Enzymes Flashcards
Which blood group has increased intestinal ALP after consumption of a fatty meal?
B or O blood group
A nonspecific liver function test
Alkaline Phosphatase
True or False: Bone ALP is not normally elevated in children and geriatric patients.
False. It is normally found in healthy serum of pediatric and geriatric patients.
Major tissue sources of ALP.
Liver, Bone, Placenta and Intestine
Specific enzyme classification of ALP.
Hydrolase (Esterase)
Most abundant phosphatases.
Bone and Liver ALP
High ALP, high GGT. Diagnosis?
Obstructive Jaundice.
High ALP, normal GGT. Diagnosis?
Paget’s Disease (Osteitis deoformans)
Primary test for obstructive jaundice.
Alkaline Phosphatase
Secondary test for obstructive jaundice.
Gamma Glutamyl Transamine Peptidase
Bone ALP isoform detected in serum of dialysis patients.
B1x
B1x, a bone ALP isoform, is used to study what disease?
Low Bone Mineral Disease
Carcinoplacental ALP that rises in cases of breast, ovarian and gynecological cancers.
Regan ALP
Regan ALP migrates with what ALP isoenzyme?
Bone ALP
Which ALP isoenzyme is inhibited by phenylalanine?
Placental, Intestinal, Regan and Nagao ALP
Which ALP isoenzyme is inhibited by levamisole?
Bone and Liver ALP
Which ALP isoenzyme is inhibited by 3M urea?
Bone ALP
Which ALP isoenzyme is inhihited by L-leucine?
Nagao ALP
Carcinoplacental ALP found in adenocarcinoma of pancreas and bile duct and pleural cancer.
Nagao ALP
What reagent inhibits Nagao ALP?
Phenylalanine and L-leucine
What reagent inhibits bone ALP?
Levamisole and 3M urea
Most anodal ALP isoenzyme.
Liver ALP
Least anodal ALP isoenzyme.
Intestinal ALP
What reagents improve the isolation of ALP isoenzymes in electrophoresis?
Neuraminidase and wheat germ lectin
Arrange ALP isoenzyme from most anodal to least anodal.
Liver ALP
Bone ALP
Placental ALP
Intestinal ALP
It is a test performed at 56 C for 10 to 15 minutes to differentiate heat stability of ALP isoenzymes.
Heat Fractionation Test or Heat Stability Test
Most heat stable ALP isoenzyme.
Placental ALP
Most heat labile ALP isoenzyme.
Bone ALP
This method uses different concentrations of phenylalanine, synthetic urea and levamisole solutions.
Chemical Inhibition Test
Other name for Bowers and Mc Comb Method.
Szasz modification
Considered as the most specific method for ALP.
Bowers and Mc Comb
Substrate used in Bowers and Mc Comb and Bessy, Lowry and Brock.
a. For what enzyme?
b. End product?
c. End color?
p-nitrophenyl phosphate
a. ALP
b. p-nitrophenol or nitrophenoxide ion
c. Yellow
What is the enzyme activator of ALP?
Magnesium
What will be the effect of hemolysis and fatty meals in serum ALP?
Falsely elevated
True or False: ALP is sensitive if stored at low temperature and leads to decreased serum level.
False. It will lead to increased serum level.
What is the purpose of 2-amino-2-methyl-1-propanol (AMP) buffer in Bowers-McComb method?
Binds phosphorus that may inhibit ALP
What is the buffer added to bind the phosphorus in order to avoid ALP inhibition?
2-amino-2-methyl-1-propanol (AMP)
Level of ALP in zinc deficiency.
Decreased
ALP isoenzyme that is a useful tumor marker in serum and CSF for most germ cell tumors.
Placental ALP
Prolonged ______ levels of ALP occur in hypophosphatasia.
low or decreased
ACP activity >50 IU/L indicates the presence of what body fluid in the sample?
Seminal fluid
Tissue sources of ACP.
Prostate (major) RBCs Platelets Liver Bone
Ideal pH for ACP reaction.
pH 5.0
Diagnostic significance of ACP.
a. Detection of prostatic adenocarcinoma
b. Forensic clinical chemistry (rape cases)
How long can seminal fluid ACP persist in vaginal washings?
7 days
ACP methodology commonly used.
Roy and Hillman
Substrate used in Roy and Hillman? For what enzyme?
Thymolphthalein monophosphate
ACP
With acidification, ACP is stable for how many days at room temperature?
2 days
This inhibits prostatic ACP.
20mM L-tartrate ions
These inhibit red cell ACP.
1mM cupric sulfate and 2% formaldehyde ions
Tartrate-Resistant Acid Phosphatase (TRAP) is present in what disease?
Hairy Cell Leukemia
What is used together with prostatic acid phosphatase to monitor recurrence of prostate cancer?
Prostate Specific Antigen (PSA)
Specific enzyme classification of ACP.
Hydrolase (Esterase)
Substrates and products of AST.
Substrate: aspartate and alpha-keto acids
Products: oxaloacetate and glutamate
2 isoenzyme fractions of AST?
Cytoplasm and Mitochondrial AST
Major tissue sources of AST.
Cardiac tissue, liver and skeletal muscle.
Other: kidney, pancreas and RBC
Rise, peak and normal of AST in AMI.
Rise: 6-8 hours
Peak: 24 hours
Normal: within 5 days
Used for monitoring therapy with potentially hepatotoxic drugs.
AST (3x above normal limit may mean cessation of therapy)
Method used in AST determination.
Karmen method
pH and wavelength in Karmen Method.
pH 7.5 at 340 nm
Other enzyme used in Karmen method aside from AST.
Malate dehydrogenase
In myocardial infarction, hepatocellular disorders and skeletal muscle involvement, AST is _________.
a. decreased
b. increased
c. normal
d. cannot be determined
b. increased
Substrates and products of ALT.
Substrates: alanine and alpha-ketoglutarate
Products: glutamate and pyruvate
Specific liver enzyme test.
ALT
Major tissue source of ALT.
Liver
Other tissue sources of ALT aside from liver.
Kidney Pancreas RBC Heart Skeletal muscles Lungs
Enzyme used to screen blood donors.
ALT
Sensitive and specific screening test for posttransfusion hepatitis and occupational toxic exposure
ALT
Method for ALT.
Couple enzymatic reaction
Reitman and Frankel
Cofactor of aminotransferases.
Pyridoxal phosphate (vitamin B6)
Disease that causes highest elevation of transferases.
Acute hepatitis
Disease that produces elevations of transferase up to 20x the normal limits.
Viral and toxic hepatitis
Increased: ALP, ALT, AST, Bilirubin (both)
Normal: TPAG
Diagnosis?
Acute injury (Hepatitis) or Necrosis
Secondary markers for acute hepatitis and necrosis.
Increased LDH 4 and 5
Increased: ALP, Bilirubin (B2)
Slightly increased: ALT, AST
Normal: TPAG
Diagnosis?
Biliary Tract Obstruction (Obstructive Jaundice)
Secondary markers for obstructive jaundice.
Increased GGT, LAP and 5’NT
Increased: bilirubin (both), Globulin (IgA)
Slightly increased/decreased: ALT, AST, ALP
Decreased: TPA
Diagnosis?
Cirrhosis
Secondary markers for cirrhosis.
Increased NH3 and slightly increased or normal LDH 4 and 5
This enzyme catalyzes te breakdown of starch and glycogen.
Amylase
It is the smallest enzyme in size.
Amylase
It is the earliest pancreatic marker.
Amylase
What are the amylase isoenzymes?
Ptyalin (S-type)
Amylopsin (P-type)
True or False: Both ptyalin and amylopsin are present in normal sera.
True
Major tissue sources of amylase.
Acinar cells of the pancreas and the salivary glands
Other tissue sources of amylase.
Adipose Tissue
Fallopian tubes
Small intestine
Skeletal muscles
Most anodal amylase isoenzyme.
Ptyalin
What do you call the pancreatic amylase?
Amylopsin
What do you call the salivary amylase?
Ptyalin
Rise, peak and normal of amylase in acute pancreatitis.
Rise: 2-12 hours
Peak: 24 hours
Normal: within 3-5 days
In acute pancreatitis, increased AMS blood levels are accompanied by increased urinary excretion. How long does amylase stay elevated in urine?
Up to 7 days
Increased plasma amylase, decreased urinary amylase.
Diagnosis?
Renal failure
What is the expected amylase level in parotitis?
Increased
Samples with high activity of AMS shiuld be diluted with what?
NaCl (to prevent inactivation)
Morphine and other opiates will cause falsely _______ serum AMS levels.
Elevated
Substrate for all amylase methodologies.
Starch
Reference method of amylase determination. (State also the units used)
Saccharogenic (Somogyi units)
It measures the amount of reducing sugars produced by the hydrolysis of starch by the usual glucose methods.
Saccharogenic method (AMS)
It measures amylase activity by following the decreases in substrate concentration.
Amyloclastic
