(F) L1: Enzymes (Part 3) Flashcards
Major Clinical Enzymes
- This contains zinc and is part of the glycolytic pathway
- Is usually coupled with amylase in the degradation of carbohydrates
Lactate Dehydrogenase (LD)
Lactate Dehydrogenase (LD)
It requires what coenzyme?
NAD+
Lactate Dehydrogenase (LD)
LD has several isoenzymes which are considered what type of molecule?
Tetrameric (has 4 subunits of H and M)
Lactate Dehydrogenase (LD) Isoenzymes
Sourced from the heart, RBCs, and kidneys
LD 1 and 2
Lactate Dehydrogenase (LD) Isoenzymes
Sourced from the lungs, pancreas, and spleen
LD 3
Lactate Dehydrogenase (LD) Isoenzymes
Sourced from the skeletal muscle, liver, and intestine
LD 4 and 5
Lactate Dehydrogenase (LD)
- Reference value for forward reaction (enzyme is related to the substrate)
- Reference value for reverse reaction (enzyme is related to the product)
A. 100-225 U/L
B. 80-280 U/L
- A
- B
Lactate Dehydrogenase (LD) Isoenzymes
These are considered to be the most abundant, most anodal, and heat stable
LD 1 and 2
LD 1: 17-27% while LD 2: 27-37%
Lactate Dehydrogenase (LD) Isoenzymes
These are considered to be the least concentrated, least anodal, and heat labile
LD 4 and 5
LD 4: 3-8% while LD 5: 0-5%
Lactate Dehydrogenase (LD) Isoenzymes
100% made up of H subunits
LD 1
Lactate Dehydrogenase (LD) Isoenzymes
Made up of 25% M subunits and 75% H subunits
LD 2
Lactate Dehydrogenase (LD) Isoenzymes
Made up of 50% M subunits and 50% H subunits
LD 3
Lactate Dehydrogenase (LD) Isoenzymes
Made up of 75% M subunits and 25% H subunits
LD 4
Lactate Dehydrogenase (LD) Isoenzymes
100% made up of M subunits
LD 5
Lactate Dehydrogenase (LD) Other Isoenzymes
- This is seen in individuals who frequently drink alcohol which contains methanol or ethylene glycol
- This appears as a 6th band in an electrophoresis medium
- Is increased in cases of drug hepatotoxicity and obstructive jaundice
- It also appears in patients with arteriosclerotic failure
Alcohol Dehydrogenase (LD 6)
Lactate Dehydrogenase (LD) Other Isoenzymes
- This consists of C subunits
- Only seen in spermatozoa and semen and never in serum or in cases of seminoma
LD with 4C Subunits
Lactate Dehydrogenase (LD) Specimen Considerations
- This substrate provides greater specificity
- This substrate is used for detecting LD 1 activity
A. Pyruvate
B. A-hydroxybutyrate
C. Lactate
- C
- B
Lactate Dehydrogenase (LD) Specimen Considerations
An increase in α-hydroxybutyrate activity indicates an increase in what 2 isoenzymes?
LD 1 and 2
Lactate Dehydrogenase (LD) Specimen Considerations
LD is stable at what temperature for how many hours and is considered to be cold-labile, most especially LD 5?
RT for 48 hours
Lactate Dehydrogenase (LD) Specimen Considerations
In cases of pleural fluid, transudates will exhibit a/an (increased/decreased) amount of LD
Decreased
Caused by physiologic reasons such as the changes in oncotic and hydrostatic pressure within the lungs; minimal intervention may be done, and is somewhat irreversible
Lactate Dehydrogenase (LD) Specimen Considerations
In cases of pleural fluid, exudates will exhibit a/an (increased/decreased) amount of LD
Increased
Caused by pathologic reasons such as inflammation, infection, and the like; may easily be solved with multiple medical interventions
Lactate Dehydrogenase (LD) Specimen Considerations
An increase in LD may also occur after what procedure as whole blood may contain LD?
Note: This may return to normal after 24 hours
Blood transfusion
Lactate Dehydrogenase (LD) Specimen Considerations
Frozen samples will have a decreased value for what isoenzyme as this is cold labile?
LD 5
Lactate Dehydrogenase (LD) Specimen Considerations
TOF: Samples should be processed within 48 hours after collection and stored at 30ºC
False (24 hours at 25ºC)
Lactate Dehydrogenase (LD) Methods of Analysis
- It is a forward reaction with lactate as the substrate
- The lactate + NAD will be acted upon by LD which produces pyruvate and NADH
- Is read at 340nm at a pH of 8.8
- Most commonly used since it produces positive rates of NADH and is not affected by inhibitors
Wacker Method (Forward or Direct Reaction)
Lactate Dehydrogenase (LD) Methods of Analysis
- It is a reverse reaction with lactate as the product
- Pyruvate with NADH is acted upon by LD to produce lactate and NAD
- Is read at 340nm at a pH of 7.2
- The reaction is 2x faster than the other method
- Is preferred for dry slide technology
- Has less costly cofactors and only requires a small specimen volume
Wrobleuski and La Due Method (Reverse or Indirect Reaction)
Lactate Dehydrogenase (LD) Diagnostic Significance
Arrange the isoenzymes from most numerous to least numerous (LD 1 to 5)
- LD 2
- LD 1
- LD 3
- LD 4
- LD 5
Lactate Dehydrogenase (LD) Diagnostic Significance
What is the order of isoenzymes in cases of acute myocardial infarctions (most numerous to least numerous)?
LD 1 to 5 (chronological or flipped pattern)
Lactate Dehydrogenase (LD) Diagnostic Significance
LD can be used to diagnose the severity or presence of AMI depending on the levels of the enzyme in the body which will exhibit the following in the presence of the disease:
- A rise in how many hours?
- A peak in how many hours?
- A continuous elevated level up to how many days?
A. 12 to 24
B. 10 to 14
C. 48 to 72
- A (12-24 hours)
- C (48-72 hours)
- B (10-14 days)
Lactate Dehydrogenase (LD) Diagnostic Significance
LD can also be used as a marker for what to aid in the diagnosis of acute leukemia, germ cell tumors, and breast and lung malignancies?
Cancer marker
Lactate Dehydrogenase (LD) Diagnostic Significance
What isoenzymes are elevated in cases of acute leukemia, germ cell tumors, and breast and lung malignancies?
LD 2, 3, and 4
LD 3 is the predominant isoenzyme
Lactate Dehydrogenase (LD) Diagnostic Significance
A moderate increase in the levels of what isoenzyme indicates acute viral hepatitis and cirrhosis?
LD 5
Lactate Dehydrogenase (LD) Diagnostic Significance
A marked increase in the levels of what isoenzyme indicates acute hepatic carcinoma and toxic hepatitis?
LD 5
LD Level Indications
- Pernicious anemia, hemolytic disorder, and megaloblastic anemia
- Hepatic carcinoma and toxic hepatitis
- Viral hepatitis and cirrhosis
- Non-specific
A. 10x increase in serum LD
B. Elevated total serum LD
C. Slight increase of serum LD
D. Highest serum LD
- D
- A
- C
- B
Increased or Decreased LD?
- Anemias
- AMI
- Leukemia
- Renal Infarction
- Hepatitis
- Hepatic Cancer
- Muscular Dystrophy
- Delirium Tremens
- Malignancy
- Pneumocystis Jerovecii Pneumonia
Increased
Major Clinical Enzymes
- Aka “ATP-creatine-N-phosphotransferase”
- Creatine with ATP will be acted upon by this enzyme to produce creatinine phosphate and ADP
- Serves as storage for creatinine phosphate in the muscle
- A dimeric molecule (2 pairs of monomers: M and B)
Creatine Kinase (CK)
Creatine Kinase (CK)
What are the 4 tissue sources?
- Brain
- Smooth muscle
- Skeletal muscle
- Cardiac muscle
Creatine Kinase (CK)
Reference values for total CK:
1. For males
2. For females
A. 15 to 130 U/L
B. 15 to 160 U/L
- B
- A