RBC FREEZING

glycerol Frozen preservation of RBCs with _________is primarily used for storing units with rare blood types and autologous units

Frozen cells can be effectively stockpiled for military mobilization or civilian disasters, but the high cost and the 24-hour shelf life after deglycerolization make them less useful for routine inventory management Recently, an effectively closed system was approved with a 2-week postthaw shelf life when the blood is collected CPDA-1, frozen within6 days and stored at -80 degrees C in_____________________________________ ____________________________________

RBC FREEZING
Addition of cryoprotective agent to RBCs that are less than 6 days old

Cryoprotective agent is added _______________________________ slowly to the RBCs with vigorous shaking ______________________thereby enabbling the cryoprotective agent to permeate the RBCs

Cells are then rapidly frozen and stored in the freezer

RBC FREEZING

Deglycerolization process ( Done if unit is going to be used for transfusion) -wash RBCs with decreasing percentages of saline

a) 12% saline b) 1.6% saline c) 0.2% dextrose in saline -removal of glycerol or cryoprotective agent

(commonly used)

RBC FREEZING
ADVANTAGES Long-term storage Maintenance of RBC viability and function
low residual leukocytes and platelets

DISADVANTAGES A time-consuming process

higher cost of equipments and materials

Storage requirements (-65C )

higher cost of product

Removal of significant amounts of plasma proteins

REJUVENATION
Principle
-rejuvenation is a process to restore depleted metabolites and improve the function and post transfusion survival of stored red cells

Rejuvenating solution is not intended for intravenous administration; after warm incubation with the solution, the red cells are washed and either glycerolized for frozen storage or kept at
1 to 6 degrees C for transfusion within 24 hours

REJUVENATION
The rejuvenating solution approved by the Food and Drug Administration contains pyruvate, inosine, phosphate, and adenine Its use is permitted only with RBCs prepared from Whole Blood collected into CPD, CP2D, or CPDA-1, and it may be added at any time between 3 days after collection of the blood and 3 days after the expiration of the unit The use of the rejuvenation solution with RBC units before 14 days of storage is not routinely accepted because the treated cells may develop supranormal levels of 2,3-diphosphoglycerate, which impairs oxygen uptake.

ABO Blood Types

Hemolytic Disease of Newborns (HDN) or Erythroblastosis Fetalis

*1st pregnancy-exposure *infuse-rhogam (prevent mother form creating Abs.)

IMPROVED ADDITIVE SOLUTIONS

Longer storage periods could improve the logistics of providing RBCs for clinical use Hess and Greenwalt and coworkers :
Customized solutions containing sodium bicarbonate, sodium phosphate, adenine, dextrose, mannitol, and sodium chloride and the use of higher pH (alkaline range) is providing for better retention of ATP levels

RBC Substitutes
Hemoglobin based oxygen carriers and the PFCs Function: Carry and transfer oxygen in the absence of intact RBCs In recent years, referred to as oxygen therapeutics

Advantages and disadvantages of Stroma-Free Hemoglobin solutions

ADVANTAGES
Long shelf life

DISADVANTAGES
Short intravascular half-life

Very stable
No antigenicity (unless bovine) No requirement for blood typing procedures

Possible toxicity
Increased oxygen affinity Increased oncotic effects

Hemoglobin-based oxygen carriers

In advanced clinical testing are chemicaly modified hemoglobin solutions One product: Polymerized bovine hemoglobin Others are prepared from modified human hemoglobin

PFCs

(perfluorochemicals)

Hydrocarbon structures in which all the H+ atoms have been replaced with fluorine Chemically inert Excellent gas solvents Carry O2 and CO2 by dissolving them

Advantages and Disadvantages of PFCs

ADVANTAGES Biologic inertness Lack of immunogenicity Easily synthesized
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DISADVANTAGES Adverse clinical effects High O2 affinity Retention in tissues Requirement for O2 administration when infused Deep-freeze storage temperatures

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Platelet Anatomy and Function

Disc-shaped : 2-4 microns in diameter of blood
150,000 to 400,000 microliter

Alpha Granules:

contain clotting factors, pH-derived growth factors

Dense Granules: contain ADP, ATP,

calcium, serotonin, fibro-stabilizing factor

Steps in Vascular Damage and Clotting Responses

Vascular Spasm: contraction of smooth muscle in arteriole walls to reduce blood flow. Platelet plug formation: 1. Platelet adhesion: Platelets contact and stick to free collagen fibers of the damaged blood vessel 2. Platelet release reaction: Activated platelets extend many projections that enable them to contact and interact with one another. They then liberate their granules.

Steps in Vascular Damage and Clotting Responses

Liberated ADP and thromboxane A2 help activate other platelets. Serotonin and thromboxane A2 function as vasoconstrictors helping to decrease blood flow. 3. Platelet aggregation: Liberated ADP makes new platelets sticky; these newly-recruited and activated platelets adhere to the originallyactivated platelets. The process causes the formation of a platelet plug.

The Blood Clotting Cascade

Extrinsic Pathway (Fast acting) 1. tissue factor (TF) or thromboplastin ________________________ ______is released by tissue cells outside of the damaged vessel. 2. TF begins a chemical reaction pathway that activates FX Thrombokinase (___). FX ____combines with Proaccelerin (_____) to form the enzyme FV Prothrombinase.
*FX-common pathway

The Blood Clotting Cascade

Intrinsic Pathway (slow acting) Activated by factors within the blood or vessels Antihemophilic factor D or Hageman factor (F12) is activated by contact with collagen fibers. F12 starts a chemical cascade that ultimately activates F10 or Thrombokinase. F10 combines with Proaccelerin (F5) to form the enzyme Prothrombinase.

The Blood Clotting Cascade

The Common Pathway Prothrombinase catalyzes the conversion of Prothrombin (F2) to Thrombin. Thrombin converts the soluble plasma protein fibrinogen in the insoluble protein fibrin (loose threads). Thrombin also activates Fibrin Stabilizing Factor (F13) which converts the loose threads into stable threads.

The Problems with Clotting Cascade

Hemophilia A: Deficiency of Factor VIII accounts for 85% cases.
Almost exclusively in males. Females are usually carriers caused by a gene mutation on the X chromosome. Occurs in about 1/10,000 male births

Other Hemophilias account for another 15%

Hemophilia B (Factor IX) Hemophilia C (Factor XI) Hemophilia D (Factor XII)

The Problems with Platelets and Abnormal Clotting

Thrombocytopenia:
Abnormally low levels of platelets. Usually below 50,000/ul of blood.
In many cases, specific antibodies are produced against platelets destroying them

Thrombus:
Abnormal clot that develops in a blood vessel.

Embolus:
Free thrombic clots carried in the blood that usually get caught in arterioles in the brain, kidney, and lungs.