will average the TNSS at 3 and 
hour as the analysis outcome to stabilize the outcome measure and simplify the analysis.  
 
We are interested in detecting the difference in the outcomes among subjects receiving a 
placebo tablet  vs. active tablet  , with and without nasal corticosteroid.  The primary endpoint is 
the TNSS.  ANCOVA (Analysis of covariance) model will be used to evaluate the difference in 
TNSS after treatment between placebo tablet vs active tablet  , adjusting for their baseline 
values.  Analysis will be carried out based on intention to treat principal with and without 
covariate adjustment.  In addition to treatment assignment (4 groups), other fixed effects in the 
model may include baseline value, subject-level covariates (e.g., age, sex, baseline disease 
severity, medical history, etc.).  Estimates of difference between placebo vs. active tablet  will be 
calculated based on appropriate contrasts.  Variable transformation (e.g., log transformation) 
will be used if the outcome distribution is skewed and normality assumption is imperative or a 
better model fit is suggested with transformed covariates.  Secondary analysis will include the 
change in PNIF and biomarkers, and similar ANCOVA model will be used.  Correlation analysis 
will also be carried out to examine the associations among outcomes.     
 
Sample size consideration: Similar to the study design in Bareille 2012, we used standard 
deviation (SD) of TNSS of 2.08 to calculate the sample size needed.  With a total of 152 
subjects (36 subjects per arm) and the adjustment for multiple comparisons, we will have 80% 
power to detect a difference of 1.5 in TNSS between mango juice vs mango juice + BS groups 
at the 2.5% level with or without nasal corticosteroid.  This calculation is based on 2 sample t-
test with sample size of 36  per group, common standard deviation of 2.08 and alpha=0.025. 
The difference of 1.5 in TNSS is right in the range of difference between various treatments in 
Bareille 2012. Since TNSS outcome is taking average over 3, 4, 5 and 6 hours results, we 
expect the variability will be reduced and sample size calculation will be conservative.  Similar 
calculation can be carried out for PNIF and Biomarkers. According to (Lipworth 2012), we 
estimate the standard deviation % change in PNIF is about 12.7, with the sample size of 36 per 
arm, we will have 80% power to detect a difference of 9.1 in % change in PNIF between 
placebo tablet vs active tablet at the 2.5% level with or without nasal corticosteroid. According to 
(Baroody 2013), we estimate the SD of Tryptase is about 5 in their FF/PL (fluticasone 


 
Version  date: 11/2/2018 
Page 18 of 18 
 
furoate/placebo) group, and we will have 80% power to detect a difference of 3.6 at the 2.5% 
level. 
 
We will use multiple imputation method to handle the missing data problem such that each 
missing value will be replaced with a set of plausible values that represent the uncertainty about 
the right value to impute.  There multiply imputed data sets will be analyzed by the proposed 
analytic methods and the results from these analyses will be combined based on Rubin’s 
formula (Rubin 1987, Multiple Imputation for Nonresponse in Surveys, New York: John Wiley & 
Sons, Inc.).    
 
We will recruit 475 subjects that meet the inclusion/exclusion criteria for screening.  We expect 
237 subjects will test positive to grass and negative to perennial allergens, 190 subjects will 
have adequate positive nasal challenge and will be randomized.  The expected dropout rate is 
20%.     
 
Scatterplots and Spearman correlation will be produced to assess correlations between 
increase/decrease levels of the above variables.  We are looking for change biomarker 
variables that reflect inflammation, TNSS and PNIF.  We will assume effective anti-inflammatory 
treatment will lead to a significant decrease in inflammatory markers (IL4, IL5, IL13 and ECP), a 
significant increase in Total Nasal Symptom Scores (TNSS) and Peak Nasal Inspiratory Flow 
(PNIF).      
 
 
 
 

ICF_001.pdf: 
 
 
RESEARCH CONSENT FORM 
VA Greater Los Angeles 
Participant Name: ________________________________________________ Date:  ____________ 
Study Title: “Effects of Broccoli Sprout Extract on Allergic Rhinitis” 
Principal Investigator:  Zhaoping Li, MD, PhD 
Co-Principal Investigator:  Joseph Yusin, MD 
Phone: 310-268-3528 
Phone: 310-478-3711 x40230 
 
 
 
Version Date: November 1, 2018 
Page 1 of 15 
 
INTRODUCTION 
 
You are being invited to take part in a research study at the VA Greater Los Angeles Healthcare 
System under the direction of Dr. Zhaoping Li, Dr. Joseph Yusin and their research team. Before you 
decide to take part, it is important for you to know why the research is being done and what it will 
involve.  This includes any potential risks to you, as well as any potential benefits to you and/or to the 
future population of individuals you represent.  
 
This document describes the purpose, procedures, benefits, risks, discomforts and precautions of the 
study.  It also describes the alternative procedures that are available to you and your right to withdraw 
from the study at any time.  No guarantees or assurances can be made as the results of the study. 
You must be completely truthful with your study doctor regarding your health history, including past and 
present usage of both prescription and non-prescription medications.  If you are not completely truthful 
with your study doctor you might harm yourself by participating in this study. 
 
Read the information below closely, and discuss it with family and friends if you wish.  Ask one of the 
study staff if there is anything that is not clear or if you would like more details.  Take your time to 
decide.  If you do decide to take part, your signature on this consent form will show that you received all 
of the information below, and that you were able to discuss any questions and concerns you had with a 
member of the study team.   
 
Your participation in this study is voluntary. If you don’t take part, you can still receive all usual care that 
is available to you. Your decision not to take part will not affect the relationship you have with your 
doctor or other staff, and it will not affect the usual care that you receive as a patient. If you decide to 
take part, you may still withdraw at any time.  If you do not wish to be in this study or leave the study 
early, you will not lose any benefits to which you are entitled.  
 
If you are a VA employee or student, your refusal to take part in the study will in no way influence your 
employment, ratings, subsequent recommendations, or academic progress as applicable. 
 
Only you can decide if you want to take part in this study. You should only make your decision after 
reading all the questions and answers in this form.  You may talk to your family, friends and/or your 
family doctor to help make your decision. You can take as much time as you like to decide.  
 
After you have read the entire form, you will be given the chance to ask any questions that you may 
have. When you have had the chance to ask any questions and they have been answered to your 


 
RESEARCH CONSENT FORM 
VA Greater Los Angeles 
Study Title: “Effects of Broccoli Sprout Extract on Allergic Rhinitis” 
Principal Investigator:  Zhaoping Li, MD, PhD 
Co-Principal Investigator:  Joseph Yusin, MD 
Phone: 310-268-3528 
Phone: 310-478-3711 x40230 
 
 
 
Version Date: November 1, 2018 
Page 2 of 15 
satisfaction, if you decide to take part, sign the pages at the end of this form to show that you agree to 
be part of the study. This is called “giving your consent”.  
 
Even after you have signed this study consent form you can change your mind and decide not to 
participate in the study. You do not have to give a reason. 
 
BACKGROUND AND PURPOSE 
 
Allergic rhinitis, commonly referred to as “hay fever” is suffered by many people, especially those living 
in high pollutant areas, despite conventional treatment. We believe that some people who suffer from 
allergic rhinitis may improve their nasal health if they consume broccoli sprout extracts alone or in 
combination with conventional medications.  The purpose of this study is to see if taking a pill which 
contains broccoli sprout extract (BSE) will increase the levels of natural helpful antioxidant enzymes in 
your nose and sinuses. Antioxidant enzymes are proteins produced by the body to protect cells against 
the harmful effects of chemicals such as those found in air pollution. We will also be measuring certain 
cells that are related to inflammation in the airway.  We would like to see if taking broccoli sprout extract 
decreases inflammation caused by nasal exposure to grass. We propose to study the outcomes of 
broccoli sprout extract on allergic rhinitis alone or in combination with standard of care nasal 
corticosteroids   
 
You have been invited to participate in this study because you are 18 years of age or older and you 
may be allergic to grass. 
 
DURATION OF THE RESEARCH 
   
The study is intended to recruit 475 people with the expectation of having 152 people complete the 
study. Your participation in this study could last up to 55 days.  
 
STUDY PROCEDURES  
 
Overview:  If you agree to take part in the study, and you qualify to continue the study after visit 1, you 
will be asked to come into the clinic for a total of 2 additional visits.  
 
The study schedule is summarized in a table at the end of this section. If you qualify, it is important that 
you attend each scheduled visit. Visit 2 and 3 (Day 14 and 35) will last about 5 hours. We will record 
any possible side effects throughout the entire study.  
 
After you have consented by signing this form, the study team will ask you about the medications that 
you’re currently taking (including over-the-counter medications). If certain allergy medications are 
prescribed to you (such as loratadine, cetirizine, fluticasone, triamcinolone, etc.) we’ll confirm with you 
the date that you last took these medications, and ask that you hold certain allergy medications during 


 
RESEARCH CONSENT FORM 
VA Greater Los Angeles 
Study Title: “Effects of Broccoli Sprout Extract on Allergic Rhinitis” 
Principal Investigator:  Zhaoping Li, MD, PhD 
Co-Principal Investigator:  Joseph Yusin, MD 
Phone: 310-268-3528 
Phone: 310-478-3711 x40230 
 
 
 
Version Date: November 1, 2018 
Page 3 of 15 
your participation in the study. After you and the research team have gone over your medications you 
will be given a list of these medications (including new vitamins and supplements) sheet to remind you 
to avoid taking them for the duration of the study. Three days prior to visit 1, you should avoid the 
following foods: arugula, bok choy, broccoli, broccoflower, brussel sprouts, cabbage, cauliflower, chard, 
chinese cabbage, collard greens, daikon, kale, kohlrabi, mustard greens, radishes, rutabagas, turnips 
and watercress.  You will be given a list of these foods to remind you to avoid eating them for the 
duration of the study. 
 
You will be scheduled for visit 1 within 20 days after consenting to undergo an allergy skin test that will 
determine if you have nasal allergies to grass pollen and will then undergo a nasal challenge test.  If the 
nasal challenge test causes nasal symptoms (runny nose, itching, nasal congestion) you will qualify to 
continue with the other visits..    
 
Once you have successfully completed the screening phase of the study, you will return for Visit 2 in 
two weeks.   
  
You will need to have the following examinations, tests or procedures done during the study: 
 
• 
General Physical Examination (Day 1, 14 and 35):   
You will undergo a routine physical examination before any nasal challenge or nasal irrigation is 
performed.  The examination will include looking into your nose, listening to your heart and 
lungs and obtaining your weight, height, blood pressure and heart rate.   
 
• 
Urine Test (Day 1): 
If you are a woman and can have children, a urine (and also if needed a blood) pregnancy test 
will be done to confirm that you are not pregnant. The study doctor or study staff will tell you if 
the pregnancy test results are positive. The results of the pregnancy testing must be negative in 
order for you to be in the study.  
 
• 
Spiro metry  (Day 1, 14 and 35):    
You will be asked to exhale forcefully into a tube for about 6 seconds.  You will be guided by the 
technician.  This test measures your lung function.   
 
• 
Allergy Skin Testing for sensitivity to Common Aeroallergens (Day 1): 
To determine if you will be able to participate in the entire study, we will need to determine if you 
have an allergy to only grass pollen. You will be skin tested to common grass pollen along with 
cat, dog, dust mite and mold.  Allergy skin testing is a routine procedure allergists perform when 
evaluating patients with allergies.  It involves cleaning a small area on your arm with alcohol, 
drying the area, then scratching the skin with a small needle containing the allergen (grass, cat, 
dog, dust mite or mold) on your arm we will also apply histamine and saline on your arm.  After 


 
RESEARCH CONSENT FORM 
VA Greater Los Angeles 
Study Title: “Effects of Broccoli Sprout Extract on Allergic Rhinitis” 
Principal Investigator:  Zhaoping Li, MD, PhD 
Co-Principal Investigator:  Joseph Yusin, MD 
Phone: 310-268-3528 
Phone: 310-478-3711 x40230 
 
 
 
Version Date: November 1, 2018 
Page 4 of 15 
20 minutes, we will check the areas to see which area has a response. If you have a response it 
will feel like a mosquito bite with a small bump with itching and redness at the site. If you test 
positive to grass pollen and based on the skin test results, the study doctor determines you do 
not have a true allergy to cat, dog, dust mite or mold you will qualify to continue the study.   
 
• 
Nose (nasal) lavage (Day 14 and 35):   
A nose lavage is a rinsing of the nose and sinuses with a sinus rinse (sterile salt water). A nasal 
lavage is performed before each nasal challenge. For the nose wash you will lean over a sink 
with your head titled at an angle, and breathe out of your mouth while you gently pour 8 ounces 
(1 cup) of sinus rinse into one nostril, and let the sinus rinse drip out of the other nostril until the 
sinus rinse is gone.  
  
• 
Nasal Challenge (Day 1, 14 and 35):   
There will be two types of nasal challenges performed: the “up-dosing” and the “actual” nasal 
challenge. 
 
The up-dosing nasal challenge completed at Visit 1: The up-dosing nasal challenge is done to 
determine how much grass pollen solution is needed for you to experience allergy nasal 
symptoms. You will need to have a certain level of symptoms in order to continue in the study.     
The technician will apply three sprays of the grass pollen solution into each nostril.  10 minutes 
after the first dose, you will fill out a questionnaire that will indicate to the technician your nasal 
symptom level. Based on the level of your symptoms, the technician will decide to continue the 
challenge with a stronger dose or to stop with that particular dose.  If the technician decides to 
continue, you will receive one spray of a higher concentration grass pollen solution in each 
nostril and then complete the nasal symptom form 10 minutes following the spray.  The nasal 
spray followed by completing the questionnaire will be repeated with gradually higher 
concentrations of the grass pollen solution until you experience a certain level of symptoms. 
You will need to have a certain level of symptoms before the technician reaches the maximum 
concentration.  If you do not, you will not be able to continue with the study.     
 
The actual nasal challenge completed at visits 2 and 3:  The actual nasal challenge will involve 
spraying a grass pollen solution into both nostrils.  The grass pollen concentration used will be 
determined by the up-dosing nasal challenge performed at the first visit. As with the up-dosing 
nasal challenge, you will perform a nasal lavage 30 minutes prior to beginning the nasal 
challenge.  Over the next 4 hours, the following three items will be collected: 
1. Total nasal symptom score (TNSS)  
2. Peak Nasal Inspiratory Flow (PNIF)  
3. Mucus collection to analyze inflammatory cells and cell markers.   
 
Mucus collection procedure: To collect mucus, a small nasal sponge, smaller than the size of 
your nostril opening will be placed inside your nose, after two minutes, the sponge will be 


 
RESEARCH CONSENT FORM 
VA Greater Los Angeles 
Study Title: “Effects of Broccoli Sprout Extract on Allergic Rhinitis” 
Principal Investigator:  Zhaoping Li, MD, PhD 
Co-Principal Investigator:  Joseph Yusin, MD 
Phone: 310-268-3528 
Phone: 310-478-3711 x40230 
 
 
 
Version Date: November 1, 2018 
Page 5 of 15 
removed and then sent to the lab to look at the cells and cell markers.  This mucus collection 
technique will occur at the same time intervals as the TNSS and PNIF collections 30 minutes 
after nasal lavage, 5, 15, 30 minutes and then 1,2,3 and 4 hours following the nasal challenge. 
 
• 
Blood draw (Day 1, 14 and 35):  
We will draw 10mL (about 2 teaspoons) of blood during the screening phase of the study. We 
will check your blood for different types of cell markers found in allergy driven cells and genes 
that are related to your ability to produce antioxidants. Your blood will also be measured for 
sulforaphane (a substance found in broccoli sprouts) levels.  Total blood drawn during this study 
is 30mL (about 2 tablespoons) over the 5-week study period. 
  
• 
Questionnaire (Day 1, 14 and 35):   
You will be asked to complete a questionnaire called the Total Nasal Symptom Score (TNSS) 
where you will be asked to rate the symptoms of nasal congestion/postnasal drainage, itching, 
runny nose or sneezing.  As mentioned above, you will complete this questionnaire when you 
undergo nasal challenges.    
 
• 
Peak Nasal Inspiratory Flow (Day 1, 14 and 35):  
You will also undergo a procedure called Peak Nasal Inspiratory Flow (PNIF) where you will be 
asked to exhale fully and quickly while having an airtight seal that covers your mouth and nose.  
After this is done, the technician will record the reading.  You will repeat this study 2 more times.  
 
• 
Dietary Supplement: Avmacol® (Day 14 and 35):  We will ask you to take 4 tablets of a 
dietary supplement with your evening meal containing broccoli sprout extract (BSE). The 
supplement will be taken about the same time each day.  We will ask you not to eat certain 
cruciferous vegetables (such as broccoli, kale, cauliflower, and brussel sprouts) for 3 days 
before visit 1, and while on the study. A complete list of these vegetables will be given to you by 
the study coordinator.  
 
 
The study visit for Day 1 (screen) will last approximately 1 to 3 hours (based on how long the nasal 
challenge will take); study visits for Days 14 and 35 will last up to 5 hours.  Please plan accordingly.   
 
Subjects who participate in this study will be randomized to receive a three-week supply of one of the 
following regimens. Note, there is no active substance in placebo.    
1. nasal corticosteroid   
2. placebo nasal spray + dietary supplement containing BSE  
3. nasal corticosteroid  + dietary supplement containing BSE  
4. placebo nasal spray  
 
This will be determined randomly, in a process similar to flipping a coin. You will have an equal (25%) 
chance of being assigned to each study drug group.  This is a double-blind study, which means that 


 
RESEARCH CONSENT FORM 
VA Greater Los Angeles 
Study Title: “Effects of Broccoli Sprout Extract on Allergic Rhinitis” 
Principal Investigator:  Zhaoping Li, MD, PhD 
Co-Principal Investigator:  Joseph Yusin, MD 
Phone: 310-268-3528 
Phone: 310-478-3711 x40230 
 
 
 
Version Date: November 1, 2018 
Page 6 of 15 
neither you nor the study doctor will know to which of these study drug groups you have been assigned.  
In case of an emergency, the study doctor can get this information. Visit 1 would take place once you 
have avoided consuming these vegetables for three days.     
 
 
Visit 1 
Day 1 
Visit 2 
Day 14 
Visit 3 
Day 35 
Informed Consent 
X 
 
 
Medical History, Physical Exam 
(including weight and vitals) 
X 
X 
X 
Urine (WOCBP) 
X 
 
 
Genetic Testing  
X 
 
 
Blood Draw (SFN)  
X 
X 
X 
Allergy Skin test 
X 
 
 
Nasal Exam 
X 
X 
X 
Nasal Challenge 
X 
X 
X 
Nasal Irrigation 
 
X 
X 
Spirometry 
X 
X 
X 
 
POSSIBLE RISKS OR DISCOMFORTS 
 
You may experience new symptoms or a change in your medical condition while taking part in this 
study. These symptoms or changes in your medical condition may or may not be due to study 
drugs/supplement being used.   
 
Please tell the study doctor or study staff right away if you have any side effects. Please tell them if you 
have any other problems with your health or the way you feel during the study, whether or not you think 
these problems are related to the study.  
You will receive either nasal fluticasone or a nasal placebo spray.  Placebo is a substance that looks 
like a drug but has no drug in it. If you receive placebo during the study, it is possible that your allergies 
may get worse. Please ask the study doctor or study staff if you have any questions about placebo. 
Risks of the usual care you receive are not risks of the research.  Those risks are not included in this 
consent form.  You should talk with your health care providers if you have any questions about the risks 
of usual care. 
 
General Risk 
There is a risk of experiencing allergy symptoms (itchy eyes and runny nose) if you stop taking 
antihistamines during the study. 
 


 
RESEARCH CONSENT FORM 
VA Greater Los Angeles 
Study Title: “Effects of Broccoli Sprout Extract on Allergic Rhinitis” 
Principal Investigator:  Zhaoping Li, MD, PhD 
Co-Principal Investigator:  Joseph Yusin, MD 
Phone: 310-268-3528 
Phone: 310-478-3711 x40230 
 
 
 
Version Date: November 1, 2018 
Page 7 of 15 
POSSIBLE RISKS ASSOCIATED WITH STUDY PROCEDURES/TESTS  
 
Allergy skin testing: 
There may be some mild redness and/or itching at the sites where the needle scratches the skin.  This 
is expected and a normal part of this test when it is positive.  If present, these symptoms will disappear 
over a period of less than one hour.  There is an extremely small (less than 0.02%) chance you may get 
a severe allergic reaction to this test which may include hives (itchy rash) that may last several hours, 
“hay fever-like” symptoms such as sneezing or runny nose, chest tightness or even a drop in blood 
pressure.  Qualified staff will perform skin testing and a board certified allergist will be available to 
address any complications.  Medications such as epinephrine and Benadryl will be readily available.   
 
Nose Lavage: 
The sensation of water in the nose may be mildly uncomfortable initially. There is a possibility of 
swallowing the saline, which has a salt-water taste but contains no other substances.  Additional 
potential complications include inhaling the fluid, which is uncomfortable, but the amount of fluid is so 
small that there is no known possibility of any respiratory complication.  
 
Nasal Challenge: 
Since we are placing grass pollen solution that you are allergic to in your nostril, you may experience  
“hay fever-like” symptoms (such as sneezing, nasal congestion, runny nose, or watery eyes).  There is 
a very small chance that you could experience chest tightness or even a drop in blood pressure, known 
as “anaphylaxis”.  If this does occur, the test would be stopped and you would be treated.  The reason 
why we perform a physical exam and have you perform spirometry (mentioned above) is to determine if 
you may be at slightly higher risk of developing anaphylaxis.  If you are at risk to have such a reaction 
based on your exam and spirometry readings, you will not undergo the test.     
 
Dietary Supplement:  Avmacol® 
The reported risks associated with taking broccoli sprout extract are mild gastrointestinal discomfort 
(including mild stomach pain, mild diarrhea, soft, loose or watery stools), nausea, belching, bloating, or 
flatulence. If you find the flavor of broccoli sprout extract mixture is unpleasant, you may have 
discomfort associated with swallowing it. No other risks have been associated with broccoli with the 
ingestion of broccoli sprout ingestion and no serious adverse events have been observed or reported.   
 
Risks associated with blood sampling:  
When giving blood you may feel dizzy or faint, or experience mild pain, bruising, irritation or redness at 
the site. In rare cases, you may get an infection.  
 
Loss of Confidentiality:  
There is a risk of loss of confidentiality of your information. You will read more about the protection of 
your information later in this form. Please ask the study doctor or study staff if you would like to know 
more about how your information will be protected while you are in this study.  


 
RESEARCH CONSENT FORM 
VA Greater Los Angeles 
Study Title: “Effects of Broccoli Sprout Extract on Allergic Rhinitis” 
Principal Investigator:  Zhaoping Li, MD, PhD 
Co-Principal Investigator:  Joseph Yusin, MD 
Phone: 310-268-3528 
Phone: 310-478-3711 x40230 
 
 
 
Version Date: November 1, 2018 
Page 8 of 15 
Risk of Pregnancy: 
Since we do not know if the broccoli sprout extract mixture is safe for an embryo or fetus, females must 
not be pregnant while in the study. Females of childbearing potential will have a urine pregnancy test at 
the start of study. You must use an effective method of contraception throughout the study.  Examples 
of acceptable effective methods are as follows: total abstinence (no intercourse), an intrauterine device 
(IUD), a double barrier method (such as a condom plus diaphragm with spermicide), a contraceptive 
implant, an oral contraceptive, or have a vasectomized partner with confirmed azoospermia (no sperm).  
If you become pregnant or suspect that you became pregnant while on the study, you must 
notify your research doctor immediately. If you are scheduled to participate, you will be immediately 
taken out of the study. If you are already taking study drug, you will be immediately taken off study drug 
and continue participation through follow-up visits. The research doctor or a member of the research 
team may ask for information about the pregnancy outcome (limited to “live birth, miscarriage or 
termination of pregnancy”).  
 
POTENTIAL BENEFITS 
 
We cannot promise that you will get any benefits from taking part in this research study.  However,  
taking part in this study may or may not make your allergic rhinitis better, and may or may not have a 
direct benefit to you. You may get placebo, which is a substance that looks like a drug but has no drug 
in it.  
 
The research may increase our understanding of the healthful effects of vegetables such as broccoli 
sprouts on the respiratory and immune system.  This information may be important in improving nasal 
health for people with allergies and asthma, and for developing ways to protect people from the harmful 
effects of air pollution. 
 
ALTERNATIVES TO PARTICIPATING IN THIS RESEARCH 
 
You will continue to get regular care from your own regular doctor(s) if you choose not to participate in 
this study. You can get fluticasone without participating in the study. You might decide to take part in 
another study instead. Talk with your study doctor about your choices. In addition, you may discuss 
your options with your regular health care provider. 
 
CONFIDENTIALITY 
 
Your data will be combined with data from other people taking part in this study.  We will write medical 
articles about the combined data we have gathered.  Any talks or papers about this study will not 
identify you.   
 
We will not share your records or identify you unless we have to by law.  There are times when we 
have to show your records to other people.  For example, someone from the State or Federal 


 
RESEARCH CONSENT FORM 
VA Greater Los Angeles 
Study Title: “Effects of Broccoli Sprout Extract on Allergic Rhinitis” 
Principal Investigator:  Zhaoping Li, MD, PhD 
Co-Principal Investigator:  Joseph Yusin, MD 
Phone: 310-268-3528 
Phone: 310-478-3711 x40230 
 
 
 
Version Date: November 1, 2018 
Page 9 of 15 
Regulatory Agencies, the Food and Drug Administration (FDA), the Office of Human Research 
Protections (OHRP), the Government Accountability Office (GAO), the VA Office of Research Oversight 
(ORO), the Institutional Review Board (IRB) – VAGLAHS and any of its delegated sub commissions, 
our local Research and Development Committee, the Research Compliance Officer and other study 
monitors may look at portions of records that identify you.  
 
Your participation in this study is entirely confidential and will not affect any medical care to which you 
are otherwise entitled.   
 
In the event your study doctor is unable to locate you (e.g. after making 3 phone call attempts and 
sending a certified letter) for the limited purpose of collecting necessary follow-up data, you authorize 
your study doctor to contact your private physician/General practitioner for the sole purpose of updating 
your study doctor with your current contact information.   
 
Participation in this study will involve a loss of privacy, but information about you will be handled as 
confidentially as possible. While the VA and the researchers on the team will do everything to prevent 
any loss of information, in the event that there is a breach of information, you will be contacted by the 
VA directly. 
 
All records containing personal information, research data, and related records will be stored in locked 
files at a VA Room at Building 500.  All data that will be collected will be assigned with a personal 
identification code to protect personal privacy.  It will be accessible only by members of the research 
team named above.  The required records, including the investigator’s research records, must be 
retained for 6 years after the study is closed by the National Archives and Records Administration 
(NARA.gov) guidance as implemented by VHA Records Schedule (DAA-0015-2015-0004). These 
records may be retained longer if required by other Federal regulations.   
 
We will keep confidential all research records that identify you to the extent allowed by law.  However, 
you should know that there are some circumstances in which we may have to show your information to 
other people.  For example, someone from the Food and Drug Administration, the Office of Human 
 
Research Protections, the Government Accounting Office, the VA Office of Research Oversight, our 
Institutional Review Board, our Data and Safety Monitoring Board and the Research Compliance Office 
may look at or copy portions of records that identify you, including your medical record; the sponsor and 
its authorized agents may look at, but not copy, portions of the records that identify you, including your 
medical record. In addition, please note that VA policy requires that a note be written in your medical 
record concerning the consent process and your enrollment in this study.  Thus non-research VA staff 
will have access to that information in the course of clinical care. 
 


 
RESEARCH CONSENT FORM 
VA Greater Los Angeles 
Study Title: “Effects of Broccoli Sprout Extract on Allergic Rhinitis” 
Principal Investigator:  Zhaoping Li, MD, PhD 
Co-Principal Investigator:  Joseph Yusin, MD 
Phone: 310-268-3528 
Phone: 310-478-3711 x40230 
 
 
 
Version Date: November 1, 2018 
Page 10 of 15 
In addition, in accordance with California law, confidentiality cannot be guaranteed if the investigator 
becomes aware that you may be a danger to yourself or to others, or becomes aware that acts of child 
abuse or elder abuse may have occurred.  
 
We may write about the combined information we have gathered, however, any presentations or 
publications from this information will not identify you. 
 
At the end of this form you will be asked for your permission to allow the VA to share your Personal 
Health Information with the research team for use in this study.  Your separate signature on that part of 
the consent form will reflect your official authorization.   
 
COSTS TO PARTICIPANTS AND PAYMENT 
 
Costs to Participants: 
You will not be charged for any treatments or procedures that are part of this research study.  If you 
usually pay co-payments for VA care and medications, you will still pay these co-payments for VA care 
and medications that are not part of this study.   
 
PAYMENT OFFERED FOR PARTICIPATION 
 
You will be paid up to $300 if you complete the entire study.  If you do not complete the study, your 
payments will be pro-rated as follows: Twenty dollars ($20.00) will be paid to participants who complete 
up to the skin test procedure at visit 1, and do not meet study qualifications (skin test results determine 
that you not allergic to grass) to continue participation in the study. Forty-five dollars ($45.00) will be 
paid to participants who meet study qualifications and complete visit 1. Ninety dollars ($90.00) will be 
paid to participants who complete visit 2. Ninety dollars ($90.00) will be paid to participants who 
complete visit 3. Additionally, those participants who will comply with study protocol and complete all 
three study visits will receive the completion of study payment in amount of $75 at the end of the study. 
These payments are to cover your travel expenses and for the time and effort you have expended. You 
will be paid in cash at the completion of your study visit.  If you decide to withdraw or you are withdrawn 
by the study doctor, your payment with be prorated for the visits you completed.     
 
In addition, it is VA policy that the amount you receive from this study is more than $550.00 it will be 
reported to the Internal Revenue Service (IRS) and may be considered taxable income.  This will be 
reported using a 1099 (Miscellaneous Income) form. This form will be issued to you and a copy mailed 
to the IRS.   
 
As part of the study, you will receive study drug and all the study tests and procedures at no cost to 
you.   
 


 
RESEARCH CONSENT FORM 
VA Greater Los Angeles 
Study Title: “Effects of Broccoli Sprout Extract on Allergic Rhinitis” 
Principal Investigator:  Zhaoping Li, MD, PhD 
Co-Principal Investigator:  Joseph Yusin, MD 
Phone: 310-268-3528 
Phone: 310-478-3711 x40230 
 
 
 
Version Date: November 1, 2018 
Page 11 of 15 
You and the VA or your insurance company/the National Health Service will continue to pay for your 
regular health care.   
 
MEDICAL TREATMENT AND COMPENSATION FOR INJURY   
 
Every reasonable safety measure will be used to protect your well-being.  If you are injured as a result 
of taking part in this study, the VA will provide necessary medical treatment at no cost to you unless the 
injury was due to your not following the study procedures.  However, no plans or funds for additional 
compensation have been set aside.  You have not waived any legal rights or released the hospital or its 
agents from liability for negligence by signing this form.   
 
If you should have a medical concern or get hurt or sick as a result of taking part in this study, call:   
 
DURING THE DAY: 
 
Dr. Zhaoping Li, MD, PhD              at       310-268-3528                    
   
 
Dr. Joseph Yusin, MD  
 
 
 
at  
310-478-3711 x 40230    
and,  
 
AFTER HOURS: 
 
Dr. Zhaoping Li, MD, PhD              at       310-261-5024                           
                                   
 
Emergency and ongoing medical treatment will be provided as needed.   
 
YOUR RIGHT TO TERMINATE PARTICIPATION 
 
It is up to you to decide whether or not to take part in this study. If you decide to take part you may still 
withdraw at any time.  If you do not wish to be in this study or leave the study early, you will not lose 
any benefits to which you are entitled.  If you don’t take part, you can still receive all usual care that is 
available to you.  Your decision not to take part will not affect the relationship you have with your doctor 
or other staff, and it will not affect the usual care that you receive as a patient.   
 
At the end of the study, the study staff will contact you, your family or your friends to ask about your 
health status even if you withdraw from the study.  If you withdraw from the study and revoke your 
Authorization to Use and Disclose your Protected Health Information, the study doctor will not call you 
at the end of the study and your information will no longer be used by the study doctor and staff for this 
study, except to the extent the parties to the research have already taken action or need the information 
to complete analysis and reports for this research. 
 


 
RESEARCH CONSENT FORM 
VA Greater Los Angeles 
Study Title: “Effects of Broccoli Sprout Extract on Allergic Rhinitis” 
Principal Investigator:  Zhaoping Li, MD, PhD 
Co-Principal Investigator:  Joseph Yusin, MD 
Phone: 310-268-3528 
Phone: 310-478-3711 x40230 
 
 
 
Version Date: November 1, 2018 
Page 12 of 15 
The sponsor will still use the study information that was collected before you withdrew from the study.  
If you stop being part of this study, the study doctor or one of the staff members will talk to you about 
any medical issues regarding the stopping of your participation. 
 
RIGHT OF INVESTIGATOR TO TERMINATE YOUR PARTICIPATION 
 
The investigator may withdraw you from participating in this research if circumstances arise that 
warrant doing so. The study doctor or the sponsor can stop your participation at any time without your 
consent for reasons such as: 
• 
If it appears to be medically harmful to you; 
• 
If you fail to follow directions for participating in the study; 
• 
If it is discovered that you do not meet the study requirements; 
• 
If the study is canceled; or  
• 
For administrative reasons, including competitive enrollment - the target number of subjects  
has enrolled.   
 
New information may become available that might affect your choice to take the study drug or your 
willingness to continue in the trial. This new information may include safety issues with the study drug. 
Such information will be shared with and discussed with you. 
 
Should you become ill during the research, you may have to withdraw, even if you would prefer to 
continue. The investigator Zhaoping, Li, M.D. or Dr. Joseph Yusin, will make the decision and inform 
you if your continued participation is not possible.  The decision may be made either to protect your 
health and safety, or because it is part of the research plan that people who develop certain conditions 
may not continue to participate. 
 
What happens if I do not want to stay in the study?  
 
You should discuss your decision with your study doctor. It is important for the study that you provide 
follow-up information on your health status until the end of the study. If you stop participating in the 
study (that is you stop taking study drug and you decide not to attend clinic or allow us to contact you 
by telephone) information about your overall health will continue to be collected until the end of the 
study.  
 
Any information that is available to the public and allowed by local law and is important for the study, 
including whether you are alive, may be used, even if you stop participating in the study.  
 
If you have provided blood (or urine, if you are a woman of child bearing potential) samples, you may 
withdraw your consent for these to be used at any time. Your samples will be destroyed at that time. 
However, all the data and samples collected before you left the study will still be used for the study. 


 
RESEARCH CONSENT FORM 
VA Greater Los Angeles 
Study Title: “Effects of Broccoli Sprout Extract on Allergic Rhinitis” 
Principal Investigator:  Zhaoping Li, MD, PhD 
Co-Principal Investigator:  Joseph Yusin, MD 
Phone: 310-268-3528 
Phone: 310-478-3711 x40230 
 
 
 
Version Date: November 1, 2018 
Page 13 of 15 
You may ask that your samples be destroyed but any data already collected about your samples will be 
used for the study.  
 
PERSONS TO CONTACT ABOUT THIS STUDY 
 
In the event that you have a question about the research or experience a research related injury or 
adverse reaction, please immediately contact one of the investigators on this study: Dr. Zhaoping Li at 
310-261-5024 or Dr. Joseph Yusin at 310-478-3711 x40230. 
 
If you have questions about your rights as a study participant, or you want to make sure this is a valid 
VA study, you may contact the VA Greater Los Angeles Institutional Review Board (IRB). This is the 
Board that is responsible for overseeing the safety of human participants in this study.  You may call 
the VA Greater Los Angeles IRB toll free at 1-310-268-4437 if you have questions, complaints or 
concerns about the study or if you would like to obtain information or offer input.   
 
SIGNIFICANT NEW FINDINGS  
 
Sometimes during the course of a research study, new information becomes available about the BSE 
that is being studied that might change a person’s decision to stay in the study.  If this happens, your 
research doctor will tell you about it and discuss with you whether you want to continue in the study.  If 
you decide to withdraw from the study, your research doctor will arrange for your medical care to 
continue.  If you decide to continue in the study, you might be asked to sign an updated informed 
consent form.  Your research doctor could also decide it to be in your best interests to withdraw you 
from the study.  If so, he or she will explain the reasons and arrange for your usual medical care to 
continue.   
 
GENETIC INFORMATION IN YOUR SAMPLE: POSSIBLE LIMITS TO INDIVIDUAL 
CONFIDENTIALITY 
 
Every tissue or fluid sample contains genetic information.  In this study we are identifying the genetic 
variation of an antioxidant enzyme that has been found to modify the effect of broccoli sprouts on 
allergic rhinitis.   Every precaution will be taken to maintain your confidentiality now and in the future 
and to safeguard your genetic material. 
 
We have learned from past research that we will not always be able to predict future research findings 
and new technologies.  You should be aware that unforeseeable problems may arise from new 
developments. Possible problems include insurance or employment discrimination based on genetic 
information. 
 
Sometimes genetic information suggesting different parentage is obtained during research.  In this 
study we do not plan to determine parentage. 


 
RESEARCH CONSENT FORM 
VA Greater Los Angeles 
Study Title: “Effects of Broccoli Sprout Extract on Allergic Rhinitis” 
Principal Investigator:  Zhaoping Li, MD, PhD 
Co-Principal Investigator:  Joseph Yusin, MD 
Phone: 310-268-3528 
Phone: 310-478-3711 x40230 
 
 
 
Version Date: November 1, 2018 
Page 14 of 15 
Within the limits imposed by technology and the law, every effort will be made to maintain the privacy of 
your genetic information. 
 
 
 
AGREEMENT TO PARTICIPATE IN THE RESEARCH STUDY 
 
Dr./Mr./Ms___________________________ has explained the research study to you.  You have been 
told of the risks or discomforts and possible benefits of the study. You have been told of other choices 
of treatment available to you.  You have been given the chance to ask questions and obtain answers.   
 
You voluntarily consent to participate in this study.  You also confirm that you have read this consent, or 
it has been read to you.  You will receive a copy of this consent after you sign it.  A copy of this signed 
consent will also be put in your medical record if applicable.   
 
I agree to participate in this research study as has been explained in this document. 
 
 
 
_________________________ 
Participant’s Name 
____________________________ 
Participant’s Signature 
___________ 
Date 
 
I agree to participate in the GENETICS part of this research study as has been explained in 
this document. 
 
 
 
_________________________ 
Participant’s Name 
____________________________ 
Participant’s Signature 
___________ 
Date 
 
_________________________ 
Name of person obtaining consent 
 
 
 
____________________________ 
Signature of person obtaining consent 
___________ 
Date 
 
 
 
 


 
RESEARCH CONSENT FORM 
VA Greater Los Angeles 
Study Title: “Effects of Broccoli Sprout Extract on Allergic Rhinitis” 
Principal Investigator:  Zhaoping Li, MD, PhD 
Co-Principal Investigator:  Joseph Yusin, MD 
Phone: 310-268-3528 
Phone: 310-478-3711 x40230 
 
 
 
Version Date: November 1, 2018 
Page 15 of 15 
 
RIGHTS OF HUMAN SUBJECTS IN MEDICAL EXPERIMENTS 
 
Any person who is requested to consent to participate as a subject in a research study involving a 
medical experiment or who is requested to consent on behalf of another has the right to: 
 
1. Be informed of the nature and purpose of the experiment. 
 
2. Be given an explanation of the procedures to be followed in the medical experiment, and any drug or 
device to be utilized. 
 
3. Be given a description of any attendant discomforts and risks reasonably to be expected from the 
experiment. 
 
4. Be given an explanation of any benefits to the subject reasonably to be expected from the 
experiment, if applicable. 
 
5. Be given a disclosure of any appropriate alternative procedures, drugs or devices that might be 
advantageous to the subject, and their relative risks and benefits. 
 
6. Be informed of the avenues of medical treatment, if any, available to the subject after the experiment 
if complications should arise. 
 
7. Be given an opportunity to ask any questions concerning the experiment or the procedure involved. 
 
8. Be instructed that consent to participate in the medical experiment may be withdrawn at any time and 
the subject may discontinue participation in the medical experiment without prejudice. 
 
9. Be given a copy of any signed and dated written consent form used in relation to the experiment. 
 
10. Be given an opportunity to decide to consent or not to consent to a medical experiment without the 
intervention of any element of force, fraud, deceit, duress, coercion, or undue influence on the subject's 
decision.