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CC 2 LAB > CREATINE KINASE & LACTATE DEHYDROGENASE > Flashcards

CREATINE KINASE & LACTATE DEHYDROGENASE Flashcards

1
Q

CREATINE KINASE (CK)

EC

A

EC 2.7.3.2

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2
Q

CK
Also known as:

A

Creatine Phosphokinase
ATP:Creatine N-phosphotransferase

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3
Q

CK function

A

ATP regeneration in contractile or transport system

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4
Q

CK

In muscle cells, it is involved in the storage of_______

Each contraction cycle uses _____to produce ATP

A

high-energy creatine phosphate

creatine phosphate

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5
Q

TISSUE SOURCE
CREATINE KINASE (CK)
• Highest activities in:

A

Skeletal muscle
Heart muscle
Brain tissue

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6
Q

TISSUE SOURCE
CREATINE KINASE (CK)

Present in much smaller quantities in:
BKPPP GLLUTS

A

Bladder
Placenta
Gastrointestinal tract
Thyroid
Uterus
Kidney
Lung
Prostate
Spleen
Liver
Pancreas

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7
Q

CREATINE KINASE (CK)
• More specific indicator of disorders than total CK levels

A

ISOENZYMES

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8
Q

• CK is a dimer; 2 subunits:
“B” for____ and “M” for____

A

Brain

Muscle

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9
Q

CK exists as three isoenzymes:

A

CK-BB (brain type)
CK-MB (hybrid type)
CK-MM (muscle type)

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10
Q

СК-BB (СК1)

A

Brain
gastrointestinal tract
uterus during pregnancy

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11
Q

not usually measurable in serum due to blood-brain barrier

A

СК-BB (СК1)

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12
Q

CK-MB (CK2)

A

Heart muscle

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13
Q

elevated levels indicate myocardial damage

A

CK-MB (CK2)

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14
Q

CK-MM (CK3)

A

Striated muscle and normal
serum

primary isoenzyme in skeletal muscle

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15
Q

The major isoenzyme in healthy individuals

Significant in myocardial tissue and a good indicator of myocardial damage

A

CK-MM (CK3)

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16
Q

ISOENZYMES
CREATINE KINASE (CK)
Atypical CK-isoenzymes

A

Macro-CK

CK-Mi

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17
Q

Complexes of CK-BB with immunoglobulins, often IgG;
age and sex-related

A

Macro-CK

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18
Q

Found in muscle, brain, liver;
indicates severe illness

A

CK-Mi

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19
Q

Which travels fastest to the ANODE

A

BB

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20
Q

Arrange from most to least fastest

A

BB
MB
Macro
MM

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21
Q

DIAGNOSTIC SIGNIFICANCE
CREATINE KINASE (CK)
• Elevated in disorders of…

A

cardiac and skeletal muscle

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22
Q

Increased CK
• Common conditions:

A

Myocardial infarction (MI)
rhabdomyolysis
muscular dystrophy

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23
Q

CK
• Source of greatest elevation:

A

Duchenne muscular dystrophy

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24
Q

METHODS OF CK ISOENZYME MEASUREMENT
CREATINE KINASE (CK)

A

1.) ELECTROPHORESIS

2.) ION-EXCHANGE CHROMATOGRAPHY

3.) IMMUNOINHIBITION

4.) IMMUNOASSAYS

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25
Q

CK
• Reference method

A

ELECTROPHORESIS

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26
Q

• Advantages:
• Can separate atypical bands, including strongly fluorescent bands near the CK-BB form.

A

ELECTROPHORESIS

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27
Q

Detects unsatisfactory separations and allows visualization of adenylate kinase (AK), which can interfere with chemical or immunoinhibition methods.

A

ELECTROPHORESIS

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28
Q

2.)ION-EXCHANGE CHROMATOGRAPHY
Procedure:
This method uses_____ to separate____

A

ion-exchange columns

CK isoenzymes.

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29
Q

Advantages:
Potentially more sensitive and precise than electrophoresis.

A

ION EXCHANGE CHROMATOGRAPHY

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30
Q

IEC

Disadvantages:
On an unsatisfactory column, _____may merge into_____, and _____may elute with_____.

Additionally,_____ can also elute with____, causing inaccuracies.

A

CK-MM -> CK-MB

CK-BB -> CK-MB

macro-CK -> CK-MB

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31
Q

Antibodies target M and B subunits to measure CK-MB activity.

A

IMMUNOINHIBITION

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32
Q

Anti-M antibodies inhibit M activity but not B activity.

A

IMMUNOINHIBITION

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33
Q

IMMUNOINHIBITION
CK activity is measured_____

Remaining activity post-inhibition indicates B subunit activity.

Residual activity is doubled to account for MB (50% inhibited).

BB activity detection can falsely elevate MB results.

A

before and after M inhibition

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34
Q

IMMUNOINHIBITION

_________ are not inhibited, causing potential errors.

A

Atypical forms (CK-Mi, macro-CK)

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35
Q

This leads to the possibility of permitting detection of infarction earlier than other methods.

A

IMMUNOASSAYS

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36
Q

Measure the concentration of enzyme protein rather than enzymatic activity and can, therefore, detect enzymatically inactive CK-MB.

A

IMMUNOASSAYS

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37
Q

Detect CK-MB reliably with minimal cross-reactivity.

A

IMMUNOASSAYS

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38
Q

ASSAY FOR ENZYME ACTIVITY
CREATINE KINASE (CK)
For analysis, CK activity is coupled with other enzyme systems and measured by______ at ______

A

absorbance change at 340 nm.

39
Q

Coupled with the PK-LD-NAD+ System
pH 9.0

A

FORWARD/DIRECT
Tanzer-Gilvarg

40
Q

Most common method
2-6x faster
Less interference

A

REVERSE/INDIRECT
Oliver-Rosalki

41
Q

Coupled with the HK-G6PD-NADP
System

pH 6.7 or 6.8

The starting chemicals are more expensive

A

REVERSE/INDIRECT
Oliver-Rosalki

42
Q

STABILITY
CREATINE KINASE (CK)
• CK activity in serum is unstable due to oxidation of______

A

sulfhydryl groups.

43
Q

CK

Inactivation can be partially reversed by the addition of sulfhydryl compounds to the assay reagent.

A

N-acetylcysteine
Mercaptoethanol
Thioglycerol
Dithiothreitol

44
Q

STORAGE
CREATINE KINASE (CK)
• CK activity in_____ is relatively unstable and is rapidly lost during storage.

A

serum

45
Q

CK

Average stabilities are:
at RT
at 4°C
at -20°C.

A

<8 hours

48 hours

1 month

46
Q

SOURCE OF ERROR
CREATINE KINASE (CK)
• Hemolysis of serum samples can cause….

A

elevated CK activity

47
Q

• Erythrocytes lack CK but contain _______which produces ATP.

Increased ATP can cause falsely_____ CK levels in assays.

Significant hemolysis (______) releases enough AK to overwhelm inhibitors.

A

adenylate kinase (AK)

elevated

above
320 mg/L hemoglobin

48
Q

SOURCE OF ERROR
CREATINE KINASE (CK)
• _______ and ______affect CK levels:

• Physically trained individuals may have higher____

Prolonged bed rest may lead to_____

A

Muscular activity and muscle mass

baseline levels

decreased CK activity.

49
Q

REFERENCE RANGE
CREATINE KINASE (CK)
Total CK:
• Males:
• Females:
• CK-MB:

A

46 to 171 U/L (37°C) (0.8 to 2.9 ukat/L)

34 to 145 U/L (37°C) (0.6 to 2.4 ukat/L)

<5% total CK

50
Q

LACTATE DEHYDROGENASE (LDH)

EC

A

E.C. 1.1.1.27

L-Lactate NAD+ Oxidoreductase

51
Q

LDH function

A

• Catalyzes the interconversion of lactic and pyruvic acids

• Hydrogen-transfer enzyme that uses the coenzyme NAD+

52
Q

LDH Reference rangee

A

Reference range: 125 - 220 U/L

53
Q

DIAGNOSTIC SIGNIFICANCE
LACTATE DEHYDROGENASE (LDH)

A

• Pernicious Anemia & Hemolytic Disorders
• Liver Disorders
• Acute Myocardial Infarction (AMI)
• Skeletal Muscle Disorders & Leukemias

54
Q

• Highest levels of total LD due to erythrocyte destruction.

A

Pernicious Anemia & Hemolytic Disorders

55
Q

LDH

• Liver Disorders
• Viral____ and ____:_______

A

hepatitis and cirrhosis

Slight elevation (2-3x ULN).

56
Q

LDH

Acute Myocardial Infarction (AMI)
______.

LD levels rise______

peak in______

may remain elevated for up to____

A

Slight elevation (2-3x ULN)

12-24 hours post-infarction

48-72 hours

10 days

57
Q

Skeletal Muscle Disorders & Leukemias

Marked_____, especially in_____

A

elevations

acute lymphoblastic leukemia

58
Q

LACTATE DEHYDROGENASE (LDH)
Structure

A

has 4 subunits

either H (heart) or M (muscle) in five arrangements

59
Q

LDH-1 (HHHH)

A

Heart - Myocardial infarction
Red blood cells - Hemolytic anemia

60
Q

Disorder
Myocardial infarction
Hemolytic anemia

A

LDH-1 (HHHH)

61
Q

LDH-2 (HHHM)

A

Heart - Megaloblastic anemia
Red blood cells - Acute renal infarct
Hemolyzed specimen

62
Q

Megaloblastic anemia
Acute renal infarct
Hemolyzed specimen

A

LDH-2 (HHHM)

63
Q

LDH-3 (HHMM)

A

Lung - Pulmonary embolism
Lymphocytes - Extensive
Spleen - Pulmonary pneumonia
Pancreas - Lymphocytosis
Acute pancreatitis
Carcinoma

64
Q

Pulmonary embolism
Extensive
Pulmonary pneumonia
Lymphocytosis
Acute pancreatitis
Carcinoma

A

LDH-3 (HHMM)

65
Q

LDH-4 (HMMM)

A

Liver - Hepatic injury or inflammation

66
Q

Hepatic injury or inflammation

A

LDH-4 (HMMM)

67
Q

LDH

Liver

A

LDH-4 (HMMM)

68
Q

LDH-5 (MMMM)

A

Skeletal muscle - Skeletal muscle injury

69
Q

LDH
Skeletal muscle injury

A

LDH-5 (MMMM)

70
Q

Relative Percentage of LD Isoenzymes in Various Tissues

Serum

A

Highest - LD2 (35)
Lowest - LD5 (5)

71
Q

Relative Percentage of LD Isoenzymes in Various Tissues

Heart

A

H - LD1(45)
L - LD5(0)

72
Q

Relative Percentage of LD Isoenzymes in Various Tissues

RBC

A

H - LD1(40)
L - LD5(0)

73
Q

Relative Percentage of LD Isoenzymes in Various Tissues

Renal cortex

A

H - LD1(35)
L - LD5(0)

74
Q

Relative Percentage of LD Isoenzymes in Various Tissues

Lung

A

H - LD3(40)
L - LD5(5)

75
Q

Relative Percentage of LD Isoenzymes in Various Tissues

Skeletal muscle

A

H - LD5(60)
L - LD1/2(0)

76
Q

Relative Percentage of LD Isoenzymes in Various Tissues

Liver

A

H - LD5(70)
L - LD1(0)

77
Q

ISOENZYMES
LACTATE DEHYDROGENASE (LDH)
• LD Flipped Pattern

• Normal LD pattern:______
•_______ indicates AMI.
• May also occur in hemolyzed serum samples.

A

LD-2 > LD-1

LD-1 > LD-2

78
Q

LDH

Normal serum

A

LDH2 > LDH1

79
Q

LDH

Acute hepatitis

A

> LDH5

80
Q

MEASUREMENT OF ISOENZYMES
LACTATE DEHYDROGENASE (LDH)

A

• Electrophoresis
• mmunoinhibition
• Chemical inhibition
• Differences in substrate affinity

81
Q

MEASUREMENT OF ISOENZYMES
LACTATE DEHYDROGENASE (LDH)
• ELECTROPHORESIS
• _____migrates fastest,____ migrates slowest.

A

LD-1

LD-5

82
Q

MEASUREMENT OF ISOENZYMES
LACTATE DEHYDROGENASE (LDH)

Measures atypical bands
(LDH complexed with immunoglobulin)

A

• ELECTROPHORESIS

83
Q

Migrates between LD-3 and LD-4

After separation -> fluorometry or
colorimetry

Not associated with clinical abnormalities

A

LD-IgA/IgG

84
Q

LD-IgA/IgG

Migrates between_______

After separation ->_______

Not associated with clinical abnormalities

A

LD-3 and LD-4

fluorometry or
colorimetry

85
Q

LDH

Differences in substrate affinity
Measurement of________

A

hydroxybutyrate dehydrogenase activity

86
Q

Differences in substrate affinity
Measurement of hydroxybutyrate dehydrogenase activity

Substrate:
___________
(with increased affinity to H subunit)
a-HBD represents LD-1 activity but not specific

A

a-hydroxybutyrate

87
Q

ASSAY FOR ENZYME ACTIVITY
LACTATE DEHYDROGENASE (LDH)

A

FORWARD/DIRECT - Wacker

REVERSE/INDIRECT - Wrobleuski-Ladue

88
Q

FORWARD/DIRECT
• Wacker
• Optimal pH:

A

8.3 - 8.9

89
Q

LDH

• Most common method because it is not affected by product inhibition

A

FORWARD/DIRECT
• Wacker

90
Q

REVERSE/INDIRECT
• Wrobleuski-Ladue

Optimal pH:

A

7.1 - 7.4

91
Q

LDH

• This is ~3x faster but more susceptible to substrate exhaustion and loss of linearity

A

REVERSE/INDIRECT
• Wrobleuski-Ladue

92
Q

LDH errors

Erythrocyte Contamination
LD concentration in erythrocytes:

________higher than in serum
Any degree of_____ renders a sample unacceptable for analysis.

A

100-150 times

hemolysis

93
Q

LDH error

Stability of LD Activity
LD activity is unstable in______ regardless of storage temperature.
Ideal storage:_____, analyze within _____hours.

A

serum

25°C

48 hours

94
Q

LDH Errors

Most labile isoenzyme.
Loses activity quicker at_____ compared to_____.

For LD isoenzyme analysis: Store serum samples at____, analyze within____ hours of collection.

A

LD-5 Isoenzyme

4°C; 25°C

25°C; 24 hours

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