I General Description of EnzymesA Chemical composition 1 Enzymes are proteins compounds of high molecular weight containing Carbon (C) hydrogen (H) oxygen (O) sulfur (S) nitrogen (N) 2 Possess primary secondary tertiary (sometimes quaternary) structure 3 Same properties as other proteins

a Charge (isoelectric point electrophoresis) b Solubility c Denaturation (extremes in heat pH chemical) d Non-specific or specific (antigenic) binding 4 Some enzymes have iosenzyme forms

a Different form of an enzyme due to differing arrangements of the polypeptide units in the quaternary structure

b Maintains specificity for the same chemical reaction c Possess unique diagnostic value show specificity to certain organ

systems 1) Isoenzymes of creatine kinase (CK) CK-MM to skeletal muscle CK-MB to cardiac tissue

CK-BB 2) Isoenzymes of lactate dehydrogenase (LD) LD1 and LD2 to cardiac tissue LD4 and LD5 to liver and skeletal muscle

B Biochemical Function

1 Catalyze chemical reactions accelerates specific chemical reactions

toward equilibriumS PSubstrate (S) + Enzyme (ES Product (P) +Enzyme

2 Lowers the energy of activation required for the reaction to proceed

3 Properties of enzymes

a Effective in small concentrationsb Remain unchanged in the reaction not

consumed in the reactionc Effect speed of reaction (not final

concentration of substrate orproducts)d Show greater degree of specificity than

chemical catalystse Unstable and easily destroyed

4 Cofactors substances associated with some enzymes that are needed for

maximal enzyme activity

a Coenzyme organic cofactors (NAD NADH) serve as second

substrates for enzyme reactionsb Activators inorganic cofactors (Ca++ Fe++

Zn++ K+ Mg++)

C Importance of Enzymes in Health and Disease

1 Enzymes are intracellular2 Enzyme concentration differs dependent upon

type of tissuea AST high concentration in liverb Amylase high concentration in pancreas and

salivary glands

Enzymes of Clinical SignificancebullCreatine Kinase

bull1 Specimen collection and handlingbulla Serum is preferred especially if fractionationelectrophoresis also

bullorderedbullb Heparinized plasma may be acceptable

bullMany anticoagulants (fluoride EDTA sodium citrate) inhibitbullenzyme activity and required activators (Ca2+ Mg2+)

bullc No hemolysis intracellular components interfere with chemicalbullreaction in laboratory measurements (NADH ATP HK etc)bulld Avoid lipemia especially for spectrophotometric analysisbulle Relatively unstable analyze within 4 hours of collection elsebullstable at 2-6oC for up to one week stability increases when stored

bullat lower temps (-20oC)

Wide tissue distribution injury tissue necrosis will cause elevation of thisenzyme in plasmaseruma Skeletal muscleb Heart tissuec Brain nerve tissue

Method of Measurment

3 Physiologic reaction CK is a transferase ATP storage and generation incontractile or transport systems predominantly in muscle cells

Creatine + ATP CK1048774 creatine phosphate + ADP4 Method of measurement

Most common method modified Rosalki-Oliver methodcreatine phosphate + ADP CK creatine + ATPATP + glucose HK ADP + glucose-6-phosphate

Glucose-6-phosphate + NADP+ G6PD 6-phosphogluconate +NADPH

(Increased ABS at 340 nm)

Isoenzymes clinical relevance of CK activity often depends more onisoenzyme fractionation than on total levelsa Isoenzymes occur as a dimer1) B polypeptide units2) M polypeptide unitsb Three isoenzyme forms numbered by their anodal (+)electrophoresis mobility with lsquofraction 1rsquo migrating fastest towardsthe anode1) CK1 = CK-BB (brain type) Normal ~02) CK2 = CK-MB (hybrid heart) Normal lt53) CK3 = CK-MM (muscle type) Normal 95-100

c Immunologic measurement of CK-MB1 )Immunoinhibition of subunit M (anti-M)

2 )Measure CK before and after inhibition3 )Disadvantage falsely elevated if CK-BB activity present

d Enzyme immunoassay measurement of CK-MB (mass)

A lsquosandwich techniquersquo that uses 2 antibodies one directedagainst the lsquoMrsquo subunit the other against the lsquoBrsquo subunit which isalso labeled with a reagent enzyme that catalyzes the conversion ofsubstrate to product

6 Reference Ranges affected by gender

bullmuscle mass exercise agebullTotal CK

bullMale 15-160 IULbullFemale 15-130 IULbullCK-MB lt5 of total CK

7 Clinical significancea Acute Myocardial Infarction (AMI)1) Increased total CK increased CK-MB2) Total CK increase 7-12 times normal withrise apparent at 4-6 hours post MIpeaks at 18-24 hours andreturns to normal by 72 hours3) The rise peak and fall of CK-MB parallels that of total CKbut a little sooner4) Other cardiac events (traumainflammation) variable results

b Skeletal muscle disease1) Increased total CK normal CK-MB (increased CK-MM)2) Muscular dystrophy especially Duchenne type may seetotal CK 50x ULN3) Inflammation of muscle especially viral also malignanthyperthermia4) Normal CK activity in neurogenic muscle disordersmultiple sclerosis myasthenia gravis poliomyelitis

c Central nervous system disorders

1) Increased total CK normal CK-MB (increased CK-BB)

2) Trauma or pathology indicated when CK-BB is present

3) Cerebral vascular disease encephalopathy stroke (CK-BB )may be normal

d Other

1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased

CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due

to CKactivity in RBC

B Lactate dehydrogenase (LD) 1 Specimen collection and handling

a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants

(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative

2 Wide cellulartissue distribution

a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells

3 Physiologic reaction

bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +

H+

4 Method of measurement

Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)

Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)

Both the forward and reverse reactions have been used in the clinical lab

with the rate of the reverse reaction being ~3 times faster allowing

measurement of smaller sample volumes and shorter reaction times

However the forward reaction has better linearity at this time

5 Isoenzymes clinical relevance of total LD activity often depends more on

isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)

b Five isoenzyme forms numbered by their anodal electrophoresis

6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(

1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

7 Clinical significance

a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver

disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver

c Acute myocardial infarction (AMI)

1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable

results

d Hemolytic anemia

1) Increased total LD 20-50 times normal increased LD-1 and

LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate

MagaloblasticAnemia3) Hemolytic anemia

C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)

1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable

other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric

analysisd Relatively stable analyze within 8 hours of

collection else store at2-4oC for up to 4 days

2 Wide cellulartissue distribution

a Skeletal muscle

b Cardiac muscletissue

c Liver

d Kidney pancreas RBC

3 Physiologic reaction

AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the

synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy

Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate

4 Method of measurement Karmen method

Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate

Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)

5 Reference Range 0-56 IUL

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

B Biochemical Function

1 Catalyze chemical reactions accelerates specific chemical reactions

toward equilibriumS PSubstrate (S) + Enzyme (ES Product (P) +Enzyme

2 Lowers the energy of activation required for the reaction to proceed

3 Properties of enzymes

a Effective in small concentrationsb Remain unchanged in the reaction not

consumed in the reactionc Effect speed of reaction (not final

concentration of substrate orproducts)d Show greater degree of specificity than

chemical catalystse Unstable and easily destroyed

4 Cofactors substances associated with some enzymes that are needed for

maximal enzyme activity

a Coenzyme organic cofactors (NAD NADH) serve as second

substrates for enzyme reactionsb Activators inorganic cofactors (Ca++ Fe++

Zn++ K+ Mg++)

C Importance of Enzymes in Health and Disease

1 Enzymes are intracellular2 Enzyme concentration differs dependent upon

type of tissuea AST high concentration in liverb Amylase high concentration in pancreas and

salivary glands

Enzymes of Clinical SignificancebullCreatine Kinase

bull1 Specimen collection and handlingbulla Serum is preferred especially if fractionationelectrophoresis also

bullorderedbullb Heparinized plasma may be acceptable

bullMany anticoagulants (fluoride EDTA sodium citrate) inhibitbullenzyme activity and required activators (Ca2+ Mg2+)

bullc No hemolysis intracellular components interfere with chemicalbullreaction in laboratory measurements (NADH ATP HK etc)bulld Avoid lipemia especially for spectrophotometric analysisbulle Relatively unstable analyze within 4 hours of collection elsebullstable at 2-6oC for up to one week stability increases when stored

bullat lower temps (-20oC)

Wide tissue distribution injury tissue necrosis will cause elevation of thisenzyme in plasmaseruma Skeletal muscleb Heart tissuec Brain nerve tissue

Method of Measurment

3 Physiologic reaction CK is a transferase ATP storage and generation incontractile or transport systems predominantly in muscle cells

Creatine + ATP CK1048774 creatine phosphate + ADP4 Method of measurement

Most common method modified Rosalki-Oliver methodcreatine phosphate + ADP CK creatine + ATPATP + glucose HK ADP + glucose-6-phosphate

Glucose-6-phosphate + NADP+ G6PD 6-phosphogluconate +NADPH

(Increased ABS at 340 nm)

Isoenzymes clinical relevance of CK activity often depends more onisoenzyme fractionation than on total levelsa Isoenzymes occur as a dimer1) B polypeptide units2) M polypeptide unitsb Three isoenzyme forms numbered by their anodal (+)electrophoresis mobility with lsquofraction 1rsquo migrating fastest towardsthe anode1) CK1 = CK-BB (brain type) Normal ~02) CK2 = CK-MB (hybrid heart) Normal lt53) CK3 = CK-MM (muscle type) Normal 95-100

c Immunologic measurement of CK-MB1 )Immunoinhibition of subunit M (anti-M)

2 )Measure CK before and after inhibition3 )Disadvantage falsely elevated if CK-BB activity present

d Enzyme immunoassay measurement of CK-MB (mass)

A lsquosandwich techniquersquo that uses 2 antibodies one directedagainst the lsquoMrsquo subunit the other against the lsquoBrsquo subunit which isalso labeled with a reagent enzyme that catalyzes the conversion ofsubstrate to product

6 Reference Ranges affected by gender

bullmuscle mass exercise agebullTotal CK

bullMale 15-160 IULbullFemale 15-130 IULbullCK-MB lt5 of total CK

7 Clinical significancea Acute Myocardial Infarction (AMI)1) Increased total CK increased CK-MB2) Total CK increase 7-12 times normal withrise apparent at 4-6 hours post MIpeaks at 18-24 hours andreturns to normal by 72 hours3) The rise peak and fall of CK-MB parallels that of total CKbut a little sooner4) Other cardiac events (traumainflammation) variable results

b Skeletal muscle disease1) Increased total CK normal CK-MB (increased CK-MM)2) Muscular dystrophy especially Duchenne type may seetotal CK 50x ULN3) Inflammation of muscle especially viral also malignanthyperthermia4) Normal CK activity in neurogenic muscle disordersmultiple sclerosis myasthenia gravis poliomyelitis

c Central nervous system disorders

1) Increased total CK normal CK-MB (increased CK-BB)

2) Trauma or pathology indicated when CK-BB is present

3) Cerebral vascular disease encephalopathy stroke (CK-BB )may be normal

d Other

1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased

CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due

to CKactivity in RBC

B Lactate dehydrogenase (LD) 1 Specimen collection and handling

a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants

(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative

2 Wide cellulartissue distribution

a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells

3 Physiologic reaction

bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +

H+

4 Method of measurement

Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)

Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)

Both the forward and reverse reactions have been used in the clinical lab

with the rate of the reverse reaction being ~3 times faster allowing

measurement of smaller sample volumes and shorter reaction times

However the forward reaction has better linearity at this time

5 Isoenzymes clinical relevance of total LD activity often depends more on

isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)

b Five isoenzyme forms numbered by their anodal electrophoresis

6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(

1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

7 Clinical significance

a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver

disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver

c Acute myocardial infarction (AMI)

1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable

results

d Hemolytic anemia

1) Increased total LD 20-50 times normal increased LD-1 and

LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate

MagaloblasticAnemia3) Hemolytic anemia

C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)

1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable

other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric

analysisd Relatively stable analyze within 8 hours of

collection else store at2-4oC for up to 4 days

2 Wide cellulartissue distribution

a Skeletal muscle

b Cardiac muscletissue

c Liver

d Kidney pancreas RBC

3 Physiologic reaction

AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the

synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy

Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate

4 Method of measurement Karmen method

Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate

Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)

5 Reference Range 0-56 IUL

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

2 Lowers the energy of activation required for the reaction to proceed

3 Properties of enzymes

a Effective in small concentrationsb Remain unchanged in the reaction not

consumed in the reactionc Effect speed of reaction (not final

concentration of substrate orproducts)d Show greater degree of specificity than

chemical catalystse Unstable and easily destroyed

4 Cofactors substances associated with some enzymes that are needed for

maximal enzyme activity

a Coenzyme organic cofactors (NAD NADH) serve as second

substrates for enzyme reactionsb Activators inorganic cofactors (Ca++ Fe++

Zn++ K+ Mg++)

C Importance of Enzymes in Health and Disease

1 Enzymes are intracellular2 Enzyme concentration differs dependent upon

type of tissuea AST high concentration in liverb Amylase high concentration in pancreas and

salivary glands

Enzymes of Clinical SignificancebullCreatine Kinase

bull1 Specimen collection and handlingbulla Serum is preferred especially if fractionationelectrophoresis also

bullorderedbullb Heparinized plasma may be acceptable

bullMany anticoagulants (fluoride EDTA sodium citrate) inhibitbullenzyme activity and required activators (Ca2+ Mg2+)

bullc No hemolysis intracellular components interfere with chemicalbullreaction in laboratory measurements (NADH ATP HK etc)bulld Avoid lipemia especially for spectrophotometric analysisbulle Relatively unstable analyze within 4 hours of collection elsebullstable at 2-6oC for up to one week stability increases when stored

bullat lower temps (-20oC)

Wide tissue distribution injury tissue necrosis will cause elevation of thisenzyme in plasmaseruma Skeletal muscleb Heart tissuec Brain nerve tissue

Method of Measurment

3 Physiologic reaction CK is a transferase ATP storage and generation incontractile or transport systems predominantly in muscle cells

Creatine + ATP CK1048774 creatine phosphate + ADP4 Method of measurement

Most common method modified Rosalki-Oliver methodcreatine phosphate + ADP CK creatine + ATPATP + glucose HK ADP + glucose-6-phosphate

Glucose-6-phosphate + NADP+ G6PD 6-phosphogluconate +NADPH

(Increased ABS at 340 nm)

Isoenzymes clinical relevance of CK activity often depends more onisoenzyme fractionation than on total levelsa Isoenzymes occur as a dimer1) B polypeptide units2) M polypeptide unitsb Three isoenzyme forms numbered by their anodal (+)electrophoresis mobility with lsquofraction 1rsquo migrating fastest towardsthe anode1) CK1 = CK-BB (brain type) Normal ~02) CK2 = CK-MB (hybrid heart) Normal lt53) CK3 = CK-MM (muscle type) Normal 95-100

c Immunologic measurement of CK-MB1 )Immunoinhibition of subunit M (anti-M)

2 )Measure CK before and after inhibition3 )Disadvantage falsely elevated if CK-BB activity present

d Enzyme immunoassay measurement of CK-MB (mass)

A lsquosandwich techniquersquo that uses 2 antibodies one directedagainst the lsquoMrsquo subunit the other against the lsquoBrsquo subunit which isalso labeled with a reagent enzyme that catalyzes the conversion ofsubstrate to product

6 Reference Ranges affected by gender

bullmuscle mass exercise agebullTotal CK

bullMale 15-160 IULbullFemale 15-130 IULbullCK-MB lt5 of total CK

7 Clinical significancea Acute Myocardial Infarction (AMI)1) Increased total CK increased CK-MB2) Total CK increase 7-12 times normal withrise apparent at 4-6 hours post MIpeaks at 18-24 hours andreturns to normal by 72 hours3) The rise peak and fall of CK-MB parallels that of total CKbut a little sooner4) Other cardiac events (traumainflammation) variable results

b Skeletal muscle disease1) Increased total CK normal CK-MB (increased CK-MM)2) Muscular dystrophy especially Duchenne type may seetotal CK 50x ULN3) Inflammation of muscle especially viral also malignanthyperthermia4) Normal CK activity in neurogenic muscle disordersmultiple sclerosis myasthenia gravis poliomyelitis

c Central nervous system disorders

1) Increased total CK normal CK-MB (increased CK-BB)

2) Trauma or pathology indicated when CK-BB is present

3) Cerebral vascular disease encephalopathy stroke (CK-BB )may be normal

d Other

1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased

CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due

to CKactivity in RBC

B Lactate dehydrogenase (LD) 1 Specimen collection and handling

a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants

(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative

2 Wide cellulartissue distribution

a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells

3 Physiologic reaction

bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +

H+

4 Method of measurement

Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)

Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)

Both the forward and reverse reactions have been used in the clinical lab

with the rate of the reverse reaction being ~3 times faster allowing

measurement of smaller sample volumes and shorter reaction times

However the forward reaction has better linearity at this time

5 Isoenzymes clinical relevance of total LD activity often depends more on

isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)

b Five isoenzyme forms numbered by their anodal electrophoresis

6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(

1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

7 Clinical significance

a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver

disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver

c Acute myocardial infarction (AMI)

1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable

results

d Hemolytic anemia

1) Increased total LD 20-50 times normal increased LD-1 and

LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate

MagaloblasticAnemia3) Hemolytic anemia

C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)

1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable

other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric

analysisd Relatively stable analyze within 8 hours of

collection else store at2-4oC for up to 4 days

2 Wide cellulartissue distribution

a Skeletal muscle

b Cardiac muscletissue

c Liver

d Kidney pancreas RBC

3 Physiologic reaction

AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the

synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy

Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate

4 Method of measurement Karmen method

Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate

Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)

5 Reference Range 0-56 IUL

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

3 Properties of enzymes

a Effective in small concentrationsb Remain unchanged in the reaction not

consumed in the reactionc Effect speed of reaction (not final

concentration of substrate orproducts)d Show greater degree of specificity than

chemical catalystse Unstable and easily destroyed

4 Cofactors substances associated with some enzymes that are needed for

maximal enzyme activity

a Coenzyme organic cofactors (NAD NADH) serve as second

substrates for enzyme reactionsb Activators inorganic cofactors (Ca++ Fe++

Zn++ K+ Mg++)

C Importance of Enzymes in Health and Disease

1 Enzymes are intracellular2 Enzyme concentration differs dependent upon

type of tissuea AST high concentration in liverb Amylase high concentration in pancreas and

salivary glands

Enzymes of Clinical SignificancebullCreatine Kinase

bull1 Specimen collection and handlingbulla Serum is preferred especially if fractionationelectrophoresis also

bullorderedbullb Heparinized plasma may be acceptable

bullMany anticoagulants (fluoride EDTA sodium citrate) inhibitbullenzyme activity and required activators (Ca2+ Mg2+)

bullc No hemolysis intracellular components interfere with chemicalbullreaction in laboratory measurements (NADH ATP HK etc)bulld Avoid lipemia especially for spectrophotometric analysisbulle Relatively unstable analyze within 4 hours of collection elsebullstable at 2-6oC for up to one week stability increases when stored

bullat lower temps (-20oC)

Wide tissue distribution injury tissue necrosis will cause elevation of thisenzyme in plasmaseruma Skeletal muscleb Heart tissuec Brain nerve tissue

Method of Measurment

3 Physiologic reaction CK is a transferase ATP storage and generation incontractile or transport systems predominantly in muscle cells

Creatine + ATP CK1048774 creatine phosphate + ADP4 Method of measurement

Most common method modified Rosalki-Oliver methodcreatine phosphate + ADP CK creatine + ATPATP + glucose HK ADP + glucose-6-phosphate

Glucose-6-phosphate + NADP+ G6PD 6-phosphogluconate +NADPH

(Increased ABS at 340 nm)

Isoenzymes clinical relevance of CK activity often depends more onisoenzyme fractionation than on total levelsa Isoenzymes occur as a dimer1) B polypeptide units2) M polypeptide unitsb Three isoenzyme forms numbered by their anodal (+)electrophoresis mobility with lsquofraction 1rsquo migrating fastest towardsthe anode1) CK1 = CK-BB (brain type) Normal ~02) CK2 = CK-MB (hybrid heart) Normal lt53) CK3 = CK-MM (muscle type) Normal 95-100

c Immunologic measurement of CK-MB1 )Immunoinhibition of subunit M (anti-M)

2 )Measure CK before and after inhibition3 )Disadvantage falsely elevated if CK-BB activity present

d Enzyme immunoassay measurement of CK-MB (mass)

A lsquosandwich techniquersquo that uses 2 antibodies one directedagainst the lsquoMrsquo subunit the other against the lsquoBrsquo subunit which isalso labeled with a reagent enzyme that catalyzes the conversion ofsubstrate to product

6 Reference Ranges affected by gender

bullmuscle mass exercise agebullTotal CK

bullMale 15-160 IULbullFemale 15-130 IULbullCK-MB lt5 of total CK

7 Clinical significancea Acute Myocardial Infarction (AMI)1) Increased total CK increased CK-MB2) Total CK increase 7-12 times normal withrise apparent at 4-6 hours post MIpeaks at 18-24 hours andreturns to normal by 72 hours3) The rise peak and fall of CK-MB parallels that of total CKbut a little sooner4) Other cardiac events (traumainflammation) variable results

b Skeletal muscle disease1) Increased total CK normal CK-MB (increased CK-MM)2) Muscular dystrophy especially Duchenne type may seetotal CK 50x ULN3) Inflammation of muscle especially viral also malignanthyperthermia4) Normal CK activity in neurogenic muscle disordersmultiple sclerosis myasthenia gravis poliomyelitis

c Central nervous system disorders

1) Increased total CK normal CK-MB (increased CK-BB)

2) Trauma or pathology indicated when CK-BB is present

3) Cerebral vascular disease encephalopathy stroke (CK-BB )may be normal

d Other

1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased

CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due

to CKactivity in RBC

B Lactate dehydrogenase (LD) 1 Specimen collection and handling

a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants

(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative

2 Wide cellulartissue distribution

a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells

3 Physiologic reaction

bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +

H+

4 Method of measurement

Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)

Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)

Both the forward and reverse reactions have been used in the clinical lab

with the rate of the reverse reaction being ~3 times faster allowing

measurement of smaller sample volumes and shorter reaction times

However the forward reaction has better linearity at this time

5 Isoenzymes clinical relevance of total LD activity often depends more on

isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)

b Five isoenzyme forms numbered by their anodal electrophoresis

6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(

1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

7 Clinical significance

a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver

disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver

c Acute myocardial infarction (AMI)

1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable

results

d Hemolytic anemia

1) Increased total LD 20-50 times normal increased LD-1 and

LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate

MagaloblasticAnemia3) Hemolytic anemia

C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)

1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable

other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric

analysisd Relatively stable analyze within 8 hours of

collection else store at2-4oC for up to 4 days

2 Wide cellulartissue distribution

a Skeletal muscle

b Cardiac muscletissue

c Liver

d Kidney pancreas RBC

3 Physiologic reaction

AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the

synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy

Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate

4 Method of measurement Karmen method

Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate

Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)

5 Reference Range 0-56 IUL

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

4 Cofactors substances associated with some enzymes that are needed for

maximal enzyme activity

a Coenzyme organic cofactors (NAD NADH) serve as second

substrates for enzyme reactionsb Activators inorganic cofactors (Ca++ Fe++

Zn++ K+ Mg++)

C Importance of Enzymes in Health and Disease

1 Enzymes are intracellular2 Enzyme concentration differs dependent upon

type of tissuea AST high concentration in liverb Amylase high concentration in pancreas and

salivary glands

Enzymes of Clinical SignificancebullCreatine Kinase

bull1 Specimen collection and handlingbulla Serum is preferred especially if fractionationelectrophoresis also

bullorderedbullb Heparinized plasma may be acceptable

bullMany anticoagulants (fluoride EDTA sodium citrate) inhibitbullenzyme activity and required activators (Ca2+ Mg2+)

bullc No hemolysis intracellular components interfere with chemicalbullreaction in laboratory measurements (NADH ATP HK etc)bulld Avoid lipemia especially for spectrophotometric analysisbulle Relatively unstable analyze within 4 hours of collection elsebullstable at 2-6oC for up to one week stability increases when stored

bullat lower temps (-20oC)

Wide tissue distribution injury tissue necrosis will cause elevation of thisenzyme in plasmaseruma Skeletal muscleb Heart tissuec Brain nerve tissue

Method of Measurment

3 Physiologic reaction CK is a transferase ATP storage and generation incontractile or transport systems predominantly in muscle cells

Creatine + ATP CK1048774 creatine phosphate + ADP4 Method of measurement

Most common method modified Rosalki-Oliver methodcreatine phosphate + ADP CK creatine + ATPATP + glucose HK ADP + glucose-6-phosphate

Glucose-6-phosphate + NADP+ G6PD 6-phosphogluconate +NADPH

(Increased ABS at 340 nm)

Isoenzymes clinical relevance of CK activity often depends more onisoenzyme fractionation than on total levelsa Isoenzymes occur as a dimer1) B polypeptide units2) M polypeptide unitsb Three isoenzyme forms numbered by their anodal (+)electrophoresis mobility with lsquofraction 1rsquo migrating fastest towardsthe anode1) CK1 = CK-BB (brain type) Normal ~02) CK2 = CK-MB (hybrid heart) Normal lt53) CK3 = CK-MM (muscle type) Normal 95-100

c Immunologic measurement of CK-MB1 )Immunoinhibition of subunit M (anti-M)

2 )Measure CK before and after inhibition3 )Disadvantage falsely elevated if CK-BB activity present

d Enzyme immunoassay measurement of CK-MB (mass)

A lsquosandwich techniquersquo that uses 2 antibodies one directedagainst the lsquoMrsquo subunit the other against the lsquoBrsquo subunit which isalso labeled with a reagent enzyme that catalyzes the conversion ofsubstrate to product

6 Reference Ranges affected by gender

bullmuscle mass exercise agebullTotal CK

bullMale 15-160 IULbullFemale 15-130 IULbullCK-MB lt5 of total CK

7 Clinical significancea Acute Myocardial Infarction (AMI)1) Increased total CK increased CK-MB2) Total CK increase 7-12 times normal withrise apparent at 4-6 hours post MIpeaks at 18-24 hours andreturns to normal by 72 hours3) The rise peak and fall of CK-MB parallels that of total CKbut a little sooner4) Other cardiac events (traumainflammation) variable results

b Skeletal muscle disease1) Increased total CK normal CK-MB (increased CK-MM)2) Muscular dystrophy especially Duchenne type may seetotal CK 50x ULN3) Inflammation of muscle especially viral also malignanthyperthermia4) Normal CK activity in neurogenic muscle disordersmultiple sclerosis myasthenia gravis poliomyelitis

c Central nervous system disorders

1) Increased total CK normal CK-MB (increased CK-BB)

2) Trauma or pathology indicated when CK-BB is present

3) Cerebral vascular disease encephalopathy stroke (CK-BB )may be normal

d Other

1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased

CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due

to CKactivity in RBC

B Lactate dehydrogenase (LD) 1 Specimen collection and handling

a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants

(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative

2 Wide cellulartissue distribution

a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells

3 Physiologic reaction

bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +

H+

4 Method of measurement

Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)

Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)

Both the forward and reverse reactions have been used in the clinical lab

with the rate of the reverse reaction being ~3 times faster allowing

measurement of smaller sample volumes and shorter reaction times

However the forward reaction has better linearity at this time

5 Isoenzymes clinical relevance of total LD activity often depends more on

isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)

b Five isoenzyme forms numbered by their anodal electrophoresis

6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(

1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

7 Clinical significance

a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver

disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver

c Acute myocardial infarction (AMI)

1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable

results

d Hemolytic anemia

1) Increased total LD 20-50 times normal increased LD-1 and

LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate

MagaloblasticAnemia3) Hemolytic anemia

C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)

1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable

other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric

analysisd Relatively stable analyze within 8 hours of

collection else store at2-4oC for up to 4 days

2 Wide cellulartissue distribution

a Skeletal muscle

b Cardiac muscletissue

c Liver

d Kidney pancreas RBC

3 Physiologic reaction

AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the

synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy

Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate

4 Method of measurement Karmen method

Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate

Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)

5 Reference Range 0-56 IUL

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

C Importance of Enzymes in Health and Disease

1 Enzymes are intracellular2 Enzyme concentration differs dependent upon

type of tissuea AST high concentration in liverb Amylase high concentration in pancreas and

salivary glands

Enzymes of Clinical SignificancebullCreatine Kinase

bull1 Specimen collection and handlingbulla Serum is preferred especially if fractionationelectrophoresis also

bullorderedbullb Heparinized plasma may be acceptable

bullMany anticoagulants (fluoride EDTA sodium citrate) inhibitbullenzyme activity and required activators (Ca2+ Mg2+)

bullc No hemolysis intracellular components interfere with chemicalbullreaction in laboratory measurements (NADH ATP HK etc)bulld Avoid lipemia especially for spectrophotometric analysisbulle Relatively unstable analyze within 4 hours of collection elsebullstable at 2-6oC for up to one week stability increases when stored

bullat lower temps (-20oC)

Wide tissue distribution injury tissue necrosis will cause elevation of thisenzyme in plasmaseruma Skeletal muscleb Heart tissuec Brain nerve tissue

Method of Measurment

3 Physiologic reaction CK is a transferase ATP storage and generation incontractile or transport systems predominantly in muscle cells

Creatine + ATP CK1048774 creatine phosphate + ADP4 Method of measurement

Most common method modified Rosalki-Oliver methodcreatine phosphate + ADP CK creatine + ATPATP + glucose HK ADP + glucose-6-phosphate

Glucose-6-phosphate + NADP+ G6PD 6-phosphogluconate +NADPH

(Increased ABS at 340 nm)

Isoenzymes clinical relevance of CK activity often depends more onisoenzyme fractionation than on total levelsa Isoenzymes occur as a dimer1) B polypeptide units2) M polypeptide unitsb Three isoenzyme forms numbered by their anodal (+)electrophoresis mobility with lsquofraction 1rsquo migrating fastest towardsthe anode1) CK1 = CK-BB (brain type) Normal ~02) CK2 = CK-MB (hybrid heart) Normal lt53) CK3 = CK-MM (muscle type) Normal 95-100

c Immunologic measurement of CK-MB1 )Immunoinhibition of subunit M (anti-M)

2 )Measure CK before and after inhibition3 )Disadvantage falsely elevated if CK-BB activity present

d Enzyme immunoassay measurement of CK-MB (mass)

A lsquosandwich techniquersquo that uses 2 antibodies one directedagainst the lsquoMrsquo subunit the other against the lsquoBrsquo subunit which isalso labeled with a reagent enzyme that catalyzes the conversion ofsubstrate to product

6 Reference Ranges affected by gender

bullmuscle mass exercise agebullTotal CK

bullMale 15-160 IULbullFemale 15-130 IULbullCK-MB lt5 of total CK

7 Clinical significancea Acute Myocardial Infarction (AMI)1) Increased total CK increased CK-MB2) Total CK increase 7-12 times normal withrise apparent at 4-6 hours post MIpeaks at 18-24 hours andreturns to normal by 72 hours3) The rise peak and fall of CK-MB parallels that of total CKbut a little sooner4) Other cardiac events (traumainflammation) variable results

b Skeletal muscle disease1) Increased total CK normal CK-MB (increased CK-MM)2) Muscular dystrophy especially Duchenne type may seetotal CK 50x ULN3) Inflammation of muscle especially viral also malignanthyperthermia4) Normal CK activity in neurogenic muscle disordersmultiple sclerosis myasthenia gravis poliomyelitis

c Central nervous system disorders

1) Increased total CK normal CK-MB (increased CK-BB)

2) Trauma or pathology indicated when CK-BB is present

3) Cerebral vascular disease encephalopathy stroke (CK-BB )may be normal

d Other

1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased

CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due

to CKactivity in RBC

B Lactate dehydrogenase (LD) 1 Specimen collection and handling

a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants

(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative

2 Wide cellulartissue distribution

a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells

3 Physiologic reaction

bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +

H+

4 Method of measurement

Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)

Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)

Both the forward and reverse reactions have been used in the clinical lab

with the rate of the reverse reaction being ~3 times faster allowing

measurement of smaller sample volumes and shorter reaction times

However the forward reaction has better linearity at this time

5 Isoenzymes clinical relevance of total LD activity often depends more on

isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)

b Five isoenzyme forms numbered by their anodal electrophoresis

6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(

1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

7 Clinical significance

a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver

disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver

c Acute myocardial infarction (AMI)

1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable

results

d Hemolytic anemia

1) Increased total LD 20-50 times normal increased LD-1 and

LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate

MagaloblasticAnemia3) Hemolytic anemia

C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)

1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable

other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric

analysisd Relatively stable analyze within 8 hours of

collection else store at2-4oC for up to 4 days

2 Wide cellulartissue distribution

a Skeletal muscle

b Cardiac muscletissue

c Liver

d Kidney pancreas RBC

3 Physiologic reaction

AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the

synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy

Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate

4 Method of measurement Karmen method

Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate

Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)

5 Reference Range 0-56 IUL

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

Enzymes of Clinical SignificancebullCreatine Kinase

bull1 Specimen collection and handlingbulla Serum is preferred especially if fractionationelectrophoresis also

bullorderedbullb Heparinized plasma may be acceptable

bullMany anticoagulants (fluoride EDTA sodium citrate) inhibitbullenzyme activity and required activators (Ca2+ Mg2+)

bullc No hemolysis intracellular components interfere with chemicalbullreaction in laboratory measurements (NADH ATP HK etc)bulld Avoid lipemia especially for spectrophotometric analysisbulle Relatively unstable analyze within 4 hours of collection elsebullstable at 2-6oC for up to one week stability increases when stored

bullat lower temps (-20oC)

Wide tissue distribution injury tissue necrosis will cause elevation of thisenzyme in plasmaseruma Skeletal muscleb Heart tissuec Brain nerve tissue

Method of Measurment

3 Physiologic reaction CK is a transferase ATP storage and generation incontractile or transport systems predominantly in muscle cells

Creatine + ATP CK1048774 creatine phosphate + ADP4 Method of measurement

Most common method modified Rosalki-Oliver methodcreatine phosphate + ADP CK creatine + ATPATP + glucose HK ADP + glucose-6-phosphate

Glucose-6-phosphate + NADP+ G6PD 6-phosphogluconate +NADPH

(Increased ABS at 340 nm)

Isoenzymes clinical relevance of CK activity often depends more onisoenzyme fractionation than on total levelsa Isoenzymes occur as a dimer1) B polypeptide units2) M polypeptide unitsb Three isoenzyme forms numbered by their anodal (+)electrophoresis mobility with lsquofraction 1rsquo migrating fastest towardsthe anode1) CK1 = CK-BB (brain type) Normal ~02) CK2 = CK-MB (hybrid heart) Normal lt53) CK3 = CK-MM (muscle type) Normal 95-100

c Immunologic measurement of CK-MB1 )Immunoinhibition of subunit M (anti-M)

2 )Measure CK before and after inhibition3 )Disadvantage falsely elevated if CK-BB activity present

d Enzyme immunoassay measurement of CK-MB (mass)

A lsquosandwich techniquersquo that uses 2 antibodies one directedagainst the lsquoMrsquo subunit the other against the lsquoBrsquo subunit which isalso labeled with a reagent enzyme that catalyzes the conversion ofsubstrate to product

6 Reference Ranges affected by gender

bullmuscle mass exercise agebullTotal CK

bullMale 15-160 IULbullFemale 15-130 IULbullCK-MB lt5 of total CK

7 Clinical significancea Acute Myocardial Infarction (AMI)1) Increased total CK increased CK-MB2) Total CK increase 7-12 times normal withrise apparent at 4-6 hours post MIpeaks at 18-24 hours andreturns to normal by 72 hours3) The rise peak and fall of CK-MB parallels that of total CKbut a little sooner4) Other cardiac events (traumainflammation) variable results

b Skeletal muscle disease1) Increased total CK normal CK-MB (increased CK-MM)2) Muscular dystrophy especially Duchenne type may seetotal CK 50x ULN3) Inflammation of muscle especially viral also malignanthyperthermia4) Normal CK activity in neurogenic muscle disordersmultiple sclerosis myasthenia gravis poliomyelitis

c Central nervous system disorders

1) Increased total CK normal CK-MB (increased CK-BB)

2) Trauma or pathology indicated when CK-BB is present

3) Cerebral vascular disease encephalopathy stroke (CK-BB )may be normal

d Other

1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased

CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due

to CKactivity in RBC

B Lactate dehydrogenase (LD) 1 Specimen collection and handling

a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants

(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative

2 Wide cellulartissue distribution

a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells

3 Physiologic reaction

bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +

H+

4 Method of measurement

Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)

Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)

Both the forward and reverse reactions have been used in the clinical lab

with the rate of the reverse reaction being ~3 times faster allowing

measurement of smaller sample volumes and shorter reaction times

However the forward reaction has better linearity at this time

5 Isoenzymes clinical relevance of total LD activity often depends more on

isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)

b Five isoenzyme forms numbered by their anodal electrophoresis

6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(

1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

7 Clinical significance

a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver

disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver

c Acute myocardial infarction (AMI)

1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable

results

d Hemolytic anemia

1) Increased total LD 20-50 times normal increased LD-1 and

LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate

MagaloblasticAnemia3) Hemolytic anemia

C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)

1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable

other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric

analysisd Relatively stable analyze within 8 hours of

collection else store at2-4oC for up to 4 days

2 Wide cellulartissue distribution

a Skeletal muscle

b Cardiac muscletissue

c Liver

d Kidney pancreas RBC

3 Physiologic reaction

AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the

synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy

Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate

4 Method of measurement Karmen method

Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate

Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)

5 Reference Range 0-56 IUL

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

Wide tissue distribution injury tissue necrosis will cause elevation of thisenzyme in plasmaseruma Skeletal muscleb Heart tissuec Brain nerve tissue

Method of Measurment

3 Physiologic reaction CK is a transferase ATP storage and generation incontractile or transport systems predominantly in muscle cells

Creatine + ATP CK1048774 creatine phosphate + ADP4 Method of measurement

Most common method modified Rosalki-Oliver methodcreatine phosphate + ADP CK creatine + ATPATP + glucose HK ADP + glucose-6-phosphate

Glucose-6-phosphate + NADP+ G6PD 6-phosphogluconate +NADPH

(Increased ABS at 340 nm)

Isoenzymes clinical relevance of CK activity often depends more onisoenzyme fractionation than on total levelsa Isoenzymes occur as a dimer1) B polypeptide units2) M polypeptide unitsb Three isoenzyme forms numbered by their anodal (+)electrophoresis mobility with lsquofraction 1rsquo migrating fastest towardsthe anode1) CK1 = CK-BB (brain type) Normal ~02) CK2 = CK-MB (hybrid heart) Normal lt53) CK3 = CK-MM (muscle type) Normal 95-100

c Immunologic measurement of CK-MB1 )Immunoinhibition of subunit M (anti-M)

2 )Measure CK before and after inhibition3 )Disadvantage falsely elevated if CK-BB activity present

d Enzyme immunoassay measurement of CK-MB (mass)

A lsquosandwich techniquersquo that uses 2 antibodies one directedagainst the lsquoMrsquo subunit the other against the lsquoBrsquo subunit which isalso labeled with a reagent enzyme that catalyzes the conversion ofsubstrate to product

6 Reference Ranges affected by gender

bullmuscle mass exercise agebullTotal CK

bullMale 15-160 IULbullFemale 15-130 IULbullCK-MB lt5 of total CK

7 Clinical significancea Acute Myocardial Infarction (AMI)1) Increased total CK increased CK-MB2) Total CK increase 7-12 times normal withrise apparent at 4-6 hours post MIpeaks at 18-24 hours andreturns to normal by 72 hours3) The rise peak and fall of CK-MB parallels that of total CKbut a little sooner4) Other cardiac events (traumainflammation) variable results

b Skeletal muscle disease1) Increased total CK normal CK-MB (increased CK-MM)2) Muscular dystrophy especially Duchenne type may seetotal CK 50x ULN3) Inflammation of muscle especially viral also malignanthyperthermia4) Normal CK activity in neurogenic muscle disordersmultiple sclerosis myasthenia gravis poliomyelitis

c Central nervous system disorders

1) Increased total CK normal CK-MB (increased CK-BB)

2) Trauma or pathology indicated when CK-BB is present

3) Cerebral vascular disease encephalopathy stroke (CK-BB )may be normal

d Other

1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased

CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due

to CKactivity in RBC

B Lactate dehydrogenase (LD) 1 Specimen collection and handling

a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants

(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative

2 Wide cellulartissue distribution

a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells

3 Physiologic reaction

bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +

H+

4 Method of measurement

Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)

Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)

Both the forward and reverse reactions have been used in the clinical lab

with the rate of the reverse reaction being ~3 times faster allowing

measurement of smaller sample volumes and shorter reaction times

However the forward reaction has better linearity at this time

5 Isoenzymes clinical relevance of total LD activity often depends more on

isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)

b Five isoenzyme forms numbered by their anodal electrophoresis

6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(

1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

7 Clinical significance

a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver

disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver

c Acute myocardial infarction (AMI)

1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable

results

d Hemolytic anemia

1) Increased total LD 20-50 times normal increased LD-1 and

LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate

MagaloblasticAnemia3) Hemolytic anemia

C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)

1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable

other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric

analysisd Relatively stable analyze within 8 hours of

collection else store at2-4oC for up to 4 days

2 Wide cellulartissue distribution

a Skeletal muscle

b Cardiac muscletissue

c Liver

d Kidney pancreas RBC

3 Physiologic reaction

AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the

synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy

Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate

4 Method of measurement Karmen method

Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate

Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)

5 Reference Range 0-56 IUL

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

Method of Measurment

3 Physiologic reaction CK is a transferase ATP storage and generation incontractile or transport systems predominantly in muscle cells

Creatine + ATP CK1048774 creatine phosphate + ADP4 Method of measurement

Most common method modified Rosalki-Oliver methodcreatine phosphate + ADP CK creatine + ATPATP + glucose HK ADP + glucose-6-phosphate

Glucose-6-phosphate + NADP+ G6PD 6-phosphogluconate +NADPH

(Increased ABS at 340 nm)

Isoenzymes clinical relevance of CK activity often depends more onisoenzyme fractionation than on total levelsa Isoenzymes occur as a dimer1) B polypeptide units2) M polypeptide unitsb Three isoenzyme forms numbered by their anodal (+)electrophoresis mobility with lsquofraction 1rsquo migrating fastest towardsthe anode1) CK1 = CK-BB (brain type) Normal ~02) CK2 = CK-MB (hybrid heart) Normal lt53) CK3 = CK-MM (muscle type) Normal 95-100

c Immunologic measurement of CK-MB1 )Immunoinhibition of subunit M (anti-M)

2 )Measure CK before and after inhibition3 )Disadvantage falsely elevated if CK-BB activity present

d Enzyme immunoassay measurement of CK-MB (mass)

A lsquosandwich techniquersquo that uses 2 antibodies one directedagainst the lsquoMrsquo subunit the other against the lsquoBrsquo subunit which isalso labeled with a reagent enzyme that catalyzes the conversion ofsubstrate to product

6 Reference Ranges affected by gender

bullmuscle mass exercise agebullTotal CK

bullMale 15-160 IULbullFemale 15-130 IULbullCK-MB lt5 of total CK

7 Clinical significancea Acute Myocardial Infarction (AMI)1) Increased total CK increased CK-MB2) Total CK increase 7-12 times normal withrise apparent at 4-6 hours post MIpeaks at 18-24 hours andreturns to normal by 72 hours3) The rise peak and fall of CK-MB parallels that of total CKbut a little sooner4) Other cardiac events (traumainflammation) variable results

b Skeletal muscle disease1) Increased total CK normal CK-MB (increased CK-MM)2) Muscular dystrophy especially Duchenne type may seetotal CK 50x ULN3) Inflammation of muscle especially viral also malignanthyperthermia4) Normal CK activity in neurogenic muscle disordersmultiple sclerosis myasthenia gravis poliomyelitis

c Central nervous system disorders

1) Increased total CK normal CK-MB (increased CK-BB)

2) Trauma or pathology indicated when CK-BB is present

3) Cerebral vascular disease encephalopathy stroke (CK-BB )may be normal

d Other

1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased

CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due

to CKactivity in RBC

B Lactate dehydrogenase (LD) 1 Specimen collection and handling

a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants

(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative

2 Wide cellulartissue distribution

a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells

3 Physiologic reaction

bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +

H+

4 Method of measurement

Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)

Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)

Both the forward and reverse reactions have been used in the clinical lab

with the rate of the reverse reaction being ~3 times faster allowing

measurement of smaller sample volumes and shorter reaction times

However the forward reaction has better linearity at this time

5 Isoenzymes clinical relevance of total LD activity often depends more on

isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)

b Five isoenzyme forms numbered by their anodal electrophoresis

6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(

1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

7 Clinical significance

a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver

disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver

c Acute myocardial infarction (AMI)

1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable

results

d Hemolytic anemia

1) Increased total LD 20-50 times normal increased LD-1 and

LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate

MagaloblasticAnemia3) Hemolytic anemia

C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)

1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable

other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric

analysisd Relatively stable analyze within 8 hours of

collection else store at2-4oC for up to 4 days

2 Wide cellulartissue distribution

a Skeletal muscle

b Cardiac muscletissue

c Liver

d Kidney pancreas RBC

3 Physiologic reaction

AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the

synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy

Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate

4 Method of measurement Karmen method

Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate

Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)

5 Reference Range 0-56 IUL

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

Isoenzymes clinical relevance of CK activity often depends more onisoenzyme fractionation than on total levelsa Isoenzymes occur as a dimer1) B polypeptide units2) M polypeptide unitsb Three isoenzyme forms numbered by their anodal (+)electrophoresis mobility with lsquofraction 1rsquo migrating fastest towardsthe anode1) CK1 = CK-BB (brain type) Normal ~02) CK2 = CK-MB (hybrid heart) Normal lt53) CK3 = CK-MM (muscle type) Normal 95-100

c Immunologic measurement of CK-MB1 )Immunoinhibition of subunit M (anti-M)

2 )Measure CK before and after inhibition3 )Disadvantage falsely elevated if CK-BB activity present

d Enzyme immunoassay measurement of CK-MB (mass)

A lsquosandwich techniquersquo that uses 2 antibodies one directedagainst the lsquoMrsquo subunit the other against the lsquoBrsquo subunit which isalso labeled with a reagent enzyme that catalyzes the conversion ofsubstrate to product

6 Reference Ranges affected by gender

bullmuscle mass exercise agebullTotal CK

bullMale 15-160 IULbullFemale 15-130 IULbullCK-MB lt5 of total CK

7 Clinical significancea Acute Myocardial Infarction (AMI)1) Increased total CK increased CK-MB2) Total CK increase 7-12 times normal withrise apparent at 4-6 hours post MIpeaks at 18-24 hours andreturns to normal by 72 hours3) The rise peak and fall of CK-MB parallels that of total CKbut a little sooner4) Other cardiac events (traumainflammation) variable results

b Skeletal muscle disease1) Increased total CK normal CK-MB (increased CK-MM)2) Muscular dystrophy especially Duchenne type may seetotal CK 50x ULN3) Inflammation of muscle especially viral also malignanthyperthermia4) Normal CK activity in neurogenic muscle disordersmultiple sclerosis myasthenia gravis poliomyelitis

c Central nervous system disorders

1) Increased total CK normal CK-MB (increased CK-BB)

2) Trauma or pathology indicated when CK-BB is present

3) Cerebral vascular disease encephalopathy stroke (CK-BB )may be normal

d Other

1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased

CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due

to CKactivity in RBC

B Lactate dehydrogenase (LD) 1 Specimen collection and handling

a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants

(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative

2 Wide cellulartissue distribution

a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells

3 Physiologic reaction

bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +

H+

4 Method of measurement

Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)

Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)

Both the forward and reverse reactions have been used in the clinical lab

with the rate of the reverse reaction being ~3 times faster allowing

measurement of smaller sample volumes and shorter reaction times

However the forward reaction has better linearity at this time

5 Isoenzymes clinical relevance of total LD activity often depends more on

isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)

b Five isoenzyme forms numbered by their anodal electrophoresis

6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(

1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

7 Clinical significance

a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver

disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver

c Acute myocardial infarction (AMI)

1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable

results

d Hemolytic anemia

1) Increased total LD 20-50 times normal increased LD-1 and

LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate

MagaloblasticAnemia3) Hemolytic anemia

C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)

1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable

other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric

analysisd Relatively stable analyze within 8 hours of

collection else store at2-4oC for up to 4 days

2 Wide cellulartissue distribution

a Skeletal muscle

b Cardiac muscletissue

c Liver

d Kidney pancreas RBC

3 Physiologic reaction

AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the

synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy

Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate

4 Method of measurement Karmen method

Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate

Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)

5 Reference Range 0-56 IUL

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

c Immunologic measurement of CK-MB1 )Immunoinhibition of subunit M (anti-M)

2 )Measure CK before and after inhibition3 )Disadvantage falsely elevated if CK-BB activity present

d Enzyme immunoassay measurement of CK-MB (mass)

A lsquosandwich techniquersquo that uses 2 antibodies one directedagainst the lsquoMrsquo subunit the other against the lsquoBrsquo subunit which isalso labeled with a reagent enzyme that catalyzes the conversion ofsubstrate to product

6 Reference Ranges affected by gender

bullmuscle mass exercise agebullTotal CK

bullMale 15-160 IULbullFemale 15-130 IULbullCK-MB lt5 of total CK

7 Clinical significancea Acute Myocardial Infarction (AMI)1) Increased total CK increased CK-MB2) Total CK increase 7-12 times normal withrise apparent at 4-6 hours post MIpeaks at 18-24 hours andreturns to normal by 72 hours3) The rise peak and fall of CK-MB parallels that of total CKbut a little sooner4) Other cardiac events (traumainflammation) variable results

b Skeletal muscle disease1) Increased total CK normal CK-MB (increased CK-MM)2) Muscular dystrophy especially Duchenne type may seetotal CK 50x ULN3) Inflammation of muscle especially viral also malignanthyperthermia4) Normal CK activity in neurogenic muscle disordersmultiple sclerosis myasthenia gravis poliomyelitis

c Central nervous system disorders

1) Increased total CK normal CK-MB (increased CK-BB)

2) Trauma or pathology indicated when CK-BB is present

3) Cerebral vascular disease encephalopathy stroke (CK-BB )may be normal

d Other

1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased

CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due

to CKactivity in RBC

B Lactate dehydrogenase (LD) 1 Specimen collection and handling

a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants

(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative

2 Wide cellulartissue distribution

a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells

3 Physiologic reaction

bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +

H+

4 Method of measurement

Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)

Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)

Both the forward and reverse reactions have been used in the clinical lab

with the rate of the reverse reaction being ~3 times faster allowing

measurement of smaller sample volumes and shorter reaction times

However the forward reaction has better linearity at this time

5 Isoenzymes clinical relevance of total LD activity often depends more on

isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)

b Five isoenzyme forms numbered by their anodal electrophoresis

6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(

1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

7 Clinical significance

a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver

disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver

c Acute myocardial infarction (AMI)

1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable

results

d Hemolytic anemia

1) Increased total LD 20-50 times normal increased LD-1 and

LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate

MagaloblasticAnemia3) Hemolytic anemia

C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)

1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable

other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric

analysisd Relatively stable analyze within 8 hours of

collection else store at2-4oC for up to 4 days

2 Wide cellulartissue distribution

a Skeletal muscle

b Cardiac muscletissue

c Liver

d Kidney pancreas RBC

3 Physiologic reaction

AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the

synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy

Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate

4 Method of measurement Karmen method

Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate

Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)

5 Reference Range 0-56 IUL

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

d Enzyme immunoassay measurement of CK-MB (mass)

A lsquosandwich techniquersquo that uses 2 antibodies one directedagainst the lsquoMrsquo subunit the other against the lsquoBrsquo subunit which isalso labeled with a reagent enzyme that catalyzes the conversion ofsubstrate to product

6 Reference Ranges affected by gender

bullmuscle mass exercise agebullTotal CK

bullMale 15-160 IULbullFemale 15-130 IULbullCK-MB lt5 of total CK

7 Clinical significancea Acute Myocardial Infarction (AMI)1) Increased total CK increased CK-MB2) Total CK increase 7-12 times normal withrise apparent at 4-6 hours post MIpeaks at 18-24 hours andreturns to normal by 72 hours3) The rise peak and fall of CK-MB parallels that of total CKbut a little sooner4) Other cardiac events (traumainflammation) variable results

b Skeletal muscle disease1) Increased total CK normal CK-MB (increased CK-MM)2) Muscular dystrophy especially Duchenne type may seetotal CK 50x ULN3) Inflammation of muscle especially viral also malignanthyperthermia4) Normal CK activity in neurogenic muscle disordersmultiple sclerosis myasthenia gravis poliomyelitis

c Central nervous system disorders

1) Increased total CK normal CK-MB (increased CK-BB)

2) Trauma or pathology indicated when CK-BB is present

3) Cerebral vascular disease encephalopathy stroke (CK-BB )may be normal

d Other

1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased

CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due

to CKactivity in RBC

B Lactate dehydrogenase (LD) 1 Specimen collection and handling

a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants

(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative

2 Wide cellulartissue distribution

a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells

3 Physiologic reaction

bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +

H+

4 Method of measurement

Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)

Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)

Both the forward and reverse reactions have been used in the clinical lab

with the rate of the reverse reaction being ~3 times faster allowing

measurement of smaller sample volumes and shorter reaction times

However the forward reaction has better linearity at this time

5 Isoenzymes clinical relevance of total LD activity often depends more on

isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)

b Five isoenzyme forms numbered by their anodal electrophoresis

6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(

1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

7 Clinical significance

a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver

disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver

c Acute myocardial infarction (AMI)

1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable

results

d Hemolytic anemia

1) Increased total LD 20-50 times normal increased LD-1 and

LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate

MagaloblasticAnemia3) Hemolytic anemia

C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)

1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable

other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric

analysisd Relatively stable analyze within 8 hours of

collection else store at2-4oC for up to 4 days

2 Wide cellulartissue distribution

a Skeletal muscle

b Cardiac muscletissue

c Liver

d Kidney pancreas RBC

3 Physiologic reaction

AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the

synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy

Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate

4 Method of measurement Karmen method

Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate

Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)

5 Reference Range 0-56 IUL

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

6 Reference Ranges affected by gender

bullmuscle mass exercise agebullTotal CK

bullMale 15-160 IULbullFemale 15-130 IULbullCK-MB lt5 of total CK

7 Clinical significancea Acute Myocardial Infarction (AMI)1) Increased total CK increased CK-MB2) Total CK increase 7-12 times normal withrise apparent at 4-6 hours post MIpeaks at 18-24 hours andreturns to normal by 72 hours3) The rise peak and fall of CK-MB parallels that of total CKbut a little sooner4) Other cardiac events (traumainflammation) variable results

b Skeletal muscle disease1) Increased total CK normal CK-MB (increased CK-MM)2) Muscular dystrophy especially Duchenne type may seetotal CK 50x ULN3) Inflammation of muscle especially viral also malignanthyperthermia4) Normal CK activity in neurogenic muscle disordersmultiple sclerosis myasthenia gravis poliomyelitis

c Central nervous system disorders

1) Increased total CK normal CK-MB (increased CK-BB)

2) Trauma or pathology indicated when CK-BB is present

3) Cerebral vascular disease encephalopathy stroke (CK-BB )may be normal

d Other

1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased

CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due

to CKactivity in RBC

B Lactate dehydrogenase (LD) 1 Specimen collection and handling

a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants

(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative

2 Wide cellulartissue distribution

a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells

3 Physiologic reaction

bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +

H+

4 Method of measurement

Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)

Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)

Both the forward and reverse reactions have been used in the clinical lab

with the rate of the reverse reaction being ~3 times faster allowing

measurement of smaller sample volumes and shorter reaction times

However the forward reaction has better linearity at this time

5 Isoenzymes clinical relevance of total LD activity often depends more on

isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)

b Five isoenzyme forms numbered by their anodal electrophoresis

6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(

1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

7 Clinical significance

a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver

disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver

c Acute myocardial infarction (AMI)

1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable

results

d Hemolytic anemia

1) Increased total LD 20-50 times normal increased LD-1 and

LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate

MagaloblasticAnemia3) Hemolytic anemia

C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)

1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable

other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric

analysisd Relatively stable analyze within 8 hours of

collection else store at2-4oC for up to 4 days

2 Wide cellulartissue distribution

a Skeletal muscle

b Cardiac muscletissue

c Liver

d Kidney pancreas RBC

3 Physiologic reaction

AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the

synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy

Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate

4 Method of measurement Karmen method

Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate

Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)

5 Reference Range 0-56 IUL

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

7 Clinical significancea Acute Myocardial Infarction (AMI)1) Increased total CK increased CK-MB2) Total CK increase 7-12 times normal withrise apparent at 4-6 hours post MIpeaks at 18-24 hours andreturns to normal by 72 hours3) The rise peak and fall of CK-MB parallels that of total CKbut a little sooner4) Other cardiac events (traumainflammation) variable results

b Skeletal muscle disease1) Increased total CK normal CK-MB (increased CK-MM)2) Muscular dystrophy especially Duchenne type may seetotal CK 50x ULN3) Inflammation of muscle especially viral also malignanthyperthermia4) Normal CK activity in neurogenic muscle disordersmultiple sclerosis myasthenia gravis poliomyelitis

c Central nervous system disorders

1) Increased total CK normal CK-MB (increased CK-BB)

2) Trauma or pathology indicated when CK-BB is present

3) Cerebral vascular disease encephalopathy stroke (CK-BB )may be normal

d Other

1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased

CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due

to CKactivity in RBC

B Lactate dehydrogenase (LD) 1 Specimen collection and handling

a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants

(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative

2 Wide cellulartissue distribution

a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells

3 Physiologic reaction

bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +

H+

4 Method of measurement

Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)

Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)

Both the forward and reverse reactions have been used in the clinical lab

with the rate of the reverse reaction being ~3 times faster allowing

measurement of smaller sample volumes and shorter reaction times

However the forward reaction has better linearity at this time

5 Isoenzymes clinical relevance of total LD activity often depends more on

isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)

b Five isoenzyme forms numbered by their anodal electrophoresis

6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(

1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

7 Clinical significance

a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver

disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver

c Acute myocardial infarction (AMI)

1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable

results

d Hemolytic anemia

1) Increased total LD 20-50 times normal increased LD-1 and

LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate

MagaloblasticAnemia3) Hemolytic anemia

C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)

1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable

other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric

analysisd Relatively stable analyze within 8 hours of

collection else store at2-4oC for up to 4 days

2 Wide cellulartissue distribution

a Skeletal muscle

b Cardiac muscletissue

c Liver

d Kidney pancreas RBC

3 Physiologic reaction

AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the

synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy

Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate

4 Method of measurement Karmen method

Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate

Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)

5 Reference Range 0-56 IUL

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

b Skeletal muscle disease1) Increased total CK normal CK-MB (increased CK-MM)2) Muscular dystrophy especially Duchenne type may seetotal CK 50x ULN3) Inflammation of muscle especially viral also malignanthyperthermia4) Normal CK activity in neurogenic muscle disordersmultiple sclerosis myasthenia gravis poliomyelitis

c Central nervous system disorders

1) Increased total CK normal CK-MB (increased CK-BB)

2) Trauma or pathology indicated when CK-BB is present

3) Cerebral vascular disease encephalopathy stroke (CK-BB )may be normal

d Other

1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased

CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due

to CKactivity in RBC

B Lactate dehydrogenase (LD) 1 Specimen collection and handling

a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants

(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative

2 Wide cellulartissue distribution

a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells

3 Physiologic reaction

bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +

H+

4 Method of measurement

Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)

Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)

Both the forward and reverse reactions have been used in the clinical lab

with the rate of the reverse reaction being ~3 times faster allowing

measurement of smaller sample volumes and shorter reaction times

However the forward reaction has better linearity at this time

5 Isoenzymes clinical relevance of total LD activity often depends more on

isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)

b Five isoenzyme forms numbered by their anodal electrophoresis

6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(

1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

7 Clinical significance

a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver

disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver

c Acute myocardial infarction (AMI)

1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable

results

d Hemolytic anemia

1) Increased total LD 20-50 times normal increased LD-1 and

LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate

MagaloblasticAnemia3) Hemolytic anemia

C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)

1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable

other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric

analysisd Relatively stable analyze within 8 hours of

collection else store at2-4oC for up to 4 days

2 Wide cellulartissue distribution

a Skeletal muscle

b Cardiac muscletissue

c Liver

d Kidney pancreas RBC

3 Physiologic reaction

AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the

synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy

Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate

4 Method of measurement Karmen method

Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate

Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)

5 Reference Range 0-56 IUL

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

c Central nervous system disorders

1) Increased total CK normal CK-MB (increased CK-BB)

2) Trauma or pathology indicated when CK-BB is present

3) Cerebral vascular disease encephalopathy stroke (CK-BB )may be normal

d Other

1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased

CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due

to CKactivity in RBC

B Lactate dehydrogenase (LD) 1 Specimen collection and handling

a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants

(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative

2 Wide cellulartissue distribution

a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells

3 Physiologic reaction

bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +

H+

4 Method of measurement

Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)

Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)

Both the forward and reverse reactions have been used in the clinical lab

with the rate of the reverse reaction being ~3 times faster allowing

measurement of smaller sample volumes and shorter reaction times

However the forward reaction has better linearity at this time

5 Isoenzymes clinical relevance of total LD activity often depends more on

isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)

b Five isoenzyme forms numbered by their anodal electrophoresis

6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(

1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

7 Clinical significance

a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver

disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver

c Acute myocardial infarction (AMI)

1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable

results

d Hemolytic anemia

1) Increased total LD 20-50 times normal increased LD-1 and

LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate

MagaloblasticAnemia3) Hemolytic anemia

C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)

1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable

other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric

analysisd Relatively stable analyze within 8 hours of

collection else store at2-4oC for up to 4 days

2 Wide cellulartissue distribution

a Skeletal muscle

b Cardiac muscletissue

c Liver

d Kidney pancreas RBC

3 Physiologic reaction

AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the

synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy

Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate

4 Method of measurement Karmen method

Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate

Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)

5 Reference Range 0-56 IUL

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

d Other

1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased

CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due

to CKactivity in RBC

B Lactate dehydrogenase (LD) 1 Specimen collection and handling

a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants

(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative

2 Wide cellulartissue distribution

a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells

3 Physiologic reaction

bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +

H+

4 Method of measurement

Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)

Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)

Both the forward and reverse reactions have been used in the clinical lab

with the rate of the reverse reaction being ~3 times faster allowing

measurement of smaller sample volumes and shorter reaction times

However the forward reaction has better linearity at this time

5 Isoenzymes clinical relevance of total LD activity often depends more on

isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)

b Five isoenzyme forms numbered by their anodal electrophoresis

6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(

1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

7 Clinical significance

a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver

disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver

c Acute myocardial infarction (AMI)

1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable

results

d Hemolytic anemia

1) Increased total LD 20-50 times normal increased LD-1 and

LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate

MagaloblasticAnemia3) Hemolytic anemia

C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)

1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable

other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric

analysisd Relatively stable analyze within 8 hours of

collection else store at2-4oC for up to 4 days

2 Wide cellulartissue distribution

a Skeletal muscle

b Cardiac muscletissue

c Liver

d Kidney pancreas RBC

3 Physiologic reaction

AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the

synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy

Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate

4 Method of measurement Karmen method

Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate

Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)

5 Reference Range 0-56 IUL

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

B Lactate dehydrogenase (LD) 1 Specimen collection and handling

a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants

(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative

2 Wide cellulartissue distribution

a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells

3 Physiologic reaction

bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +

H+

4 Method of measurement

Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)

Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)

Both the forward and reverse reactions have been used in the clinical lab

with the rate of the reverse reaction being ~3 times faster allowing

measurement of smaller sample volumes and shorter reaction times

However the forward reaction has better linearity at this time

5 Isoenzymes clinical relevance of total LD activity often depends more on

isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)

b Five isoenzyme forms numbered by their anodal electrophoresis

6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(

1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

7 Clinical significance

a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver

disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver

c Acute myocardial infarction (AMI)

1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable

results

d Hemolytic anemia

1) Increased total LD 20-50 times normal increased LD-1 and

LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate

MagaloblasticAnemia3) Hemolytic anemia

C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)

1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable

other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric

analysisd Relatively stable analyze within 8 hours of

collection else store at2-4oC for up to 4 days

2 Wide cellulartissue distribution

a Skeletal muscle

b Cardiac muscletissue

c Liver

d Kidney pancreas RBC

3 Physiologic reaction

AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the

synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy

Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate

4 Method of measurement Karmen method

Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate

Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)

5 Reference Range 0-56 IUL

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

2 Wide cellulartissue distribution

a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells

3 Physiologic reaction

bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +

H+

4 Method of measurement

Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)

Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)

Both the forward and reverse reactions have been used in the clinical lab

with the rate of the reverse reaction being ~3 times faster allowing

measurement of smaller sample volumes and shorter reaction times

However the forward reaction has better linearity at this time

5 Isoenzymes clinical relevance of total LD activity often depends more on

isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)

b Five isoenzyme forms numbered by their anodal electrophoresis

6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(

1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

7 Clinical significance

a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver

disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver

c Acute myocardial infarction (AMI)

1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable

results

d Hemolytic anemia

1) Increased total LD 20-50 times normal increased LD-1 and

LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate

MagaloblasticAnemia3) Hemolytic anemia

C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)

1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable

other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric

analysisd Relatively stable analyze within 8 hours of

collection else store at2-4oC for up to 4 days

2 Wide cellulartissue distribution

a Skeletal muscle

b Cardiac muscletissue

c Liver

d Kidney pancreas RBC

3 Physiologic reaction

AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the

synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy

Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate

4 Method of measurement Karmen method

Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate

Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)

5 Reference Range 0-56 IUL

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

3 Physiologic reaction

bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +

H+

4 Method of measurement

Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)

Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)

Both the forward and reverse reactions have been used in the clinical lab

with the rate of the reverse reaction being ~3 times faster allowing

measurement of smaller sample volumes and shorter reaction times

However the forward reaction has better linearity at this time

5 Isoenzymes clinical relevance of total LD activity often depends more on

isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)

b Five isoenzyme forms numbered by their anodal electrophoresis

6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(

1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

7 Clinical significance

a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver

disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver

c Acute myocardial infarction (AMI)

1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable

results

d Hemolytic anemia

1) Increased total LD 20-50 times normal increased LD-1 and

LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate

MagaloblasticAnemia3) Hemolytic anemia

C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)

1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable

other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric

analysisd Relatively stable analyze within 8 hours of

collection else store at2-4oC for up to 4 days

2 Wide cellulartissue distribution

a Skeletal muscle

b Cardiac muscletissue

c Liver

d Kidney pancreas RBC

3 Physiologic reaction

AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the

synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy

Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate

4 Method of measurement Karmen method

Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate

Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)

5 Reference Range 0-56 IUL

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

4 Method of measurement

Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)

Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)

Both the forward and reverse reactions have been used in the clinical lab

with the rate of the reverse reaction being ~3 times faster allowing

measurement of smaller sample volumes and shorter reaction times

However the forward reaction has better linearity at this time

5 Isoenzymes clinical relevance of total LD activity often depends more on

isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)

b Five isoenzyme forms numbered by their anodal electrophoresis

6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(

1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

7 Clinical significance

a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver

disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver

c Acute myocardial infarction (AMI)

1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable

results

d Hemolytic anemia

1) Increased total LD 20-50 times normal increased LD-1 and

LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate

MagaloblasticAnemia3) Hemolytic anemia

C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)

1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable

other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric

analysisd Relatively stable analyze within 8 hours of

collection else store at2-4oC for up to 4 days

2 Wide cellulartissue distribution

a Skeletal muscle

b Cardiac muscletissue

c Liver

d Kidney pancreas RBC

3 Physiologic reaction

AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the

synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy

Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate

4 Method of measurement Karmen method

Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate

Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)

5 Reference Range 0-56 IUL

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

Both the forward and reverse reactions have been used in the clinical lab

with the rate of the reverse reaction being ~3 times faster allowing

measurement of smaller sample volumes and shorter reaction times

However the forward reaction has better linearity at this time

5 Isoenzymes clinical relevance of total LD activity often depends more on

isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)

b Five isoenzyme forms numbered by their anodal electrophoresis

6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(

1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

7 Clinical significance

a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver

disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver

c Acute myocardial infarction (AMI)

1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable

results

d Hemolytic anemia

1) Increased total LD 20-50 times normal increased LD-1 and

LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate

MagaloblasticAnemia3) Hemolytic anemia

C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)

1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable

other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric

analysisd Relatively stable analyze within 8 hours of

collection else store at2-4oC for up to 4 days

2 Wide cellulartissue distribution

a Skeletal muscle

b Cardiac muscletissue

c Liver

d Kidney pancreas RBC

3 Physiologic reaction

AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the

synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy

Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate

4 Method of measurement Karmen method

Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate

Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)

5 Reference Range 0-56 IUL

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

5 Isoenzymes clinical relevance of total LD activity often depends more on

isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)

b Five isoenzyme forms numbered by their anodal electrophoresis

6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(

1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

7 Clinical significance

a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver

disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver

c Acute myocardial infarction (AMI)

1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable

results

d Hemolytic anemia

1) Increased total LD 20-50 times normal increased LD-1 and

LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate

MagaloblasticAnemia3) Hemolytic anemia

C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)

1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable

other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric

analysisd Relatively stable analyze within 8 hours of

collection else store at2-4oC for up to 4 days

2 Wide cellulartissue distribution

a Skeletal muscle

b Cardiac muscletissue

c Liver

d Kidney pancreas RBC

3 Physiologic reaction

AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the

synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy

Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate

4 Method of measurement Karmen method

Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate

Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)

5 Reference Range 0-56 IUL

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

b Five isoenzyme forms numbered by their anodal electrophoresis

6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(

1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

7 Clinical significance

a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver

disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver

c Acute myocardial infarction (AMI)

1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable

results

d Hemolytic anemia

1) Increased total LD 20-50 times normal increased LD-1 and

LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate

MagaloblasticAnemia3) Hemolytic anemia

C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)

1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable

other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric

analysisd Relatively stable analyze within 8 hours of

collection else store at2-4oC for up to 4 days

2 Wide cellulartissue distribution

a Skeletal muscle

b Cardiac muscletissue

c Liver

d Kidney pancreas RBC

3 Physiologic reaction

AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the

synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy

Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate

4 Method of measurement Karmen method

Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate

Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)

5 Reference Range 0-56 IUL

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(

1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD

7 Clinical significance

a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver

disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver

c Acute myocardial infarction (AMI)

1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable

results

d Hemolytic anemia

1) Increased total LD 20-50 times normal increased LD-1 and

LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate

MagaloblasticAnemia3) Hemolytic anemia

C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)

1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable

other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric

analysisd Relatively stable analyze within 8 hours of

collection else store at2-4oC for up to 4 days

2 Wide cellulartissue distribution

a Skeletal muscle

b Cardiac muscletissue

c Liver

d Kidney pancreas RBC

3 Physiologic reaction

AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the

synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy

Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate

4 Method of measurement Karmen method

Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate

Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)

5 Reference Range 0-56 IUL

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

7 Clinical significance

a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver

disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver

c Acute myocardial infarction (AMI)

1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable

results

d Hemolytic anemia

1) Increased total LD 20-50 times normal increased LD-1 and

LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate

MagaloblasticAnemia3) Hemolytic anemia

C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)

1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable

other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric

analysisd Relatively stable analyze within 8 hours of

collection else store at2-4oC for up to 4 days

2 Wide cellulartissue distribution

a Skeletal muscle

b Cardiac muscletissue

c Liver

d Kidney pancreas RBC

3 Physiologic reaction

AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the

synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy

Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate

4 Method of measurement Karmen method

Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate

Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)

5 Reference Range 0-56 IUL

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

c Acute myocardial infarction (AMI)

1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable

results

d Hemolytic anemia

1) Increased total LD 20-50 times normal increased LD-1 and

LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate

MagaloblasticAnemia3) Hemolytic anemia

C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)

1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable

other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric

analysisd Relatively stable analyze within 8 hours of

collection else store at2-4oC for up to 4 days

2 Wide cellulartissue distribution

a Skeletal muscle

b Cardiac muscletissue

c Liver

d Kidney pancreas RBC

3 Physiologic reaction

AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the

synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy

Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate

4 Method of measurement Karmen method

Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate

Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)

5 Reference Range 0-56 IUL

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

d Hemolytic anemia

1) Increased total LD 20-50 times normal increased LD-1 and

LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate

MagaloblasticAnemia3) Hemolytic anemia

C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)

1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable

other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric

analysisd Relatively stable analyze within 8 hours of

collection else store at2-4oC for up to 4 days

2 Wide cellulartissue distribution

a Skeletal muscle

b Cardiac muscletissue

c Liver

d Kidney pancreas RBC

3 Physiologic reaction

AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the

synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy

Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate

4 Method of measurement Karmen method

Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate

Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)

5 Reference Range 0-56 IUL

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)

1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable

other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric

analysisd Relatively stable analyze within 8 hours of

collection else store at2-4oC for up to 4 days

2 Wide cellulartissue distribution

a Skeletal muscle

b Cardiac muscletissue

c Liver

d Kidney pancreas RBC

3 Physiologic reaction

AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the

synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy

Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate

4 Method of measurement Karmen method

Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate

Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)

5 Reference Range 0-56 IUL

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

2 Wide cellulartissue distribution

a Skeletal muscle

b Cardiac muscletissue

c Liver

d Kidney pancreas RBC

3 Physiologic reaction

AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the

synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy

Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate

4 Method of measurement Karmen method

Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate

Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)

5 Reference Range 0-56 IUL

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

3 Physiologic reaction

AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the

synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy

Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate

4 Method of measurement Karmen method

Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate

Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)

5 Reference Range 0-56 IUL

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

4 Method of measurement Karmen method

Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate

Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)

5 Reference Range 0-56 IUL

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

5 Reference Range 0-56 IUL

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

6 Clinical significance

bull a Liver disease hepatocellular parenchymal disease

bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x

normalbull 2) Obstructive conditions (cholestasis some

stages ofbull cirrhosis) generally show normal or slightly

increased AST

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease

b Skeletal muscle disease

1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant

hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple

sclerosismyasthenia gravis poliomyelitis show normal AST

• Slide 1
• B Biochemical Function
• 2 Lowers the energy of activation required for the reaction to
• 3 Properties of enzymes
• 4 Cofactors substances associated with some enzymes that are
• C Importance of Enzymes in Health and Disease
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Enzymes of Clinical Significance
• Slide 13
• Method of Measurment
• Slide 15
• Slide 16
• Slide 17
• 6 Reference Ranges affected by gender
• Slide 19
• Slide 20
• c Central nervous system disorders
• d Other
• B Lactate dehydrogenase (LD) 1 Specimen collection and handl
• 2 Wide cellulartissue distribution
• 3 Physiologic reaction
• 4 Method of measurement
• Slide 27
• Slide 28
• b Five isoenzyme forms numbered by their anodal electrophores
• mobility with lsquofraction 1rsquo migrating fastest towards the anod
• Slide 31
• 7 Clinical significance
• c Acute myocardial infarction (AMI)
• d Hemolytic anemia
• C Aspartate Transaminase (AST) Historical Serum glutamate ox
• 2 Wide cellulartissue distribution (2)
• 3 Physiologic reaction (2)
• 4 Method of measurement Karmen method
• 5 Reference Range 0-56 IUL
• 6 Clinical significance
• b Skeletal muscle disease