LEC2 - ENZYMES - ALP, ACP, AST, ALT Flashcards
Characterized by its ability to hydrolyze a large variety of organic phosphate esters with the formation of an alcohol and a phosphate
ion
Phosphatases
alkaline phosphatase whole name
Alkaline Orthophosphoric Monoester
Phosphohydrolase
2 clinically significant enzyme under phosphatases
alp and acp
alkaline phosphatase
acid phosphatase
Alkaline Phosphatase reference value
Reference Value: 30-90 ul
Alkaline Phosphatase optimal pH
8.6-10 pH
acid phosphatase optimal pH
3-5 pH
class of phosphatases
class 3
is an enzyme involved in the cleavage of phosphate-containing compounds in alkaline pH.
Alkaline Phosphatase (EC 3.1.3.1) or ALP
It facilitates movement of substances
across cell membranes
Alkaline Phosphatase (EC 3.1.3.1) or ALP
in healthy serum, ALP mendles and usually derived from which organs
liver and bone
this enzyme functions to liberate inorganic phosphate from an organic phosphate ester with the concomitant production of an alcohol
phosphatase
Alkaline Phosphatase (EC 3.1.3.1) or ALP
Several isoenzymes exist which include
- Placental isoenzyme
- Intestinal isoenzyme
- Liver isoenzyme
- Bone isoenzymes.
mnemonic for the chromosome number of the isoenzymes of Alkaline phosphatase
1 KiLo ng Baboy
2 Palengke ng India
chromosome 1
Ki- kidney
Lo- liver
Baboy - Bone
chromosome 2
Palengke - placenta
India - intestinal
aside from serum ALP, what endogenous substance increases as well during period of growth and muscle development
creatinine level
kasi muscle diba.
at normal pregnancy, increased ALP activity /Placental isoenzyme is peak at what week of pregnancy
16th to 20th week of normal pregnancy
Alkaline Phosphatase isoenzyme which can be used to determine any abnormalities in the placenta
Placental isoenzyme
Placental isoenzyme is lower in pregnant of blood groups ____
blood A or AB in low levels only
Intestinal isoenzyme
it increases after consuming fatty meal within ___
2-3 hrs
Intestinal isoenzyme
it increases after consuming fatty meal within 2-3 hrs, specially for what group
type B or O blood group
not inly the intestinal ALP isoenzyme is higher in population with B and O blood group, but as well as the general ALP enzyme
what could be the reason for this one?
because of the differences in intestinal ALP levels
isoenzyme that is very helpful in detection of post hepatic and liver disease
Liver isoenzyme
when total ALP activity is elevated, mostly it is contributed by what isoenzyme
LIVER ISOENZYME
ALP isoenzyme that increases in osteoblastic activities
Bone isoenzymes
in children, elevated as they are still growing
in older peeps, elevated as the bones are damaged causing the leaked in the blood
how to differentiate the alp isoenzymes from each other
electrophoresis
heat fractionation
MNEMONIC
in electrophoresis, which ALP isoenzyme is the fastest and the slowest
I Promise (to) Be Loved
fastest - Liver
Slowest - Intestine
MNEMONIC
heat denaturation
arranged the ALP isoenzymes from the most heat stable to most labile
Promise Ikaw Lang Baby
placenta - most stable
Intestine
Liver
Bone - most labile
in heat fractionation, isoenzymes are can be denatured above what temp
60*C
Liver ALP is inhibited by
Levamisole
is the iso enzyme of alp that is the most heat labile
Bone ALP
chemical inhibition test that inhibits the bone amp
Urea
Levamisole
is the iso enzyme of alp that is the most heat stable
Placental ALP
chemical inhibition test that inhibits the placental alp
Phenylalanine Rgt
Phenylalanine reagent will inhibit what enzymes
placental ALP
Intestinal ALP
Urea will inhibit which isoenzyme
bone
Levamisole will inhibit which ALPisoenzyme
liver and bone isoenzyme
We know that if mataas si total ALP, mostly ang major contributor is si liver isoenzyme, but what might be the condition?
obstructive jaundice - ALP is increased due to greater rate of secretion
In biliary tract obstruction, serum ALP increases are primarily result of increased synthesis of ALP enzyme by induced __
cholestasis
***nakita sa twitter
enzyme marker kapag hepatocellular and hepatobilliary
hepatocellular - AST and ALT
hepatobiliary - GGT and ALP
for bone disorders, highest elevations of ALP occur in what disease
Paget’s disease (osteitis deformans)
anticoagulant that will inactivate ALP
EDTA, citrate, oxalate - falsely lowers actvity (chelators kasi sila eh)
serum is the preferred sample
what ion or electrolyte that ALP requires for activation
magnesium ions
in PPT can be ZINC as well is an activator but it is actually a component of ALP according to the Book
why do Carcinoplacental ALP is called as Carcinoplacental ALP
present only in the presence of cancer or carcinoma, placental alp because it has the same chemical composition with placental alp but they are not the same
Carcinoplacental ALP types
Regan ALP
Nagao ALP
Kasahara ALP
Regan ALP is a carcinoplacental ALP that is found in
lung, breast, ovarian and gynecological cancers;
bone ALP co-migrator
regan - carcinoplacental ALP
the most heat stable carcinoplacental ALP
Regan ALP
Regan ALP (carcinoplacental ALP) is inhibited by
phenylalanine
reagent
Nagao ALP (carcinoplacental ALP ) is found in
adenocarcinoma of the pancreas and bile duct,
a carcinoplacental ALP that is can be seen in pleural cancer
Nagao ALP
variant of Regan ALP
Nagao ALP
Nagao ALP
(carcinoplacental alp) is inhibited by
L-leucine and phenylalanine
Kasahara ALP (carcioplancental ALP) is predominant in what condition
hepatoma/hepatocellular carcinoma
2 isoenzymes of liver sample
kasahara ALP as a tumor marker
liver isoenzyme for detection of post hepatic biliary disease
methods for ALP determination
electrophoresis
heat fractionation
chemical inhibition test
bowers-mccomb
using electrophoresis, the use of ___ and ___ improves the separation of bone and liver ALP
neuraminidase and wheat germ lectin
Heat fractionation for ALP is done at what temp and how many mns
56*C at 10-15 mns
most specific Methods of Determination of ALP and is IFCC-RECOMMENDED method
Bowers and Mc Comb(continuous-monitoring technique)/ SZASZ modification
what is the constant level of pH of Bowers and Mc Comb for determining ALP
– pH 10.15 or alkaline
it is currently the routine method for ALP
Bowers-McComb method
aside from the constant pH for determining ALP using Bowers-McComb, what is the wavelength required
405 nm
substrate of Bowers and Mc comb (continuous-monitoring technique)
p-nitrophenylphosphate
Bowers and Mc Comb
ALP will act on the substrate
p-nitrophenylphosphate resulting to products which are
p-nitrophenol + phosphate ion
(alcohol and inorganic phosphate)
the substrate recommended for ALP
p-nitrophenyl phosphate
methods that are using Beta-glycerophosphate as their substrate
bodansky
shinowara
jones
reinhart
methods that are using Phenylphosphate as their substrate
King and Armstrong
methods that are using Phenolphthalein
diphosphate as their substrate
huggins and talalay
methods that are using Alpha-naphthol
phosphate as their substrate
moss
methods that are using Buffered phenolphthalein
phosphate as their substrate
Klein, Babson and
Reid
a method King and Armstrong and a substrate of Phenylphosphate has a product of
phenol
Activators for ALP are ____
zinc, magnesium.
and other cations,
In ALP determination, Chelators(such as _____) can falsely lower activity
EDTA, citrate, oxalate as they have the mechanism of binding
Activity of enzyme ____ slightly on storage due
to loss of inhibitors.
increases
ALP is relatively stable at____ for up to one week.
4 * C
optimum pH of ALP
Optimum pH: 8.6-10
ALP is inhibited by phosphorus, using Bowers-McComb, what is added to bind phosphorus?
AMP - 2 amino-2-methyl-1-propanol
Increased ALP is seen on the ff. conditions
- Osteitis deformans/Paget’s disease
- Osteomalacia
- Rickets
- Osteoblastic bone tumors
- Bone Cancer
- Sprue
- Hyperparathyroidism
- Obstructive jaundice
- Hepatitis and cirrhosis
most important activator for ALP is
magnesium
most common causes of ALP elevations
liver and bone diseases
Decreased ALP are seen in what cirmcumstances and conditions
After blood transfusions or cardiopulmonary bypass (transiently decrease)
Malnutrition
Hypophosphatemia (prolonged, severely low levels)
Zinc deficiency (prolonged, severely low levels.
Acid Phosphatase is active at what pH
pH 5.0
what are the 5 isoenzymes of Acid phosphatase
prostatic ACP
erythrocyte ACP
lysosomal ACP
testicular ACP
macrophagic ACP - (bone)
among the 5 isoenzymes of ACP, what are the significant
bone and prostatic
Diagnostic significance of acid phosphatase
detection of prostatic carcinoma
Acid Phosphatase
major tissue sources
prostate
other tissue source of acid phosphatase aside from prostate
rbc, platelet, liver, and bone
Reference values for ACP
2.5-11.7 (total ACP male)
0-3.5 ng/mL (prostatic ACP)
is a hydrolytic enzyme secreted by a number of cells
Acid Phosphatase (EC 3.1.2.3) or ACP
There are several isoenzymes of ACP, each with
tissue specificity. Isoenzymes may be fractionated
into how many bands
five bands
the band 1 of ACP, its major source is the
prostate gland
ACP Isoenzymes
* Band 1, is inhibited by
tartrate
ACP isoenzymes
Band 2 and 4 isoenzymes are from ____.
granulocytes
ACP isoenzymes
Band 3 is the major form present in ___.
plasma
This isoenzyme (band 3) is derived from ____.
platelets, erythrocytes, and monocytes
Band 5 of ACP soenzyme is found mainly in ___.
osteoclasts
the only resistant in tartrate inhibition among the 5 bands of ACP isoenzyme
band 5
the method with highest specificity among the method of determination of ACP is the
Roy and Hillman
Tartrate-inhibited ACP (Prostatic Isoenzyme)
disease associated
- Prostatic Cancer
- Benign prostatic hyperplasia
- Prostatic infarction
- Urinary tract obstruction, carcinoid tumors of the rectum, and prostatic massage
___also has implications in suspected rape
ACP
Positive ACP is
evident in vaginal swab if semen is present for the first ___ hrs to ___ days from
the incident
12 hours up to four days
difference of TRAP 5b to ALP isoforms of B/I, B1, AND B2
TRAB 5B is for bone resorption (osteoclasts)
ALP isoforms are for bone deposition (osteoblasts)
is considered to be the marker of osteoclasts
TRAP 5B
Tartrate-resistant ACP (Bone Isoenzyme)
disease correlation
- Active osteoclast-mediated bone resorption
- Gaucher’s cells
- Hairy cell leukemia
- chronic leukemia,
- lymphoma
how to differentiate the prostatic and non prostatic acp
through Chemical Inhibition
Technique
in Chemical Inhibition
Technique
prostatic acp is resistant in ___
copper
in Chemical Inhibition
Technique
rbc acp is resistant in ___
tartrate
Moderate elevation of Total ACP are seen in what conditions
- Female Breast CA
- Paget’s disease
- Hyperparathyrodism
Non-prostatic ACP elevations
- Neimann-Pick disease
- Gaucher’s disease
- Myelocytic leukemia
Catalyzes the transfer of an amino group of one amino acid to a
hydrocarbon to form a different amino acid
Aminotransferases
Aminotransferases are of two
- Aspartate Aminotransferase (EC 2.6.1.1) or AST
- Alanine Aminotransferase (EC 2.6.1.2) or ALT
Cofactor of aminotrasnferase
vitamin b6 or pyridoxal-S’-phosphate or P-5’-P
isoenzyme of AST (SGOT )
cytoplasmic AST
mitochondrial AST
tissue source of AST
cardiac tissue, liver and skeletal muscle
SGPT means
Serum Glutamate Pyruvate Transaminase i
SGOT means
serum glutamic-oxaloacetic transaminase
what isoenzyme of AST is the abundant fraction in healthy serum
cytoplasmic AST
in terms of tissue necrosis, which AST isoenzyme is abundant
mitochondrial AST
which means normal si cystoplasmic abnormal naman si mitochondrial
choices : AST, ALT, LDH
___ and ____ have higher activity in the liver than ____, but both are non specific enzymes
AST and LDH have higher activity in the liver than ALT, but both are non specific enzymes
choices : AST, ALT, LDH
both __ and ___ have equal concentrations in the hepatocytes
AST and LDH
choices : AST, ALT, LDH
the kidney has higher __ compared to __ and ___
the kidney has higher AST activity compared to ALT and LDH
ref range of AST
5-37 u/l
The half-life of AST is __
17+- 5 hrs
ALT has a half-life of
47 +- 10 hours
AST is stable in serum at refrigerator
temperature for up to ____,
three weeks
ALT has the same
stability but markedly decreases with ___.
freezing
Specimens for AST and ALT are stable in whole
blood for up to 12 to 24 hours, but increase
with time due to release from red blood cells
tue or fa;se
true
optimum ph of ast and alt
Optimum pH: 7.4
- Involved in the transfer of an amino group
between aspartate and α-ketoacids with the
formation of oxaloacetate and glutamate
AST (Aspartate
Aminotransferase)
AST (Aspartate
Aminotransferase)
Has 2 isoenzymes fractions:
cytoplasm and
mitochondrial
AST (Aspartate
Aminotransferase)
Major tissue source:
cardiac tissue, liver and skeletal muscles
AST other sources
Other sources: kidney, pancreas and RBC
ast ref values
Reference values: (5-37 U/L)
method of determiation for AST
- Karmen Method – pH 7.5; 340 nm
AST is used for monitoring therapy with potentially hepatotoxic drugs; a result more than _____ the upper border of normal should signal cessation of therapy
3X
Method of determination of AST
karmen method
Uses ___ and monitors the change in absorbance
malate dehydrogenase
significance of (Increased
AST activity
In the evaluation of
myocardial infarction,
hepatocellular disorders and skeletal muscle
involvement
MI AST level is usually ___ times the upper
limit of normal
4-10
Clinical
Significance
(Decreased AST
activity)
- Decreased level is seen during
___
pregnancy
ALT (Alanine
aminotransferase)
highest concentration is in
liver
Other sources of ALT
kidney, pancreas,
RBC, heart, skeletal muscles,
lungs
highest concentration is in the liver, hence, increased level of ALT means
hepatic injury
ALT (Alanine
aminotransferase ref range
Reference Values: 6-37 U/L
this transferase monitors the course of liver treatment and the effects of drug therapy
ALT
this transferase’s level is used to screen blood donors
ALT
AST or ALT
____ measurement is a more sensitive and specific screening test for post-transfusion hepatitis or occupational toxic exposure
ALT
limit of interference of hemolysis with hemoglobin for ALT, GGT, INORGANIC PHOSPHATE, and CHOLESTEROL
2.5 g/L to 4.5 g/l
Method of determination of ALT
Coupled Enzymatic reaction: pH 7.5; 340 nm
Reitman-Frankel Method
Aminotransferase activity measurement
is done by coupled enzymatic reactions,
using ___ as the final reaction product
NADH
Reagents with NH4 (ammonium) will give falsely __ ALT and AST owing to the
conversion of NADH to NAD by the
ammonium ion
increased
+++recommended that
methods should include P-5’-P (vitamin b6) in the reagent for aminotransferases
International Federation of Clinical
Chemistry (IFCC)
Diagnosis of acute or chronic viral
hepatitis ALT increases to a
greater degree than AST
tru or false
true
Aminotransferase levels
are altered in
- Hepatocyte injury (increase in AST, and ALT but to a lesser degree)
- Muscle injury (increase in both enzymes)
- Kidney infarcts (increase in both enzymes)
- Renal failure (falsely lowered)
which hepatitis is responsible for acute liver injury resulting in increased plasma levels of ALT and AST
according to the book, it’s A, B, and C
severe viral or toxic hepatitis can elevate transferases up to ___
20x the normal limits
high AST/ALT ratio suggest what condition
advanced alcoholic liver disease
