Mechanism and Repository Study for the Red Cell Storage Duration Study

MARS will enroll approximately 250 of the subjects participating in the RECESS study, and an
additional 50 healthy volunteers. The RECESS subjects participating in MARS will have blood
drawn before their cardiac surgery, and again 2 days after their cardiac surgery. If they
have received any red blood cell (RBC) transfusions during their surgery or in the 96 hours
following the end of their surgery, they will also have blood drawn at approximately 6 days,
28 days, and 180 days after their surgery, and will answer a health questionnaire at the
28-day and 180-day visits. The healthy volunteers participating in MARS will each have a
single blood draw.

All MARS subjects will have blood samples sent to a repository for future study. A subset
of the RECESS subjects participating in MARS, and all the healthy volunteers participating
in MARS, will also have a number of laboratory tests performed as part of the MARS study.
These tests will include

1. laboratory measures of vascular signaling, including NO content, NO disposition;
oxidative modification to RBC membrane thiols; and the flux in vasoactive
S-nitrosothiols (RSNOs).

2. laboratory measures of coagulation, including thrombin generation; prothrombin fragment
1+2, fibrinopeptide A; soluble thrombomodulin, protein C, PAI-1, tissue plasminogen
activator, Factor V, Factor VII, Factor VIII, D-Dimer, antithrombin III, soluble
endothelial protein C receptor, TFPI, Xia, INR, PT, and PTT (all using standard testing
methods); and clot formation measured by rotation thrombo-elastometry.

3. laboratory measures of microparticle counts, including CD3(T cells),
CD66b(Granulocytes), CD 14(Monocytes), CD42a(Platelets), CD235a(RBCs),
CD41a(Platelets), CD44(Surface glycoprotein), CD35(Complement receptor1),
CD108(Semaphorin 7A), Annexin V, CD154(CD40-ligand), CD62p(p-selectin),
CD59(Protectin), CD55(complement DAF), CD58(LFA-3).

4. laboratory measures of inflammation and immune function, including regulatory T
cells,Th17 cells, MMP-9, MPO, Pi-1 (total), sE-Selectin, sICAM-1, sVCAM-1, IL-1β, IL-2,
IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12 (p70), IL-13, IFN-γ, GM-CSF, and TNF-α.

At one hospital, samples from RBC units transfused to MARS subjects will also be analyzed
for nitric oxide parameters.

The primary objective of MARS is to compare post-operative values of these laboratory
parameters between RBC-transfused RECESS subjects randomized to receive RBCs stored 10 days
or less and RBC-transfused RECESS subjects randomized to receive RBCs stored 21 days or
more.

Secondary objectives include

1. Determining whether values of the above laboratory parameters are associated with
clinical outcomes among RECESS participants, including change in the multi-organ
dysfunction score (MODS) from pre-surgery through death, hospital discharge, or
post-operative day 7 (whichever occurs first); change in the MODS from pre-surgery
through death, hospital discharge, or post-operative day 28 (whichever occurs first);
and all-cause mortality.

2. Determining whether there are correlations between the above laboratory parameters in
RECESS subjects at each time point (pre-surgery and post-operative days 2, 6, and 180).

3. Comparing changes in the above laboratory parameters from pre-surgery through Day 2 for
randomized RECESS subjects who undergo surgery but do not receive any RBC transfusions
versus those in each randomized treatment group who do receive RBC transfusions.

4. Comparing values of the above laboratory parameters in healthy volunteers to
pre-surgery values in RECESS subjects scheduled to undergo cardiac surgery.

5. Assessing how nitric oxide parameters in RBC units differ depending on storage duration
of the units.

6. Assessing how post-transfusion nitric oxide parameters in transfused subjects differ
depending on the pre-transfusion nitric oxide parameters and the parameters in the
transfused units.

Inclusion Criteria for RECESS subjects participating in MARS:

1. Randomized in Red Cell Storage Duration Study (RECESS)

2. Patients ≥ 18 years old

Exclusion Criteria for RECESS subjects participating in MARS:

- There are no exclusion criteria.

Inclusion Criteria for healthy volunteers participating in MARS:

1. At least 18 years old

2. In generally good health

Exclusion Criteria for healthy volunteers participating in MARS:

1. History of red blood cell transfusion within the previous 6 months.

2. History of surgery within the previous 6 months.

3. Currently treated with inhaled nitric oxide, nitroglycerin in any form, or
nitroprusside.

4. Currently treated with prednisone, corticosteroids, cyclosporine, chemotherapy,
remicade, methotrexate, Enbrel and/or any antibody based therapy to modulate the
immune system.

5. Currently treated with heparin or other anticoagulants (for example coumadin,
pradaxa, and low molecular weight heparin).

6. Received any non-steroidal anti-inflammatory drug within the previous 24 hours.

7. Received aspirin within the previous 5 days.