CC 2 LAB > PHOSPHATASES > Flashcards

PHOSPHATASES Flashcards

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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
Most heat labile
Bone
26
ISOENZYME ANALYSIS ALKALINE PHOSPHATASE (ALP) • Chemical Inhibition
Phenylalanine 3M urea Levamisole Leucine
27
" Phenylalanine • inhibits (2)
intestinal, and placental ALP
28
Phenylalanine also inhibits the…
carcinoplacental isoenzyme
29
3M Urea Inhibits ____ ALP
Bone
30
Levamisole Inhibits (2) ALP
Bone and Liver
31
Leucine Inhibits the…
Nagao isoenzyme
32
NEOPLASTIC ISOENZYMES ALKALINE PHOSPHATASE (ALP)
Regan isoenzymes Nagao isoenzymes
33
• carcinoplacental ALPs
Regan and Nagao isoenzymes
34
ALP Isoenzymes migrates to the same position as the bone fraction most heat stable of all ALP isoenzymes
Regan
35
ALP Isoenzyme found in lung, breast, ovarian, and colon cancers
Regan
36
variant of Regan isoenzyme found in metastatic carcinoma and adenocarcinoma
Nagao
37
Associated with hepatoma and gastrointestinal tract tumors
Kasahara
38
DIAGNOSTIC SIGNIFICANCE ALKALINE PHOSPHATASE (ALP) Significant in…
hepatobiliary and bone disorders
39
T or F ALP is more prominent in obstructive conditions than hepatocellular disorders
True
40
ALP Observed with____ involvement in bone disorders
osteoblast
41
Hepatobiliary Disorders: Biliary tract obstruction:
ALP levels 3 to 10 times the ULN
42
Hepatobiliary disorders Hepatitis and cirrhosis: increases by..
slight increases (usually <3 times the ULN)
43
DIAGNOSTIC SIGNIFICANCE ALKALINE PHOSPHATASE (ALP) • Bone Disorders: Highest elevation seen in this condition
Paget's disease • Highest elevations in Paget's disease (Osteitis deformans)
44
DIAGNOSTIC SIGNIFICANCE ALKALINE PHOSPHATASE (ALP) Other disorders:
osteomalacia, rickets, hyperparathyroidism, osteogenic sarcoma
45
Bone disorders ALP Elevated during ——-and periods of physiologic——-
healing bone fractures bone growth
46
ALKALINE PHOSPHATASE (ALP) " Pregnancy: • Increased ALP activity (______at 16-20 weeks; _____ in third trimester) • Returns to normal within ___after labor
1.5 times ULN 2-3 times ULN 3-6 days
47
DIAGNOSTIC SIGNIFICANCE ALKALINE PHOSPHATASE (ALP) Pregnancy: Complications:
elevated in hypertension, preeclampsia, eclampsia, and threatened abortion
48
DIAGNOSTIC SIGNIFICANCE ALKALINE PHOSPHATASE (ALP) Decreased ALP: inherited condition with subnormal ALP activity
Hypophosphatasia
49
DIAGNOSTIC SIGNIFICANCE OF ALP ISOENZYMES ALKALINE PHOSPHATASE (ALP) • Bone Isoenzyme: • Increased due to_____ activity • Elevated in…
osteoblastic children during growth and adults over 50
50
DIAGNOSTIC SIGNIFICANCE OF ALP ISOENZYMES ALKALINE PHOSPHATASE (ALP) Intestinal Isoenzyme: • Dependent on… • Elevated after____, in digestive tract diseases, and chronic hemodialysis
blood group and secretor status fatty meal
51
ASSAY FOR ENZYME ACTIVITY ALKALINE PHOSPHATASE (ALP)
Continuous-Monitoring Technique: Based on Bowers and McComb method
52
ALP • Most specific method
Bowers and McComb method
53
Bowers and McComb Calculates ALP activity using molar absorptivity of…
p-nitrophenol
54
Bowers and McComb Substrate = Positive result = Optimal pH:
para Nitrophenylphosphate (PNPP); colorless Yellow (p-nitrophenol) 10.2
55
Bowers and McComb Measurement: Increase in absorbance at____ Absorbance directly proportional to____ activity
405 nm ALP
56
SOURCE OF ERROR ALKALINE PHOSPHATASE (ALP)
Hemolysis Timing of assays Dietary influences
57
SOURCE OF ERROR ALKALINE PHOSPHATASE (ALP) Hemolysis: Can cause slight elevations in ALP levels ALP concentration:
~6 times higher in erythrocytes than in serum
58
SOURCE OF ERROR ALKALINE PHOSPHATASE (ALP) Timing of Assays: ALP assays should be performed_____ Serum activity increases by _______when left standing at __________
immediately after collection 3% to 10% 25°C or 4°C for several hours
59
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_____
Blood group B and O 25% higher values high-fat meal
60
NOMENCLATURE & FUNCTION ACID PHOSPHATASE (ACP) EC Systematic name
E.C. 3.1.3.2 Orthophosphoric Monoester Phosphohydrolase (Acid Optimum)
61
ACP • Optimal pH = • Activator =
5.0 Magnesium & Zinc
62
TISSUE SOURCE ACID PHOSPHATASE (ACP) : Richest source, highest activity Prostate
Prostate
63
ACP • Other sources:
Bone, liver, spleen, kidney, erythrocytes, platelets
64
DIAGNOSTIC SIGNIFICANCE ACID PHOSPHATASE (ACP) •_________: • Historically used for detecting metastatic carcinoma of the prostate • Less sensitive than newer markers like _______
Prostatic Carcinoma Detection Prostate-Specific Antigen (PSA)
65
Other Conditions with ACP Elevations:
Hyperplasia of the prostate Prostatic surgery, rectal examination, and prostate massage (conflicting reports)
66
Other Conditions with ACP Elevations: bone diseases: lagets disease, breast cancer with bone metastases,
Gauchers disease
67
ACP elevation Forensic applications: Elevated ACP in _____as evidence of rape
vaginal washings
68
ACP Elevation Platelet damage: Elevations in_____ due tc excessive platelet destruction
thrombocytopenia
69
METHODS OF PROSTATIC ACP DETECTION ACID PHOSPHATASE (ACP)
• Chemical Inhibition • Immunologic Techniques
70
METHODS OF PROSTATIC ACP DETECTION ACID PHOSPHATASE (ACP) • Chemical Inhibition: " Uses substrates like______ Differentiates prostatic ACP using_____ inhibition
thymolphthalein monophosphate tartrate
71
METHODS OF PROSTATIC ACP DETECTION ACID PHOSPHATASE (ACP) • Equation: • Not entirely specific, as lysosomal ACP is also inhibited by tartrate
Total ACP - ACP after tartrate inhibition = Prostatic ACP
72
ACP Immunologic Techniques: • Uses antibodies specific for prostatic ACP Improved sensitivity but not useful for screening non- metastasized cancer
73
ASSAY FOR ENZYME ACTIVITY ACID PHOSPHATASE (ACP) • Substrate: Thymolphthalein monophosphate
Roy and Hillman (Quantitative Endpoint)
74
ASSAY FOR ENZYME ACTIVITY ACID PHOSPHATASE (ACP) Substrate: • Babson, Read & Phillips (Continuous Monitoring) • Substrate:
Thymolphthalein monophosphate Alpha-naphthyl phosphate
75
ASSAY FOR ENZYME ACTIVITY ACID PHOSPHATASE (ACP) • Immunochemical Techniques for prostatic ACP:
• Radioimmunoassay (RIA) • Counterimmunoelectrophoresis • Immunoprecipitation • Immunoenzymatic Assay (Tandem E)
76
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
prostatic ACP p-nitrophenylphosphate p-Nitrophenol
77
SOURCE OF ERROR ACID PHOSPHATASE (ACP) • Sample Handling: • Serum should be separated from__________________ to prevent leakage of erythrocyte and platelet ACP.
red cells immediately after clotting
78
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.
1 to 2 hours; room temperature
79
ACP To Prevent Decrease: Freeze or acidify serum to a______ Acidified serum remains stable at room temperature for up to______
pH < 6.5 2 days
80
Hemolysis: • Avoid hemolysis as it leads to contamination with…
erythrocyte ACP
81
• RIA Procedures: • Require_____ serum samples. • Stable for up to____ at _____
nonacidified 2 days at 4°C.
82
REFERENCE RANGE ACID PHOSPHATASE (ACP) • Reference Range • Prostatic ACP,_____ • Tartrate-resistant ACP • adults: children:
0 to 3.5 ng/mL 1.5-4.5 U/L (37°C) 3.5-9.0 U/L (37°C)
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