Mitochondrial Dysfunction in Diabetes

The investigators' prior research has focused on defining the changes in expression of
nuclear encoded mitochondrial genes that predict changes in insulin sensitivity in skeletal
muscle, with the goal of defining the molecular mechanisms underlying the connection between
mitochondrial dysfunction and insulin resistance in skeletal muscle.

Three groups of subjects will be studied: lean, healthy control subjects (n=12) obese
non-diabetic subjects (n=12) and patients with type 2 Diabetes Mellitus (n=12) for a total
of 36 subjects. Twenty subjects have completed the study at Arizona State University; the
remaining 16 subjects will be accrued at the Mayo Clinic in Arizona.

Subjects will come to the Mayo Clinic 4 times. On Study Day 1 Subjects will be screened
with a medical history & physical and a 75 g oral glucose tolerance test to determine if
they are eligible for the study. On Study Day 2, subjects will report to the clinic after an
overnight fast, undergo a euglycemic clamp (using deuterated glucose) to determine insulin
sensitivity, and in addition have a muscle biopsy (basal biopsy for comparison). On Study
Day 3, subjects will report to the clinic after an overnight fast, and undergo a Volume
Oxygen Maximum (VO_2 max) determination. On Study Day 4, subjects will report to the clinic
after an overnight fast to perform an exercise bout with muscle biopsy 30 minutes after the
end of exercise. This will be followed by an overnight fast, and an additional muscle
biopsy at 24 hours post exercise.

Inclusion Criteria

1. Subjects must be able to communicate meaningfully with the investigator and must be
legally competent to provide written informed consent.

2. Subjects may be of either sex with age as described in each protocol. Female subjects
must be non-lactating and will be eligible only if they have a negative pregnancy
test throughout the study period.

3. Subjects must range in age as described in each specific protocol.

4. Subjects must have the following laboratory values:

1. Hematocrit ≥ 35 vol%

2. Serum creatinine ≤ 1.6 mg/dl

3. Aspartate Aminotransferase (AST) (SGOT) < 2 times upper limit of normal

4. Alanine Aminotransferase (ALT) (SGPT) < 2 times upper limit of normal

5. Alkaline phosphatase < 2 times upper limit of normal

6. Triglycerides < 150 mg/dl.

7. Prothrombin Time (PT) 11.7 -14.3 seconds (during Intralipid/heparin infusion,PT
will be determined to insure that it is < 1.5-2.0 times the normal value.)

8. Partial thromboplastin time (PTT) 23.0-37.0 seconds.

Exclusion Criteria

1. Subjects must not be receiving any of the following medications: thiazide or
furosemide diuretics, beta-blockers, or other chronic medications with known adverse
effects on glucose tolerance levels unless the patient has been on a stable dose of
such agents for the past three months before entry into the study. Subjects may be
taking a stable dose of estrogens or other hormonal replacement therapy, if the
subject has been on these agents for the prior three months. Subjects taking systemic
glucocorticoids are excluded.

2. Subjects with a history of clinically significant heart disease (New York Heart
Classification greater than grade II; more than non-specific ST-T wave changes on the
EKG), peripheral vascular disease (history of claudication), or pulmonary disease
(dyspnea on exertion of one flight or less; abnormal breath sounds on auscultation)
will not be studied.

3. Recent systemic or pulmonary embolus, untreated high-risk proliferative retinopathy,
recent retinal hemorrhage, uncontrolled hypertension, systolic BP>180, diastolic
BP>105, autonomic neuropathy, resting heart rate >100, electrolyte abnormalities.