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CC 2 LAB > PHOSPHATASES > Flashcards

PHOSPHATASES Flashcards

1
Q

ALKALINE PHOSPHATASE (ALP)
• E.C.

A

3.1.3.1

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2
Q

Systematic name of ALP

A

Orthophosphoric Monoester Phosphohydrolase (Alkaline optimum)

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3
Q

ALP function

A

Catalyzes hydrolysis of phosphomonoesters at alkaline pH

  • Liberates inorganic phosphate from organic phosphate ester
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4
Q

ALP

• Produces an_____ as a byproduct

A

alcohol

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5
Q

ALP
Optimal pH

A

= 9.0 to 10.0

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6
Q

ALP
Activator:

A

Mg2+& Zinc

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7
Q

ALP reaction

A

Phosophomonoester -> Alcohol +
Phosphate ion

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8
Q

TISSUE SOURCE
ALKALINE PHOSPHATASE (ALP)

Highest Concentrations:

A

Intestine
Liver (sinusoidal and bile canalicular membranes)
Placenta
Bone (osteoblasts)
Spleen
Kidney

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9
Q

ISOENZYME ANALYSIS
ALKALINE PHOSPHATASE (ALP)

A

Electrophoresis
Heat stability
Chemical Inhibition
Abnormal reactions

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10
Q

Electrophoresis mnemonics ( I Promise 2 Be Loyal)

A

Intestinal
Placental
Bone
Liver

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11
Q

Heat stability (Promise Ikaw Lang Baby)

A

Placental
Intestinal
Liver
Bones

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12
Q

ALP

• Major isoenzymes:

A

intestine, liver, bone, placenta

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13
Q

most useful technique for isoenzyme analysis

A

Electrophoresis

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14
Q

ALP

ISOENZYMES fastest migration

A

Liver isoenzyme:

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15
Q

Liver isoenzyme: 2 types

Mother Fucker

A

Major liver band/fraction
Fast liver fraction (a1 liver)

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16
Q

Bone ALP
-increases due,to…

A

osteoblastic activity

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17
Q

BONE ALP normally elevated in…

A

children during periods of growth and in adults older than age 50

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18
Q

Intestinal ALP

-depends on the______ and ______ of the individual’
-increases after consumption of______
-bound by erythrocytes of “_____” group
-diseases of the_____ and _____

A

blood group and secretor status

fatty meal

A

digestive tract and cirrhosis

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19
Q

-chronic hemodialysis

A

Intestinal ALP

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20
Q

LIVER isoenzymes

  • hepatobillary conditions
  • metastatic carcinoma (obstructive liver disease)
A

Major liver fraction

Fast liver fraction

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21
Q

ISOENZYME ANALYSIS
ALKALINE PHOSPHATASE (ALP)

• Used to identify isoenzyme source
• ALP activity is measured before and after heating the serum at____ for____

A

Heat Stability

56°C

10 minutes

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22
Q

Residual activity <20% after heating:

A

bone phosphatase

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23
Q

Residual activity > 20% after heating:

A

liver phosphatase

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24
Q

: most heat stable

A

Placental ALP

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25
Q

Most heat labile

A

Bone

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26
Q

ISOENZYME ANALYSIS
ALKALINE PHOSPHATASE (ALP)
• Chemical Inhibition

A

Phenylalanine
3M urea
Levamisole
Leucine

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27
Q

” Phenylalanine
• inhibits (2)

A

intestinal, and placental ALP

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28
Q

Phenylalanine also inhibits the…

A

carcinoplacental isoenzyme

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29
Q

3M Urea
Inhibits ____ ALP

A

Bone

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30
Q

Levamisole
Inhibits (2) ALP

A

Bone and Liver

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31
Q

Leucine
Inhibits the…

A

Nagao isoenzyme

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32
Q

NEOPLASTIC ISOENZYMES
ALKALINE PHOSPHATASE (ALP)

A

Regan isoenzymes
Nagao isoenzymes

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33
Q

• carcinoplacental ALPs

A

Regan and Nagao isoenzymes

34
Q

ALP Isoenzymes

migrates to the same position as the bone fraction

most heat stable of all ALP isoenzymes

A

Regan

35
Q

ALP Isoenzyme found in lung, breast, ovarian, and colon cancers

A

Regan

36
Q

variant of Regan isoenzyme
found in metastatic carcinoma and adenocarcinoma

A

Nagao

37
Q

Associated with hepatoma and gastrointestinal tract
tumors

A

Kasahara

38
Q

DIAGNOSTIC SIGNIFICANCE
ALKALINE PHOSPHATASE (ALP)

Significant in…

A

hepatobiliary and bone
disorders

39
Q

T or F

ALP is more prominent in obstructive conditions than hepatocellular disorders

A

True

40
Q

ALP

Observed with____ involvement in bone disorders

A

osteoblast

41
Q

Hepatobiliary Disorders:
Biliary tract obstruction:

A

ALP levels 3 to
10 times the ULN

42
Q

Hepatobiliary disorders

Hepatitis and cirrhosis: increases by..

A

slight increases
(usually <3 times the ULN)

43
Q

DIAGNOSTIC SIGNIFICANCE
ALKALINE PHOSPHATASE (ALP)
• Bone Disorders:

Highest elevation seen in this condition

A

Paget’s disease

• Highest elevations in Paget’s disease (Osteitis deformans)

44
Q

DIAGNOSTIC SIGNIFICANCE
ALKALINE PHOSPHATASE (ALP)

Other disorders:

A

osteomalacia,
rickets, hyperparathyroidism,
osteogenic sarcoma

45
Q

Bone disorders

ALP

Elevated during ——-and periods of physiologic——-

A

healing bone fractures

bone growth

46
Q

ALKALINE PHOSPHATASE (ALP)
“ Pregnancy:
• Increased ALP activity
(______at 16-20 weeks; _____ in third trimester)

• Returns to normal within ___after labor

A

1.5 times ULN

2-3 times ULN

3-6 days

47
Q

DIAGNOSTIC SIGNIFICANCE
ALKALINE PHOSPHATASE (ALP)
Pregnancy:

Complications:

A

elevated in hypertension,
preeclampsia, eclampsia, and threatened abortion

48
Q

DIAGNOSTIC SIGNIFICANCE
ALKALINE PHOSPHATASE (ALP)

Decreased ALP: inherited condition with subnormal ALP activity

A

Hypophosphatasia

49
Q

DIAGNOSTIC SIGNIFICANCE OF ALP ISOENZYMES
ALKALINE PHOSPHATASE (ALP)
• Bone Isoenzyme:
• Increased due to_____ activity
• Elevated in…

A

osteoblastic

children during growth and adults over 50

50
Q

DIAGNOSTIC SIGNIFICANCE OF ALP ISOENZYMES
ALKALINE PHOSPHATASE (ALP)

Intestinal Isoenzyme:
• Dependent on…
• Elevated after____, in digestive tract diseases, and chronic hemodialysis

A

blood group and secretor status

fatty meal

51
Q

ASSAY FOR ENZYME ACTIVITY
ALKALINE PHOSPHATASE (ALP)

A

Continuous-Monitoring Technique:

Based on Bowers and McComb method

52
Q

ALP

• Most specific method

A

Bowers and McComb method

53
Q

Bowers and McComb

Calculates ALP activity using molar absorptivity of…

A

p-nitrophenol

54
Q

Bowers and McComb

Substrate =
Positive result =
Optimal pH:

A

para Nitrophenylphosphate
(PNPP); colorless

Yellow (p-nitrophenol)

10.2

55
Q

Bowers and McComb

Measurement:
Increase in absorbance at____
Absorbance directly proportional to____ activity

A

405 nm

ALP

56
Q

SOURCE OF ERROR
ALKALINE PHOSPHATASE (ALP)

A

Hemolysis
Timing of assays
Dietary influences

57
Q

SOURCE OF ERROR
ALKALINE PHOSPHATASE (ALP)

Hemolysis:

Can cause slight elevations in ALP levels

ALP concentration:

A

~6 times higher in erythrocytes than in
serum

58
Q

SOURCE OF ERROR
ALKALINE PHOSPHATASE (ALP)

Timing of Assays:

ALP assays should be performed_____

Serum activity increases by _______when left standing at
__________

A

immediately after collection

3% to 10%

25°C or 4°C for several hours

59
Q

SOURCE OF ERROR
ALKALINE PHOSPHATASE (ALP)

Dietary Influences:

Diet can induce elevations in ALP activity

Blood group ________ individuals who are secretors may show:
Up to_______ after consuming a_____

A

Blood group B and O

25% higher values

high-fat meal

60
Q

NOMENCLATURE & FUNCTION
ACID PHOSPHATASE (ACP)

EC
Systematic name

A

E.C. 3.1.3.2

Orthophosphoric Monoester Phosphohydrolase (Acid Optimum)

61
Q

ACP

• Optimal pH =
• Activator =

A

5.0

Magnesium & Zinc

62
Q

TISSUE SOURCE
ACID PHOSPHATASE (ACP)

: Richest source, highest activity
Prostate

A

Prostate

63
Q

ACP

• Other sources:

A

Bone, liver, spleen, kidney, erythrocytes, platelets

64
Q

DIAGNOSTIC SIGNIFICANCE
ACID PHOSPHATASE (ACP)
•_________:
• Historically used for detecting metastatic carcinoma of the prostate
• Less sensitive than newer markers like _______

A

Prostatic Carcinoma Detection

Prostate-Specific Antigen (PSA)

65
Q

Other Conditions with ACP Elevations:

A

Hyperplasia of the prostate

Prostatic surgery, rectal examination, and prostate massage (conflicting reports)

66
Q

Other Conditions with ACP Elevations:

bone diseases: lagets disease, breast cancer with bone metastases,

A

Gauchers disease

67
Q

ACP elevation

Forensic applications: Elevated ACP in _____as evidence of rape

A

vaginal washings

68
Q

ACP Elevation Platelet damage: Elevations in_____ due tc excessive platelet destruction

A

thrombocytopenia

69
Q

METHODS OF PROSTATIC ACP DETECTION ACID PHOSPHATASE (ACP)

A

• Chemical Inhibition

• Immunologic Techniques

70
Q

METHODS OF PROSTATIC ACP DETECTION ACID PHOSPHATASE (ACP)

• Chemical Inhibition:
“ Uses substrates like______

Differentiates prostatic ACP using_____ inhibition

A

thymolphthalein monophosphate

tartrate

71
Q

METHODS OF PROSTATIC ACP DETECTION ACID PHOSPHATASE (ACP)

• Equation:
• Not entirely specific, as lysosomal ACP is also inhibited by tartrate

A

Total ACP - ACP after tartrate inhibition = Prostatic ACP

72
Q

ACP

Immunologic Techniques:
• Uses antibodies specific for prostatic ACP Improved sensitivity but not useful for screening non- metastasized cancer

A
73
Q

ASSAY FOR ENZYME ACTIVITY
ACID PHOSPHATASE (ACP)
•
Substrate: Thymolphthalein monophosphate

A

Roy and Hillman (Quantitative Endpoint)

74
Q

ASSAY FOR ENZYME ACTIVITY
ACID PHOSPHATASE (ACP)

Substrate:
• Babson, Read & Phillips (Continuous Monitoring)
• Substrate:

A

Thymolphthalein monophosphate

Alpha-naphthyl phosphate

75
Q

ASSAY FOR ENZYME ACTIVITY
ACID PHOSPHATASE (ACP)
• Immunochemical Techniques for prostatic ACP:

A

• Radioimmunoassay (RIA)
• Counterimmunoelectrophoresis
• Immunoprecipitation
• Immunoenzymatic Assay (Tandem E)

76
Q

Immunoenzymatic Assay (Tandem E):
• Incubates sample with an antibody to______

Followed by washing and incubation with______

________formed is measured photometrically and is proportional to prostatic ACP in the sample

A

prostatic ACP

p-nitrophenylphosphate

p-Nitrophenol

77
Q

SOURCE OF ERROR
ACID PHOSPHATASE (ACP)
• Sample Handling:
• Serum should be separated from__________________ to prevent leakage of erythrocyte and platelet ACP.

A

red cells immediately after clotting

78
Q

Sources of errors

ACP

Serum activity decreases within ______hours at______ if no preservative is added.

Decrease is due to CO2 loss and subsequent increase in pH.

A

1 to 2 hours; room temperature

79
Q

ACP

To Prevent Decrease:
Freeze or acidify serum to a______

Acidified serum remains stable at
room temperature for up to______

A

pH < 6.5

2 days

80
Q

Hemolysis:
• Avoid hemolysis as it leads to contamination with…

A

erythrocyte ACP

81
Q

• RIA Procedures:
• Require_____ serum samples.
• Stable for up to____ at _____

A

nonacidified

2 days at 4°C.

82
Q

REFERENCE RANGE
ACID PHOSPHATASE (ACP)
• Reference Range
• Prostatic ACP,_____

• Tartrate-resistant ACP
• adults:
children:

A

0 to 3.5 ng/mL

1.5-4.5 U/L (37°C)

3.5-9.0 U/L (37°C)

CC 2 LAB (35 decks)

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