Dietary Nitrate and Muscle Power With Aging

This study consists of two parts separated by a 14 day wash-out. Subjects will be randomly
assigned to receive either the BRJ or placebo during the first part. During the second part
they will receive the opposite of what they had during the first part. This is considered a
double-blind study, which means neither the subjects nor the investigators will know what
form (BRJ or placebo) a subject is receives.

Study Visit One (Screening) 1-2 hours

The purpose of the screening visit is to explain all aspects of the study. It will also
determine if subjects can participate in the study. Subjects will provide a complete medical
history and undergo a physical exam. They will have their blood drawn (about 1.5 teaspoons)
and provide a urine sample. They will also practice the entire neuromuscular function
exercise test. During this test, the strength of their muscles will be determined by having
them kick, push and/or pull back as hard as you can while their leg is strapped to an
exercise device. A vital signs monitor will measure your blood pressure, heart rate, and
rhythm.

During the remainder of the study subjects will be instructed to consume their normal diet.
However, they will be asked to avoid eating foods high in nitrate such as beets, spinach, and
collard greens the evening before each visit. They will be asked to refrain from the use of
antibacterial mouthwash, such as Listerine or Cepacol, during the study. Chewing gum,
alcohol, and food and drinks containing caffeine (coffee, tea, chocolate, and soft drinks)
should be avoided 24 hours prior to each visit. They will be asked to fast for 12 hour prior
to each study visit.

Study Visit Two - Approximately 5 hours

At the beginning of this visit a catheter (small, flexible, sterile plastic tube) will be
placed through a vein in one of the subject's arms. This is for collection of blood samples.
Blood will be drawn four times during this visit. Each draw will be 6 mL or about 1.2
teaspoons. The first blood draw will check nitrate and nitrite levels. They will then have a
breath test to check nitric oxide. They will then drink 140 mL (about 2/3 of a cup) of BRJ
(or placebo). Blood and breath samples will be obtained every hour. Heart rate and blood
pressure will be measured at the same times the blood and breath samples are obtained.
Subjects will then rest quietly in a private room for about 2 hours after ingestion of BRJ
(or placebo) then will perform:

- The neuromuscular function test that was practiced during the screening visit.

- After completing the exercise test they will undergo a muscle biopsy of their thigh
muscle. First, the area will be sterilized and numbed with an injection. Next, a small
incision (about ¼ inch) will be made and a small biopsy needle will be used to collect a
small sample of your muscle (about the size of a pencil eraser). This will be performed
on the leg opposite of that used during the neuromuscular test.

- One final blood and breath sample will then be obtained.

- Subjects will then be provided a 7 day supply of BRJ (or placebo) and will continue to
drink two bottles (about 5 oz) every morning.

- Subjects will also be provided with an activity tracker. This will be worn at the hip
during all waking hours except when bathing or swimming. This monitor will measure total
activity time and sedentary time.

Study Visit Three - 1-2 hours

Subjects will return to the research center with the empty BRJ bottles. At this visit a
single blood (1.2 teaspoon) and breath sample will be collected. Subjects will then complete
a questionnaire to rate how tired they felt during the previous week. They will be provided
another 7 day supply of BRJ (or placebo) and will continue to drink two bottles every
morning.

Study Visit Four - Approximately 5 hours Subjects will return to the research center and
undergo the same procedures as they did in Study Visit Two. They will also complete a
questionnaire that will rate how tired they felt during the previous week.

Washout After visit four subjects will undergo a 14 day washout period where they will not
drink the BRJ (or placebo). There are no dietary restrictions at this time; subjects may
consume caffeine, alcohol, gum, mouthwash, etc. Subjects will then repeat Study Visits Two,
Three, and Four again. However, during this they will be given the opposite form of BRJ you
had received before.

Study Visit Five Subjects will repeat the procedures completed during Study Visit Two.

Study Visit Six Subjects will repeat the procedures completed during Study Visit Three.

Study Visit Seven Subjects will repeat the procedures completed during Study Visit Four.

Inclusion Criteria:

In good health, as determined by the investigator's review of history (provided by subject
at screening visit), physical examination, and routine blood and urine tests (done at
screening visit)

Exclusion Criteria:

Men and women <65 or >79 years of age

- Unable to provide informed consent

- Currently pregnant or lactating (given the age range for the study, verbal
confirmation by subject is believed to be sufficient)

- Current smokers

- Significant orthopedic limitations or other contraindications to strenuous exercise

- Those taking phosphodiesterase inhibitors (e.g., Viagra)

- Those taking proton pump inhibitors, antacids, xanthine oxidase inhibitors, or on
hormone replacement therapy

- Those taking anti-coagulants (e.g., Coumadin) or on anti-platelet therapy

- History of neuromuscular disease (e.g., cervical spondylotic radiculomyelpathy, lumbar
spondylosis, amyotrophic lateral sclerosis, Guillain-Barré syndrome, and acquired
demyelinating polyneuropathies), cardiovascular disease (e.g., > stage I hypertension,
heart failure, myocardial infarction/ischemia, significant myocardial or pericardial
diseases (e.g. amyloidosis, constriction), moderate or severe valvular disease, renal
disease, liver disease, or anemia