CC2 Lab Enzymology 1 & 2 Flashcards
1
Q
reacts at a designated time
A
Fixed time
2
Q
multiple measurements of absorbance changes
are made during the reaction
A
Kinetic assay
3
Q
An enzyme important in the regeneration of adenosine triphosphate
A
Creatine Kinase - “ATP-Creatine-N Phosphotransferase”
4
Q
Predominantly found in skeletal muscles, heart muscles, brain tissues
A
Creatine Kinase - “ATP-Creatine-N Phosphotransferase”
5
Q
Creatine Kinase is most commonly used in the diagnosis of ___,___ and ____.
A
acute myocardial infarction, muscular dystrophy, and central nervous system disorders
6
Q
Abundant in Cardiac and Skeletal muscles
A
CK-MM - “Muscle Type”, “CK-3”
7
Q
Major isoenzyme in sera of healthy people
A
CK-MM - “Muscle Type”, “CK-3”
8
Q
Normally found in neonatal sera (rare in adult serum)
A
CK-BB - “Brain Type”, “CK-1”
9
Q
Elevated in brain injury and carcinomas
A
CK-BB - “Brain Type”, “CK-1”
10
Q
With significant amount in the heart
A
CK-MB - “hybrid type”, “CK-2”
11
Q
Most specific for Myocardial damage (AMI)
A
CK-MB - “hybrid type”, “CK-2”
12
Q
Falsely elevated in hemolysis
A
CK-MB - “hybrid type”, “CK-2”
13
Q
CK-MB - “hybrid type”, “CK-2”
Elevates in ______
Peaks at ______
Normalize in __________
A
Elevates in 4-8 hours
Peaks at 12-24 hours
Normalize in 48-72 hours
14
Q
Common in older women
A
Macro-CK
15
Q
Migrates middle of MM and MB
A
Macro-CK
16
Q
Migrates cathodal to CK-MM
A
Mitochondrial CK (CK-Mi)
17
Q
Indicates severe illness
A
Mitochondrial CK (CK-Mi)
18
Q
Test Methods under CK
A
- Forward - Tanzer and Gilvarg Assay
- pH: 9.0
- Absorbance: 340nm - Reverse - Oliver Rosalki
- pH: 6.8
- Absorbance: 340 nm - Electrophoresis
19
Q
proteins within cells
A
Enzymes
20
Q
General Properties of Enzymes:
A
Active Site: Water Free
Allosteric site: Cavity other than the active site
21
Q
non-protein that must bond to a particular enzyme before a reaction occurs
A
Cofactors
22
Q
Enzyme Storage
A
-20C
23
Q
A transferase that hastens the interconversion of lactic acid and pyruvic acid
A
Lactate Dehydrogenase - L-lactate: NAD oxidoreductase
24
Q
Highest levels on Lactate Dehydrogenase are detected in ____ & ______
A
pernicious anemia and hemolytic disorders
25
Other disease associated with Lactate Dehydrogenase elevations are _____, ______, _______, ________.
hepatic disorders, acute myocardial infarction, pulmonary infarct, acute lymphoblastic leukemia
26
Isoenzymes: LDH1 (HHHH)
Heart, RBC, Kidneys
27
Isoenzymes: LDH2 (HHHM)
Major isoenzyme in healthy people
Most abundant and heat stable
28
Isoenzymes: LDH3 (HHMM)
lungs, pancreas, spleen, lymphocytes
29
Isoenzymes: LDH4 (HMMM)
skeletal muscle, liver, intestine
30
Isoenzyme: LDH5 (MMMM)
liver, skeletal muscle, intestine
31
Isoenzymes: LDH6
an arterioslcerotic Cardiovascular failure marker
32
LDH Clinical Significance
1. Myocardial Infarction
- Elevates at 12-24 hours after onset
- Peaks at 48-72 hours
- Remains elevated for 10 days
2. Hepatitis
3. Hemolysis
4. Lung and muscle disorders
33
LDH Methods
1. Wacker method - Forward or direct
- pH: 8.8
- Absorbance: 340 nm
2. Wroblewski Ladue - Reverse or indirect
- pH: 7.2
- Abrosbanc: 340 nm
34
Transport and stores oxygen to intracellular respiratory enzymes of contractile cells (With high affinity to O2)
Myoglobin
35
Found in myocardium and with greater cardiac specificity
Troponin I
36
A Heart Failure Biomarker
Brain-type natriuretic peptide
37
Derived from a pro-hormone, “Pro-BNP”
Brain-type natriuretic peptide
38
Myoglobin:
Elevates _______
Peaks at ______
Normalizes _________
Elevates 2-3 hours after onset
Peaks at 8-12 hours
Normalizes 18-30 hours
39
Troponin I:
Elevates _____
Peaks at ______
Normalize in ______
Elevates 3-6 hours after onset
Peaks at 12-18 hours after onset
Normalize in 6 days
40
Considered as the smallest enzyme
Amylase - “Alpha 1-4 Glucan 4 Glucohydrolase”
41
Amylase Activators
Calcium and Chloride
42
Amylase Substrate
Starch or Glycogen
43
Amylase Major sources:
1. Acinar cells of the pancreas - releases Amylopsin
2. Salivary glands - releases Ptyalin
44
Amylase used for the breakdown of ___ & ___
Glycogen and Starch
45
Amylase Clinical Significance
1. Acute Pancreatitis - first to elevate
- Elevates 2-12 hours after onset
- Peaks at 24 hours
- Normalizes in 3-5 days
2. Intestinal obstruction
3. Cholecystitis
4. Acute appendicitis
46
Amylase Methods: measures reducing sugar from starch breakdown
Saccharogenic
47
Amylase Methods: also known as Iodometric method
Amyloclastic
48
Amylase Methods: Measure amylase activity by following decrease in substrate concentration
Amyloclastic
49
Indicator used of Amyloclastic
Iodide
50
Amylase Methods: uses dye to check amylase activity
Chromogenic
51
Amylase Methods: Follows breakdown of starch
Amyloclastic
52
A single-chain glycoprotein with a molecular weight of 48 kDa
Lipase - Triacylglycerol acylhydrolase
53
Lipase needs the presence of ____ & ____
Bile salts and colipase
54
Lipase Function:
Hydrolyze glycerol esters of long-chain fatty acids
55
Most lipase activity found in serum are usually from _______
the pancreas and some
are secreted by gastric and intestinal mucosa
56
molecular weight of Lipase
48 kDa
57
Lipase Clinical Significance
1. Acute Pancreatitis
2. Perforated or duodenal ulcer
3. Intestinal obstruction
4. Mesenteric vascular obstruction
58
Lipase Reference Method
Cherry Crandal
59
What substance is utilized on Cherry Crandal for hydrolysis
Olive oil
60
Cherry Crandal indicator
Phenolphthalein
61
Liberates inorganic phosphate from an organic esters with the concomitant production of alcohol
Alkaline phosphatase
62
Alkaline phosphatase Also known as
Alkaline Orthophosphoric Monoester Phosphohydrolase
63
Alkaline phosphatase requires __ & __ as activator
Magnesium and Manganese
64
Alkaline phosphatase Major Sources:
Liver
Kidneys
Placenta
Intestine
Spleen
Bone
65
Alkaline phosphatase Optimum pH
9.0-10.5
66
Alkaline phosphatase Elevated in:
Osteoblastic activity in kids during growth
>50 years old
Pregnancy (16-18 weeks)
67
Alkaline phosphatase Clinical Significance
Obstructive jaundice
Pregnancy
Paget’s disease
Rickets
Bone Cancer
Other bone disease
68
Alkaline phosphatase Methods
Electrophoresis
Liver ALP
Bone ALP
Placental ALP
Intestinal ALP
Regan
Nagao
Heat Stability
Placental
Intestinal
Liver
Bone
Chemical Inhibition
Phenylalanine
3M Urea
L-leucine
Levamisole
69
Give the substrate and the product of Bodansky, Shinowara, Jones, Reinhart
Substrate: B-glycero phosphate
Product: Inorganic Phosphate Glycerol
70
Give the substrate and the product of King Armstrong
Substrate: Phenylphosphate
Product: Phenol
71
Give the substrate and the product of Bessy, Lowry, Brock, Bowers, Mccomb
Substrate: P-nitrophenyl phosphate
Product: P-nitrophenol
72
Transfer of amino group between aspartate and alpha-ketoacids resulting to Oxaloacetate and Glutamate
Aspartate aminotransferase
73
Aspartate aminotransferase also known as
Serum Glutamic Oxaloacetic Transaminase
74
Aspartate aminotransferase major & other source
Major:
Liver
Cardiac Tissue
Skeletal Muscle
Other Source:
Kidneys
Pancreas
RBCs
75
Aspartate aminotransferase Significance:
Hepatocellular disorders (Hepatitis, Cirrhosis, Bile duct obstruction, Hepatic cancer)
Skeletal muscle injury
Gangrene
Myocarditis
Reye’s syndrome
AMI (Acute Myocardial Infarction)
Elevates 6-8 hours after onset
Peaks at 24 hours
Normalize in 5 days
76
Aspartate aminotransferase methods
1. Karmen method
utilizes Malate dehydrogenase to monitor change in absorbance at 340nm
pH: 7.3-7.8
2. Reitman Frankel
77
Transfer of amino group from alanine to alpha-ketoglutarate
Alanine aminotransferase
78
Alanine aminotransferase also known as
Serum Glutamic Pyruvic Transaminase
79
Alanine aminotransferase Abundant in the
liver
80
Alanine aminotransferase other source
Kidney
Pancreas
RBCs
Heart
Skeletal Muscle
Lungs
81
Alanine aminotransferase needs this vitamin as coenzyme
vitamin B6
82
Alanine aminotransferase Significance:
Elevated in Hepatic Disorders
Monitor course of hepatitis treatment
Used to screen blood donors
83
Alanine aminotransferase METHODS
Coupled Enzymatic Reaction
utilizes LDH to monitor change in absorbance at 340nm
pH: 7.3-7.8
84
Principle of Coupled Enzymatic Reaction under ALT Method
Alanine + alpha ketoglutarate ---ALT---> Pyruvate + glutamate
Pyruvate + NADH + H+ ----LDH----> Lactate + NAD+
85
Catalyzes the transfer or glutamyl groups between peptides through linkage at a gamma carboxyl group
gamma glutamyl transferase
86
gamma glutamyl transferase also known as
Gamma glutamyl transpeptidase
87
gamma glutamyl transferase main source
Hepatic Cells
Kidneys
Prostate
Pancreas
Brain
88
gamma glutamyl transferase sensitive indicator of
alcoholism
89
gamma glutamyl transferase Significance:
Hepatic disorders
Alcoholism
Drug overdose
90
gamma glutamyl transferase method
Spectrophotometric method
91
Spectrophotometric principle under ggt method
Gamma glutamyl-p-nitroaniline ------> p-nitroaniline
92
the absorbance of Spectrophotometric method is measured at
405nm
93
Catalyzes the hydrolysis of most ribonucleoside 5'-monophosphate and deoxynucleoside 5'-monophosphate to the corresponding nucleoside and orthophosphate
5'nucleotidase
94
5'nucleotidase also known as
Phosphoric monoester hydrolase
95
5'nucleotidase main source
Liver
96
5'nucleotidase is a Marker of
Hepatobillary disease and Infiltrative lesions of the liver
97
Catalyzes the hydrolysis of several orthophosphoric monoesters to yield the corresponding alcohol and inorganic phosphate
acid phosphatase
98
acid phosphatase also known as
Acid Orthophosphoric Monoester Phosphohydrolase
99
acid phosphatase main source
Prostate
Liver
Kidneys
RBCs
Platelets
Osteoclastic Cells
100
acid phosphatase optimum pH
4.5-7.0
101
acid phosphatase Clinical Significance:
Prostatic
marker for prostatic carcinoma
Prostatic hypertrophy
Non-prostatic: Gaucher’s disease, Neimann-pick disease
Osteoclastic - hyperthyroidism, Paget’s disease, Multiple myeloma
Hematologic
Granulocytic leukemia
chronic lymphocytic leukemia
plasma cell leukemia
hairy cell leukemia
102
acid phosphatase method
Bodansky method
King-Armstrong
Bessey-Lowry-Brock
Roy and Hillman
Babson, Read and Philips
