Blood Physiology and Hemoglobin Function
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Questions and Answers

Which of the following proteins is an integral membrane protein crucial for RBC structure?

  • Actin
  • Spectrin
  • Glycophorin A-D (correct)
  • Ankyrin
  • Band 3 protein only functions as an anion transporter in red blood cells.

    False (B)

    What is the primary function of hemoglobin?

    gas transport

    A right shift in the Hemoglobin-Oxygen Dissociation Curve indicates decreased affinity for oxygen and is more likely to ______ it to tissues.

    <p>release</p> Signup and view all the answers

    Match each stage of embryonic/fetal development with the correct globin gene activated.

    <p>Early embryonic = Zeta Late embryonic/early fetal = Alpha Early fetal = Epsilon Late fetal/adult = Beta</p> Signup and view all the answers

    Why does decreased oxygen unloading in tissues lead to increased cardiac output?

    <p>To deliver more oxygenated blood to tissues and compensate for reduced oxygen release. (A)</p> Signup and view all the answers

    Forward grouping (cell typing) involves using serum to identify antigens on red blood cells.

    <p>False (B)</p> Signup and view all the answers

    Why is 0.85% saline solution used for red blood cell washing?

    <p>It is isotonic to red blood cells</p> Signup and view all the answers

    In blood banking, a purple top tube contains ______, which is used for forward grouping.

    <p>RBC</p> Signup and view all the answers

    Match the following blood typing procedures with the component primarily used:

    <p>Forward Grouping = Antisera + RBC Reverse Grouping = Serum + A/B cells Cell Suspension = 0.85% Saline</p> Signup and view all the answers

    Which condition promotes a right shift in the oxygen dissociation curve?

    <p>Acidosis (C)</p> Signup and view all the answers

    What is the primary purpose of adding glycerol to RBCs before freezing?

    <p>To protect the cells from damage during the freezing process. (C)</p> Signup and view all the answers

    The relaxed (R) state of hemoglobin has a lower affinity for oxygen compared to the tense (T) state.

    <p>False (B)</p> Signup and view all the answers

    What change in oxygen saturation is expected when blood enters tissues, transitioning from arterial to venous blood?

    <p>Decreases from 98% to 75%</p> Signup and view all the answers

    RBCs frozen in PVC bags are more resistant to breakage at low temperatures.

    <p>False (B)</p> Signup and view all the answers

    What is the maximum storage duration for frozen RBCs?

    <p>10 years</p> Signup and view all the answers

    Stored blood often has low levels of _______, which shifts the oxygen dissociation curve to the left.

    <p>2,3-DPG</p> Signup and view all the answers

    Match the following blood preservatives with their corresponding storage durations:

    <p>Acid Citrate Dextrose (ACD) = 21 days Citrate Phosphate Dextrose (CPD/CP2D) = 21 days Citrate Phosphate Dextrose-Adenine (CPDA) = 35 days</p> Signup and view all the answers

    An additive solution extends the shelf-life of RBCs to ______ days by adding nutrients.

    <p>42</p> Signup and view all the answers

    Which change is NOT typically observed in stored blood?

    <p>Increased glucose (B)</p> Signup and view all the answers

    Match the process with its description:

    <p>Freezing = Rapidly cooling RBCs with a cryoprotective agent for long-term storage. Deglycerolization = Removing the cryoprotective agent from RBCs before transfusion. Additive Solution = Adding nutrients to extend the shelf-life of RBCs.</p> Signup and view all the answers

    Why is it important for the plastic material of blood storage bags to be permeable to CO2?

    <p>To help maintain higher pH levels during storage (C)</p> Signup and view all the answers

    What is the effect of low 2,3-DPG levels on hemoglobin's affinity for oxygen?

    <p>Increased affinity, causing decreased oxygen release to tissues. (D)</p> Signup and view all the answers

    Choose the option that is NOT considered one of the benefits of additive solutions used in blood storage?

    <p>Increases the viscosity of the RBC concentrate. (C)</p> Signup and view all the answers

    What is the primary role of adenine in the CPDA-1 preservative solution?

    <p>To increase ADP levels and ATP synthesis (A)</p> Signup and view all the answers

    Why is vigorous shaking during the addition of glycerol important when preparing RBCs for freezing?

    <p>To ensure even distribution and penetration of glycerol into the cells. (D)</p> Signup and view all the answers

    Which potentiator is used to enhance Agglutination (AGG)?

    <p>LISS (D)</p> Signup and view all the answers

    Polyclonal antibodies target a single, specific antigen, while monoclonal antibodies target multiple antigens.

    <p>False (B)</p> Signup and view all the answers

    Why does IgG, an incomplete antibody, sometimes require the Antihuman Globulin (AHG) phase for detection?

    <p>IgG facilitates sensitization but cannot achieve lattice formation alone.</p> Signup and view all the answers

    The Coombs Control Cells (CCC) are used to validate a ______ AHG test.

    <p>negative</p> Signup and view all the answers

    What does a 2+ agglutination reaction observed after adding AHG to Coombs Control Cells (CCC) indicate?

    <p>AHG validation is correct (C)</p> Signup and view all the answers

    What is the primary reason for washing cells in agglutination tests following the antigen/antibody reaction?

    <p>To dilute the cell suspension to achieve optimal antigen-antibody proportions for agglutination. (B)</p> Signup and view all the answers

    Agglutination reactions graded as 4+ indicate small agglutinates with a turbulent background.

    <p>False (B)</p> Signup and view all the answers

    Why do ABO reactions typically show stronger agglutination (3+/4+) compared to other antigen-antibody reactions?

    <p>ABO antigens are larger polymers, resulting in more available binding sites for antibodies.</p> Signup and view all the answers

    The _ effect occurs when there is an excess of antibodies, leading to a false negative reaction.

    <p>prozone</p> Signup and view all the answers

    Match the agglutination reaction grade with its description:

    <p>4+ = One solid agglutinate 3+ = Medium to large agglutinates 2+ = Small to medium agglutinates with a clear background 1+ = Small agglutinates with a turbulent background</p> Signup and view all the answers

    What causes a false positive result due to over-centrifugation?

    <p>Compacted cells forced together. (B)</p> Signup and view all the answers

    Platelets in the U.S. can be stored indefinitely as long as they are properly refrigerated.

    <p>False (B)</p> Signup and view all the answers

    In a mixed field agglutination reaction, such as when an A individual is transfused with O cells, what macroscopic result would you expect when testing the recipient's A cells immediately after transfusion?

    <p>Mixed field agglutination with a turbid background. (B)</p> Signup and view all the answers

    Flashcards

    Integral membrane proteins

    Proteins embedded in the red blood cell membrane essential for function.

    Peripheral proteins

    Proteins attached to the surface of RBC membranes, supporting structure.

    Hemoglobin function

    Primary role is transporting oxygen to tissues and CO2 to lungs.

    Oxygen Dissociation Curve

    Graph showing the relationship between O2 pressure and hemoglobin saturation.

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    2,3-DPG role

    A molecule that binds to hemoglobin, reducing O2 affinity in T state.

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    Relaxed (R) state

    The state of hemoglobin with higher oxygen affinity.

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    Oxygen dissociation curve shift

    The change in hemoglobin's ability to release oxygen due to various factors.

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    Right shift conditions

    Factors that decrease hemoglobin's oxygen affinity, promoting oxygen release.

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    Left shift conditions

    Factors that increase hemoglobin's oxygen affinity, hindering oxygen release.

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    2,3-DPG

    A molecule that decreases hemoglobin's affinity for oxygen when increased.

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    Stored blood changes

    Alterations in stored blood, causing lower oxygen delivery.

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    Preservatives for blood storage

    Substances added to blood to extend its viability during storage.

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    RBC storage container importance

    The type of container affects the viability of red blood cells during storage.

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    Increased Cardiac Output

    Compensatory response to deliver more oxygen to tissues by increasing heart rate and stroke volume.

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    Decreased Mixed Venous Oxygen Tension (PvO₂)

    Lower oxygen content in venous blood returning to the heart after tissue oxygenation.

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    0.85% Saline Solution

    Isotonic solution used to wash RBCs, preventing cell damage.

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    Forward Grouping (Cell Typing)

    Procedure matching RBCs using antisera to determine blood type.

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    Reverse Grouping (Serum Typing)

    Procedure matching serum to determine blood type by testing against A/B cells.

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    Potentiators for AGG

    Substances used to enhance antigen-antibody reactions in serological tests.

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    Antibody detection phase

    IgG is detected in the antihuman globulin phase due to its incomplete nature.

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    Polyclonal antibodies

    Antibodies produced from multiple clones targeting different antigens.

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    Direct Indirect Test (DAT)

    Test to evaluate the presence of antibodies attached to the surface of RBCs.

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    CCC in AHG validation

    A procedure validating a negative AHG test to check for sensitization levels.

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    Coombs Check Cells (CCC)

    A test used to check the presence of antibodies in blood through agglutination.

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    Grading Reactions

    Method of assessing the size and clarity of agglutination in blood tests.

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    Prozone Effect

    False negative reaction occurring when excess antibodies prevent agglutination.

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    Corrected Count Increment (CCI)

    Measures the rise in platelet levels post transfusion to evaluate success.

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    Lattice Formation Stage 2

    Formation of a structure by immune complexes as antibodies bind to antigens.

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    Rouleaux Formation

    Stacking of red blood cells due to increased plasma proteins, resembling coins.

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    Mix Field Agglutination

    Reaction seen when recent transfusion leads to mixed agglutination results.

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    ABO Reaction Strength

    Typically results in 3+/4+ agglutination due to large antibody polymers.

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    PVC brittle at low temperatures

    PVC materials can break or shatter when exposed to cold environments. This is especially important when handling components stored in PVC bags that may be frozen.

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    Additive solution benefits

    Additives extend the shelf-life of red blood cells (RBCs) to 42 days and improve plasma and platelet harvesting while reducing the viscosity of RBC concentrates.

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    Cryoprotective agent

    Glycerol is a cryoprotective agent added to RBCs less than 6 days old; it protects the cells during freezing by penetrating them.

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    RBC freezing procedure

    RBCs are mixed with glycerol and frozen quickly at temperatures below -65°C to preserve them for long-term storage.

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    Deglycerolization process

    Deglycerolization is required to remove glycerol before RBC transfusion; it is effective but can be time-consuming and expensive.

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    Advantages of frozen RBCs

    Frozen RBCs can be stored for up to 10 years, maintaining RBC viability and low leukocyte and platelet counts, but have strict storage requirements.

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    Impact of 2,3-DPG

    Low levels of 2,3-DPG can increase cardiac output but decrease mixed venous oxygen tension by making hemoglobin hold onto oxygen more tightly.

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    Function of 2,3-DPG

    2,3-DPG helps regulate hemoglobin's affinity for oxygen; lower levels result in less oxygen release to tissues, affecting oxygen delivery.

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    Study Notes

    Red Blood Cell (RBC) Structure and Function

    • RBCs are crucial for survival due to their chemical composition and structure of the membrane, which is a semipermeable lipid bilayer supported by a protein cytoskeleton.
    • Integral membrane proteins span the entire membrane, including glycophorin A-D, which makes up 20% of the membrane and gives cells their negative charge for repulsion in circulation.
    • Peripheral proteins are limited to the cytoplasmic surface of the membrane, such as spectrin, ankyrin, actin, and protein 4.1 (SAP).
    • The composition of the RBC membrane is approximately 52% protein, 40% lipid, and 8% carbohydrate.
    • The membrane's deformability is crucial for viability; ATP loss decreases spectrin phosphorylation, increasing membrane rigidity and causing spherocytes or bite cells.

    Permeability

    • RBC membranes are permeable to water and anions (like Cl- and HCO3-), but impermeable to cations (like Na+ and K+).
    • The intracellular-to-extracellular ratios for Na+ and K+ are specifically 1:12 and 25:1, respectively.
    • Calcium is actively pumped out of the RBC to maintain flexibility and prevent rigidity, controlled by calcium-ATPase and calmodulin.

    RBC Metabolism

    • RBC metabolic pathways are primarily anaerobic to deliver oxygen, not consume it.
    • The pentose phosphate pathway generates 10% of the ATP and protects hemoglobin during oxidative stress through NADPH, as deficiency leads to Heinz bodies.
    • The methemoglobin reductase pathway maintains hemoglobin in its ferrous state (Fe2+) and is dependent on NADPH; a deficiency can lead to methemoglobin accumulation and hypoxia.
    • The Luebering-Rapoport shunt produces 2,3-DPG, which plays a role in hemoglobin-oxygen binding and oxygen release in tissues.

    Effect of 2,3-DPG

    • 2,3-DPG lowers hemoglobin's oxygen affinity, facilitating oxygen release in tissues.
    • When hemoglobin releases oxygen, it shifts to a less stable (T) form.
    • When it binds oxygen, it shifts to a more stable (R) form, increasing affinity for oxygen.
    • The hemoglobin-oxygen dissociation curve illustrates the relationship between partial oxygen pressure and hemoglobin saturation.

    Conditions Affecting the Curve

    • Conditions causing a rightward shift (decreased affinity):
      • Anemia
      • Acidosis
      • Increased 2,3-DPG
      • Fever
    • Conditions causing a leftward shift (increased affinity):
      • Multiple transfusions of stored blood
      • Alkalosis
      • Decreased 2,3-DPG
      • Hypothermia

    RBC Storage

    • Stored blood has low 2,3-DPG levels, leading to a leftward shift and making it harder for hemoglobin to release oxygen, leading to inadequate tissue oxygenation.
    • Viable cell count decreases, glucose, ATP, and pH also decrease during storage, while lactic acid and K+ increase.
    • Preservatives are used to extend the shelf-life of stored RBCs, such as CPD, CPDA-1, and ACD.
    • The FDA requires over 75% of transfused RBCs to survive for at least 24 hours.

    RBC Freezing

    • RBCs can be frozen for long-term storage for rare blood types and autologous units.
    • Glycerol is typically used as a cryoprotective agent.
    • RBCs are frozen at temperatures below -65°C, and the freezing process involves deglycerolization after thawing.
    • The freezing procedure can be done using high or low glycerol concentrations.

    Washing Steps

    • Washing RBCs involves using saline solutions.
      • First wash with 12% saline.
      • Second wash with 1.6% saline.
      • Final wash with 0.2% dextrose in normal saline.
    • Washing removes unbound antibodies and serum proteins that interfere with agglutination reactions.

    Reagent Preparation

    • 10% bleach solution is used for disinfection.
    • A 0.85% saline solution is used for washing RBCs and is isotonic to RBCs, thus preventing lysis.

    Antibody/Antigen Reactions

    • Antibody excess leads to a prozone effect (false negative)
    • Proper dilution of cells and antibodies is important to increase reaction likelihood.
    • Antibody-Antigen reactions can be graded to identify different antibodies and their reactions (eg: 1+, 2+, 3+, 4+).

    Platelet Structure and Storage

    • Platelets contain granules (alpha, dense, and lysosomes) with important components involved in their function.
    • Platelets are stored for 5 days due to bacterial contamination concerns.
    • During storage, platelets become activated, releasing granules and losing ATP, potentially clumping into large sheets.
    • Loss of platelet function can affect the structural integrity and lead to a swelling of the platelets.

    Blood Typing

    • ABO blood typing determines the presence of A and B antigens on RBCs.
    • Known antisera identifies unknown antibodies.
    • Agglutination occurs if the antigen is present

    Other Relevant Information

    • Different blood types need to be tested for appropriate blood transfusion.
    • Various conditions affect RBC and platelet function and viability during storage.
    • Preservatives help maintain RBC viability, specifically extending refrigerated storage time.
    • Proper procedures for blood and platelet handling and preparation are important to ensure accurate testing.

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    Immunohematology Notes 2025 PDF

    Description

    This quiz covers essential topics related to blood physiology, red blood cell (RBC) structure and function, and the role of hemoglobin. Questions focus on membrane proteins, oxygen dissociation curves, and blood typing procedures. Test your knowledge on these critical aspects of hematology and their relevance in clinical settings.

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