LEC2 - ENZYMES - ALP, ACP, AST, ALT Flashcards by CHARLENE SAN JOSE

Characterized by its ability to hydrolyze a large variety of organic phosphate esters with the formation of an alcohol and a phosphate
ion

Phosphatases

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alkaline phosphatase whole name

Alkaline Orthophosphoric Monoester
Phosphohydrolase

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2 clinically significant enzyme under phosphatases

alp and acp

alkaline phosphatase
acid phosphatase

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Alkaline Phosphatase reference value

Reference Value: 30-90 ul

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Alkaline Phosphatase optimal pH

8.6-10 pH

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acid phosphatase optimal pH

3-5 pH

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class of phosphatases

class 3

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is an enzyme involved in the cleavage of phosphate-containing compounds in alkaline pH.

Alkaline Phosphatase (EC 3.1.3.1) or ALP

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It facilitates movement of substances
across cell membranes

Alkaline Phosphatase (EC 3.1.3.1) or ALP

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in healthy serum, ALP mendles and usually derived from which organs

liver and bone

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this enzyme functions to liberate inorganic phosphate from an organic phosphate ester with the concomitant production of an alcohol

phosphatase

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Alkaline Phosphatase (EC 3.1.3.1) or ALP

Several isoenzymes exist which include

Placental isoenzyme
Intestinal isoenzyme
Liver isoenzyme
Bone isoenzymes.

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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

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aside from serum ALP, what endogenous substance increases as well during period of growth and muscle development

creatinine level

kasi muscle diba.

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at normal pregnancy, increased ALP activity /Placental isoenzyme is peak at what week of pregnancy

16th to 20th week of normal pregnancy

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Alkaline Phosphatase isoenzyme which can be used to determine any abnormalities in the placenta

Placental isoenzyme

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Placental isoenzyme is lower in pregnant of blood groups ____

blood A or AB in low levels only

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Intestinal isoenzyme

it increases after consuming fatty meal within ___

2-3 hrs

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Intestinal isoenzyme

it increases after consuming fatty meal within 2-3 hrs, specially for what group

type B or O blood group

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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

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isoenzyme that is very helpful in detection of post hepatic and liver disease

Liver isoenzyme

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when total ALP activity is elevated, mostly it is contributed by what isoenzyme

LIVER ISOENZYME

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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

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how to differentiate the alp isoenzymes from each other

electrophoresis
heat fractionation

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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

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