CC1 Flashcards
1
Q
Enumerate Prostate Enzymes
A
ACP
G-6-PDH
2
Q
Enumerate Miscellaneous Enzymes
A
5’ nucleotidase
Cholinesterase/pseudocholinesterase
Angiotensin Converting Enzyme (ACE)
Ceruloplasmin
Ornithine Carbamoyl Transferase (OCT)
G-6-PD
3
Q
a phosphoric monoester hydrolase
A
5’ N
4
Q
5’ N source
A
predominantly secreted by the liver
5
Q
Marker for hepatobiliary disease and infiltrative lesions of the liver
A
5’ N
6
Q
5’ N is a marker for these conditions
A
Hepatobiliary disease
Infiltrative lesions of the liver
7
Q
reference range for 5’N
A
0-1.6 units
8
Q
index of parenchymal function of the liver
A
Cholinesterase/
Pseudocholinesterase
9
Q
Cholinesterase/Pseudocholinesterase source
A
Liver
10
Q
Monitor effects of muscle relaxants (succinylcholine) after surgery
A
Cholinesterase/
Pseudocholinesterase
11
Q
Marker for insecticide/pesticide poisoning (organophosphate – poisonous agent)
A
Cholinesterase/
Pseudocholinesterase
12
Q
reference range for cholinesterase/pseudocholinesterase
A
0.5-1.3 units (plasma)
13
Q
a.k.a Peptidyldipeptidase A or Kininase II
A
Angiotensin Converting Enzyme (ACE)
14
Q
aka Angiotensin Converting Enzyme (ACE)
A
Peptidyldipeptidase A or Kininase II
15
Q
Converts angiotensin I to angiotensin II within the lungs (RAAS)
A
ACE
16
Q
Primary enzyme of RAAS
A
Angiotensin Converting Enzyme (ACE)
17
Q
Promotes vasoconstriction of the renal arterioles to increase blood pressure and stimulates the adrenal cortex to release aldosterone so that aldosterone will promote Na reabsorption
A
Angiotensin II
18
Q
Events that activates RAAS
A
Blood pressure/volume is decrease
Low plasma sodium level
19
Q
source of ACE
A
Macrophage and epithelioid cells of the lungs
20
Q
Possible indicator of neuronal dysfunction
(Alzheimer’s/neurodegenerative disease)
A
ACE
21
Q
ACE is increased in these conditions
A
Sarcoidosis
Acute and chronic bronchitis
Leprosy
22
Q
Copper-carrying protein which acts as an enzyme
A
Ceruloplasmin
23
Q
Marker for Wilson’s disease (hepatolenticular disease)
A
Ceruloplasmin
24
Q
Clin. significance of Ornithine Carbamoyl Transferase (OCT)
A
Hepatobiliary diseases
25
Reference range for Ornithine Carbamoyl Transferase (OCT)
8-20 mU/mL
26
Maintain NADPH in the reduced form in the RBCs
G-6-PD
27
Specimen for G6PD
Red cell hemolysate
Serum
28
Responsible for maintaining and stabilizing
the membrane integrity of RBCs
reduced NADH (maintained by G6PD)
29
Protects RBCs from toxic agents that can
induce hemolytic reactions
Reduced NADH (maintained by G6PD)
30
Sources of G6PD
• Adrenal cortex
• Spleen
• RBC
• Lymph nodes
31
Enzyme used as a newborn screening marker
G6PD
32
G6PD is INCREASED in these conditions
MI
Megaloblastic anemia
33
G6PD is DECREASED in these conditions
Drug-induced hemolytic anemia (intake of primaquine, antimalarial drugs)
34
Reference range for G6PD
10-15 U/g Hgb
1200-2000 mU/mL pRBC
35
Biologic intracellular proteins that catalyze biochemical reactions
Enzymes
36
Affects the reaction of the organic matter
Enzymes
37
NOT consumed/changed in composition (only substrate → product)
Enzymes
38
Increased enzymes in serum is due to the following conditions
Cell injury/degradation
Increased membrane permeability
Organ damage (severely increased enzyme in serum/plasma)
39
What happen when there is an increased membrane permeability?
INCREASED ENZYMES in serum, allowing these proteins to move out easily from the cells
40
Enzymes are found in _____________
all body tissue (intracellular)
41
Concentration in serum is very low as it is very abundant in cytoplasm
Enzymes
42
Indication when there is a SEVERELY INCREASED enzyme in serum/plasma
Organ damage
43
FUNCTION OF ENZYMES
1. Hydration of Carbon Dioxide (respiration)
2. Nerve Induction (fast nerve impulse transmission)
3. Muscle Contraction (locomotion)
4. Nutrient Degradation (digestion)
5. Growth and Reproduction (work with hormones)
6. Energy Storage and Use
44
Enzymes with similar catalytic activity, but differ in their physical, biochemical, and immunologic properties
Isoenzymes
45
What are the difference between enzymes and its isoenzymes?
physical, biochemical, and immunologic properties
46
What is the similarity between enzymes and its isoenzymes?
catalytic activity
47
Type of cofactor that serve as a second substrate for enzyme
COENZYME
48
coenzyme tightly bound to an enzyme
Prosthetic group
49
T/F
If there is a coenzyme in the reaction, enzyme involved is oxidoreductase – ends with “dehydrogenase” (promotes redox reaction)
T
50
Part of the reagent since the primary enzyme is the target in the serum
Secondary coupling/indicator enzyme
51
Requires a coenzyme: pyridoxal phosphate/Vit B6
Aminotransferases
52
Coenzyme required by aminotransferases
pyridoxal phosphate/Vit B6
53
A cofactor that enhances enzyme activity by altering the spatial configuration of the active site of enzyme or
enhanced substrate finding
Activator
54
Types of cofactor
Coenzyme
Activator
55
These can be grouped as metallic or nonmetallic
Activators
56
Metallic activators
Mg, Iron, Zinc, Ca
57
Nonmetallic activators
Cl, Br
58
when used as coenzyme, it is converted into two forms when acted upon by oxidoreductase
NAD
59
Oxidized form acted upon by Oxidoreductase
NAD
60
Reduced form acted upon by Oxidoreductase
NADH
61
Measured absorbance if the product is OXIDIZED form
NAD
Measured: DECREASE in absorbance
62
Measured absorbance if the product is REDUCED form
NADH
Measured: INCREASE (HIGH) in absorbance
63
Enumerate liver enzymes
AST/SGOT
ALT/SGPT
GGT
ALP/Alkaline Orthophosphoric Monoester Phosphohydrolase
ACP/Acid Orthophosphoric Monoester Phosphohydrolase
5’ N
64
Enumerate cardiac enzymes
CK
LDH
AST
65
Enumerate non-enzymatic cardiac markers
Myoglobin
Troponins
66
Enumerate pancreatic enzymes
AMS
LPS
67
Type of enzyme:
Aspartate Aminotransferase (AST)/
Serum glutamic-oxaloacetic
transaminase (SGOT)
TRANSFERASE
68
Type of enzyme:
Alanine Aminotransferase (ALT)/
Serum glutamic-pyruvic
transaminase (SGPT)
TRANSFERASE
69
Type of enzyme:
Gamma-Glutamyltransferase
(GGT)
TRANSFERASE
70
Type of enzyme:
Acid Phosphatase (ACP)/
Acid Orthophosphoric Monoester
Phosphohydrolase
HYDROLASE
71
Type of enzyme:
Alkaline Phosphatase (ALP)/
Alkaline Orthophosphoric
Monoester Phosphohydrolase
HYDROLASE
72
Type of enzyme:
CK/CKP
TRANSFERASE
73
Type of enzyme:
LDH
OXIDOREDUCTASE
74
Type of enzyme:
5’ nucleotidase (5’N)
phosphoric monoester HYDROLASE
75
Type of enzyme:
Amylase (AMS)
HYDROLASE
76
Type of enzyme:
Lipase (LPS)
HYDROLASE
77
Transfer of an amino group between aspartate and α-keto glutaric acid
Aspartate Aminotransferase (AST)/
Serum glutamic-oxaloacetic transaminase (SGOT)
78
Involved in the synthesis and degradation of
AA (protein catabolism, deamination)
Aspartate Aminotransferase (AST)/
Serum glutamic-oxaloacetic
transaminase (SGOT)
79
Major organ affected of AST/SGOT
HEART
80
Substrate for AST/SGOT
Aspartic acid (aspartate)
a-ketoglutaric acid (a-ketoglutarate)
81
End products of AST/SGOT
Glutamic acid (glutamate)
Oxaloacetic acid (oxaloacetate)
82
Color developer for AST/SGOT
2,4 DNPH
(2,4-Dinitrophenylhydrazine)
83
Color intensifier for AST/SGOT
0.4 N NaOH
84
Colorimetric method
for aminotransferase
Reitman and Frankel
85
Major source of AST/SGOT
Heart
86
T/F
AST/SGOT is widely distributed
T
87
Tissue sources of AST/SGOT with increased activities
* Cardiac tissue
* Liver
* Skeletal muscle
88
AST/SGOT is increased in these conditions
* AMI
* Hepatocellular disorder: chronic liver disorder with progressive damage
* Skeletal muscle disorder: muscular dystrophy (Duchenne)
* Trichinosis
89
Enzyme increased in
chronic liver disorder with progressive damage
AST/SGOT
(a hepatocellular disorder)
90
Enzyme increased in muscular dystrophy (Duchenne)
AST/SGOT
91
Enzyme increased in Trichinosis
AST/SGOT
92
Isoenzymes of AST/SGOT
Cytoplasmic AST
Mitochondrial AST
93
most abundant AST isoenzyme in normal serum
Cytoplasmic AST
94
AST isoenzyme present in mitochondrial membrane
Mitochondrial AST
95
AST isoenzyme that is increased in cell necrosis
Mitochondrial AST
96
Assay for AST/SGOT
KARMEN METHOD
97
2°/indicator enzyme used in Karmen method for AST
Malate Dehydrogenase
98
Coenzyme used in Karmen method for AST
Pyridoxal phosphate/Vit B6
99
Monitored in Karmen method for AST and the wavelength used
decrease in absorbance at 340 nm
(measures oxidized NAD)
100
Variables in Karmen method? What will be the effect?
Hemolysis (False ↑) – very sensitive
101
Reference range for AST/SGOT using Karmen method
5 – 30 U/L
102
T/F
Product formed by the 1° enzyme (AST) will become the substrate for the 2° enzyme (MD)
T
103
In Karmen method, what is the substrate used by MD which is also the product formed by the primary enzyme AST?
oxaloacetate
104
In Karmen method, what are the products formed acted upon by MD?
malate + NAD
105
Transfer of an amino group between alanine
and α-ketoglutarate
Alanine Aminotransferase (ALT)/
Serum glutamic-pyruvic transaminase (SGPT)
106
More liver specific than AST
Alanine Aminotransferase (ALT)/
Serum glutamic-pyruvic
transaminase (SGPT)
107
Major organ affected by ALT/SGPT
LIVER
108
Substrates used by ALT/SGPT
Alanine
a-ketoglutaric acid (a-ketoglutarate)
109
End products of ALT/SGPT
Glutamic acid (glutamate)
Pyruvic acid (pyruvate)
110
Color developer for ALT/SGPT
2,4 DNPH
111
Color intensifier for ALT/SGPT
0.4 N NaOH
112
Major source of ALT/SGPT
LIVER
113
Minor sources of ALT/SGPT
* Kidneys
* Pancreas
* RBC
* Heart
* Skeletal muscle
* Lungs
114
ALT/SGPT is increased in these conditions
* Hepatocellular disorders
(liver specific enzyme)
* Acute liver inflammation
115
Enzyme used to monitor hepatitis treatment
and drug therapy effects
ALT/SGPT
116
Enzyme used to screen post transfusion hepatitis
ALT/SGPT
117
Screens blood donors (not routine; only with jaundice)
ALT/SGPT
118
Sensitive test for
occupational toxic exposure
ALT/SGPT
119
Assay for ALT/SGPT
LACTATE DEHYDROGENASE (LD)
120
2°/indicator enzyme for ALT/SGPT assay
LACTATE DEHYDROGENASE (LD)
121
Coenzyme used in LD for ALT
Pyridoxal phosphate/Vit B6
122
Monitored in LD for ALT? What is the wavelength?
decrease in absorbance at 340 nm
(measures oxidized NAD)
123
Reference range for ALT using LD assay
6 – 37 U/L
124
In LD assay, what is the substrate used by LD which is also the product formed by the primary enzyme ALT?
pyruvate
125
In LD assay for ALT, what are the products formed by LD?
lactate + NAD
126
aka De Ritis Ratio
AST/ALT Ratio
SGOT/SGPT Ratio
127
Used to differentiate the cause of hepatic disorder
De Ritis Ratio (AST/ALT Ratio)
128
De Ritis Ratio (AST/ALT Ratio):
> 1
nonviral origin
129
De Ritis Ratio (AST/ALT Ratio):
<1
viral origin
130
what is the cause of hepatic disorder when the AST is higher than ALT?
non-viral
>1 (high AST: low ALT)
131
what is the cause of hepatic disorder when the ALT is higher than AST?
viral
<1 (low AST: high ALT)
132
T/F
In De Ritis Ratio, a ratio of exactly 1 is possible.
FALSE!
Ratio of 1 is not possible because AST has MANY tissue sources
133
Catalyze transfer of γ-glutamyl residue from γ-glutamyl peptides to amino acids, H20, etc.
Gamma-Glutamyltransferase
(GGT)
134
Common donor (biologic system) in GGT
glutathione
135
Substrates used in GGT
Glutathione + AA
136
Products formed when GGT is used as an enzyme
glutamyl-peptide +
L-cysteinylglycine
137
Source of GGT
Canaliculi of hepatic cells, specifically in the
epithelial lining of biliary ductulus
138
GGT is used to diagnose these conditions
Hepatobiliary disorders (obstructive jaundice)
Chronic alcoholism (ethanol intoxication)
139
Marker for occult alcoholism
GGT
140
Most sensitive marker for acute alcoholic hepatitis
GGT
141
Assay for GGT
SZAZ ASSAY
142
measures the absorbance of p-nitroaniline at 405-420 nm
SZAZ ASSAY for GGT
143
Wavelength used in Szaz assay to measure p-nitroaniline
405-420 nm (visible light region)
144
T/F
Wavelength requirement may be a clue to the detected product
T
400-700 nm – visible light region
Nonvisible regions:
<400 – UV region
>700 – IR region
144
substrates used in Szaz assay using GGT
y-glutamyl-p-nitroanilide +
glycylglycine
145
products formed in Szaz assay using GGT
y-glutamyl-glycylglycine +
p-Nitroaniline
146
Catalyze hydrolysis of phosphomonoesters
at an acid pH (5.0)
Acid Phosphatase (ACP)/
Acid Orthophosphoric Monoester
Phosphohydrolase
147
Liberate inorganic PO4 from an organic PO4
ester with alcohol production at an acid pH
Acid Phosphatase (ACP)/
Acid Orthophosphoric Monoester
Phosphohydrolase
148
pH requirement in ACP
5.0
149
NOT prostate specific
Acid Phosphatase (ACP)/
Acid Orthophosphoric Monoester
Phosphohydrolase
150
Incorporated in the prostatic fluid secreted by
the prostate gland (normally present in
seminal fluid)
Acid Phosphatase (ACP)/
Acid Orthophosphoric Monoester
Phosphohydrolase
151
Substrates used in ACP
phosphomonoester + H2O
152
Products formed by ACP
alc + phosphate ion
153
Sources of ACP
* Prostate (male)
* RBCs
* Platelets
* Bone (osteoclast – for bone resorption)
154
Detects metastatic prostatic cancer
ACP
155
Significant in forensic rape investigation
ACP
156
Sample used in forensic rape investigation using ACP
vaginal washings from rape victim
157
Detectability of ACP in forensic rape investigation
4 days or less
(Options if >4 days: vaginal laceration)
158
Added to differentiate
prostatic form (specific) from nonspecific
form like RBC ACP
Inhibitor
159
Inhibitors used for ACP
L-tartrate ions
Formaldehyde,
Cupric ions
160
L-tartrate ions inhibits these ACP isoenzymes
prostatic ACP
lysosomal ACP
161
Formaldehyde,
Cupric ions inhibits this ACP isoenzyme
RBC ACP
162
Reference range for Prostatic ACP
0-3.5 ng/ml
163
In Shinowara method, what is the substrate used and the products formed?
substrate: p-nitrophenyl-phosphate (PNPP) - COLORLESS
products: p-nitrophenol -YELLOW
phosphate ion
164
general methods for ACP
Quantitative end point
Continuous monitoring
165
Substrate used is Thymolphthalein monophosphate (most commonly used; sensitive and specific)
Quantitative end point
166
Substrate used is a-naphthyl phosphate for ACP
Continuous monitoring
167
Catalyze hydrolysis of phosphomonoesters at an Alk pH (9-10) (cleaves monoester bonds present in substrates)
Alkaline Phosphatase (ALP)/
Alkaline Orthophosphoric Monoester Phosphohydrolase
168
Liberate inorganic PO4 from an organic PO4
ester with alcohol production at an Alk pH
Alkaline Phosphatase (ALP)/
Alkaline Orthophosphoric
Monoester Phosphohydrolase
169
pH required in ALP
9-10
170
Hydrolase enzyme that requires an activator
ALP
171
Activator used in ALP
Mg2+
172
T/F
ALP is liver specific
F
173
Predominant ALP isoenzyme in normal serum
Liver ALP
Bone ALP
174
Sources of ALP
* Liver
* Bone (osteoblast)
* Placenta
* Intestine
* Renal tissues (not measured)
175
Enzyme significantly ↑ in Paget's disease/Osteitis deformans
ALP
176
ALP is significantly increased in this condition
Paget’s disease/
Osteitis deformans
177
Important for evaluation of hepatobiliary (obstructive types) and bone disorders
ALP
178
ALP Isoenzymes
1. Liver ALP
2. Bone ALP
3. Placental ALP
4. Intestinal ALP
179
Assay for ALP
Bowers and McComb
180
Principle: molar absorptivity of p-Nitrophenol, Absorbance is measured at 405 nm (colorimetric measurement of yellow-colored p-Nitrophenol), pH 10.2
Bowers and McComb for ALP
181
optimal pH used in Bowers and McComb
10.2
182
In Bowers and McComb, absorbance is measured at this wavelength
405 nm
183
colorimetric measurement of yellow-colored p-Nitrophenol as product of ALP
Bowers and McComb
184
ALP Reference range in ADULTS
30 – 90 U/L
185
ALP Reference range in 0-3 months
70 – 220 U/L
186
ALP Reference range in 3-10 years
50 – 260 U/L
187
ALP Reference range in 10 yr - puberty
60 – 295 U/L
highest due to active bone development; involves osteoblast resulting to release of ALP in serum)
188
Fastest ALP isoenzyme
Liver ALP
189
2 fractions of Liver ALP
Major liver band
Fast liver (a1) band
190
responsible for predominant liver ALP level in normal serum
Major liver band
191
responsible for fast-migrating liver ALP in electrophoresis
Fast liver (a1) band
192
Liver ALP is increased PATHOLOGICALLY in
Liver diseases
193
Most anodal (fastest) ALP isoenzyme
Liver ALP
194
3rd most heat stable ALP isoenzyme
Liver ALP
195
ALP residual activity after heating:
decreased to >20%
Liver ALP
196
ALP isoenzyme inhibited by Levamisole
Liver ALP
Bone ALP
197
Heat labile ALP isoenzyme (If the temp is ↑, activity is markedly ↓)
Bone ALP
198
Bone ALP is increased PHYSIOLOGICALLY in
Bone growth
199
Bone ALP is increased PATHOLOGICALLY in
* Bone disease
* Healing of bone fractures
200
2nd most anodal ALP isoenzyme
Bone ALP
201
Least heat stable ALP isoenzyme
Bone ALP
202
ALP residual activity after heating:
decreased to <20%
Bone ALP
203
ALP isoenzyme inhibited by 3M urea
Bone ALP
204
Most heat stable ALP isoenzyme (withstand heating at 65ºC for 30 minutes)
Placental ALP
205
Placental ALP can withstand this temperature for how many minutes
65ºC for 30 minutes
206
Placental ALP is increased PHYSIOLOGICALLY in
Pregnancy (16th & 20th week of gestation)
207
Placental ALP is increased PATHOLOGICALLY in
Malignancy/cancer
(carcinoplacental ALP)
208
3rd most anodal ALP isoenzyme
Placental ALP
209
Inhibited by Phenylalanine
Placental ALP
Intestinal ALP
Regan ALP
Nagao ALP
210
Slowest moving ALP fraction
Intestinal ALP
211
Intestinal ALP is increased PHYSIOLOGICALLY in
* Blood group B and O
* Fatty meal consumption
212
Intestinal ALP is increased PATHOLOGICALLY in
GIT disorders
213
Least anodal ALP isoenzyme
Intestinal ALP
214
2nd most heat stable ALP isoenzyme
Intestinal ALP
215
Total ALP elevations by Liver or Bone ALP are
differentiated by heating of serum at 56°C for 10 mins.
(focuses on liver and bone due to predominance)
HEAT STABILITY
216
Adding of chemical reagent to the
sample to inhibit the activity of certain
isoenzyme
CHEMICAL INHIBITION
217
CARCINOPLACENTAL ALP
REGAN ALP
NAGAO ALP
218
Most heat stable ALP
REGAN ALP
219
Bone ALP co-migrator
REGAN ALP
(2nd most anodal)
220
Regan ALP is associated with these conditions
Lung, breast, gynecological cancers
221
Nagao ALP is associated with these conditions
Adenocarcinoma of the pancreas and bile duct, pleural cancer
222
Carcinoplacental ALP inhibited by Phenylalanine only
REGAN ALP
223
Carcinoplacental ALP inhibited by both Phenylalanine and L-leucine
NAGAO ALP
224
CARDIAC ENZYMES (MI PROFILE)
CK/CPK
LDH
AST
225
Involved in the storage of high-energy
creatine phosphate in muscle cells
Creatine Kinase (CK)/
Creatine Phosphokinase
(CPK)
226
transferase enzyme that catalyzes
the transfer of PO4 group betw. substrates
Kinase
(CPK – inappropriate term)
227
high energy
reservoir in muscle cells; utilized by muscle
cells to form waste product - creatinine
Creatine phosphate
228
originates in the liver from the amino acid arginine, glycine and methionine
Creatine
229
amino acids where creatine originates
methionine
arginine
glycine
230
formed from muscle metabolism; excreted in the nephrons of the kidney at a constant rate
Creatinine
231
T/F
From the liver, creatine is transferred to the
muscles. In the muscles, creatine is converted
to creatine phosphate through the action of
the enzyme CK
T
232
substrates used in CK
creatine + ATP
233
products formed in CK
creatine phosphate + ADP
234
T/F
CK is widely distributed
T
235
CK has inc. activities in these tissues
* skeletal muscle
* heart
* brain
236
First cardiac enzyme to elevate after AMI
CK2 (CK-MB)
(>6% of total CK)
237
Percentage of CK-MM
94-98%
238
Percentage of CK-MB
2-6%
239
Percentage of CK-BB
<1%
240
Reference range of Total CK in MALE
15-160 U/L
(higher than female due to inc. activity & muscle mass)
241
Reference range of Total CK in FEMALE
15-130 U/L
242
Reference range of CK-MB
<6% of total CK
243
Dimeric enzyme
CK
M - muscle, B - brain
244
CK isoenzymes
CK 1 (CK-BB)
CK 2 (CK-MB)
CK 3 (CK-MM)
Macro-CK
CK-Mi
245
CK isoenzyme that migrate fastest toward anode (most anodal)
CK1
CK-BB
Brain type
246
CK isoenzyme: 2nd fastest to migrate toward anode
CK2
CK-MB
Hybrid type
247
CK isoenzyme that migrate slowest toward anode (least anodal)
CK3
CK-MM
Muscle type
248
CK1 (CK-BB) - Brain type has inc. concentration in these sites
CNS
GI tract
Uterus (pregnancy)
249
Large CK molecule – cannot pass BBB
CK1
CK-BB
Brain type
250
CK isoenzyme ↑ in heart tissue
(cardiac muscle specific)
CK2
CK-MB
Hybrid type
251
↑ CK-MB denotes?
AMI
252
Major isoenzyme in striated muscles and normal serum
CK3
CK-MM
Muscle type
253
Macro-CK composition
CK-BB + antibodies (IgG/IgA)
CK-MM + lipoproteins
254
Migrate midway betw. CK-MM and CK-MB
Macro-CK
255
Migrates cathodal to CK-MM
Mitochondrial CK (CK-Mi)
256
CK isoenzyme located in mitochondrial membrane; used to diagnose cell necrosis/severe damage
Mitochondrial CK (CK-Mi)
257
Assay for CK
TANZER-GILVARG (FORWARD)
OLIVER-ROSALKI (REVERSE)
258
Measured in TANZER-GILVARG (FORWARD)
↓ in absorbance at 340 nm
259
Optimum pH in TANZER-GILVARG (FORWARD
9.0
260
Measured in OLIVER-ROSALKI (REVERSE)
↑ in absorbance at 340 nm
261
Optimum pH in OLIVER-ROSALKI (REVERSE)
6.8
262
SOURCES OF ERROR for CK measurement
Hemolysis (false inc.)
Physical activity and IM injections (false inc.)
Photosensitive (false dec.)
Immobilized/bedridden (false dec.)
263
Abundant enzyme inside the RBC that mimics the activity of CK causing hemolysis as a source of error
Adenylate kinase
264
Catalyzes interconversion of lactic and pyruvic acids
Lactate Dehydrogenase (LDH)
265
T/F
LDH is tissue specific
False
*Widely distributed
266
This may affect LDH activity
Storage
↓: frozen/low temp.
Maintained: RT for 2 days
267
Substrate for LD
Lactate + NAD
268
Products of LD
Pyruvate + NADH + H+
269
Inc. activities of LD are found in:
* Heart
* Liver
* Skeletal muscle
* RBC
270
Late cardiac marker (not practical for AMI dx)
LDH
271
SOLE PURPOSE: Monitor px response to therapy for cardiac diseases (i.e. AMI)
LDH
272
LDH reference range
100-225 U/L
273
Tetramer enzyme
LDH
(4 subunits/monomers of two active sub-unit forms [H & M]) H - heart, M - muscle
274
Tissue source of LDH 1
Heart,
RBC
275
Tissue source of LDH 2
Heart,
RBC
276
Tissue source of LDH 3
Lung,
Spleen,
Pancreas
277
Tissue source of LDH 4
Liver
278
Tissue source of LDH 5
Skeletal muscle,
Liver
279
Disorder associated with LDH 1
AMI,
Hemolytic anemia
280
Disorder associated with LDH 2
Renal Infarction,
Megaloblastic anemia
281
Disorder associated with LDH 3
Pulmonary embolism
282
Disorder associated with LDH 4
Hepatic injury
283
Disorder associated with LDH 5
Muscle dystrophy
Hepatic disorders
284
percentage of LDH 1 in total LDH
20-30%
285
percentage of LDH 2 in total LDH
30-40%
286
percentage of LDH 3 in total LDH
20-25%
287
percentage of LDH 4 in total LDH
7-15%
288
percentage of LDH 5 in total LDH
5-15%
289
LDH isoenzyme NOT normally seen in healthy people
LDH 6 / Alcohol dehydrogenase
290
LDH 6 / Alcohol dehydrogenase is present in these conditions
Drug hepatotoxicity
Obstructive jaundice
Atherosclerotic failure
291
has high affinity to α-hydroxybutyrate
LDH 1
292
LDH Normal Electrophoretic migration pattern
LDH 1>2>3>4>5
293
LDH Relative conc. in normal serum
LDH 2>1>3>4>5
294
LDH pattern in AMI/IV hemolysis
LDH 1>2>3>4>5
(FLIPPED PATTERN)
295
Used to differentiate AMI from IV hemolysis with flipped pattern
LDH 1
CK-MB
296
Differentiate AMI from IV hemolysis
AMI: high LDH1, high CK-MB
IV hemolysis: high LDH1, normal CK-MB
297
Assays for LDH
WACKER METHOD (FORWARD)
WROBLEUSKI LA DUE (REVERSE)
α-hydroxybutyrate dehydrogenase (α-HBD)
298
Assay: Lactate —> Pyruvate
WACKER METHOD (FORWARD)
299
Assay: Pyruvate —> Lactate
WROBLEUSKI LA DUE (REVERSE)
300
Measured in WACKER METHOD (FORWARD)
↑ in absorbance at 340 nm
301
Optimal pH in WACKER METHOD (FORWARD)
8.3 – 8.9
302
Commonly used LDH measurement due to:
o Production of positive rate
o Not affected by product inhibition
WACKER METHOD (FORWARD)
303
Measured in WROBLEUSKI LA DUE (REVERSE)
↓ in absorbance at 340 nm
304
Optimal pH in WROBLEUSKI LA DUE (REVERSE)
7.1 – 7.4
305
3x faster but more susceptible to substrate exhaustion
WROBLEUSKI LA DUE (REVERSE) for LDH
306
Has greater affinity of H subunit
α-hydroxybutyrate dehydrogenase (α-HBD)
307
α-hydroxybutyrate dehydrogenase (α-HBD) represent this LDH isoenzyme
LDH 1
(only isoenzyme with complete 4 subunits)
308
CK-MB (>6%) in AMI
Appearance (rise):
Peak:
Normalize:
Appearance (rise): 4-8 hours (EARLIEST)
Peak: 12-24 hours
Normalize: 3 days
309
AST in AMI
Appearance (rise):
Peak:
Normalize:
Appearance (rise): 6-8 hours
Peak: 24 hours
Normalize: 5 days
310
LDH in AMI
Appearance (rise):
Peak:
Normalize:
Appearance (rise): 10-24 hours (late to rise)
Peak: 48-72 hours
Normalize: 10 days (longest to persist)
Normalize after 10 days
311
Secreted by the acinar cells of the pancreas, useful in the digestion process
PANCREATIC ENZYMES
312
Catalyzes breakdown of starch and glycogen via α, 1-6 branching linkages
Amylase (AMS)
313
First enzyme to elevate in acute pancreatitis but nonspecific
Amylase (AMS)
314
Smallest enzyme in terms of molecular weight
Amylase (AMS)
315
Reaction eq. of AMS
CHO --(AMS)--> Maltodextrins
316
Major source of AMS
* Pancreas (acinar cells)
* Salivary gland
317
Minor source of AMS
* Fallopian tube
* Adipose tissues
* Small intestine
* Skeletal muscle
318
AMS is inc. in these conditions
* Acute pancreatitis
* Parotitis
* Renal failure
* Macroamylasemia
319
Earliest nonspecific marker of acute pancreatitis
Amylase (AMS)
320
Inflammation of carotid gland (viral infections), AMS hypersecretion in serum
Parotitis
321
T/F
AMS is normally filtered in glomerulus due to its low mol. weight; normally present in urine. During renal failure (altered kidney filtration) → AMS cannot pass through → reabsorbed → returned back to serum.
T
322
AMS + Ab/Ig; not filtered by glomerulus; reabsorbed
Macroamylasemia
323
aka Salivary Amylase
Ptyalin
324
fast moving; more anodal AMS isoenzyme (lower conc. in serum)
Ptyalin (salivary amylase)
325
slow moving AMS isoenzyme (highest conc. in serum)
Amylopsin (Pancreatic amylase)
326
aka Pancreatic amylase
amylopsin
327
AMYLASE METHODOLOGIES
AMYLOCLASTIC
SACCHAROGENIC
CHROMOGENIC
CONTINUOUS MONITORING
328
Measures disappearance of starch substrate
AMYLOCLASTIC
329
Indicator used in AMYLOCLASTIC method for AMS
Iodine (only react with polysaccharide)
330
Color of Starch-Iodine complex
dark-blue
331
Color of Glycogen-Iodine complex
mahogany-brown (a substitute for starch)
332
T/F
Decrease in color intensity is due to the conversion by AMS of polysaccharide substrates to simpler form (Thus, absorbance measured is also decreased)
T
(amyloclastic method)
333
T/F
In AMYLOCLASTIC method for amylase,
AMS activity = Absorbance
F
AMS activity ∝ Absorbance
334
Measured in this method is the product appearance (liberated reducing sugar)
SACCHAROGENIC
Starch → reducing sugars
335
T/F
In SACCHAROGENIC method for amylase,
AMS = Reducing sugar formed
T
336
Measures the increasing color from production of product acted by AMYLASE
CHROMOGENIC
(uses chromogenic dye fragment)
337
T/F
In CHROMOGENIC method for amylase,
AMS activity = soluble starch-dye fragment formed
T
338
Coupling of several enzyme to monitor AMS activity
CONTINUOUS
MONITORING
339
Measured in CONTINUOUS MONITORING for amylase
↑ in absorbance at 340 nm
340
Several enzymes used in cont. monitoring for ams
AMS
a-glucosidase
HK
G6PD
341
Target of LPS
ester bonds (present in lipids/fats)
342
Hydrolyzes ester linkages of fats to produce alcohols and fatty acids
Lipase (LPS)
343
Hydrolysis of dietary TAG in the intestine to 2-monoglyceride and fatty acids (enhances fat absorption)
Lipase (LPS)
344
Larger molecule of pancreatic enzyme
Lipase
345
Pancreatic enzyme that remains longer in circulation
Lipase
346
Reaction eq. of LPS
Triacylglycerol + 2H2O ---LPS---> 2-monoglyceride + 2 FA
347
Only source of LPS
Pancreas (acinar cells)
348
Early and specific marker of acute pancreatitis
Lipase
(but not as fast as AMS)
349
Assays for lipase
CHERRY CRANDALL
TIETZ
Turbidimetric methods
350
Substrate in Cherry Crandall
50% olive oil (triolein – a purer form of fat reagent)
351
Substrate in Tietz
50% olive oil (triolein – a purer form of fat reagent)
352
Titrating agent in Cherry Crandall
0.4N NaOH (fatty acid titration is done)
353
Titrating agent in Tietz
0.4N NaOH (fatty acid titration is done)
354
Indicator and end color in Cherry Crandall
Phenolphthalein
Pink
355
Indicator and end color in Tietz
Thymolphthalein + Veronal
Blue
356
End point of both Cherry Crandall and Tietz
Fatty Acid (Oleic Acid)
357
Estimation of liberated fatty acids by measuring the amount of light blocked by the insoluble particles in the sample
Turbidimetric methods for Lipase
358
Reagent used in turbidimetric methods for LPS
TAG (hydrophobic, nonpolar, insoluble – turbid soln)
359
T/F
When LPS act
on TAG, it will be converted to a more polar, soluble – clear soln)
**Measured absorbance is DECREASED (inversely proportional)
T
360
AMYLASE in acute pancreatitis
Rise:
Peak:
Persists:
Rise: 2-12 hours (earliest)
Peak: After 24 hours
Persists: 3-5 days
361
LIPASE in acute pancreatitis
Rise:
Peak:
Persists:
Rise: 6 hours
Peak: After 24 hours
Persists: 7 days (longest to persist)
