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Clinical Pathology > Protein > Flashcards

Protein Flashcards

1
Q

Plasma Proteins Background

A

Most abundant components

Chains of amino acids, most proteins are combined with ohter substances

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2
Q

Plasma Protein Functions

A

Blood coagulation (Fibrinogen, Coagulaition factors)

Maintaining oncotiv presure (Albumin)

Host defense ( immunoglobulins, complement)

Transport of substances ( albumin, transferrin)

Provide mitrogen balance for nutrition ( albumin)

Regulation of cellular metabolism (hormones)

Prevention of proteolysis (a-1 antitrypsin)

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3
Q

Two major categories of Plasma Proteins:

A

Albumin

Globulins

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4
Q

Albumin

A

One of the smallest proteins

Single most abundant protein

Synthesized in the liver

Accounts for 75% of colloidal osmotic pressure

Important carrier molecule

Transports FFA, bile acids, bilirubin, calcium, hormones, drugs

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5
Q

Acute Phase Response

A
  • Time Course:
    • major Apps:
      • Increase rapidly within 2-24 hrs, and decrease rapidly
    • Moderate APPs:
      • increase over several days and decrease more slowly
        • Species differences in major and moderate APPs
    • APPs often increased prior to presence of inflammatory leukogram
    • Persist until insult resolved
    • May be useful for disease monitoring in the future
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6
Q

Globulins

A
  • All non-albumin Proteins:
    • immunoglobulins
    • Acute phase proteins
    • Complement proteins
    • Lipoproteins
  • Over 1000 different proteins characterized, very heterogenous group
    • about 10 of these contribute significantly to globulin concentration
    • Multitude of functions
    • most combined with other substances
      • lipid, carbohydrate
  • Most synthesized in Liver
    • except immunoglobulins, which are secreted by plasma cells and B lymphocytes
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7
Q

Acute Phase Proteins

A

Proteins that change concentration by >25% in response to inflammatory cytokines

Postive AAPs → increase in concentration

Negative AAPs → decrease in concentration

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8
Q

Plasma

A

liquid portion of unclotted blood

Contails all proteins,

Mst be collected with anticoagulant

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9
Q

Serum

A

liquid portion of clotted blood

Serum contains all protien exceopt fibrinogen (Consumed during clotting)

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10
Q

Analytical Principles:

Total Protein Refractometry

A
  • TP b refractometer
    • estimation of protein in plasma
    • reported as part of the hemogram
    • Light refraction proportional to solids in solution
  • Calibration Scales
    • [total solids] = proteins + glucose + electrolytes
    • [total protein] = proteins (albumin, globulins)
  • Assumes concentrations of glucose, electrolytes, urea, and lipids are normal
  • Interferences for [TPref]
    • hyperglycemia
    • increaed urea
    • hypercholesterolemia
    • lipemia
    • excess EDTA in blood
      • Hyperbilirubinemia and hemolysis do not interfere with the refractive index, however they may make the demarcation difficult to read
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11
Q

Analytical Principles:

Measurement of Fibrinogen

A
  • [fibrenogen] by heat percipitation
    • crude measurement used in large animal species
    • Often more sensitive to inflammation than leukogram changes in LA
    • Reported on the hemogram
    • [TPref] measured → plasma heated for 3 min at 56C → [TPref] re-measured after centrifugation
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12
Q

Analytical Principles:

Total Protein Spectrometry

A
  • Total protein via spectrophotometry - Biuret reaction
    • detects peptide bonds in proteins
    • Most common method to measure TP in serum
    • Reported on the serum chemistry profile
    • TPref usually > then TP biuret
      • non-protien solids measured by refractometry
      • Fibrinogen in plasma not found in serum
  • Biuret Reaction:
    • Cu binding to peptide binds → creates voilet complex
    • color change read spectrophotometrically
    • Interferences:
      • hemolysis: + interference in some assays
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13
Q

Analytical Principles:

Albumin Measurements

A
  • Bromocresol Green dye-binding
    • Bromcresol Green dye perferentially binds to albumin, causes color change, read spectrphotopmetrically
    • Reported on the serum chemistry profile
    • Interferences:
      • will bind to some globulins, esp if albumin is low
      • Leads to falsely increaesd albumin
      • Hemolysis + intergerence in some assays
    • Inaccurate for measurement in rabbits and old world monkeys
    • Unrelianle in most species of bird
      • Obtain [TPbiuret] then measure albumin via serum protein eletrophoresis
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14
Q

Analytical principles:

Globulins Measurement

A

Indirect (calculated) measurement on serum chemistry profile

[TP] - [albumin] = [globulins]

Gractionate globulins

Serum protein electrophoresis (SPE)

Indications: hyperglobulinemia

Inflammatory vs. neoplasia

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15
Q

Analytical Principles:

Serum Protein Electrophoresis

A

serum is applied to an agarose gel and proteins migrate either towards the cathode or the anode

Separates proteins based on size, shape and charge

  • Densitometer translates bands into peaks
    • area under each peak is proportional to the % of the fraction in serum
    • Absolute quantity of each fraction = [TP] x %
  • Acute phase proteins
    • alpha and beta globulin
  • Immunoglobulins
    • gamma globulin
    • Occasional beta globulin
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16
Q

Analytical Principles:

Summary:

CBC

A

[TPref] → estimate of albumin + globulins via reftractometry

[Fibrinogen] → large animla cbc by heat precipitation

17
Q

Analytical Principles:

Summary

Chemistry Profile

A

[TPbiuret] → measurement of albumin + globulims by spectrophotometry.

No fibrinogen → removed during clotting process

{Albumin] → measures by BCG

unreliable in birds, rabbits, and old world monkeys

[Globulin] → calculated by [TPbiuret] - [Albumin]

18
Q

Analytical Principles:

Summary:

Ancillary Tests

A

Serum protien electrophoesis

Fractionate globulins

Quantify albumin in birds

19
Q

Dysproteinemia

A

Presence of normal protein at abnormal concentrations, or abnormal protein in blood

Hypoalbumenimia

Hyperalbumemia

Hypoglobinemia

Hyperglobinemia

Major Mechanisms:

Decreased production, increased production, loss, relative (Shifting of water)

20
Q

Abnormal proteins, and abnormal protein concentrations

A

TP, albumin, Globulin concentrations should increase together

Patterns are important

Nonselective: both albumin and globulin affected. Panhypoprteinemia

Selective: only albumin or globulin affected

Interpret in context of relevant history, physical exam findings, CBC results etc.

21
Q

Nonselective Hyperproteinemia:

Dehydration

A

increased [TP], concurrent increased in [albumin] and [globulin] usually due to dehydration

Usually accompanied by increased Hct

Loss of plasma water results in relatve increase in albumin and globulins

22
Q

Nonselective Hypoproteinemia:

Overhydration

A

Panhypoproteinimaia: Concurrent hypoalbuminemia and Hypoglobulinemia

Overhydration or hemodilution

Uncommon causes of hypoproteinemia

Iatrogenic: excess administration of IV fluids

Edematous Disorders

congestive heart failure, cirrhosis, nephrotic syndrome

Excess ADH secretions: SIDH

23
Q

Nonselective Hypoproteinemia:

Blood Loss

A

Hemorrhage

decreased blood volume

Fluid shifts from extravascular space to intravascular

Hemodilution

Hypoprotenemia and anemia

24
Q

Nonselective Hypoproteinemia:

Protein losing enteropathy

A
  • Protein rich intestinal secretions typically are digested into amino acids and absorbed in the small intestine, then transported to the portal system and lymphatic vessels
  • When intestinal disease prohibits absorption or transport of the proteins, the proteins are lost in feces
  • When the rate of protein loss exceeds ability of the livere and B lymphocytes to produce proteins, Hypoproteinemia occurs
25
Q

Nonselectie hypoproteinemia:

Protein losing dermatopathy

A
  • Thermal/chemical burn
  • Plasma proteins oozing out of vessels
  • globulins may be increased with inflammation
  • Early:
    • nonselective hypoproteinemia
  • Late:
    • nonselective hypoproteinemia masked by inflammation
26
Q

Nonselective Hypoproteinemia:

Effusive Disease

A
  • Pleuritis and Peritonitis:
    • extravasation of plasma proteins into pleural and peritoneal cavities
  • Vasculitis:
    • extravasation of proteins into interstital space
27
Q

Hypoalbuminemia:

Decreased Productin:

Inflammation

A
  • Albumin is the major negative acute phase protein
  • changes in production occur soon after onset of inflammation
    • changes can be detected within hours to days
  • Increased production of Positive Acute Phase Proteins necessitates decreased albumin production to conserve amino acids for upregulated production of positive acute phase proteins
  • Often accompanied by Increase in globulins
  • Inflammatory dysproteinemia = Increased [globulin] + decreased [albumin]
    • [TP] may be WRI
  • Often accompanied by inflammatory Leukogram
  • The most common dysproteinemia you will see
28
Q

Hypoalbuminemia:

Decreased Production:

Hepatic Failure

A
  • 60-80% of functional liver lost before hypoproteinemia occurs
  • Albumins and globulin production may both me decreased, however immunoglobulin is not decreased; therefore globulins are often normal
  • Other signs of hepatic failure present
    • decreased UN, glucose, cholesterol etc)
29
Q

Hypoalbuminemia:

Decreased Production:

Starvation / Cachexia

A
  • Starvation or cachetic states
    • decrease intake of nutrients
      • decrease protein production
    • Increase protein degradation
    • Hypoproteinemia dut to hypoablunemia
      • hypoglobulinemai rarely occurs
30
Q

Hypoalbuminemia:

Decreased Production:

Malabsorption / maldigestion

A

Malabsorption or maldigestion

decreased intake of nutrients = to few amino acids for protein production

Most commonly a selective hypoalbuminemia; rarely accompanied by hypoglobulinemia

31
Q

Hypoalbuminemia:

increased loss

Protein-losing nephropathy

A
  • Albumin:
    • small – therefore passes more readily than most globulins through damaged glomerular membranes
  • Expect concurrent proteinuria and increased urine protein / creatininie ratio
  • Hypoglobulinemia may occur in severe cases
  • Might be evidence of renal insufficiency – depends on number of functional nephrons
32
Q

Selective Hypoglobinemia

A

Generally due to decreased beta or gamma globulin

Usually due to:

Failure of transfer of passive immunity

Inherited or acquired immune deficiency

33
Q

Selective Hyperglobinemia

A

Increased [globulin] without concurrent increase in albumin

Inflammation: acute response, delayed response, accompanied by decreased albuminand often an inflammatory leukogram

Antigenic stimulaitons

B-lymphocyte neoplasia

Multiple myeloma, extramedullary plasmacytoma, lymphosarcoma, chronic lymphocytic leukemia

Serum prtien electrophoresis: investigate hyperglobulinemia pathophysiology

Clinical Pathology (20 decks)

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