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Clinical Pathology > Proteins > Flashcards

Proteins Flashcards

1
Q

General concepts

A

> 1,000 characterized in the serum

- plasma: albumin, globulins (fibrinogen, clotting factors)

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2
Q

Serum does not contain ______

A

Fibrinogen

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3
Q

Most of the plasma proteins are synthesized by ________

A

Hepatocytes

  • exceptions: Ig (B cells and plasma cells)
  • proteins contribute to colloid oncotic pressure
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4
Q

Protein dyscrasia

A

Presence of abnormal protein

- abnormal structure

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5
Q

Dysproteinemia

A

Presence of normal protein at abnormal concentration

- selective or nonselective dysproteinemia

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6
Q

Nonselective hyperproteinemia

A

All protein concentrations are increased at the same rate

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7
Q

Selective hyperproteinemia

A

Total protein concentration is increased and some protein concentration are increased more than others

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8
Q

Nonselective hypoproteinemia

A

Total protein concentration is decreased and all proteins are decreased

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9
Q

Selective hypoproteinemia

A

Total protein concentration is decreased and some protein concentration are decreased more than others

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10
Q

Selective electrophoresis

A

To determine if the condition is selective or nonselective

  • albumin and globulins decreased or increased at the same rate (nonselective)
  • albumin and globulins decreaed or increased but not proportional (selective)
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11
Q

Other than dehydration, the most common cause of dysproteinemia is _____

A

Inflammation

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12
Q

Positive acute phase proteins

A
  • SSA and CRP are increased in less than a day

- major APPs: fibrinogen, CRP, SAA, haptoglobin, a1 acid glycoprotein, ferritin, ceruloplasmin

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13
Q

Negative APP

A

Decrease plasma or serum concentration due to inflammatory process
- major negative APPs: albumin, transferrin

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14
Q

Delayed response proteins

A

Serum or plasma concentration increases 1-3 weeks after onset of inflammation
- major delayed response proteins: Ig, complement factors (C3)

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15
Q

Refractometer

A

Degree of water refraction in a solution is proportional to the quantity of solids
- most of the solids in plasma are proteins

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16
Q

Refractometer - interferences

A
  • other substances (urea, glucose, Na, Cl)
  • lipemia increases refractive index
  • hemolysis does not interfere
  • bilirubin does not interfere
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17
Q

Biuret reaction measures ______

A

Total protein

  • colorimetric: amount of color change is proportional to the amount of protein
  • interferences: hemolysis (positive), dextran (positive)
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18
Q

Albumin concentration

A

BCG dye binding reaction

  • colorimetric
  • interferences: binding to Ig when albumin concentration is low, hemoglobin (positive), triglycerides (negative), heparin (positive)
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19
Q

Globulin concentration

A

Determined by subtraction

  • total protein - albumin = globulins
  • is the concentration of all other proteins other than albumin
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20
Q

Serum protein electrophoresis (SPE)

A

Migration through a acetate cellulose or agarose gel

  • most proteins migrate towards the anode –> albumin is the smallest and negatively charged and migrates further, some Igs are positively charged and migrates towards the catode
  • separate in 4-6 groups of one or more band
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21
Q

Albumin is a negative acute phase protein because it _______

A

Decreases concentration in inflammation

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22
Q

SPE - species variations

A
  1. 1 mg/dL is the minimum detectable concentration
    - most dogs, cats, and horses: a1, a2, b1, b2, and y
    - cattle: a, b and y
    - concentration of proteins are calculated after the stained gel is scanned with a densitometer –> area under curve is the total stained protein
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23
Q

Concentration of each group is its specific percentage times _______

A

The total protein concentration (measured using biuret technique)

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24
Q

Diagnostic value

A
  • differentiating into selective or nonselective
  • scanning for a monoclonal gammopathy
  • helping to assess albumin concentration when globulins interfere with BCG
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25
Q

________ is the most common cause of hyperproteinemia

A

Hemoconcentration

  • caused by loss of plasma water
  • vomiting, diarrhea, impaired renal concentration, sweating, increased vascular permeability
  • if TP 7 g/dL, then in a 10% dehydrated animal it would be 7.7 g/dL
  • nonselective
  • other: erythrocytes, prerenal azotemia, hyperstenuria
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26
Q

Increased protein synthesis

A

Inflammation is a common cause of hyperproteinemia

  • does not always cause hyperproteinemia
  • increase production of positive APP and delayed response proteins (mostly by hepatocytes)
  • increased production of positive APP: within hours and persists until inflammation is resolved
  • increased plasma conc in 2 days
  • increase of [fibrinogen] and [haptoglobin] can cause hyperproteinemia
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27
Q

Hyperproteinemia due to inflammation

A

Decreased production of negative APP

  • albumin half life: 8 days in dogs, 19 days in horses
  • hypoalbuminemia due to inflammation: several days in dogs, 2 weeks in horses
  • transferrin: half life not well established, decreased conc after at least a week
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28
Q

Inflammation leads to increased production of ________

A

Delayed response proteins

  • 1-3 weeks after the onset of inflammation
  • Ig, mostly IgG and complement
  • C3 usually <1.0 mg/dL
  • IgG mild: <1.0 mg/dL to marked >4.0 mg/dL
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29
Q

Other findings with hyperproteinemia due to inflammation

A
  • anemia of inflammatory disease
  • neutrophilia, neutropenia
  • lymphocytosis, lymphopenia
  • monocytosis
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30
Q

B lymphocyte neoplasia

A

B cells may produce Ig

  • plasma cell neoplasia: multiple myeloma (most frequent cause) or extramedullary plasmacytoma
  • lymphocyte neoplasia: lymphoma or lymphocytic leukemia
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31
Q

Expected dysproteinemia pattern with B cell neoplasia

A
  • IgG migrates with g globulin fraction
  • IgM or IgA with B-y or B globulin fraction
  • atypical pattern seen due to protein degradation or complex with other protein, Ig complexes, and incomplete Ig
  • other [Ig] are decreased
  • mild-moderate hypoalbuminemia due to decreased production or neg feedback by oncotic pressure receptors in hepatocytes
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32
Q

B cell neoplasia - associated findings

A
  • Bence Jones proteinuria: light chains of Ig in the urine
  • hyperviscosity syndrome: high Ig leads to increase plasmatic viscosity
  • hypercalcemia
  • AL amyloidosis caused by Ig light chains
  • PCR for Ig variable region gene rearrangement can be used for diagnosis
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33
Q

Blood loss

A

Increased loss from vascular space, hypoproteinemia occurs when remaining blood is diluted

  • extracellular extravascular fluid shifts to intravascular
  • panhypoproteinemia (nonselective)
  • anemia (depends on severeity and duration of hemorrhage)
34
Q

Protein losing nephropathy

A

Immune complex or amyloid deposition in glomeruli –> retraction of podocytes or loss of selective permeability of glomerular basal membrane –> protein loss

  • if protein loss > protein production then hypoproteinemia
  • larger proteins are not lost, so it is a selective process
  • glomerulopathy: glomerulonephritis or amyloidosis may be present
35
Q

Protein losing nephropathy - lab findings

A
  • proteinuria, mostly albuminuria
  • moderate-marked hypoproteinemia with moderate-marked hypoalbuminemia
  • serum electrophoresis consistent with selective protein loss
  • possible renal failure
  • possibly hypercholesterolemia if nephrotic syndrome present
36
Q

Protein losing enteropathy

A

Protein rich intestinal secretions (digestion and resorbed thru lymphatics) –> if reabsorption is impaired (SI mucosal disease, or lymphatics disease) protein lost in feces

  • if protein loss > protein production, then hypoproteinemia
  • protein exudation and decreased intake
  • blood loss, intestinal parasites
37
Q

Protein losing enteropathy - lab findings

A
  • mild-marked hypoproteinemia

- serum electrophoresis: nonselective

38
Q

Protein losing dermopathy

A

Thermal or chemical burns –> protein exudation from cutaneous lesions

  • if protein loss > protein production, then hypoproteinemia
  • after 2 days findings, will include APP shifts
39
Q

Protein losing dermopathy - lab findings

A
  • early: nonselective hypoproteinemia

- later: nonselective hypoproteinemia masked by acute or chronic inflammatory dysproteinemia

40
Q

Plasma loss (peritonitis, pleuritis, vasculitis)

A
  • pleuritis/peritonitis: inflammation –> protein loss in pleural or peritoneal cavities –> hypovolemia –> shift of fluids from extracellular extravascular to intravascular –> dilution of remaining proteins (hypoproteinemia)
  • aka: 3rd space loss
  • vasculitis: similar, proteins lost to perivascular interstitial tissue adjacent to affected vessel
41
Q

Plasma loss - lab findings

A
  • early: nonselective hypoproteinemia

- late: nonselective hypoproteinemia masked by acute or chronic inflammatory dysproteinemia

42
Q

Hepatic insufficiency (hepatic failure)

A

Marked decrease in hepatic functional mass (<20% remaining) –> decreased production of nearly all plasma proteins (except Ig)

  • normal protein catabolism combined with decreased production = hypoproteinemia
  • decreaed synthesis and/or increased catabolism
43
Q

Hepatic insufficiency - disorders

A
  • cirrhosis
  • hepatic necrosis or inflammation (not acute)
  • portosystemic shunt (hepatic atrophy)
  • extensive liver damage due to neoplasms
44
Q

Hepatic insufficiency - lab findings

A
  • hypoproteinemia, hypoalbuminemia, normoglobulinemia, hypoglobulinemia
  • SE: nonselective, B2 and y may be increased
45
Q

Malabsorption or maldigestion

A

Deficient intake of carbs, lipids and proteins –> once depleted storages, normal catabolism and use of animo acids for gluconeogenesis –> hypoproteinemia

46
Q

Malabsorption

A

Small intestine diseases with generalized mucosal involvement

47
Q

Maldigestion

A

Exorine pancreatic insufficiency (chronic pancreatitis or atrophy) leads to deficiency in protease, lipase, and amylase

48
Q

Malabsorption or maldigestion - lab findings

A

Hypoproteinemia, hypoalbuminemia, normo or hypoglobulinemia

- SE: nonselective

49
Q

Cachectic states

A

Protein catabolism exceeds production –> hypoproteinemia

  • before hypoproteinemia –> protein production (mainly albumin) at expenses of body fat and muscle mass
  • chronic diseases: infectious and malignant neoplasia
  • marked malnutrition or starvation
50
Q

Cachectic state - lab findings

A
  • hypoproteinemia, hypoalbuminemia, normo or hypoglobulinemia
  • SE: nonselective
51
Q

Lymphoid hypoplasia/aplasia

A

B cells produce Ig and not other major plasma proteins

  • decreased Ig production in face of normal concentration of other proteins –> mild hypoproteinemia (hypoglobulinemia), normoalbuminemia
  • SE: decreased gamma group
  • combined immunodeficiency (arabian, appaloasa, basset hounds, corgis, jack russell)
  • infectious diseases, chemotherapy
52
Q

Failure of passive transfer

A

Failure to absorb immunoglobulins from colostrum –> decreased plasma Ig –> hypoproteinemia, hypoglobulinemia

  • inflammation can mask the hypoproteinemia
  • calves with [IgG] of less than 1,000 mg/dL
  • [serum TP] fo 5.0 g/dL or less 83%
53
Q

Hemodilution

A

Increased ECF is uncommon (by itself) to cause hypoproteinemia

  • worsens hypoproteinemia from other causes
  • due to: excess IVF, edematous disorders, excessive ADH secretion, use of plasma expanders
54
Q

Hyperalbuminemia due to hemoconcentration

A

Dehydration, decreased ECF

  • decreased water –> increased concentration of all blood contents
  • most common cause of hyperalbuminemia
55
Q

Hyperalbuminemia due to induced synthesis by _____

A

Glucocorticoid therapy

  • increase in 4-5 days and 4 weeks (dogs and cats) of treatment
  • maybe increased production or increased life span
56
Q

Falsely increased albumin using BCG method

A

BCG dye binds preferentially to albumin but also to alpha and beta globulins

  • heparinized plasma samples can lead to increase due to binding to fibrinogen
  • may mask hypoalbuminemia
57
Q

Hypoalbuminemia along with hyperproteinemia

A

Not due to hemoconcentration
- inflammatory: negative APP, takes 1-2 weeks to appear, concurrent B cell neoplasia due to inflammatory cytokines, increased colloidal osmotic pressure or damaged tissues (liver, kidney, intestines)

58
Q

Hypoalbuminemia along with hypoproteinemia

A

Decreased production or loss or both

59
Q

Hyperglobulinemia

A

Frequently with hyperproteinemia but possible with normoproteinemia due to hypoalbuminemia

  • commonly caused by hemoconcentration, inflammation (chronic), B cell neoplasia
  • may be result of increases in one or several globulins
  • SPE: differentiate hemoconcentration from inflammation and B cell neoplasia, patterns of inflammation and B cell neoplasia may overlap
60
Q

Hypoglobulinemia

A

Other than neonates, commonly with hypoproteinemia and occasionally with normoproteinemia

  • decreased production: hepatic insufficiency and lymphoid hypoplasia/aplasia
  • loss: blood loss, intestine mucosa, skin
61
Q

Positive acute phase proteins

A

Any injury that causes inflammation can increase positive APPs

  • can increase globulins and TP
  • individually, magnitude can vary
62
Q

Why use positive APPs

A
  • insensitivity of other tests (pyrexia, neutrophlia)

- provide another method of monitoring inflammation for therapeutic decisions and prognostic

63
Q

Positive APPs - factors to consider

A
  • persistence of increase of positive APPs in chronic inflammation
  • may be processes decreasing concentration of positive APPs at the same time of inflammation –> disorders that cause loss of plasma proteins, activation of plasmin or thrombin (decreasing fibringogen), intravascular hemolysis
64
Q

Fibrinogen

A

Plasma protein produced by hepatocytes

  • used for production of fibrin by thrombin
  • positive APP but consumption may take place at the same time masking increased production
65
Q

Fibrinogen - analytical process

A

Heat precipitaiton

  • measure [TP] in an aliquot of plasma sample –> heat precipitate fibrinogen in a second aliquot and centrifuge the sample to separate fibrinogen –> measure [TP] again –> [fibrinogen]=1st [TP] - 2nd [TP]
  • adequate for documenting hyperfibrinogenemia, but not hypofibrinogenamia
  • subtracting serum [protein] from plasma [protein] does not work since other solutes are released and/or consumed during coagulation
66
Q

Hyperfibrinogenemia

A

Due to: hemoconcentration or inflammation

  • TP may be increased, but also WRI
  • fibrinogen increases are more consistent than TP in inflammation
67
Q

Plasma protein to fibrinogen ratio (PP:F ratio)

A

Ratio helps to differentiate hyperfibrinogenemia of inflammation from hemoconcentration
- ratio varies among species and age groups

68
Q

Plasma protein to fibrinogen ratio (PP:F ratio) - species variation

A
  • cattle: >15 likely dehydration, <10 likely inflammation

- horses: >20 likely dehydration, <15 likely inflammation

69
Q

PP:F ratio - additional factors

A
  • concurrent inflammation and dehydration will make interpretation difficult
  • dehydration by itself will cause mild increases in [fibrinogen]
  • guidelines not adequate for calves and foals
  • guidelines not adequate for cattle and horses with pathologic process that causes hypoproteinemia
  • accuracy of refractometer is 0.1 mg/dL, so ratio will be more accurate with higher [fibrinogen]
70
Q

Except for fibrinogen, ______ are preferred

A

Serum samples

- relative stable to freezing

71
Q

Variety of tests are used depending on APP

A
  • serum C reactive protein
  • haptoglobin
  • serum amyloid A
  • a1: acid glycoprotein
  • ceruloplasmin
  • ferritin
72
Q

Causes other than inflammation for APP concentration changes

A
  • [CRP] in pregnant bitches
  • glucocorticoids can increase [haptoglobin] in dogs
  • anti-helmintic drugs can increase [haptoglobin]
  • phenobarbitol in dogs may increase
  • lactation in cattle increases [CRP]
  • increased [ferritin] with increased iron stores
  • increased [ferritin] in horses after exercising
73
Q

Reasons to measure Ig

A

Most common cause to assessing Ig is to determine passive transfer from mares to foals and cows to calves

  • congenital or acquired immunodeficiency
  • monoclonal gammopathies
74
Q

Failure of passive transfer - process

A

Placentaiton in horses and calves prevent in utero transfer of Ig

  • need to ingest colostrum before gut closure to obtain maternal Ig
  • neonate foals have no plasma IgG
  • foals and calves have limited ability to produce IgM
  • after colostrum intake, [TP] increases due to IgG, but also other APP
  • Ig uptake is mediated by Fc receptor on epithelial cells
75
Q

Failure of passive transfer causes

A

Lack of colostrum intake (too weak to nurse)

  • inadequate IgG in the colostrum (low conc or low amount of ingested colostrum)
  • failure to absorb (after gut closure) and potentially due to different haplotypes of Fc receptor in calves
76
Q

Establishing FTP - foal guidelines

A

Foal guidelines: sample from 18 to 48 hrs after birth

  • FTP when IgG < 400 mg/dL
  • [IgG] >800 mg/dL as adequate is recommended as criterion
  • [IgG] <200 mg/dL is complete FTP
  • [IgG] <800 mg/dL and >200 mg/dL is considered partial FPT
77
Q

Establishing FTP - calve guidelines

A

Sample from 1 and 8 days after birth

  • [IgG] > 1000-1600 mg/dL as adequate passive transfer
  • [IgG] <500-800 mg/dL is complete FTP
  • [IgG] <1600 mg/dL and >800 mg/dL is partial FPT
78
Q

Radial immune diffusion (RID)

A

Most used

  • not the most accurate
  • takes 24 hrs for results (time for Ig to diffuse thru the gut)
79
Q

Glutaraldehyde coagulation test

A
  • inexpensive
  • not reliable with total blood
  • for foals good to excellent agreement
  • for calves may require dilution of the sample
80
Q

Latex agglutination

A

Latex beads covered with anti equine IgG agglutinate in the presence of IgG

  • semiquantitative test good agreement with RID
  • not as good as glutaraldehyde coagulation test
  • moderately expensive but takes only 10 minutes

Clinical Pathology (9 decks)

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