Effect of Sulforaphane-rich Broccoli Sprout Homogenate on Ozone Induced Inflammation Through Modulation of NRF2

Potential subjects will be seen for a baseline screening visit at which time informed
consent will be obtained, a 12 lead ECG will be performed and spirometry with subsequent
sputum induction will be performed. Only subjects meeting the lung function criteria as
well as criteria for adequate sputum production will continue in the study. If the sample
is considered "borderline" by the study team, the subject maybe invited back to repeat
collection of another sputum sample. Subjects will be required to abstain from caffeine for
12 hours prior to all study visits and must not use/ingest non-steroidal anti-inflammatory
medications, anti-oxidant vitamins, juices/drinks fortified with extra vitamin supplement
and cruciferous vegetables 7 days prior to all visits. Subjects will be provided with a list
of cruciferous vegetables.

Qualified volunteers will next be seen for a training visit at which time they will undergo
physical exam by a study physician. After baseline spirometry is evaluated, the subject
will exercise on a treadmill, and measurements of minute ventilation will be collected.
Treadmill speed and/or elevation will be adjusted until the subject's minute ventilation is
at the target level of 30-40 L/min. The resulting speed and elevation will be used during
the ozone exposure sessions. At the training visit, participants will also undergo nasal
epithelial biopsy from one nare, and a buccal swab will be collected for genotyping.
Baseline blood samples will also be collected at this time for analyses of markers of
inflammation, cytokine assessments, SFN levels and complete blood cell count with
differential.

After completion of the training session, the subject will return for 4 sequential days.
During the first 3 days, subjects will ingest (observed ingestion) either broccoli sprout
homogenate or alfalfa sprout homogenate based on earlier randomization. One hour after the
supplement dose on the 3rd day, the volunteer will undergo exposure to 0.4 ppm ozone for a 2
hour period. During exposure, subjects will perform 15 minutes of moderate exercise on a
treadmill (minute ventilation = 30-40 L/min), each separated by 15 minutes of seated rest.
Heart rate and rhythm will be continuously monitored throughout the exposure. Spirometry
and symptom scoring will be performed both prior to and after exposure. One hour after the
end of exposure, a nasal epithelial biopsy will be collected from the nare that is
contralateral to that used at the prior visit. Four hours after completion of the exposure,
blood will be drawn for study endpoints and a sputum induction will be performed. The
following day the subject will return for a follow-up visit to include vital signs, symptom
scoring, spirometry, sputum induction and venipuncture for blood endpoints.

Subjects will be required to abstain from a list of vegetables for 1 week prior to each
study session until the completion of that session. Subjects will also be asked to maintain
a food diary for the same time.

After a 2 to 6 weeks washout period, participants will again return for a repeated sequence
of 4 days with 3 days of supplement ingestion followed by ozone exposure using the alternate
supplement and a 24 hour post exposure follow-up visit. Samples for analyses will be
collected at the same time points as those used in the initial sequence.

Inclusion Criteria:

1. Normal lung function, defined as (NHanes 2001 set):

FVC of > 80 % of that predicted for gender, ethnicity, age and height; FEV1 of > 80 %
of that predicted for gender, ethnicity, age and height; FEV1/FVC ratio of > .70.

2. Negative pregnancy test for females who are not s/p hysterectomy with oophorectomy;

3. Willing to avoid corticosteroids and non-steroidal anti-inflammatory medications for
1 week prior the baseline screening visit and again for 1 week prior to all
remaining visits;

4. Willing to avoid antioxidant vitamins and cruciferous vegetables as well as
juices/drinks with added vitamin supplements for 7 days prior the baseline screening
visit and throughout initial dosing period and ozone challenge as well as 7 days
prior to the start of the 2nd dosing period and subsequent challenge.

5. Willing to abstain from caffeine for 12 hours prior to all visits.Inclusion Criteria:

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Exclusion Criteria:

1. Any chronic medical condition considered by the PI as a contraindication to the
exposure study including, but not limited to, asthma, significant cardiovascular
disease, diabetes, chronic renal or thyroid disease;

2. Use of tricyclics and MAO inhibitors;

3. Pregnancy or nursing a baby;

4. Any current smoking;

5. Viral upper respiratory tract infection within 2 weeks of challenge;

6. Any acute infection requiring antibiotics within 2 weeks of challenge;

7. Abnormal physical findings at the baseline visit:

Abnormalities on lung auscultation Temperature > 37.8 Systolic BP>150 mm hg or < 85
mm Hg or diastolic BP>90 mm Hg or < 50 Oxygen saturation of < 94%;

8. Subjects must demonstrate the ability to produce an acceptable induced sputum sample
during the screening session. If the sample is unsatisfactory, the subject's
participation will end at that point;

9. Subjects who have been prescribed daily anti-inflammatory medications or medications
for asthma will be excluded. Oral contraceptives are acceptable. Antidepressants and
other medications may be permitted if in the opinion of the investigator the
medication will not interfere with the study procedures or compromise safety; and if
the dosage has been stable for 1 month;

10. Subjects who are unwilling to refrain from strenuous physical activity for 24 hours
before and after exposure;

11. Current nutritional disorder such as anorexia, bulimia, irritable bowel syndrome,
Crohn's disease etc;

12. Current use of immunosuppressive drugs;

13. History of intolerance of or aversion to broccoli;

14. Inability or unwillingness of a participant to give written informed consent;

15. Receipt of LAIV (Live Attenuated Influenza Vaccine), also known as FluMist®, or other
live vaccine within the prior 14 days.

16. Orthopedic injuries or impediments that would preclude treadmill exercise.