Chem PPT Flashcards Unit 2
Mass spectrometry is:
A mass spectrometer is:
A mass spectrometer consists of the following
five elements:
A vacuum system is used to prevent collision
of ions during interaction with:
It requires the use of a vacuum from 10-3 to 109
torr one of two sources:
Class of spectrometer where the ions make one
trip through the instrument and then strike the
detector, where they are destructively detected
is
3 types of the above:
Trapping mass spectrometers:
What does MS/MS stand for?
What is also called MS/MS?
What is characterized by a highly selective
detection technique used for quantitative
analyses, characterization and compound
identification?
What does GC-MS stand for?
What is used by the NIST as a definitive
method of qualifying standard reference
materials and assigning certified values to
many clinical analytes?
What is used as confirmatory tests for drug
testing presumptively found to be positive by
immunochemical analyses?
What does LC-MS stand for?
What is used for analysis of polar and non
polar analytes and large molecular weight
compounds?
What is used in screening and confirmation of
genetic disorders and inborn errors of
metabolism?
The study of matter through the formation of
gas phase ions that are characterized using
mass spectrometers by their mass, charge,
structure, and/or physico-chemical properties
An analytical instrument that first ionizes a
target molecule and then separates and
measures the mass-to-charge (m/z) ratio of
these molecules or their fragments
Ion souce, vacuum system, mass analyzer,
detector, computer
Magnetic or electrical fields
Mechanical vacuum or efficient high vacuum
pump
Beam type
Quadrupole mass spectrometer, magnetic
sensor mass spectrometer, time of flight mass
spectrometer
Are based on the trapping of ions to capture
and hold ions for an extended time in a small
region of space
Mass spectrometry/mass spectrometry
Tandem mass spectrometry
Tandem mass spectrometry
Gas chromatography mass spectrometry
GC-MS
GC-MS
Liquid chromatography mass spectrometry
LC-MS
LC-MS
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Chem PPT Flashcards Unit 2
Matrix-assisted laser desorption/ionization –
Time-of-flight mass spectrometry
What is MALDI-TOF-MS used for?
For the detection of specific compounds,
identification of proteins, or identification of
an organism
What are proteins with catalytic properties?
Enzymes
What are protein catalysts of biological origin? Enzymes
What is a substance that modifies and increases Catalyst
the rate of a chemical reaction without being
permanently changed or consumed?
How are individual enzymes identified?
By using the name of the substrate or group
upon which they act and then adding the suffix
-ase.
The type of reaction involved is identified
TRUE
when naming enzymes.
What are the different structures of proteins?
Primary, secondary, tertiary, quaternary
Biological activity is affected by denaturization Elevated temperatures, extremes of pH,
due to what factors?
changes in ionic strength, and chemical
addition
What is one of a group of enzymes catalyzing
Isoenzyme
the same reaction by different molecular
structures and characterized by varying
physical, biochemical, and immunologic
properties?
What is a substance that modifies and increases A catalyst
the rate of a particular chemical reaction
without being consumed or permanently
altered?
What are protein catalyst of biological origin?
Enzymes
What can enzymes convert?
Enzymes can convert substrate molecules that
can be used for the measurement of their
activity
What defined as the quantity of enzyme that
International unit (IU)
catalyzes the reaction of 1 µmol of substrate
per minute?
What is the International System of Units (SI)- Katal unit (KU)
derived unit for catalytic activity?
What unit defined moles converted per second? Katal unit (KU)
How enzyme concentration governs the rate of The rate of reaction is generally proportional to
Enzyme-Catalyzed Reactions?
the amount of enzyme present in the system
and is the basis for the quantitative
determination of enzymes by measurement of
reaction rates.
What is substrate concentration?
A reactant in an enzyme-catalyzed reaction that
binds to the active center of an enzyme.
What does MALDI-TOF-MS stand for?
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Chem PPT Flashcards Unit 2
What is first order kinetics?
What is zero-order kinetics?
What is a constant for a given enzyme acting
under given conditions; the experimentally
determined substrate concentration at which
the enzymatic reaction velocity equals ½ of the
maximum velocity of the enzymatic reaction?
What is a plot of the reciprocal of the velocity
of an enzyme catalyzed reaction (ordinate; yaxis) against the reciprocal of the substrate
concentration (abscissa; x-axis)?
What shows maximum activity in vitro in the
pH range of enzymes in plasma?
What are the activity values observed in pH as
low and high?
What can be controlled by the use of buffer
solutions?
When does enzymes inactivation and
denaturation occurs?
What is the accepted measuring temperature
for enzymes in the clinical laboratory?
What is a substance that reduces the rate of an
enzymatic reaction and is classified as
reversible or irreversible?
What is a structural analog of the substrate that
combines with the free enzyme and competes
with the normal substrate for binding at the
active site?
What is structurally different from the
substrate, it binds at a site on the enzyme
molecule that is different from the substratebinding site?
in an enzymatic reaction, as more substrate is
consumed, the reaction rate declines and
becomes dependent on substrate concentration;
this is a phrase of first-order dependence on
substrate concentration when the rate of the
reaction is proportional to the concentration of
substrate or when the enzyme concentration is
fixed and the substrate concentration is varied.
In an enzymatic reaction, when the reaction
rate is constant at maximum value, the reaction
rate depends only on enzyme concentration and
is independent of substrate concentration; the
rate of reaction is proportional to the zero
power of the substrate concentration.
Michaelis-Menten constant (Km)
Lineweaver-Burk plot
pH range from 7 to 8
Low as 1.5 (pepsin)
High as 10.5 (ALP)
Pronounced effects of pH on enzyme reactions.
Occurs at temperature of 60 degrees Celsius to
70 degrees Celsius.
37 degrees Celsius
Inhibitor
Competitive inhibitor
Noncompetitive inhibitor
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Chem PPT Flashcards Unit 2
What is the small molecule or ion that
increases the rate of an enzyme-catalyzed
reaction by promoting formation of the most
active state of the enzyme or of other reactants
such as the substrate?
When used as analytical reagents for
measuring metabolites, they increase the
specificity of the substance being determined
What is a Coenzyme
NAD and NADH are examples of?
What is a Prosthetic group
What is a poenzyme
H What is a oloenzyme
The rate of an enzyme catalyzed-reaction is
directly proportional to the amount of _____
present in the system
Fixed-time and continuous-monitoring
methods are used to measure ______.
Progress of an enzyme reaction is monitored
by measuring in time the decreasing
concentration of the ______ or the increasing
concentration of the _____.
The amount of substrate transformed into
products during an enzyme-catalyzed reaction
can be measured by?
Immunoassays are used to measure
______instead of _______.
What assay is capable of identifying all
molecules with the antigenic determinants
necessary for recognition by the antibody and
has been found to be significant in the
determination of some digestive enzymes, such
as trypsin, when inactive precursors and
inhibitors of catalytic activity are present in
plasma
When enzymes are used as analytical reagents
for the measurement of metabolites there is a
great increase of _____ for the substance being
determined.
Activator
Enzymes
A small molecule (some contain structures
derived from vitamins) that is required for
some enzyme-catalyzed reactions to occur; is
temporarily or permanently bound to the
enzyme
Coenzymes
a tightly bound, non-peptide structure required
for the activity of an enzyme
The protein component of an enzyme
An active enzyme formed by combination of a
coenzyme and an apoenzyme
active enzyme
reaction rates
substrate, product
Spectrophotometry
Fluorometry
Chemiluminescence
protein mass, catalytic activity
Immunoassay
Specificity
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Chem PPT Flashcards Unit 2
Enzymatic methods that allow all substrate
Equilibrium methods
present in a reaction mixture to be completely
converted into a measurable product before
measurement, are referred to as
Enzymatic methods which demonstrate that the Kinetic Methods
change in substrate concentration over a fixed
time interval is directly proportional to its
initial concentration
What is an immunoassay?
An assay based on the reaction of an antigen
with an antibody specific for the reaction.
What is antigen?
1) Any material capable of reacting with
an antibody without necessarily being
capable of inducing antibody
formation.
2) Analyte being detected by the
immunoassay.
What are antibodies?
An immunoglobulin (Ig) class of molecule
(IgA, IgG, IgM) that bins specifically to an
antigen or hapten
Which immunoglobulin is the most widely
Immunoglobulin G (IgG)
used reagent?
What is an Immunogen?
A protein of a substance coupled to a carrier
cable of introducing an immune response.
What is a hapten?
A chemically defined determinant that by itself
will not stimulate an immune response. It is an
incomplete antigen
What is affinity?
1) It refers to the thermodynamic quantity
defining the energy of interaction of a single
antibody-combining site and its corresponding
epitope on the antigen.
2) It is a property of the substance bound
(antigen)
What is avidity?
1) Refers to the overall strength of the
binding of antibody and antigen and
includes the sum of the binding
affinities of all individual combining
sites on the antibody.
It is a property of the binder (antibody)
What are factors affecting antigen-antibody
1) Binding forces
binding?
2) Reaction mechanism
3) Precipitin reaction
4) Chemical factors:
1)Ion species and ionic strength effects
2) Polymer effect
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Chem PPT Flashcards Unit 2
In the binding forces, what are 3 major
contributing forces?
1) Electrostatic van der Waals-London
dipole-dipole interactions
2) Hydrophobic interactions
3) Ionic coulombic bonding
1) Phase 1:Initial reaction of a multivalent
antigen and a bivalent anitbody
2) Phase 2: Subsequent growth of the
complexes
3) Phase 3: Precipitation of the complex
before after a critical size is reached.
In the reaction mechanism, what are the 3
phases?
These soluble complexes have all antigenic sites
that are covered with antibody, and lattice
formation is inhibited.
This insoluble complexes (optimal proportion) state
occurs when 2 to 3 antibody molecules are present
for each antigen molecule; produces maximum
lattice formation and therefore maximum
precipitate.
These soluble complexes have all antigenic sites
that are saturated by antigen. Triplets (2 antigen + 1
antibody) are maximum size attained by particles.
no precipitate is formed.
Anion and cationic salts can affect the binding of
antibodies by what 2 chemical factors?
What are 3 examples of polymer effect in terms of
having high molecular weight, high degree of
linearity and aqueous solubility that can
significantly increase the rate of immune complex
growth and enhances precipitation of immune
complex?
What are 2 types of qualitative methods regarding
passive gel diffusion that use semisolid medium
(e.g., agar agarose), and that the initial
concentration of antigen and antibody are critical?
The term for an analytical technique that combines
separations of antigens by electrophoresis with
immunodiffusion against an antiserum?
Also known as two-dimensional
immunoelectrophoresis, what is the term describing
to drive the antigen into a gel containing antibodies
specific for the antigens of interest.
This technique involves an electrophoresis step,
followed by transfer of the separated proteins onto
on overlying strip of nitrocellulose or nylon
membrane by a process called?
This electrophoresis step involves DNA blotting
onto a membrane.
This electrophoresis step involves RNA blotting
onto a membrane.
Antibody Excess
Equivalence Zone
Antigen Excess
1.
2.
1.
2.
3.
Ion species
Strength effects
Dextran
Polyvinyl alcohol
polyethylene glycol
Raidal immunodifussion (RID) (single
immunodiffusion) and Ouchterlony technique
(double diffusion)
Immunoelectrophersis (IEP)
Crossed immunoelectrophoresis
Electro-blotting.
Western/Southern blotting
Northern RNA
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Chem PPT Flashcards Unit 2
What does RID stand for?
What is the definition for RID?
Radial Immunodiffusion
A passive diffusion method which a
concentration gradient is used for a single
reactant
During RID, what happens when antibody is
A well defined ring of precipitation around the
added?
well indicates presence of antigen
What is also known as the “rocket” technique? Electroimmunoassay
What is the definition of electroimmunoassay? A type of immunoassay where a single
concentration gradient is established for the
antigen and an applied voltage is used to drive
the antigen from the application well into a
homogenous suspension of antibody in the gel
Turbidimetric and Nephelometric assays are
The rate of formation of immune complexes in
convenient techniques to measure what?
vitro using their light scattering abilities
What are some examples of radioactive and
Competitive, Noncompetitive, and
non-isotopic labels?
heterogeneous immunoassays
What is a Heterogeneous immunoassay?
Immunochemical assay that require separation
from the bound label
What does RIA stand for?
Radioimmunoassay
Define RIA
Used radioactive isotopes of iodine, 125I and
131I and tritium (3H) as labels
What does EIA stand for?
Enzyme Immunoassay
What does ALP stand for?
Alkaline phosphatase
What does HRP stand for?
Horseradish peroxidase
Give some examples of enzymes that are used ALP, HRP, Glucose-6-dehydrogenase, Bin EIA?
galactosidase
What are 3 Examples of Enzyme Immunoassay Enzyme-Linked Immunosorbent Assay
(EIA)?
(ELISA)
Enzyme Multiplied Immunoassay Technique
(EMIT)
Clone Enzyme Donor Immunoassay (CEDIA)
What does Fluoroimmunoassay (FIA) use?
Uses a fluorescent molecule
What is fluorescent used as in FIA?
as an indicator label to detect and quantify
immunological reactions
What is Chemiluminescent immunoassay?
a chemiluminescent molecule, such as
acridinium
What is Chemiluminescent immunoassay used is used as an indicator label to detect and
for?
quantify immunological reaction
What is Electrochemiluminescent
an elecrochemiluminescent molecule, such as
immunoassay?
ruthenium
What is electrochemiluminescent
is used as an indicator label to detect and
immunoassay used for?
quantify immunological reaction
What is one definition of automation?
The process whereby an analytical instrument
performs many tests with only minimal
involvement of an analyst
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Chem PPT Flashcards Unit 2
What is another definition of automation?
What is Random-access analysis?
What is Continuous-flow analysis?
What is Single-channel analysis?
What is Multiple-channel analysis?
What is multiple-channel analysis similar to?
What is Parallel analysis?
What is Discrete analysis?
How many types of the specimen identifiers?
What are the technologies used for automatic
identification and data collection?
What are the delays in preparation of
specimen?
What are the methods of delivery specimen?
How reliable is a courier service?
How does pneumatic tube system transport?
controlled operation of an apparatus, process,
or system by mechanical or electronic devices
without human intervention
analyses are performed on a collection of
specimens sequentially and each specimen is
analyzed for a different selection of tests.
it specimen in a batch passes through the same
continuous stream at the same rate and is
subjected to the same analytical reactions
each specimen is subjected to a single process
so that only results for a single analyte are
produced.
each specimen is subjected to multiple
analytical processes so that a set of test results
is obtained on a single specimen.
Similar to random access analysis
all specimens are subjected to a series of
analytical processes at the same time and in a
parallel fashion
the sample is aspirated into the sample probe
and then is delivered, often with reagent,
through the same orifice into a reaction cup or
another container.
These are: specimen labels, serial number, part
number, manufacturer, and assigned patient
number
There are technologies used for automatic
identification and data collection as barcoding,
optical character recognition, magnetic stripe
and magnetic ink character recognition, voice
identification, radiofrequency identification
touchscreen, light pen, handprint tablets,
optical mark reader, smartcards.
-Clotting of blood in specimen collection tubes
-Subsequent centrifugation
-Preparation of serum and plasma
-Transfer of serum the secondary tubes
Courier service, pneumatic tube systems,
electric track vehicles, and mobile robots.
Courier service is usually reliable but costly
and may require batches, but risk of specimen
breakage or loss
Pneumatic tube systems provides rapid
specimen transportation, risk of misrouting or
hemolysis
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Chem PPT Flashcards Unit 2
How does specimen delivery as an electric
track vehicle?
How does specimen delivery as a mobile
robot?
Once collected into a red top tube what is the
first thing one must do before specimen
preparation for routine tests can begin?
Once a red top tube is allowed to clot what is
the next step in specimen preparation for
routine testing?
Once a red top tube has been allowed to clot
and centrifuged until separated what is the next
step in preparing the specimen for routine
testing?
What are two common ways to get specimens
from site of collection to the lab?
What are the pros and cons of currier service.
What are the pros and cons of the pneumatic
tube system?
What is an electric track vehicle?
Larger capacity than pneumatic tube systems
makes use of dry ice or refrigerator packs
possible
Easily adapted and programmable, but need for
batching and personnel to load robots.
Red top tubes have no anticoagulant and
therefore must be allowed to clot before
preparing them for routine tests.
Centrifuge the specimen for at 1200g for 10
minutes
then separate serum from the cells, labeling the
secondary tube appropriately.
Currier service and Pneumatic tube system
Pros•Reliable
•best way to get specimens to the lab if from a
site across cities or even states.
Cons•costly
•require multiple specimen batch collections
and deliveries throughout the day
•risk of specimen breakage
•risk of specimens becoming too warm or cold
if left outside in collection box.
Pros•Rapid specimen transport
Cons•Risk of specimens being sent to the wrong
place and becoming lost
•tube traffic may occur causing tubes to collide
possibly shutting the system down
-this could cause hemolysis or leaking
specimens
A form of specimen delivery similar to the
pneumatic tube system in which specimens are
carried on a track, not within a tube system,
and in a specimen carrier box which can be
larger and more versatile in the type of
specimen being transported. Also can utilize
ice packs or dry ice to keep specimens under
the correct temperature conditions.
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Chem PPT Flashcards Unit 2
Mobile robots are being utilized for what
laboratory service?
Having instrumentation that is capable of
sampling directly from a primary tube or
centrifuging and aliquoting the plasma or
serum into a secondary tube on its own will
reduce what and increase what?
Three processes of removing protein and other
interferents are?
In immunochemistry, instrumentation is
capable of separating bound and free fractions
via which methods?
Define carryover in regards to specimen
transport during analytical testing
What sample introduction system utilizes a
peristaltic pump?
What sample introduction system utilizes
pipets and is highly automated.
Instrumentation must have what capability to
store reagents?
Instrumentation must have this to identify
patients and tests to be performed on each
sample.
Rate of transport of reaction mixture through
the system to the measurement station affects
what?
on-time events on reagent addition (or
activation) relative to measurement will affect
what?
What are the five main means of mixing
reactants in instrumentation?
Some temperature control and efficient heat
transfer methods seen in instrumentation are…
Mobile robots are being used for specimen
transport within hospitals. They are loaded
with a schematic blue print of the hospital and
can be programmed to go from area to area to
collect and transport specimens to the proper
lab.
Reduce-Specimen evaporation, turn around
time, risk of disease transmission, specimen
contamination
Increase-efficiency
Dialysis
Column chromatography
Filtration
•coated beads
•coated tubes
•microtiter plates
•magnetic and nonmagnetic microparticles
The transport of a quantity of analyte or
reagent from one specimen reaction into and
contaminating a subsequent one.
Continuous-flow system
Discrete systems
Refrigeration
Bar code scanner
Timing of the reaction
Timing of reaction
Forceful dispensing
magnetic stirring
vigorous lateral displacement
a rotating paddle
the use of ultrasonic energy
Air baths
water baths
contact with warm plates
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Chem PPT Flashcards Unit 2
Analyzers can employ different types of optical Photometers
measurement devices, list five.
Spectrophotometers
Reflectance photometers
Fluorometers
Luminometers
One way computers have impacted the clinical all functions are performed uniformly, in a
laboratory is:
repeatable manner and in the correct sequence
The computer controls the electromechanical
operation of the analyzer. This means?
Having the computer control the automated
reproducibility
equipment, calculation of results and
monitoring of operation will increase the ___
of results.
One way computers have impacted the clinical Computers can monitor instrument functions
laboratory is:
for correct execution and record the sites and
Improved acquisition, processing and storage
nature of a malfunction so the tech can
of operational data from the analyzers. This
troubleshoot and repair the problem.
means?
One way computers have impacted the clinical The instrument operator is alerted to changes
laboratory is:
in analyzer by the computer directly (ie
Computer interfacing with the instrument
problems, errors, low reagents, etc) so the tech
operator. Explain this.
can troubleshoot the problem, if needed, and
the issue can be fixed.
One way computers have impacted the clinical This allows for LIS and instrument to be linked
laboratory is:
so results can be transferred directly to the
Computer facilitation of communication with
patients records.
mainframe computer. Explain this.
One way computers have impacted the clinical This allows the tech to work on multiple
laboratory is:
analyzers simultaneously
Computer integration of the monitoring
functions of one or more analyzers with a
single workstation
Explain this?
Instrument cluster
When several different instruments are set up
to be controlled by one tech.
Work cell
A robotic specimen processor monitored by a
tech.
The work cell performs what tasks?
Centrifuge, aliquot, label, transport and store
specimens
Optical measurement devices includes:
- Photometers
- Spectrophotometers
- Reflectance photometers
- Fluorometers
- Luminometers
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Chem PPT Flashcards Unit 2
Impact of integration of computers into
automated analyzers and analytical systems
are:
Features of advanced analytical systems
include:
Work Cells consist of:
Automated Specimen Transport include:
Automated Speciment Processing include:
Name the three approaches that have been used
to automatically sort specimen?
Name the automated specimen modules which
store specimens refrigerated in specific
locations that are logged into a database and
Adaptation of other industries’ systems to the
laboratory?
How many types of conveyor sorting system
have been used?
What is advantage of conveyor system in the
first approach?
-
Computer control of the
electromechanical operation of the
analyzer
- Computer acquisition, processing and
storage of operational data from the
analyzers
- Computer interfacing with the
instrument operator
- Computer facilitation of
communication with mainframe
computers
- Computer integration of the monitoring
functions of one or more analyzers with
a single workstation
• Workstations
• Equipped for a defined task
• Manual specimen transport
• Built-in diagnostics
• Instrument clusters
• Control of several instruments
by one technologist
• To reduce labor costs
- Robotic specimen preparation
(Centrifuge, aliquot, label, transport
and store specimens)
- Technologist as monitor
- Conveyor belts
- Robot arms
- Stand-alone specimen-processing systems
- Integrated and modular automation system
- Role of process control software
- Workload
- 13 components of integrated systems
1) A conveyor belt,
2) automated sorter using racks and
3) stand-alone sorters.
Automated specimen storage and retrieval
Three types
Direct sampling from a conveyor track in a
loop configuration eliminates the need for
separation equipment to sort specimens.
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Chem PPT Flashcards Unit 2
What is disadvantage of conveyor system in
the first approach?
What is the second approach of automated
processing conveyor system?
What is the third approach of automated
processing conveyor system?
What are Practical Considerations that
influence a laboratory’s decision to automate
part or all of its operations?
List evaluation of requirement in practical
considerations ?
List Problems of integration in practical
considerations?
What is Point-of-Care Testing (POCT)?
What are other terms for POCT?
In what types of environments would POCT be
employed?
What are some advantages to POCT?
What are the main objectives when designing a
Point-of-Care instrument?
What are key components of POCT device
design?
Direct sampling may limit the rate of specimen
movement on the track to the sampling speed
of the slowest workstation.
It sorts specimen into group according to their
destination in the laboratory such as for
hematology or chemistry.
The sorter is integral to the conveyor system,
and specimens are sorted as they are
transported.
Evaluation of requirements and,
Problems of integration
Must be first step in decision to automate
Mapping of workflow (Box 16-4)
80% rule of thumb
Visits with vendors
CLSI standard AUTO03-A
Functional control model
Device integration
Integration of process controllers and software
It is a method of clinical laboratory testing in
which the analysis is performed at the site
close to where the healthcare is provided to the
patient.
Other terms used to describe POCT included:
bedside, near patient, physician's office, extra
laboratory, decentralized, off-site, ancillary,
alternative site and unit-use testing.
POCT would be encountered at home,
community pharmacy, retail clinics, and more
Reduced turnaround time, improved patient
management, reduction in process error and
reduction in time delays
The main objectives of the design are to:
1. Enable the required reaction to take place
that facilitates recognition of the analyte of
interest
2. Ensure reliable performance of the device
over a specified period of time
3. Minimize the risk of error associated with
the use of the device
Operator interface, bar code identification
systems, sample and reagent delivery
mechanisms, reaction cell, sensors, control and
communications systems, data management
and storage
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Chem PPT Flashcards Unit 2
What are the POCT devices classified as?
Dipsticks, complex strips, and immunostrips
are examples of what type of in vitro POCT
devices?
What are some single-use quantitative
cartridge and strip tests with a monitoring
devices?
What are the analytical principles of in vivo
POCT technology?
What are the analytical principles of ex vivo
POCT technology?
What are the analytical principles of
noninvasive POCT technology?
What are the analytes used in in vivo POCT
technology?
What are the analytes used in ex vivo POCT
technology?
What are the analytes used in noninvasive
POCT technology?
How to POCT devices interface with
laboratory information systems?
What does CIC stand for?
What is CIC?
What are POCT01-A2 user requirements?
What is the formula for an amino group?
What is the formula for a carboxyl group?
What is an amino acid?
Peptides and proteins are made of which basic
structural unit?
What is the definition for ampholyte?
What is another term for ampholyte?
Amino acid metabolism is synthesized from
what?
What is the daily requirement for protein
intake for adults?
What is the primary source of amino acids for
protein synthesis?
What is another source of free amino acids?
What happens when the is an absence of
protein uptake?
In vitro, in vivo, ex vivo, minimally invasive
Single-use qualitative strip or cartridge and/or
strip devices
Glucose monitoring device, cardiac markers,
allergy tests, fertility tests, drugs of abuse
Optical fluorescence and electrochemistry
Optical fluorescence and electrochemistry
Electrochemistry/ iontophoresis, and
multi wavelength spectrophotometry
pH, blood gases, subcutaneous glucose
pH, blood gases, electrolytes, glucose
Transcutaneous glucose, bilirubin
Electronic transfer of data from the analyzers
to the LIS and into the patient's electronic
medical record
Connectivity Industry Consortium
Developed point-of-care communication
standards which facilitate linking of devices to
information management system
CLSI connectivity standards to ensure that the
POCT devices meet critical user requirement
NH2
COOH
Organic compounds containing both amino
and carboxyl functional groups
Amino acids
A zero net charge as the negative and positive
charge balance at a neutral pH
Zwitterion
Alpha-keto acids
About 0.8 g/kg body weight
Dietary protein
Endogenous protein
Breakdown of muscle protein serves as a
source of amino acids
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Chem PPT Flashcards Unit 2
What are essential amino acids?
Where are essential amino acids usually
obtained from?
How many essential amino acids are there?
Name all the essential amino acids
What is Kwashiorkor?
How is Kwashiorkor obtained?
How is Kwashiorkor characterized?
Deficiency of both calories and protein is
known as?
What causes marasmus?
What is caused by marasmus?
What is the clinical implication known as
primary aminoaciduria?
Where can the defect be located?
What is secondary aminoaciduria?
What is the main clinical use of amino acid
analysis?
What is the most abundant amino acid?
What type of specimens are used for the
analysis of amino acids?
When do amino acids exhibit high diurnal
variations?
When do amino acids exhibit low diurnal
variations?
The analysis of amino acids require a normal
diet for how many days before collection?
True or False?
Blood and urine specimen should be collected
simultaneously
True or False?
All medications don’t have to be noted
Why should specimen be processed rapidly
and frozen rapidly?
Amino acids that are not synthesized by
humans and are essential dietary constituents
for maintaining health or growth
Meat, milk, eggs, and fish
8
Isoleucine, leucine, lysine, methionine,
phenylalanine, threonine, tryptophan, and
valine
A diet with adequate calories with low protein
intake
Through malnutrition due to protein
deficiency
Decreased serum albumin, edema, ascites,
growth failure, immune deficiency, and
apathy
Marasmus
Protein-calories malnutrition
Generalized muscle wasting
A disease due to an inherited enzyme defect
In the pathway which a specific amino acid is
metabolized or in a transport system for an
amino acid
Disease of an organ such as the liver which an
active site of an amino acid metabolism, or
generalized renal tubular dysfunction
Detection and monitoring of inherited
disorders
Glycine
Heparanized plasma, urine, and CSF
During mid-afternoon
In the early morning
2-3 days
True
False, all medications need to be noted
To preserve glutamine
15
Chem PPT Flashcards Unit 2
Why does plasma need to be separated from
whole blood rapidly?
How do most procedures entail removal of
proteins?
What does TLC stand for?
Name 3 screening tests for the analysis of
amino acids
What does LC-MS/MS stand for?
What does CE stand for?
What does GC stand for?
Name quantitative tests for the analysis of
amino acids
What is a protein?
How are amino acids linked?
Proteins contain a group of complex organic
compounds that contain which elements?
Which is the characteristic element?
What is proteome?
Proteome defines the filed of?
A peptide is a compound that consists of how
many amino acids linked in a chain?
Define a peptide bond
True or False
Proteins act as structural components
Proteins facilitate catalysis of what kind of
reaction?
How can proteins generate energy?
True or False
Proteins can serve as ion channels and pumps
What kind of molecules can proteins act as?
True or False?
Proteins can effect immune defense
Proteins can act as what for intercellular
communication?
True or False?
Proteins constitute signaling networks for
extracellular communication
So homocysteine can be measured accurately
By precipitation or ultrafiltration
Thin-layer Chromatography
TLC, Guthrie test, and liquid
chromatography-tandem mass spectrometry
Liquid chromatography-tandem mass
spectrometry
Capillary electrophoresis
Gas chromatography
Ion exchange chromatography with postcolumn reaction with ninhydrin, reversedphase high-performance liquid
chromatography, LC-MS/MS, CE, and GC
A polymer of amino acids
By peptide bonds with a specific sequence
that folds into a defined structure
Carbon, hydrogen, oxygen, nitrogen, and
usually sulfur.
nitrogen
The total complement of proteins present at
the time in a call or cell type expressed by the
genetic material of an organism
proteomics
Two or more via peptide bonds
The amide bond that is formed between the
carboxyl group of one amino acid and the
amino group of another
True
Chemical reactions including synthesis of
DNA, RNA and proteins
Through electronic transfer
True
Carrier molecules
True
Receptors, hormones, and cytokines
False, proteins constitute signaling networks
for intercellular communication
16
Chem PPT Flashcards Unit 2
Proteins are covalently linked how?
the α-amino group of one amino acid is link
covalently bonded to what?
What is the amino acid with the free amino
group in the peptide termed?
What is the amino acid with the free carboxyl
group termed?
Where does protein synthesis begin?
What are peptides named from?
What are Primary structure?
by peptide bonds
with the α-carboxyl of a second amino acid
N-terminal residue
C-terminal residue
with the N-terminal residue
the N-terminal residue
sequence of amino acids in a peptide or
protein
deoxyribonucleic acid (DNA).
What is the sequence of amino acids
comprising the polypeptide chain that is
genetically encoded?
The following are extra questions from 18:22-38
What does the secondary structure of a protein The recurring formations along the
describe?
polypeptide chain caused by intramolecular
forces such as hydrogen bonding
What are the three parts of the secondary
α-helix,β-pleated sheets, and random coil
structure?
What does random coil refer to?
The segments of protein that lack secondary
structure.
What does tertiary structure of a protein refer
The folding of the chain of amino acids into a
to?
three dimensional structure
How can the tertiary structure be described?
Folded, three-dimensional conformation of
the protein that is stabilized by both covalent
bonds and non-covalent forces mainly
involving amino acid R group
What has a great effect on a proteins tertiary
The proteins environment
structure?
What can cause denaturation of proteins?
Mechanical agitation, heat or extreme chemical
treatment
What does quaternary structure of a protein
The association of multiple peptide bonds
refer to?
Association of what forms a functional protein Two or more polypeptide chains, or subunits.
complex?
What happens to molecular size of proteins
Size increases
when they are denatured?
How can proteins be separated?
gel filtration chromatography
gradient pore gel electrophoresis
ultracentrifugation
polyacrylamide gel electrophoresis
Mass spectrometry
What factors can affect protein solubility?
pH, ionic strength, temperature, dielectric
constant of the solvent.
17
Chem PPT Flashcards Unit 2
Is albumin soluble in water?
What else is albumin soluble in?
What is albumin insoluble in?
What is globulin soluble in?
Is globulin soluble in water or saturated and
concentrated salt solution?
What is the solubility of albuminoids?
What enables proteins separation by
electrophoresis, isoelectric focusing, and ion
exchange chromatography?
On what basis does differential adsorption
separate polypeptides and other molecules?
What are two examples of differential
adsorption?
What is affinity chromatography?
What may Affinity purification of antibodies
or proteins be used as?
What is density gradient ultracentrifugation
used for?
What are the physical methods of quantitative
analysis?
What are two other types of quantitative
analysis?
What are the methods of qualitative alalysis of
proteins?
What is the Kjeldahl method in measurement
of total protein?
What is the standard reference for N2 in the
Kjeldahl method?
What is the principle of biuret method?
Yes
dilute and moderately concentrated salt
solution (ammonium sulfate)
saturated and concentrated salt solution
hydrocarbon solvents
weak salt solution
No
Insoluble in most common reagents
The variable charge and isoelectric points
On the basis of their affinity for a
hydrophobic stationary phase
Liquid chromatography, and reversed-phase
chromatography
Highly specific molecular interactions to bind
a protein or a peptide to a stationary phase,
while other proteins and peptides are removed
A preparative technique to prepare specific
antibodies or proteins as assay components
Used to separate some protein complexes that
have a different density when compared with
most proteins, such as lipoproteins with low
density but with high lipid content
Dye binding
Direct absorbance measurements
Measurement of protein ligands
Selective precipitation
Mass spectrometry
Activity measurements and immunoassays
Electrophoresis
Chromatography
Genetic analysis
Functional assays
Mass spectrometry
A standard reference method based on
nitrogen determination
1 gram N2 = 6.25 grams of protein
Cupric ions in the alkaline copper tartrate
reagent react with peptide bonds to form a
blue violet complex proportional to the
number of peptide bonds present.
18
Chem PPT Flashcards Unit 2
What does the analysis depend on in the biuret
method?
What is the composition of the biuret reagent?
What is direct spectrophotometric
measurement of total protein?
What is the principle of the dye-binding
method for measuring total protein?
What are the commonly used dyes?
For measurement of total protein, what is the
method of direct spectrophotometric?
How is the dye-binding method for
measurement of total protein?
What dyes are commonly used?
What is the Folin-Ciocalteu (Lowry) method?
What is a refractometry method for
measurement of total protein?
How are the turbidimetric and nephelometric
methods based on the formation of aggregates
or precipitation when every agent?
What are precipitation reagents?
What is the formula to calculate for normal
A/G ratio?
What is the A/G ratio if albumin is greater than
globulin?
What is the A/G ratio if globulin is greater than
albumin?
What is Serum Protein Electrophoresis (SPE)
performed at pH 8.6 in the low ionic strength
buffer?
The presence of peptide bonds
Alkaline copper tartrate as a source of cupric
ions
Rochelle salt (NaK tartrate) is a complexing
agent to keep copper in solution
Sodium hydroxide for alkalinity
Potassium iodide which prevents
autoreduction of copper
Measurement of protein content using
absorbance of ultraviolet (UV) light at 200 to
225 nm and 270 to 290 nm.
Dye binds to protonated amine group of
amino acids with absorption at 595 nm
Pyrogallol red (for urine and CSF)
Coomassie brilliant blue
Measurement of protein content using
absorbance of ultraviolet (UV) light at 200 to
225 nm and 270 to 290 nm.
Dye binds protonated amine group of amino
acids with absorption at 595 nm
Pyrogallol red (for urine and CSF)
Coomassie brilliant blue
That is a measurement of refractive index to
measure total solutes in urine specimen
That is a measurement of refractive index to
measure total solutes in urine specimen
These are added to lower protein solubility or
when an antibody is added to a protein
Trichloroacetic acid (TCA)
Sulfosalicylic acid (SSA)
Sulfosalicylic acid combined with sodium
sulfate
Benzethonium chloride
Benzalkonium salts under alkaline conditions
Normal A/G ratio = 1 – 2.5:1
A/G = Albumin/Globulin: 1
A/G = 1: Globulin/Albumin
(Inverted A/G ratio)
Proteins with a negative charge at pH 8.6 move
toward the anode (positive pole)
Proteins with a positive charge moved
towards the cathode (negative pole)
19
Chem PPT Flashcards Unit 2
How many types of supporting media in SPE
are there?
What are stains used in serum protein
electrophoresis?
What are silver stains used for?
These are papers, garose gel, cellulose acetate,
starch gel and acrylamide gel
Amido black and ponceau S
Coomassie brilliant blue
Silver stains are used for nanogram
concentrations of proteins and nucleic acid,
they are more sensitive than Coomassie blue
stain
They are also used for CSF and urine proteins,
shades of green, yellow, blue and red
What is the intensity of staining individual
It is proportional to the amount of protein in a
bands of protein generally?
band
What is a quantitative assessment in SPE
Quantitative assessment is done by
done?
densitometry of stained hands
In electrphoresis, when the protein is too high a Each band in the stained pattern should be
concentration, what is each band appearance?
uniformly colored; holes or doughnut-like
appearance within a band occurs
How is the order of migration in SPE?
The order is following Albumin, Alpha1globulin, Alpha2-globulin, Beta-globulin,
Gamma globulin
What is a serum preferred?
Serum is the preferred specimen
Where extra fraction will show if plasma is
It will show in the beta-gamma region, and
used?
fibrinogen is a beta globulin
What is the main application of SPE?
The main current application of SPE is for the
detection or quantification of monoclonal
immunoglobulins that occur in disorders, such
as multiple myeloma.
How does monoclonal immunoglobulins
Monoclonal immunoglobulins usually appear
usually appear?
as sharp bands in the gamma (γ) or beta (β)
region
How different between monoclonal
Monoclonal immunoglobulins differ from
immunoglobulins and normal
normal immunoglobulins, which migrate as a
immunoglobulin’s?
diffuse zone, mainly in the gamma region
What are 5 types of supporting media for
Paper
serum protein electrophoresis (SPE)?
Agarose gel
Cellulose acetate
Starch gel
Acrylamide gel
3 Stains used for SPE’s include ?
Amido black
Ponceau S
Coomassie brilliant blue
What silver stain used for?
- nanogram concentrations of proteins and
nucleic acid
- more sensitive than Coomassie blue stain
- used for CSF and urine proteins
- shades of green, yellow, blue and red
20
Chem PPT Flashcards Unit 2
Serum Protein Electrophoresis (SPE)
What is the order of migration?
What if plasma is used as a specimen?
What is the preferred specimen in protein
electrophoresis?
What is the main current application of SPE?
What do Monoclonal immunoglobulins usually
appear as?
What do Monoclonal immunoglobulins differ
from normal immunoglobulins?
In regards to electrophoresis, the abbreviation
IFE stands for?
With the application of specific antibodies, IFE
can be directed against which immunoglobins/
light chains?
IFE is used on what type of gel?
Electrophoretic separation of
urinary proteins is used to detect what?
What is another term for the presence of free
monoclonal immunoglobulin light chains?
The detection of Bence-Jones proteins can be
indicative of what disease?
Fill in the Blank:
relies on the separation of ions in an
electrical field in a vacuum?
In mass spectrometry, separation of molecules
is purely based on what?
-the intensity of staining individual bands of
protein generally is proportional to the amount
of protein in a band
-quantitative assessment is done by
densitometry of stained hands
-In electrophoresis, each band in the stained
pattern should be uniformly colored; holes or
doughnut-like appearance within a band occurs
when the protein is too high a concentration.
Remedy : Dilute elevated specimens before
re-running the electrophoresis
1.Albumin
2.Alpha1-globulin
3.Alpha2-globulin
4.Beta-globulin
5.Gamma globulin
- extra fraction will show in the beta-gamma
region
- Fibrinogen is a beta globulin
Serum
is for the detection or quantification of
monoclonal immunoglobulins that occur in
disorders, such as multiple myeloma.
sharp bands in the gamma(γ) or beta(β) region
normal immunoglobulins, which migrate as a
diffuse zone, mainly in the gamma region
Immunofixation Electrophoresis (IFE)
Immunoglobins: IgG, IgM, IgA
Also kappa (κ) and lambda (λ) light chains
Agarose gel
The presence of free monoclonal
immunoglobulin light chains or other lowmolecular-mass proteins
Bence-Jones Proteins
Multiple Myeloma
Mass Spectrometry
Mass-to-charge (m/z) ratio
21
Chem PPT Flashcards Unit 2
In what case would a mass spec. be ideal?
What is an Acute Phase Response (APR)?
Proteins affected or increased by APR are
known as?
Known Positive APPs include:
Proteins not affected or decreased by APR are
called?
Known Negative APPs include:
What is Prealbumin (Transthyretin)?
Ideal for highly precise qualitative analysis of
peptides
A systemic response to infection, tissue injury
or inflammatory processes resulting in fever,
increased WBC count, and changes in the
concentration of many plasma proteins.
Positive acute phase proteins (APPs)
 α1-antitrypsin
 Fibrinogen
 α1- acid glycoprotein
 C-reactive protein
 Haptoglobin
 Serum amyloid
 Ceruloplasmin
 C4, C3
Negative acute phase proteins (APPs)
 Transthyretin
 Albumin
 Transferrin
A transport protein of thyroid hormones and
RBP
A hollow core containing T3 and T4 binding
sites
Negative cooperativity
Prealbumin/ Transthyretin is composed of four
identical subunits that associate to form what?
Binding of one hormone molecule decreases
the binding affinity of the second so that only
one site is normally occupied. This is known as
what?
Where is prealbumin/ transthyretin
In the liver and to a lesser extent in the choroid
synthesized?
plexus of the central nervous system, accounting for
the relatively high concentration in CSF
Synthesis of prealbumin/ transthyretin is
stimulated by what hormones and drugs?
Prealbumin/ Transthyretin is often used as
indicator of?
Prealbumin/ Transthyretin is increased during
what conditions?
Prealbumin/ Transthyretin is decreased during
what conditions?
Synthesis is stimulated by glucocorticosteroid
hormones, androgens and many nonsteroidal
anti-inflammatory drugs (NSAIDs) including
aspirin
Protein Nutrition
 With corticosteroid or NSAID therapy
Hodgkin Disease
 Inflammation and malignancy
 Cirrhosis of the liver
 Protein-losing diseases of the gut or
kidneys
22
Chem PPT Flashcards Unit 2
What is the migration pattern for prealbumin/
transthyretin on Serum Protein Electrophoresis
(SPE)?
Name 2 methods that can measure prealbumin:
What is Retinol-Binding Protein (RBP)?
What element is required for the synthesis of
RBP?
What is required for RBP to transport out of
the Golgi apparatus?
Where is RBP absorbed in the body?
Fill in the Blank:
Serum concentration of RBP is
chronic renal disease.
RBP is increased with?
What is the most abundant protein in the
plasma from mid-gestation until death?
Albumin is a major protein component of most
extra vascular body fluids including?
Albumin is increased during?
Albumin is decreased in?
Immunoturbidimetric or
immunonephelometric methods
A transport protein bound to prealbumin
Zinc
Retinol
It is absorbed by the proximal renal tubular
cells and catabolized
Increased
in
RBP is decreased in?
Albumin is synthesized by?
Albumin is a major transporter of?
Migrates as a minor band anodal to albumin
 Corticosteroid
 NSAID therapy
 And in Hodgkin Disease
Liver disease, protein malnutrition, and acute
phase response (APR)
Albumin
 CSF
 Interstitial fluid
 Urine
 Amniotic fluid
The hepatic parenchymal cells
Free fatty acids, bilirubin, calcium, thyroid and
steroid hormones, drugs and thiol-containing
compounds
 Dehydration
 Prolonged application of the tourniquet
 Specimen evaporation before analysis
 Analbumemia – a rare genetic
deficiency with plasma albumin
concentration less than 0.5 g/L
 Inflammation – Albumin is a negative
APP. Acute and chronic inflammation
are common causes of
hypoalbuminemia
 Hepatic disease
23
Chem PPT Flashcards Unit 2
How does hepatic disease decrease the
concentration of albumin?
How is albumin decreased in urinary loss?
How is albumin decreased in Gastrointestinal
loss?
How is albumin decreased in protein energy
malnutrition (Marasmus)?
How is albumin decreased in burn injury?
How is albumin decreased in Edema and
ascites?
Whsat are two ways for Albumin
Determination?
What is High-Density Lipoprotein?

Loss of parenchymal cells function by
50%
 Nutritional deficiency
 Increased distribution into the
extravascular space
 Direct inhibition of synthesis by toxins
such as alcohol
- Small increases in albumin excretion to
>30 mg/d indicate early stages of
glomerular or tubular injury and risk of
progression to more severe kidney
disease (Microalbuminuria)
- Severe glomerular injury produces the
nephrotic syndrome, which is
characterized by excretion of >3.5 g/d
of protein (mainly albumin).
Inflammatory disease of the intestinal tract is
associated with increase gastrointestinal loss of
albumin
Albumin concentration help to detect and
monitor protein nutritional status
Burn patients experience severe loss of
albumin from wounds due to the combined
effects of epithelial losses, accelerated
catabolism and the APR
Edema and ascites are usually secondary to
increased vascular permeability, and albumin
levels are decreased as a result of
redistribution of albumin into extravascular
space
- Dye-Binding methods
• Bromcresol green (BCG)
• Bromphenol blue (BPB)
• Bromcresol purple (BCP)
• Hydroxyazobenzenebenzoic acid
(HABA)
- Immunoturbidimetric and
nephelometry
Also called alpha lipoprotein or good cholesterol
Characterized by high density because of its protein
and phospholipid content
Apolipoprotein A1 the major protein, is a
negative APP that is considered to have antiinflammatory properties
24
Chem PPT Flashcards Unit 2
What is α1-Acid Glycoprotein?
What factors does α1-Acid Glycoprotein
increases in?
What factors does α1-Acid Glycoprotein
decreases in?
How is electrophoresis used for α1-Acid
Glycoprotein Determination?
How is α1-Acid Glycoprotein Determination
quantified?
What is α1-Antitrypsin (AAT)?
-
Also known as orosomucoid
One of the highest concentration
proteins in the alpha-1 globulin religion
- Amember of the lipocalin family of
proteins that bind lipophilic substances,
such as progesterone and basic drugs
including propanolol, chlorpromazine,
quinidine, cocaine and benzodiazepine.
- It is synthesized mainly by hepatic
parenchymal cells
- A positive APP
- G.I. inflammatory disease
- Malignant neoplasms
- Corticosteroids and NSAIDs
medication
- Estrogen medication
- Protein-losing syndrome such as
nephrotic syndrome
Using periodic acid Schiff or other
carbohydrate stains
By immunochemical methods, including
turbidimetry and nephelometric.
- Is a serpin (serine proteinase inhibitor)
that inactivates serine proteases such as
neutrophil elastase
- Synthesize primarily by hepatic
parenchymal cells
- It is the most important inhibitor of
leukocyte elastase, as release by
neutrophils
- Levels are elevated by the APR and by
estrogen oral contraception
- Levels are low in individuals with
neonatal respiratory distress syndrome,
severe pancreatitis and protein-losing
disorders
- Quantified by immunoturbidimetry or
immunonephelometry
- Phenotyping of AAT is performed by
isoelectric focusing
25
Chem PPT Flashcards Unit 2
What is α -Fetoprotein (AFP)?
-
-
What is Haptoglobin (Hp)?
-
-
-
What is α2-Macroglobulin (AMG)?
-
-
-
-
-
-
Is the most abundant plasma proteins in
the early embryonic life
Homologous to albumin but differs in
having about 4% carbohydrate
Serum concentration serves as a tumor
marker for Hepatocellular and germ
cell carcinoma
Maternal serum concentrations are
analyzed as indicators of fetal trisomy
of chromosome 18 or 21
Concentrations are increased by
glucocorticosteroids, androgens and
many NSAIDs
Levels are elevated in selective proteinlosing syndrome such Nephrotic
syndrome
Significantly elevated in biliary
obstruction in the absence of severe
hepatocellular disease
Measured by immunoturbidimetry and
immunonephelometry
A major plasma proteinase inhibitor
A very large molecule that does not
diffuse from the plasma space into
extracellular fluids in significant
amounts
Homologous to complement
components C3 and C4 in inhibiting
classes of proteinases, including those
with serine, cysteine and metal ions in
their catalytic sites
Synthesized by the liver
Levels are higher in women of
childbearing age and in infants and
children
Increased in nephrotic syndrome
Decreased in severe acute pancreatitis
or with advance carcinoma of the
prostate
Ccan be measured by zone
electrophoresis in the alpha2-globulin
region
Quantified by immunoturbidimetry or
immunonephelometry
26
Chem PPT Flashcards Unit 2
What is Ceruloplasmin (Cp)?
Following are clinical significant of what type
of plasma protein?
decreased levels are due to dietary copper
insufficiency (including malabsorption),
associated with neutropenia,
thrombocytopenia, low serum iron and
hypochromic, normocytic, or microcytic
anemia that does not to respond to iron therapy
What disease is characterized by increased
total body copper and deposited in tissues
including the hepatic parenchymal cells, the
brain, and the periphery of the iris (resulting in
the characteristic Kayser-Fleisher ring)?
How measure Ceruloplasmin (Cp)?
What is separation and transportation or
holding condition for Ceruloplasmin (Cp)?
What is concentration of Ceruloplasmin (Cp)
at birth?
When is peak concentration of Ceruloplasmin
(Cp)?
What is concentration of Ceruloplasmin (Cp)
for adult?
What is other name for Transferrin?
An α2-globulin that contains
approximately 95% of the total serum
copper, giving it a blue color
- May give plasma a greenish tint when
concentrations are significantly
elevated during pregnancy
- Synthesized primarily by the liver, and
is responsible for catalysis of reduction
and oxidation reactions
- Important in regulating the ionic state
of iron, oxidizing from the ferrous
(Fe2+) to the ferric (FE3+) state, thus
permitting incorporation of the iron into
transferrin
- It is a weak, late-reacting positive APP
- Increased significantly by estrogens, as
in pregnancy or with the use of oral
contraceptives
Ceruloplasmin (Cp)
-
Wilson disease, or hepatolenticular
degeneration
by immunoturbidimetry and
immunonephelometry
serum or plasma should be separated from
blood as soon as possible
specimen should be refrigerated for up to
three days or frozen at below -70°C for
prolonged storage
Low at birth
increased to peak concentrations at 2 or 3
years of age
decline slowly until adolescence, when adult
concentrations are reached
also known as siderophilin
27
Chem PPT Flashcards Unit 2
What is the major plasma transport protein for
iron?
What is the main factor in plasma which
Transferrin accounts for it?
Where is Transferrin in the body synthesized
and how it migrates in electrophoresis?
What is relationship between Transferrin and
iron concentration?
What is concentration of Transferrin and iron
saturation in hereditary hemochromatosis
(iron-overload state)?
What is diurnal cycle for Transferrin?
What is level of Transferrin in pregnancy and
inflammation?
What is the level of synthesis of Transferrin in
following diseases?
chronic liver disease, protein malnutrition,
protein loss as seen in nephrotic syndrome or
in protein-losing enteropathy
What are laboratory considerations for
Transferrin determination?
What is the formula for TIBC (Total Iron
Binding Capacity)?
Transferrin (TRF) migrates in…………on
routine serum electrophoresis.
What is equation which TIBC is used to
estimate TRF?
What Low-Density Lipoprotein is termed on
the basis of its electrophoretic mobility?
What percentage of LDL is lipid?
What percentage of LDL is protein?
What is the main lipid component of LDL?
What is the main protein component of LDL?
Transferrin
total iron binding capacity
synthesized primarily in the liver and usually
migrates in the β1-region on routine clinical
electrophoresis of serum
plasma concentrations are inversely related to
iron concentration
concentration is normal in hereditary
hemochromatosis (iron-overload state), but
iron saturation often exceeds 55%
serum iron concentration usually is highest in
the morning and decreases markedly in the
evening
high concentrations are present in pregnancy
and during estrogen administration
is a negative APP, and low concentrations
occur in inflammation and malignancy
Decreases
Serum electrophoresis
Immunoturbidimetry and
Immunonephelometry methods
Photometric assays for TIBC (Total Iron
Binding Capacity)
TIBC = UIBC + Iron
Where: TIBC = total iron binding capacity
UIBC = unsaturated binding capacity
β1 –region
TRF (mg/dL) = 0.70 x TIBC (µg/dL)
or
Iron
% Transferrin saturation = --------------- x 100
TIBC
β-lipoprotein
80%
20%
cholesterol ester
apolipoprotein B
28
Chem PPT Flashcards Unit 2
Increase in LDL concentration confer
increased risk of ……………..?
How concentration of LDL are assessed?
Following is characteristic of what plasma
protein?
a low-molecular-mass protein found on the cell
surfaces of all nucleated cells
Which protein plasma is the light or beta chain
of the human leukocyte antigens (HLAs)?
Which protein is high in individuals with renal
failure, inflammation, and neoplasm especially
those associated with B lymphocytes
Name a protein marker for renal tubular
function?
BMG levels has been used as a staging
criterion for what type of disease?
Name a protein plasma which can deposit as
amyloid, leading to systemic amyloidosis in
many patients of long-term dialysis?
What protein is the terminal component of the
coagulation system, aggregates to form a
fibrous network when it is cleaved by the
protease thrombin?
What protein is a highly elongated protein that
consist of six polypeptide chains joined by
disulfide bonds?
Fibrinogen is positive or negative APP?
What protein is the major contributors to
plasma viscosity because of its large size?
What does occur when Fibrinogen is low?
cardiovascular disease
assessed by calculation of LDL cholesterol
that are based on other lipid measurements,
direct quantification of LDL cholesterol, or
quantification of apolipoprotein B
β2-Microglobulin (BMG)
β2-Microglobulin (BMG)
β2-Microglobulin (BMG)
urinary BMG (β2-Microglobulin)
multiple myeloma
β2-Microglobulin (BMG)
Fibrinogen
Fibrinogen
Positive
Fibrinogen
the result of consumption from extensive
bleeding or dysregulation of the coagulation
system
What is the level of Fibrinogen in disseminated Low
intravascular coagulation (DIC)?
In what disease Dysfibrinogenemia (functional Liver disease
disorders of fibrinogen) may be seen?
What high levels of Fibrinogen on a chronic
increased risk of cardiovascular disease,
basis are associated with?
which may relate to increase plasma viscosity
or the increased tendency for thrombosis
One of the assay to measure Fibrinogen is?
functional clotting assays
What method can be used to measure
measured by immunoturbidimetry methods or
Fibrinogen level?
by turbidimetric methods based on the
addition of precipitating reagents
29
Chem PPT Flashcards Unit 2
What protein may be mistaken for a
paraprotein if not completely removed from
serum on serum protein electrophoresis?
Where C-reactive Protein (CRP) is found?
Name the bacteria that CRP is able to bind the
cell wall C polysaccharide of it?
CRP is positive or negative APR?
CRP used extensively as a marker of……….?
Where CRP synthesized in the body?
What is function of CRP?
CRP binds to a variety of compounds is
dependent on the presence of …………?
What is diagnostic window for CRP?
In what disorders CRP increases?
CRP is a stronger stimulus in bacteria or viral
infection?
Name the protein marker for bacterial infection
in newborns?
What protein marker is used to assess the
activity of disorders such as rheumatoid
arthritis and Crohn disease?
What are referred of High-sensitivity CRP
(hsCRP) assay?
What types of assays are used for high
sensitivity CRP?
What type of gel is used for electrophoresis of
CRP?
What are complement proteins and where are
they found?
What complement proteins are in the Classical
Pathway?
How is the Classical Pathway activated?
What complement proteins are in the
Alternative Pathway?
Fibrinogen
in the sera of acutely ill individuals
Streptococcus pneumoniae
an early positive APR
inflammation
Liver
to protect the body against foreign organisms
and to assist in clearing tissue debris
Calcium
Levels begin to rise within 6 to 12 hours of
the onset and peaks within 48 hours
myocardial infarction, stress, inflammation,
trauma, neoplastic proliferation, infection,
surgery
Bacterial infection is usually a stronger
stimulus than viral infection
CRP
CRP
Detection limit of about 1 mg/L
levels greater than 2 mg/L is suggestive of
increased cardiovascular risk
Particle-enhanced immunoturbidimetry,
nephelometry, or sandwich immunoassays
with fluorescence or chemiluminescent
detection.
Cellulose acetate or agarose gel
Complement proteins help to complement the
ability of antibodies and phagocytic cells to
clear pathogens from an organism. They are
found in normal blood plasma and also as
cellular receptors on cell surfaces.
It includes C1, C4, C2, and C3 (in order of
activation)
It is activated primarily by complexes of
immunoglobulins with antigen or bacteria or
other ligands with CRP
Includes C3, factor B and D, and properdin
30
Chem PPT Flashcards Unit 2
How is the Alternative Pathway activated?
What complement proteins are in the Lectin
Pathway?
How is the Lectin Pathway activated?
What complement proteins are in the
Membrane Attack complex?
What protein is activated through all of the
pathways?
What is chemotaxis?
What is opsonization?
What is phagocytosis?
Where are complement components mainly
synthesized?
Of all the complement proteins, which is the
most abundant?
What does a genetic deficiency of C2 and C4
cause?
What causes hereditary angioedema?
What are coagulation proteins?
What organ primarily produces all of the
coagulation factors?
Of all the coagulation factors, which is the
most important?
What causes low concentrations of clotting
factors?
What are some hereditary disorders that cause
excessive bleeding?
What may cause a risk of thrombosis?
It is activated by bacterial
lipopolysaccharides, cellular proteases and
cobra venom
It includes mannan-binding protein (MBP),
ficolins, and proteases
It is activated by binding of ficolins and MBP
to mannose-rich oligosaccharides that are
present in the cell walls of many
microorganisms
Includes C5 through C9 and inserts into cell
membranes and lyses cells
The common step involved is the activation of
C3 to C3b.
The movement or orientation of an organism
or cell along a chemical concentration
gradient or in response to a chemical stimulus
It refers to an immune process where particles
such as bacteria are targeted for destruction by
an immune cell known as a phagocyte
The engulfing and ingestion of bacteria or
other foreign bodies by phagocytes to form an
internal vesicle known as a phagosome
The liver
C3
A genetic deficiency of C2 and C4 is
associated with autoimmune immune complex
diseases such as systemic lupus erythematosus
(SLE), polymyositis and glomerulonephritis.
Heterozygous deficiency of C1 inhibitor
results in hereditary angioedema (HAE)
A complex system designed to prevent
bleeding from injuries by forming blood clots
The liver
Fibrinogen
The result of hereditary deficiencies,
consumption by active clotting, liver disease
or anticoagulant therapy
Hemophilia A, factor VIII deficiency,
hemophilia B, and factor IX deficiency
Risk of thrombosis may be result of factor V
Leiden deficiency, antithrombin III deficiency
and protein C deficiency
31
Chem PPT Flashcards Unit 2
What are immunoglobulins (antibodies)?
What stimulates the production of
immunoglobulins, and what do they attack?
What is the structure composition of
immunoglobulins?
On what chromosomes are the light and heavy
chains encoded?
What forms the antigen-binding site on the
immunoglobulins?
What are heavy chains made up of?
What are light chains made up of?
Where are immunoglobulins synthesized?
What are the five classes of immunoglobulins?
Which of the five immunoglobulins are the
most abundant?
What does IgG do?
What is the circulating half-life of IgG?
Which, if any, of the immunoglobulins can
cross the placenta blood barrier?
What are the four subclasses of
Immunoglobulin G (IgG)?
They’re a family of proteins the contain
highly specific antigen-binding sites.
Synthesis of immunoglobulins is stimulated
by foreign immunogens, leading to
immunoglobulins that selectively bind foreign
molecules or organisms.
All immunoglobulins include one or more
basic units consisting of two identical heavy
(H) chains encoded on chromosome 14 and
two identical light (L) chains encoded on
chromosome 2 that are joined by disulfide
bonds.
Heavy chain is encoded on chromosome 14
and the light chains on chromosome 2
Two variable domains, one from the light
chain and one from the heavy chain, together
form an antigen-binding site.
They consist of gamma, alpha, mu, delta, and
epsilon
Kappa and Lambda
Immunoglobulins are synthesized by cells of
the B-lymphocyte lineage.
IgG, IgA, IgM, IgE, and IgD
The most abundant class of Ig, making up 70
to 75% of total Ig. Of this amount, 65% is
extravascular and 35% is found in plasma.
It is responsible for long-term physiologic
protection and the majority of neutralization
of bacteria and virus antigens
Approximately 22 days
IgG
IgG includes four subclasses - IgG1, IgG2,
IgG3, and IgG4
- IgG1 and IgG3 strongly activate
complement via the classical pathway
- IgG2 weekly activates complement
- IgG4 does not activate complement
- IgG1 and IgG3 bind Fc receptors on
phagocytic cells can cross the placenta
via receptor-mediated active transport
IgG1 is the major neonatal Ig.
32
Chem PPT Flashcards Unit 2
What are some characteristics of
Immunoglobulin A (IgA)?
-
What are two types of Immunoglobulin A
(IgA)?
-
Where is Immunoglobulin A (IgA) synthesized
and secreted?
-
-
-
What is Immunoglobulin M (IgM)?
-
-
What is Immunoglobulin E (IgE)?
-
Makes up 10% to 15% of serum
immunoglobulin
On electrophoresis, IgA migrates near
the junction of the beta and gamma
regions
IgA also activates the complement by
the alternative pathway
Plasma dimeric IgA1: sedimentation
coefficient of 7S
Secretory dimeric IgA2: sedimentation
coefficient of 11S and present in saliva,
tears, sweat, milk, colostrum and
secretion of nasal, gastrointestinal and
tracheobronchial origin
It is synthesized mainly by plasma cells
in the mucous membranes of the gut
and bronchi and in the ductules of the
lactating breast
This secretory component helps
transport secretory IgA across mucosal
epithelium and into secretions
Secretory IgA is more abundant than
IgG in colostrum and milk and may
help protect neonates from intestinal
infection
Produced at early stages of β-cell
development.
Comprises about 5%-10% of the total
circulating immunoglobulins
Largest immunoglobulin, a pentamer
(molecular weight > 900 kilodaltons)
Most primitive immunoglobulin and is
found on the surface of early B
lymphocytes
First or primary response
immunoglobulin to be produced
following antigen assault.
Monomeric immunoglobulin in plasma
Lowest concentration (about 0.05
mg/dL)
Attaches rapidly to the membranes of
mast cells by its Fc region
Cross-linking of two IgE molecules by
antigen causes release of histamine,
heparin and vasoactive amines by mast
cells
33
Chem PPT Flashcards Unit 2
What role does Immunoglobulin E (IgE) play
in allergic or hypersensitivity reaactions?
What is Immunoglobulin D (IgD)?
What is the significance of immunoglobulin
deficiency?
What is Polyclonal
Hyperimmunoglobulinemia?
What are some characteristics of Monoclonal
Immunoglobulins (Paraproteins)?
What malignant neoplasm is usually a single
clone of plasma cells?
What is the tumor called when the plasma cells
proliferate diffusely throughout the bone
marrow?
What is produced due to multiple myeloma?
What are decreased due to multiple myeloma?
-
Urticaria
Hay fever
Asthma
Eczema
A monomeric immunoglobulin in
plasma
- Active on the cell surface of B
lymphocytes
- Labile upon storage
- Not routinely measured
- Immunodeficiency may be the result of
a deficiency of a single factor or of a
combinations affecting multiple
systems and factors
- It results in increased susceptibility to
infections that usually do not cause
disease in people with normal immune
systems
- Polyclonal increases in plasma
immunoglobulins constitute the normal
response to infection
- IgG response predominates in most
autoimmune responses; IgA in skin,
gut, respiratory and renal infections;
and IgM in primary viral infections and
with bloodstream parasites like malaria.
- Produces a discrete band on
electrophoresis often referred to as an
M-spike or M-protein
- up to 25% of paraproteins are benign
and have been termed monoclonal
gammopathy of undetermined
significance (MGUS)
- Bence-Jones proteins: the primary
clinical interest in identifying
paraproteins involves the detection or
monitoring of proliferative disorders of
B lymphocytes
Multiple myeloma
plasmacytoma
Osteolytic bone lesions
Bone marrow cells like platelets, red cells,
neutrophils
34
Chem PPT Flashcards Unit 2
What is inhibited due to multiple myeloma?
What consequently happens?
Lymphomas or chronic lymphocytic leukemias
are examples of ___.
Where do lymphoid tumors arise?
What is a malignant proliferation of
plasmacytoid cells producing IgM?
What causes hyperviscosity that may require
treatment by plasma exchange?
What is the rare disorder in which a free Ig
heavy chain is produced?
What is the most common form of Heavy
Chain disease?
What is alpha chain disease associated with?
What can cause malabsorption in alpha chain
disease?
What are serum proteins or protein complexes
that precipitate at a temperature lower than
normal core body temperature?
What is the result of precipitation in tissues?
How do you lower a patient's cryoglobulin
concentrations?
What do type I cryoglobulins consist of?
How many days are serum usually refrigerated
for cryoglobulin analysis?
What is the next step after 7 days?
What happens when cryoglobulins that
precipitated in the cold are warmed back to
37C?
What is a pathological process in which
proteins are deposited in tissues as aggregates
(amyloid), and form b-sheet structures?
What can result from amyloidosis?
What are stable structures resistant to
proteolysis and normal clearance mechanisms?
Aggregates accumulate over time.
What happens when there is increased
production of specific proteins, genetic variants
of proteins, or fragments of proteins?
Where are the tissues obtained in order to
diagnose amyloidosis?
Normal plasma cells
Synthesis of other immunoglobulins is
reduced, recurrent infections may occur
Lymphoid tumors
From less mature stages in B-lymphocyte
development
Waldenstroms Macroglobulinemia
IgM and IgM paraproteins
Heavy Chain disease
Alpha chain disease
The heavy chain of IgA
Intestinal infiltration with clonal lymphocytes
Cryoglobulins
Vasculitis and ischemic injury to peripheral
tissues of the lower temperature
Patient must be kept in a warm environment
Monoclonal immunoglobulins, often IgM
7 days
Centrifugation
They should redissolve
Amyloidosis
Kidney failure, heart failure, and peripheral
neuropathies
Aggregates
TRUE
Formation of amyloid
From fat aspirates, rectal biopsy specimens, or
biopsy specimens of affected tissues
35
Chem PPT Flashcards Unit 2
What can the b-sheet structure of the amyloid
be stained with?
What occurs as the result of increased
concentrations of proteins including...
...Ig light chain paraproteins?
...BMG in renal failure?
...Serum amyloid A
Urine usually contains much lower protein
concentration than plasma.
How does urine maintain its concentration?
Why is analysis of urinary proteins mainly
performed?
What is the increase of urinary protein
excretion called?
When can cause proteinuria?
Why is glomerular proteinuria sometimes
referred to as albuminuria?
Glomerular proteinuria is seen in conditions
including diabetes, systemic lupus
erythemotosis, IgA nephropathy,
glomerularnephritis, hepatitis and HIV
infections.
What is characterized by hypoalbuminemia,
hypercholesterolemia, and edema
Excretion of ____ g/d protein is a criterion for
nephrotic syndrome.
What signifies early stage of glomerular injury
and a risk factor for progressionof kidney
disease in diabetes?
How is urine albumin detected?
When can functional or benign proteinuria
occur?
Postural or orthostatic proteinuria is associated
with the upright position.
First voided urine is analyzed to detect
orthostatic proteinuria.
What is the appearance of low molecular
weight proteins in the urine as a result of
decreased reabsorption by the proximal
tubules?
Congo red dye
AL amyloidosis
AB2 amyloidosis
AA amyloidosis
TRUE
Through the combined action of glomerular
filtration and tubular uptake of protein
To diagnose kidney disease or disorders with
paraprotein production
Proteinuria
Glomerular injury, postrenal proteinuria,
tubular injury, overflow proteinuria of low
molecular weight proteins
Because albumin is the major protein
TRUE
Nephrotic syndrome
>3.5
Microalbuminuria (30-300mg/d)
By dipsticks, followed by quantitative assays
like pyrogallol red dye binding assays and
benzethonium chloride turbidity assays
With exercise fever and exposure to cold
TRUE
TRUE
Tubular proteinuria
36
Chem PPT Flashcards Unit 2
What is Fanconi syndrome?
What are some of the causes of tubular
proteinuria?
What occurs when an increased load of small
proteins (hemoglobin, myoglobin, free Ig light
chain paraproteins) filtered through the
glomerulus exceed the tubular uptake capacity?
What does detection of light chains depend on
in overload proteinuria?
What color does hemoglobin or myoglobin
confers to urine, yielding positive reactions in
the dipstick test for blood?
What arises from injury, inflammation or
malignancy of the lower urinary tract?
What does Postrenal proteinuria reveal under
microscopic examination of urine?
What helps to distinguish whether a process is
occurring in the kidneys or in the lower urinary
tract?
What is an excellent screening tests for major
glomerular injury (sensitive to albumin)?
What is the extracellular fluid around the brain
and spinal column?
Analysis of CSF is mainly performed to
determine?
Which disease aids increased intrathecal
synthesis of immunoglobulins?
Methods for measuring CSF protein includes?
What is analyzed in proteins in amniotic fluid?
What represents the fluid secreted by several
salivary glands containing amylase and small
peptides?
Which immunoglobulin is tested in saliva for
evaluation of possible immunological
deficiency?
What is used in the diagnosis of
gastrointestinal protein loss?
What has been used as a measure of disease
activity in inflammatory bowel disease?
A hereditary disorder of tubular functio
Fanconi syndrome, toxicity from compounds,
poisoning from heavy metals, ischemia due to
obstruction of blood flow, decreased blood
flow in shock or heart failure, toxicity from
overload with proteins
Overload Proteinuria
Electrophoresis and immunochemical testing
Brown
Postrenal Proteinuria
Inflammatory and malignant cells and bacteria
Cast that arise in kidney tubules
Dipstick test
Cerebrospinal fluid
Increased permeability of blood-brain barrier
(BBB) or increased intrathecal synthesis of
immunoglobulin.
Multiple sclerosis
Pyrogallol red dye-binding assays
Benzethonium chloride turbidity assays
Biuret assays
α-fetoprotein (AFP)
Proteins in saliva and oral fluid
IgA
assay of feces for (α1-antitrypsin) AAT
Lactoferrin and calprotectin
37
Chem PPT Flashcards Unit 2
Refers to proteins abnormally accumulated in
the peritoneal (abdominal) and pleural (lung)
cavities
What are enzymes?
How specific are they and where are the
majority found?
What do the increases of serum enzymes
usually indicate and why are they useful?
What are Enzymes?
What is necrosis?
How many classes of Enzymes are there?
What are the Classification of enzymes
Oxidoreductase?
Proteins in Peritoneal and Pleural Fluid
Enzymes are organic catalysts than hasten a
chemical reaction without themselves being
consumed or undergoing a chemical change.
They have a high degree of specificity for a
certain substrate or class of substrates and the
majority are intracellularly found.
increased serum levels are due to tissue
damage and necrosis, therefore they are used
diagnostically to asses tissue damage.
 are organic catalysts than hasten a chemical
reaction without themselves being
consumed or undergoing a chemical change
 protein in nature with high degree of
specificity for a certain substrate or class of
substrates.
 majority are intracellularly found
 amounts found in serum are result of
normal cellular turnover; specialized tissue
or widely distributed with tissue-specific
isoenzymes or isoforms
 increased serum levels are due to tissue
damage and necrosis (markers of tissue
damage)
 thus, used diagnostically to assess tissue
damage
Necrosis is tissue damage due to elevating
serum levels.
There are 6 classes of Enzymes:
I.
Oxidoreductases
II.
Aminotransferases/Aminotransaminase
s
III.
Hydrolases
IV.
Lyases
V.
Isomerases
VI.
Ligases
Oxidase
Cytochrome oxidase
Dehydrogenase
Lactate dehydrogenase (LDH)
Malate dehydrogenase (MDH)
Isocitrate dehydrogenase (ICD)
Glucose-6-phosphate dehydrogenase
(G-6-PD)
38
Chem PPT Flashcards Unit 2
What are the Classification of enzymes
Aminotransferase/Aminotransaminase?
What are the Classification of enzymes
Hydrolase?
What are the Classification of enzymes
Lyases?
Two types of Isomerases:
Another name for Ligases enzyme:
The roles of Oxidoreductases:
The activities of Aminotransferases
/Aminotransaminases:
The factors of Hydrolases:
 Aspartate aminotransferase (AST / SGOT)
 Alanine aminotransferase (ALT / SGPT)
 Creatine kinase (CK /CPK)
 Gamma glutamyl transferase (GGT)
 Ornithine carbamyl transferase (OCT)
Esterase
Acid phosphatase (ACP)
Alkaline phosphatase (ALP)
Cholinesterase (CHS)
Lipase (LPS)
Peptidases
Typsin (PTS)
Pepsin (PPS)
Leucine aminopeptidase (LAP)
Glucosidase
Amylase (AMS)
Amylo-1,6 glycosidase
Galactosidase
 Aldolase (ALD)
 Carbonic anhydrase
 Glutamate decarboxylase
 Pyruvate decarboxylase
 Tryptophan decarboxylase
1) Glucose phosphate isomerase
2) Ribose phosphate isomerase
Glutamine synthetase
a) Catalyze an oxidation-reduction
reaction
b) Catalyze the addition or removal of
hydrogen from compounds
c) Require a coenzyme (NAD or NADP)
as a hydrogen acceptor or donor in
order to function
a) Catalyze the transfer of amino or
phosphate groups between compounds
b) Require coenzyme (pyridoxal-5’phosphate [P-5’-P]) for the amino
transfer reactions
a) Catalyze the hydrolysis of ether and
ester
b) They split molecules with the addition
of water
c) Do not usually require coenzymes but
often need activators
39
Chem PPT Flashcards Unit 2
How do Lasses operate inside a cell?
a) Catalyze the removal of groups without
hydrolysis (loss of hyroxide ion)
b) Split molecules between carbon-tocarbon bonds without the addition of
water
c) Resulting products usually contain
carbon double bonds
How do Isomerases function?
a) Catalyze the interconvertion of geometric or
optical isomers
b) Catalyze the intramolecular conversions such
as the oxidation of a functional group by an
adjacent group within the same molecule
What are the tasks of Ligases?
a) Catalyze the joining of two substrate
molecules
b) Catalyze the union of two molecules
accompanied by the breakdown of a
phosphate bond in adenosine
triphosphate (ATP)
The following are additional questions on slides 4-20
How many classes of Enzymes are there?
I.
Oxidoreductases
II.
Aminotransferases/Aminotransaminase
s
III.
Hydrolases
IV.
Lyases
V.
Isomerases
VI.
Ligases
What are the classification of enzymes
 Oxidase
Oxidoreductase?
Cytochrome oxidase
 Dehydrogenase
Lactate dehydrogenase (LDH)
Malate dehydrogenase (MDH)
Isocitrate dehydrogenase (ICD)
 Glucose-6-phosphate dehydrogenase
(G-6-PD)
What are the classification of enzymes
 Aspartate aminotransferase (AST /
Aminotransferase/Aminotransaminases?
SGOT)
 Alanine aminotransferase (ALT /
SGPT)
 Creatine kinase (CK /CPK)
 Gamma glutamyl transferase (GGT)
 Ornithine carbamyl transferase (OCT)
What are the enzymes Hydrolases actions?
Catalyze the hydrolysis of ether and ester
they split molecules with the addition of water.
Do not usually require coenzymes but often
need activators.
40
Chem PPT Flashcards Unit 2
What are the enzymes Lyases actions?
Catalyze the removal of groups without
hydrolysis (loss of hyroxide ion).
Split molecules between carbon-to-carbon
bonds without the addition of water.
Resulting products usually contain carbon
double bonds.
What are the enzymes Isomerases actions?
Catalyze the interconvertion of geometric or
optical isomers.
Catalyze the intramolecular conversions such
as the oxidation of a functional group by an
adjacent group within the same molecule.
What are the enzymes Ligases Actions?
Catalyze the joining of two substrate
molecules.
Catalyze the union of two molecules
accompanied by the breakdown of a phosphate
bond in adenosine triphosphate (ATP).
What is Phosphatase?
is an enzyme that removes a phosphate group
from its substrate by hydrolyzing phosphoric
acid monoesters into a phosphate ion and a
molecule with a free hydroxyl group.
What is Acid Phosphatase?
Promotes the hydrolysis of orthophosphate
esters.
Found in almost all body tissues, but
significantly in RBC’s and platelets.
In adult men, 50% is found in the prostate
gland.
High levels are found in semen.
What are the requirement substrates to measure p-nitrophenylphosphate (PNPP) (preferred
Total ACP Activity?
substrate for continuous monitoring)
thymolphthalein monophosphate (TMP)
(substrate of choice)
What is p-nitrophenylphosphate (PNPP)?
Is a non-proteinaceous, non-specific substrate
used to assay protein, alkaline and acid
phosphatases.
The PNPP phosphatase activity is measured
using a continuous or single-point
spectrophotometric assay based on the ability
of phosphatases to catalyze the hydrolysis of
PNPP to p-nitrophenol, a chromogenic product
with absorbance at 405 nm (1).
What is thymolphthalein monophosphate
Obtained from 10 commercial suppliers were
(TMP)?
compared spectrophotometrically at 445 and
595 nm, liquid-chromatographically with
monitoring at 254 nm, and enzymically by
measurements of activity of prostatic acid
phosphatase in human serum.
41
Chem PPT Flashcards Unit 2
What is the measurement of Total ACP
Activity?
What is isoenzymes of Acid Phosphatase
(ACP)?
What is Isoenzyme Test Include:
What is the Clinical Significance of ACP?
What is Metastatic Carcinoma?
What enzyme(s) have liver as the primary
source and can be used to detect hepatic
parenchymal disease?
What enzyme is a indicator in muscle disease ?
What enzyme(s) associated with detecting
pancreatic disease?
Where would you find enzyme lipase ?
What does ACP - Acid Phosphatase do?
Where would you find ACP?
In adult men, where is 50% of ACP found?
What condition the ACP activities were taken
placed at?
What is tartrate used in isoenzyme testing?
D-
What are the clinical significance of ACP?
What is ALP alkaline phosphatase?
What’s the optimal pH for ALP
All reactions are carried out at a pH lower than 6.0
Falsely low values are produced by improper
anticoagulant use (heparin, fluoride, oxalate)
Hemolysis produces false elevation
 Specific Prostatic Phosphatase
 Non-Specific Prostatic Phosphatase or Red
Cell Phosphatase
 inhibition of prostatic ACP by Dtartrate, followed by the determination
of the serum level of other ACP
fractions
 immunoassay with antibodies specific
to certain isoenzymes
- electrophoresis
 aids in detecting metastatic carcinoma
 other types of cancer and bone diseases
 evidence of rape
Metastatic Carcinoma is the spreading of
cancer, but with the Clinical Significant of
ACP they can detect and prevent the cancer
spreading.
Alanine aminotransferase
Aspartate aminotransferase
Creatine kinase
Amylase
Lipase
 Pancreas
Promotes the hydrolysis of orthophosphate
ester
In all body tissues, significantly in RBC’s and
platelets
found in the prostate gland, high levels are
found in semen
pH lower than 6.0
inhibition of prostatic ACP , followed by the
determination of the serum level of other ACP
fractions
aids in detecting metastatic carcinoma
Other types of cancer and bone disease
Evidence of rape
Hydrolyzed phosphate esters
At 8.6 ÷ 10
42
Chem PPT Flashcards Unit 2
What is ALP used for?
Where will you find ALP?
What is the ALP isoenzyme that is most
susceptible to heat inactivation (heat labile),?
What are bone disease patients will have
elevated level of ALP?
diagnosis of hepatobiliary disease and bone
disease associated with increased osteoblastic
 in liver, bone, kidney, intestines and
placenta
 Bone isoenzyme


Osteomalacia
Rickets
Paget’s disease
Why ALP level is high in children?
It’s from the osteoblasts during bone
growth
What are semiautomatic and automatic pipettes
 These devices are programmed and
and dispensers used for?
used to simultaneously dispense
aliquots into multiple wells.
Why are they piston driven?
 To allow the user to pipette with a few
or as many tips as necessary.
What is centrifugation?
The process of using centrifugal force to
separate lighter from heavier parts of a
solution, the correct term to describe the force
required to separate two phases in a centrifuge
is relative centrifugal force (RCF), also called
relative centrifugal field.
 Examples are horizontal head or
swinging bucket, fix angle or angle
head, ultracentrifuge and axial.
What is gravimetry?
The process used to measure the mass of a
substance using a balance to compare the mass
of an unknown to that of a known mass. The
comparison is called weighing and the absolute
standards with which masses are compared are
called weights.
What are the types of balances used?
Double pan, single pan and the electronic
balance.
The following are additional questions for slides 19:21-37
What does Alkaline phosphatase (ALP) do and Hydrolyzes phosphate esters; optimum pH =
what is its optimum pH?
8.6-10
What are the activators for Alkaline
Activators include Mg2+, Co2+ and Mn2+
phosphatase?
What are the inhibitors for Alkaline
Inhibitors include phosphate, borate, oxalate
phosphatase?
and cyanide ions
How is the Alkaline phosphatase used in
Used in the diagnosis of hepatobiliary disease
clinical diagnosis?
and bone disease associated with increased
osteoblastic activity
Where Alkaline phosphatase is located?
Found significantly in liver, bone, kidney,
intestines and placenta
43
Chem PPT Flashcards Unit 2
Does the bone isoenzymes of Alkaline
phosphatase (ALP) tolerate to heat?
How does the bone isoenzymes Alkaline
phosphatase (ALP) influence in the person
with bone disease?
What is the outbreak of Alkaline Phosphatase
(ALP) in children?
What is the B. Liver isoenzyme?
How does the B.Liver isoenzyme migrate on
electrophoresis?
What is the outcome of B.Liver isoenzyme in
person with hepatobiliary?
What is the next type of isoenzymes Alkaline
phosphatase (ALP)?
How does the C.Intestinal isoenzyme migrate
on electrophoresis?
What is the D. Placental isoenzyme?
What period does the D.Placental isoenzyme
have a higher concentration?
What is the optimum temperature for
D.Placental isoenzyme?
What is the last type of Alkaline phosphatase
(ALP)?
What is the carcinoplacental Alkaline
phosphatase?
What is the optimum temperature for
carcinoplacental Alkaline phosphatase?
What is the method used to Measure of
Alkaline phosphatase (ALP) activity?
What is the substrate used in Bessey-LowryBrock-methods?
What are the requirement cofactors in BesseyLowry-Brock-methods?
What is the time window for Serum to be
analyzed Measuring of Alkaline phosphatase
(ALP) activity?
What are the substances should be voided in
Measurement of Alkaline phosphatase (ALP)
activity?
Bone isoenzyme; The most susceptible to heat
inactivation (heat labile)
Elevated in persons with bone disease
a) Osteomalacia
b) Rickets
c) Paget’s disease (osteitis deformans)
High in children; Alkaline phosphatase (ALP)
from osteoblasts during bone growth
It is one of the Five type isoenzymes of
Alkaline phosphatase (ALP).
The most rapidly moving isoenzyme in
electrophoresis
Greatly elevated in persons with hepatobiliary
obstruction
C. Intestinal isoenzyme
The slowest moving isoenzyme in
electrophoresis
D.Placental isoenzymes is belonging to
Alkaline phosphatase (ALP) group.
Becomes elevated between the 16th and 20th
weeks of pregnancy.
The most heat stable (65°C)
E. Regan isoenzyme
An abnormal Alkaline Phosphatase (ALP); A
carcinoplacental Alkaline phosphatase (ALP)
that has similar properties to the placental
isoenzyme
heat stable (65°C)
Bessey-Lowry-Brock - methods
p-nitrophenylphosphate (PNPP)
Required cofactors:
Mg+2
Zn+2
Serum (analyzed within 4 hours after
collection) is preferred or heparinized plasma
EDTA, citrate and oxalate plasma must be
avoided
44
Chem PPT Flashcards Unit 2
Does the concentration of Alkaline
phosphatase (ALP) in serum increase at
refrigerated temperature?
What should the frozen Alkaline phosphatase
(ALP) be treated?
Does hemolysis increase results in Alkaline
phosphatase activity?
What is the most common method to
differentiate Alkaline Phosphatase (ALP)
isoenzymes?
How does Alkaline phosphatase (ALP) be
placed on Electrophoresis?
What substance will improve the separation of
bone ALP and liver isoenzymes on
Electrophoresis?
What residues must be dropped off bone ALP
and liver ALP on Electrophoresis?
Why should the terminal sialic acid residues be
removed off Electrophoresis?
How does intestinal Alkaline phosphatase
(ALP) be confirmed in serum?
Is intestinal Alkaline phosphatase (ALP)
resistant to neuraminidase?
What is the second method to differentiate
Alkaline phosphatase (ALP) isoenzymes?
Does Alkaline phosphatase (ALP) increase
during the pregnancy?
What is the concentration of Alkaline
phosphatase (ALP) in hepatitis and cirrhosis
condition?
Does Alkaline phosphatase (ALP) increase in
person with bone disease?
What will be happened to Alkaline
phosphatase (ALP) in hyperthyroidism case?
How concentration of Alkaline phosphatase
(ALP) in diabetes mellitus case?
What are the defintion for
amiotransferases/(aminotransaminases)?
Alkaline phosphatase (ALP) in serum stored at
refrigerated temperature increases slowly
(2%/day)
Frozen samples should be thawed 18-24 hours
at room temperature
Hemolysis falsely increases results
Electrophoresis
Order of anodal migration:
1) Liver ALP
2) Bone ALP
3) Placenta / Regan ALP
4) Intestinal ALP
Pre-treatment with neuramidase for 15 minutes
at 37°C improves separation of bone and liver
isoenzymes
Removes the terminal sialic acid residues
Sialic acid residues of bone ALP are more
attached than liver ALP, thus reducing
mobility
Overnight incubation of serum with
neuraminidase is used to confirm presence of
intestinal Alkaline phosphatase (ALP)
Intestinal Alkaline phosphatase (ALP) is
neuraminidase-resistant
Stability to denaturation by heat or chemical
Elevation during the 3rd trimester of pregnancy
Elevation in hepatitis and cirrhosis
Elevation in bone diseases with osteoblastic
activity
Elevation in hyperthyroidism
Elevation in diabetes mellitus
a) Aspartate Transferase (Transaminase) –
AST
b) Serum Glutamate Oxaloacetate
Transaminase –SGOT
45
Chem PPT Flashcards Unit 2
How does the chemical reaction happened on
Aspartate Transferase (AST)?
What is the first measurement of Aspartate
Transferase (AST) activity?
Transfers an amino group between as aspartate
and keto acids
The reaction with dinitrophenylhydrazine
(DNPH) couples color reagent with keto acid
product (Rietmann Frankel method)
What is the second measurement of Aspartate
The reaction with diazonium salts couples the
Transferase (AST) activity?
salt with the keto acid product and forms a
color (Babson)
What is the third measurement of Aspartate
The coupled enzyme assay (Karmen method)
Transferase (AST) activity?
involves NADH, malate dehydrogenase and
keto acid to form NAD. Malate dehydrogenase
catalyzes the oxidation of oxaloacetate to
malate in the indicator reaction.
What is the optimum temperature for Aspartate AST is stable in refrigerated temperature or
Transferase (AST)?
frozen samples
What are the acceptable reagents in collection
Serum or plasma collected with heparin,
Serum or plasma Aspartate Transferase (AST)? EDTA, citrate or oxalate is acceptable.
What substance is added to the Aspartate
Stability can be further enhanced by adding Ptransferase (AST) specimen for stability?
5’-P (pyridoxal-5’-phosphate) to the specimen
Where are Aspartate Transferase (AST)
AST is present in cardiac tissue, liver, skeletal
located?
muscles and RBC’s
Should Aspartate Transferase (AST) hemolysis Hemolyzed specimens must be avoided
specimens be avoided?
Does the alcohol affect Aspartate transferase
Alcohol lowers AST values.
(AST)?
How many hours after onset of Myocardial
Elevated in:
infarction will increase the Aspartate
a) Myocardial infarction
transferase (AST)?
b) Increases 6-8 hours after onset of pain
How fast Aspartate transferase (AST) will be
Elevated in:
elevated in Viral hepatitis?
a) Viral hepatitis
b) 100-fold increase
Does Aspartate transferase (AST) increase its
Elevated in Skeletal muscle diseases like
concentration in Skeletal muscle diseases?
Polymyositis
Rhabdomyolysis
Duchenne muscular dystrophy
Does Aspartate transferase (AST) affect Liver Elevated in:
disease?
Liver hypoxia
What will be happened to the concentration of Elevated in:
Aspartate transferase (AST) in strenuous
Strenuous physical activity
physical activity?
What is the Aspartate transferase (AST)
AST tends to be higher in infants and children
concentration in children?
than adults
How does Aspartate transferase (AST) be
Adult males exhibit higher AST activities than
classified in adult males and adult females?
adult females
46
Chem PPT Flashcards Unit 2
What are 5 clinical significance of ALP?
What does AST stand for?
What does SGOT stand for?
what do AST and SGOT do?
what are 3 major approaches involved in the
measurement of AST activity?
AST is stable at what temperature?
What specimen collection is needed for the
measurement of AST?
What can you add to the specimen for AST so
it can be more stable?
Where is AST present in the body?
What must be avoided in a specimen collected
for AST measurement?
What lowers the AST values?
What are the clinical significance of AST?
1) elevation during the 3rd trimester of
pregnancy
2) elevation in hepatitis and cirrhosis
3) elevation in bone diseases with osteoblastic
activity
4) elevation in hyperthyroidism
5) elevation in diabetes mellitus
Aspartate Transferase
Serum Glutamate Oxaloacetate transminase
It transfers an amino group between an
aspartate and keto acids.
L-aspartate + α-ketoglutarate
AST
oxaloacetate + L-glutamate
1. The reaction with dinitrophenylhydrazine
(DNPH) couples color reagent with keto acid
product (Rietmann Frankel method)
2. The reaction with diazonium salts couples
the salt with the keto acid product and forms a
color (Babson)
3. The coupled enzyme assay (Karmen
method) involves NADH, malate
dehydrogenase and keto acid to form NAD.
Malate dehydrogenase catalyzes the oxidation
of oxaloacetate to malate in the indicator
reaction.
At refrigeration or frozen temperature.
Serum or plasma collected with heparin,
EDTA, citrate or oxalate is acceptable.
Stability can be further enhanced by adding P5’-P (pyridoxal-5’-phosphate) to the specimen.
AST is present in cardiac tissue, liver, skeletal
muscles and RBC’s.
Hemolyzed specimens must be avoided.
Alcohol
AST is elevated in:
1) Myocardial infarction
* Increases 6-8 hours after onset of pain
2) Viral hepatitis
* 100-fold increase
3) Skeletal muscle disease
* Polymyositis
* Rhabdomyolysis
* Duchenne muscular dystrophy
47
Chem PPT Flashcards Unit 2
What is a micropipettes ?
Pipettes come in different sizes?
What is the purpose of a pipette ?
What is the mouthpiece of the pipette?
What is the calibration lines of a pipette?
What is Volume of the pipette ?
What is the T.D of the pipette?
What is the T. C of the pipette
Why must a distinction between T.D and T.C
pipettes be made ?
What is T.D calibrated with ?
What is T.C glassware calibrated with ?
What is the meniscus ?
What is centrifugation ?
What is gravimetric ?
What is a Double-pan, single-pan and
electronic balances
A hallow glass tube used for measuring small
volumes of liquid in microliters and transfer of
liquid from one source to anothere
Yes from single piece glass pipettes to more
complex adjustable or electronic pipettes for
various purposes with differing levels of
accuracy and precision
To draw up liquid for the purpose of measuring
and transferring a specific volume of liquid
The end of the pipette to which suction is
applied to draw up liquid into the pipette
Marks on the stem of the pipette to show the
point where liquid must be drawn to give a
specific volume
Stated in millimeters example inscription may
read 10ml in 1/10 ml and the pipette is
graduated in 1/10 ml increments allowing it to
be used to measure volumes up to 10 ml
“To deliver “pipettes designed to deliver a
specific volume of fluid
“To contain “ pipettes designed to contain a
certain volume of fluid
Because there may be a difference between the
volume a pipette will contain and a volume it
will deliver
Distilled or deionized water
Mercury
A crescent shaped structure appearing at the
surface of a liquid column it has the
appearance of a contact lens
The process of using centrifugal force to
separate the lighter portion of a solution
mixture or suspension from heavier portions.
The process used to measure the mass of a
substance
Types of balances
What are the Isoenzymes of CK?
CK1 or CKBB
CK2 or CKMB
CK3 or CKMM
CK- Mt
Macro- CK
Where is found CK1?
brain and nerve tissues
Where is found Ck2 in body?
At which level Ck2 is elevated and return to
normal in MI?
In cardiac muscles.
levels rise within 4-6 hours, peak at 12-24 hours
and return to normal within 48-72 hours
48
Chem PPT Flashcards Unit 2
Located between the inner and outer
membranes of mitochondria, and it constitutes
up to 15% of total CK activity
Located on chromosome 15
How to measure CK by ion-exchange
involves an anion exchange resin at a pH 8.0
chromatography?
isoenzymes have different charges and can be
selectively eluted
How to measure CK by immunoassay?
involves antisera employed against certain
subunits of each isoenzyme
How to measure CK by Tanzer-Gilvarg assay?
involves coupling with pyruvate kinase and
lactate dehydrogenase to produce a change in
absorbance measured spectrophotometrically.
How to measure CK by Oliver-Rosalki assay?
involves coupling with hexokinase and G6PD,
in which creatine is produced from creatine
phosphate
Which are the Clinical Significance of CK?
Increased in:
-Myocardial infarction
-Progressive Muscular Dystrophy (Duchenne
type)
-hypothyroidism
-vigorous muscular activity (cerebrovascular
accidents, stroke)
-repeated intramuscular injections
-drugs such as statins, fibrates, anti-retrovirals
and angiotensin II receptor antagonists
Why should Avoid hemolysis in Measurement
Adenylate kinase, ATP and G6P/glucose-6of CK activity?
phosphate liberated from RBCs may affect the
lag phase and may cause side reactions
The following are some additional questions on slides 38-54
Where is found CK-Mt?
In what health condition/disease, the AST level
found to be decreased ?
What does alanine aminotransferases?
What is te required coenzyme for full catalytic
activity of ALT?
How long can the sample being reserved
What is ALT:AST ratio (De Ritis ratio)used
for?
What is ALT:AST ratio increased in?
What is ALT:AST ratio decreased in?
- Autoimmune hepatitis
- Cholestasis
- Cirrhosis
- Alcohol liver disease
catalyzes the transfer of an amino group from
alanine to α-ketoglutarate with the formation of
glutamate and pyruvate
Pyridoxal-5’-phosphate (P-5’-P)
samples stored at room temperature are stable
for 3 days, 1 week if refrigerated
determine the etiology of hepatocellular
disease
infectious (viral) hepatitis
severe liver disease with necrosis (alcoholic
hepatitis, cirrhosis, hepatocellular carcinoma)
49
Chem PPT Flashcards Unit 2
What enzyme Catalyzes the reversible
phosphorylation of creatine by ATP
What is CREATINE PHOSPHOKINASE
(CPK) or
CREATINE KINASE (CK)
What are the isoenzymes of CK?
What appears in in brain damage or nerve
tissues?
Where is CK2 located?
What would be the optimal testing period for
CKMB?
Creatine kinase
a dimer that is composed of two subunits: Bproduct and M-product
- CK1 or CKBB
- CK2 or CKMB
- CK3 or CKMM
CK1 or CKBB
Heart muscle
followingh MI, levels rise within 4-6 hours,
peak at 12-24 hours and return to normal
within 48-72 hours
What type of CK will be use as an indicative of
- CK2 or CKMB
iIschemic heart disease ?
What is the indicative of myocardial damage?
levels is >6%
What enzyme is majorly found in striated and CK3 or CKMM
cardiac muscle and in normal serum?
What will CKMM find to be elevated in?
hypothyroiidism and intramuscular injections
Where will CK-Mt find in?
inner and outer membranes of mitochondria,
and it constitutes up to 15% of total CK
activity
What can be used in Measurement of CK
- Electrophoresis (agarose, cellulose
activity
acetate)
- Ion-exchange chromatography
- Immunoassay
- Tanzer-Gilvarg assay (coulormetric
method)
- Oliver-Rosalki assay
How to reserve CK for accurate result?
- Refrigerated or frozen protected from
light
- Addition of cysteine
2+
What substance is required as an activator for
Mg
full enzyme activity?
In what condition would CK level increased?
- Myocardial infarction
- Progressive Muscular Dystrophy
- Hypothyroidism
- vigorous muscular activity
(cerebrovascular accidents, stroke)
- repeated intramuscular injections
- drugs such as statins, fibrates, antiretrovirals and angiotensin II receptor
antagonists
50
Chem PPT Flashcards Unit 2
What Catalyzes the conversion of lactate to
pyruvate at pH 8.8-9.8 and pyruvate to lactate
at 7.4-7.8, mediated by NAD?
Lactate dehydrogenase (LD/LDH)
What does LD stand for?
What is the end product of LACTATE
DEHYDROGENASE (LD/ LDH)?
Lactate Dehydrogenase
pyruvate + NADH + H+
Which are the Isoenzymes of LDH?
LD1 (HHHH; H4)
LD2 (HHHM; H3M)
LD3 (HHMM; H2M2)
LD4 (HMMM; HM3)
LD5 (MMMM; M4)
predominantly in heart muscle and RBC’s
localization is similar to LD1, but also found in
the kidneys.
Where is found LD1 HHHH;H4 in body?
Where is found LD2 HHHM;H3M?
The Isoenzyme LD3 (HHMM;h2M2) is found
where ?
The Isoenzyme LD3 (HHMM;h2M2) accounts
for what percentage of total LD?
The Isoenzyme LD4 (HMMM; HM3) is found
where ?
The Isoenzyme LD4 (HMMM; HM3) accounts
for what percentage of total LD?
LD5 (MMMM; M4) is similar to?
Based on the detection of NADH the Wacker
method uses?
Based on the detection of NADH the
Wrobleuski-Ladue method employs what?
For measurement of Total LDH activity, why
is serum the preferred specimen ?
Measurement of Total LDH activity avoids
what?
For measurement of Total LDH activity, LD4
and LD5 can be stored at room temperature for
how many days?
What is the normal LD1:LD2 ratio ?
After MI, LD1 is > than LD2 (“flipped
pattern”). Also observed in what?
Immuno-inhibition for the measurement of
LD1 activity involves what?
Clinical Significance of LD is used for
diagnosis of what?
lymphocytes, spleen and pancreas
22%
Liver and Skeletal muscles
10%
LD4 in both localization and portion of total
the lactate-to-pyruvate reaction with the
formation of NADH
the reverse reaction of the Wacker method and
measures the decrease in absorbance as NADH
is consumed.
-heparinized plasma also acceptable
-oxalate has inhibitory effect on the enzyme
Hemolysis
2-3 days
0.45-0.74
Hemolyzed specimens
an antibody that binds all other isoforms,
leaving LD1 to be assayed by chemical reaction
Cardiac, hepatic, skeletal muscles and renal
disease
51
Chem PPT Flashcards Unit 2
LD is elevated in association with what
diseases ?
What are two clinical significances of LD1 and
LD2 when elevated?
What are three clinical significances of LD3
when elevated?
What are two clinical significances of LD4 and
LD5 when elevated?
The LD isoenzyme pattern in renal disease is
very similar to a normal pattern except for the
higher _____________?
Define Diagnostic Window.
When does CK activity increase?
When does CK activity peak?
When does CK activity return to normal?
When does AST activity increase?
When does AST activity peak?
When does AST activity return to normal?
When does LD activity increase?
When does LD activity peak?
How long does LD activity remain elevated?
What does AMI stand for?
What is a myoglobin?
How long does myoglobin take to increase
after infarction?
How long does myoglobin take to double its
initial volume after infarction?
When does myoglobin peak?
When does myoglobin return to normal?
What is myoglobin used in conjunction with to
assess AMI?
What is Troponin?
What are the 3 types of Troponin?
-Pernicious anemia
-Viral hepatitis
-Cirrhosis
-Crush injuries
-Renal disease
Hemolytic anemia
Myocardial infarction (MI)
Pulmonary edema
Pulmonary infarction
Pancreatitis
Liver disease
Skeletal muscle disease
Absolute values
The interval of time after an episode of injury
during which plasma concentrations of the
marker are increased, thereby demonstrating
the occurrence of injury
4-6 hours after the symptoms
12-24 hours
48-72 hours
6-8 hours after the symptoms
24-46 hours
4th of 5th day
8-12 hours after the symptoms
48-60 hours
10-14 days
Acute Myocardial Infarction
A heme protein that can bind oxygen
reversibly found in cardiac and striated
muscles
1-3 hours
1-2 hours
5-12 hours
18-30 hours
Troponin and CK-MB
A group of three proteins that function in
muscle contraction by binding to the thin
filaments of cardiac and skeletal muscles
Troponin T (TnT)
Troponin I (Tn I)
Troponin C (TnC)
52
Chem PPT Flashcards Unit 2
When does Cardiac troponin I (cTn I) increase
after an AMI?
How long does Cardiac troponin I (cTn I)
remain elevated after an AMI?
When is a serial quantification of Cardiac
troponin I (cTn I) necessary after an AMI?
Where is Cardiac troponin T (cTnT) found in
the body?
What is GAMMA GLUTAMYL
TRANSFERASE (GGT)?
What is an example of tissues containing
significant amounts of GGT (descending order
of activity per gram of tissue) ?
a. Kidney
b. Liver
c. Pancreas
d. Intestines
All of the above
How is GGT activity measured?
a. Szasz Method
b. Nonhemolyzed serum
Both A and B
How is GGT activity stored?
List the Clinical Significance of GGT:
a. primarily used as a marker for the
diagnosis of liver related disorders
(intrahepatic or post hepatic obstruction
b. detection of alcoholism and monitoring
of alcohol intake
c. urine contains significant amount of
GGT
d. elevated also in serum of patients
receiving anticonvulsant drugs such as
phenytoin and phenobarbital.
e. All of the above
Between male and female organism, who
shows the higher GGT activities?
GGT values are lowest in adolescents and rise
steadily throughout life:
a. True
b. False
4-8 hours
3-5 days
3-6 hours
6-9 hours
12-24 hours interval
cTnT is found in regenerating and diseased
skeletal muscle expressed in small quantities
GGT is an amino transferase that transfers a
terminal gamma-glutamyl from one compound
to an acceptor compound (transpeptidation)
E
C
Stable for at least one month at 4°C and for
one year at -20°C.
E
Male
A
53
Chem PPT Flashcards Unit 2
What does it usually mean when GGT and
ALP enzymes are elevated and when GGT is
normal but ALP is high?
What is Gamma Glutamyl Transferase (GGT)?
What does GGT do?
What contains a significant amount of GGT?
Name some tissues that contain GGT in
descending order of activity per gram of tissue.
What is used for the measurement of GGT?
What is the preferred specimen for GGT?
What can decrease GGT”s activity by 10%15%?
In what temperature can GGT be stable in and
for how long?
What diagnosis is GGT markers used for?
What contains significant amounts of GGT?
What can the GGT assays detect?
In what other patients can GGT be elevated?
What are some clinical significance of GGT?
Is GGT higher in males or females?
GGT values are lowest in 1) _____? and 2)
_____? steadily throughout life.
What are GGT levels often examined with?
What is suspected if both enzymes (GGT and
ALP) are elevated?
If GGT is normal, and ALP is high then what
is suspected?
What is Amylase (AMS)?
What is the optimum PH of AMS?
What is and what does AMS do?
If both enzymes are elevated, liver disorder is
suspected;
If GGT is normal, and ALP is high, suggestive
of bone disease
Is an amino transferase
Transfers a terminal gamma-glutamyl from one
compound to an acceptor compound
(transpeptidation).
Tissues
1) Kidney (proximal renal tubule)
2) Liver
3) Pancreas
4) Intestines
Szasz Method
*substrate
*gamma-Glutamyle-p-nitroanilide
Nonhemolyzed serum is the preferred
specimen, but EDTA plasma has also been
used.
Heparin may produce turbidity; citrate, oxalate
and fluoride.
Stable for at least one month at 4°C and for
one year at -20°C.
Liver related disorders (intrahepatic or post
hepatic obstruction).
Urine
Alcoholism and monitoring of alcohol intake.
In serum of patients receving acticonvulsant
drugs such as phenytoin and Phenobarbital.
1) Acute pancreatitis
2) Diabetes mellitus (in relation to triglycerides
increased)
In males (do to the contribution of GGT from
the prostate gland)
1) adolescents
2) rise
ALP
Liver disorder
Suggestive of bone disease
It is a digestive enzyme
6.9-7.0
a hydrolase that catalyzes the breakdown of
starch and glycogen producing products
consists of glucose, maltose and dextrins.
54
Chem PPT Flashcards Unit 2
AMS is a metalloenzyme that requires what as
a cofactor?
AMS is the _____? In size.
What are the mist affective activators of AMS?
Ca2+
Smallest
Chloride and bromide
The only plasma enzyme physiologically found Amylase AMS
in urine is?
What are the 2 forms of AMS
1) P-type (pancreas) = ptyalin
2) S-type (salivary glands) = amylopsin
What are 5 measurements of AMS activity?
Amyloclastic method
*measures amylase activity by following the
decrease is substrate concentration
*substrate = starch
2. Saccharogenic method
*measures the amount of reducing
sugars produced by the hydrolysis of starch by
the usual glucose methods
3. Chronometric method
*measures the time required for AMS
to hydrolyzed completely all the starch present
in a reaction mixture.
4. Amylometric method
*measures the amount of starch
hydrolyzed in a fixed period of time using the
intensity of the blue starch iodine color.
5. Coupled-enzyme methods
*measure change in absorbance of NAD.
*AMS acts on maltopentose to produce
maltose that is then reduced to form ATP. This
reaction is the coupled with NAD.
When is the end point, in the chronometric
When there is absence of any substrate capable
method, reached?
of forming the blue starch iodine color.
What are some clinical significance of AMS?
1) Acute pancreatitis
2) Salivary gland lesion
3) Opiates medication
In acute pancreatitis, when do AMS levels
5-8 hours after the onset of attack, peak at 12rise?
72 hours, and then declines to normal in 3-5
days.
What are some salivary gland lesions?
1) Mumps
2) Parotidis
Opiates medication causes pancreatic
Constrict
sphincters to do what?
What is Macroamylasemia?
An asymptomatic condition which results
when the amylase molecule and
immunoglobulins (IgG) combine to form a
complex.
55
Chem PPT Flashcards Unit 2
The complex is too large to be filtered across
the?
The lack of 1) ___ ___? leads to an 2) ____?
serum amylase and 3) _____? Urine amylase.
What is evaluated using the ACCR (amylase:
creatinine clearance ratio?)?
What is ACCR?
Macroamylasemia will show decreased urine
amylase wit?
In what is ACCR increased?
What is Lipase (LPS)
LPS is filtered by what?
LPS is totally reabsorbed by the?
What does LPS do?
What is used for the measurements of LPs
activity?
What is the preferred specimen for LPS
assays?
What is the stability of LPS and at what
temperatures?
What causes a reduction in lipase activity?
What increases lipase activity?
What are some clinical significance in LPS?
Glomerulus
1) Renal clearance
2) Increased
3) Decreased
Macroamylasemia
ACCR is the ratio of urinary clearance of
amylase to urinary clearance of creatinine,
expressed in %.
Low ACCR
In acute pancreatitis
A digestive enzyme secreted by the pancreas.
The glomerulus
The renal tubules, thus is not physiologically
detected in urine.
It catalyzes the hydrolysis of ester bonds in
triglycerides to produce fatty acids and 2acylglycerol.
*Generally, Titrimetric and Turbidimetric
methods
~ Cherry Crandall
~ Tietz Templaton
 Substrate = olive oil
 Cofactor = colipase
Serum




1 week at room temperature
3weeks refrigerated
5 months frozen
repeated freezing and thawing should
be avoided
3. Lipemic specimens
4. Opiates and morphine
1. Acute pancreatitis
 Lipase rises within 4-8 hours,
peaks at about 24 hours, and
decreases within 7-14 days.
2. Pancreatic duct obstruction
3. Tumors of the pancreas
4. Renal insufficiency
56
Chem PPT Flashcards Unit 2
From the graph below, the red line rises within
4-8 hours and peaks at about 24 hours, and
decreases within 7-14 days, what is this
digestive enzyme?
Lipase
The graph below indicates that the black line
Amylase
rise 5-8 hours after the onset of attack, peak at
12-72 hours, and then declines to normal in 3-5
days, what is this digestive enzyme?
What do you call the enzyme synthesized by
the liver that hydrolyzes the esters of choline?
What do you call an enzyme that is also known
as “pseudo-cholinesterase”?
What does Acetylcholinesterase use as a
substrate?
Where is Acetylcholinesterase found in the
body?
a. RBC, Spleen
b. Gray matter, nerve endings
c. A and B
d. Stomach and Heart
Acetylcholinesterase is the false cholinesterase
or WBC cholinesterase. Evaluate this
statement. True or False
Psuedocholinesterase uses what kind of
substrate?
Where is Psudeocholinesterase found in the
body?
a. serum, liver
b. pancreas, heart
c. gray matter
d. A and B
CHOLINESTERASE (CHS)
CHOLINESTERASE (CHS)
Acetlycholine
C.
False
Butyryl esters (butyrylcholine)
D. A and B
57
Chem PPT Flashcards Unit 2
Pusedocholinesterase has other reference
names. Which of these is false.
a. serum cholinesterase (SChE)
b. or acylcholine acylhydrolase
c. butyrylcholinesterase, choline esterase
II
d. acetylcholinesterase
Manometric techniques measure liberated
____________ from the formation of acetic
acid.
a. Carbon Dioxide
b. Carbon Monoxide
c. Phosphate Dioxin
d. Methane Gas
Electrometric techniques determine enzyme
activity by measuring the ____ decrease
resulting in the liberation of acetic acid.
a. Salt
b. Base
c. pH
d. Aerosol
In the Ellman technique the substrate is a
_______ ester that produces a ________,
which reacts with a disulfide to form a colored
compound.
a. Butryl, Butryl
b. Thiol, thiol
c. Glycol, glycol
d. Menthol, menthol
__________ is a commonly used substrate for
measuring serum cholinesterase activity.
a. Psuedocholine
b. Butrylcholine
c. Acetylcholine
d. Propionylthiocholine
The Succinyldithiocholine method uses Serum
or heparinized plasma only. The method is
inhibited by__________.
a. Oxalate
b. Fluoride
c. Citrate
d. All the above
D.
A
C
B
D
D
58
Chem PPT Flashcards Unit 2
PChe is stable for several weeks in refrigerated
samples; __ years in frozen serum but only up
to __ month(s) in frozen plasma
a. 3, 1
b. 2, 2
c. 4, 2
c. 2, 6
When PChe is decreased it is clinically
significant among other factors for
a. organic phosphorus insecticide
poisoning
b. prolonged apnea after succinylcholine
administration during surgery
c. hepatocellular disease, starvation
d. All the above
_______ is an oxidoreductase that catalyzes the
oxidation of glucose-6-phosphate to 6phosphogluconate, which is an important step
in the pentose-phosphate shunt in glucose
metabolism.
Name key organs where G6PD is found:
a. Adrenal Glands, Thymus
b. RBC’s, Lymph Nodes
c. Stomach, Pancreas
d. A and B
G6PD functions to maintain NADPH in a
reduced form to protect hemoglobin from
oxidation and prevent RBC hemolysis. True or
False?
a. True
b. False
The activity of G6PD is by the formation of
NADPH which then is measured mass
spectometrically.
True or False
a. True
b. False
G6PD is increased in Myocardial Infarction
and Megaloblastic Anemia. True or False
a. True
b. False
In Latinos G6PD is a common inherited sexlinked trait. True or False.
a. True
b.False
A
D
G6PD
D
A
B
A
A
59
Chem PPT Flashcards Unit 2
5’-NTP becomes elevated as a result of liver
disease. True or False
a. True
b. False
5’-NTP in _______ levels are more sensitive to
liver cancer.
a. Serum
b. Plasma
c. A and B
d. blood
The lab wants to keep a sample stable for 5’NTP. What is the longest temperature and time
that it can be kept.
a. 4 days at 4 degrees centigrade
b. 2 hours at room temperature
c. 4 months at minus 20 degrees
centigrade
d. None of the above
Lipoprotein-Associated Phospholipase A2 is a
_______ risk marker.
a. Cardiovascular
b. Endocrine
c. Skeletal Muscular System
d. None of the above
Which cells produce Lipoprotein-Associated
Phospholipase A2
a. Monocytes
b. Macrophages
c. Mast Cells
d. All of the Above
Lipoprotein-Associated Phospholipase A2 is
used in detecting atherosclerotic lesions,
especially in complex plaque prone to rupture.
True or False
a. True
b. False
Can increased levels of Lipoprotein-Associated
Phospholipase A2 be associated with future
coronary and cerebrovascular events?
a. Yes
b. No
Lp-PLA2 can measured by the ELISA method
and the Immunoturbidimetric method. True or
False
A. True
B. False
A
A
C
A
D
A
A
A
60
Chem PPT Flashcards Unit 2
Should EDTA plasma be used as a
recommended sample?
a. Yes
b. No
Myeloperoxidase is released in circulation
typically with inflammation.
True or False
a. True
b. False
A
Define Cancer.
it is a term used for diseases in which
abnormal cells divide without control and are
able to invade other tissues. It can spread to
other parts of the body through the blood and
lymph systems.
Is a substance produced by a tumor or by the
host in response to a tumor, which is used to
differentiate the tumor from normal tissue, or
to detect the presence of a tumor based on
measurements in the blood or secretions
It can be used for:-For diagnosis, prognosis and prediction For
monitoring the effects of therapy -as targets
for localization and therapy
It is a useful approach to evaluating multiple
tests for the same analyte or multiple markers
of the same type of cancer
Alkaline Phosphatase
Lactate Dehydrogenase
Neuron-Specific Enolase
Prostatic Acid Phosphatase
Prostate-Specific Antigen
Urokinase-Plasminogen Activator System
Cathepsins
Adrenocorticotropic hormone (ACTH)
Calcitonin
Human Chorionic Gonadotropin (CG)
α-Fetoprotein (AFP)
Carcinoembryonic Antigen (CEA)
Tissue Polypeptide Antigen (TPA)
Tissue Polypeptide Specific Antigen (TPS)
CYFRA 21-1
Squamous Cell Carcinoma Antigen (SCCA)
-Mucins
-CA 125
-CA 15-3
-HE4
-CA 27.29
-OVA1
-Urinary Bladder Tumor Markers
-Nuclear Matrix Protein (NMP22)
-Bladder Tumor-Associated (BTA) Analytes
DefineTumor Marker.
What is the use of Tumor Marker?
What is Receiver operating characteristic
curve (ROC) ?
Enzymes as Tumor Markers.
Hormones as Tumor Markers.
Oncofetal Antigens as tumor markers.
Cytokeratins as tumor markers.
Carbohydrate Markers as tumor markers.
Which are Bladder Cancer Markers?
A
61
Chem PPT Flashcards Unit 2
Proteins as tumor markers.
Receptors as tumor markers.
Genetic and Molecular Markers.
Which are the primary sources of Alkaline
Phosphatase
What can be seen in primary and secondary
liver cancer?
When does Lactate dehydrogenase release
take place?
Neuron-Specific Enolase (NSE) associated
with which tumors?
What is Prostatic Acid Phosphatase (PAP)?
How is Prostate-Specific Antigen (PSA)
produce?
Where is found Prostate-Specific Antigen
(PSA)?
Which of the greatest clinical use of PSA?
Which methods are used to measure PSA?
Which are the components mainly used in
Urokinase-Plasminogen Activator System?
-Soluble Mesothelin-Related Peptides
(SMRPs)
-Des-y-carboxy prothrombin (PIVKA-II)
S-100 Proteins
-Thyroglobulin and Antibodies
Chromogranins
-Estrogen and Progesterone Receptors
-Epidermal Growth Factor Receptor
Oncogenes:-RAS genes
-HER2
-BCR-ABL
Tumor-Suppressor Genes:-Retinoblastoma gene
-adenomatous polyposis Coli (APC) Gene
-BRCA1 and BRCA2
liver, bone and placenta
-Elevation of ALP with elevations in 5'nucleotidase or γ-glutamyltransferase
-as a result of cell damage
tumors of neuroendocrine origin like small cell
lung cancer (SCLC), neuroblastoma,
pheochromocytoma, carcinoid, medullary
carcinoma of the thyroid, melanoma and
pancreatic endocrine tumor
It is considered to be less sensitive for the
detection of early cancer but may prove useful
when combined with other markers for
improving prostate cancer detection or
predicting recurrence after radical
prostatectomy.
produce exclusively by epithelial cells of the
acini and ducts of the prostate gland and
widely use to screen, detect and monitory
treatment of prostate cancer.
concentrations of PSA has been found to
correlate with clinical stages of prostate
cancer.
PSA involves monitoring of definitive treatment
for prostate cancer which includes radical
prostatectomy, radiation therapy and antiandrogen therapy.
Sandwich immunoassays using labels such as
enzymes, fluorescence or chemiluminescence
1. Urokinase plasminogen activator (uPA)
2. uPA membrane-bound receptor (uPAR)
3. uPA inhibitors, PAI-1 and PAI-2
62
Chem PPT Flashcards Unit 2
Which is the main activity of UrokinasePlasminogen Activator ?
What is the used of Urokinase-Plasminogen
Activator System?
Which method is used to measure UrokinasePlasminogen Activator System?
What is Cathepsin?
What happened if Cathepsin level increased?
What methods are used to measure Cathepsin
concentration?
Which gland produced Adrenocorticotropic
hormone?
elevated
What happened if Adrenocorticotropic hormone
concentrations of plasma is elevated?
Which are cells are responsible to produced
Calcitonin?
What happened if Calcitonin concentration is
elevated?
Which cells are produced Human chorionic
gonadotropin (CG)?
the main activity is the conversion of
plasminogen to active plasmin
It has been used as a prognostic marker in
breast cancer, colorectal cancer, ovarian,
renal, hepatocellular, pancreatic, urinary,
bladder, lung (adenocarcinoma) and cervical
cancers as well as gliomas.
measured in the laboratory by the EnzymeLinked Immunosorbent Assay (ELISA)
Cathepsin are lysosome on protease
enzymes: Cathepsins B, D and L
Cathepsin increased levels are demonstrated
in breast, colorectal, gastric, lung, prostate
carcinoma, gliomas, melanomas and
osteoclastomas.
Cathepsin concentrations are generally
measured in tissue extracts by ELISA or
directly in the tissues by
immunohistochemistry
Adrenocorticotropic hormone produced by
corticotropic cells of the anterior pituitary gland
Elevated plasma concentration pancreatic,
breast, gastric and colon cancer, and benign
conditions such as chronic obstructive
pulmonary disease, mental depression,
obesity, hypertension, diabetes mellitus and
stress.
It is produced by the C cells of the thyroid in
response to increased serum calcium. It
inhibits the release of calcium from bone and
thus lowers the serum calcium concentration
-Elevated concentration is usually associated
with medullary carcinoma of the thyroid (MTC)
-It is also elevated in patients with carcinoid
tumors and cancers of the lung, breast, kidney
and liver
-It is also elevated in other nonmalignant
conditions such as pulmonary disease,
pancreatitis, hyperparathyroidism, pernicious
anemia, Pagets' disease of bone and
pregnancy
Human chorionic gonadotropin (CG)
is a glycoprotein secreted by the
syncytiotrophoblastic cells of the healthy
placenta
63
Chem PPT Flashcards Unit 2
What happened if Human chorionic gonadotropin
(CG) elevated?
What is the use of GC and AFP together?
What method is used to measure chorionic
gonadotropin?
What is Oncofetal antigens?
How to work Oncofetal antigens in fetal life?
Which are the Oncofetal antigens tumor markers?
What happened if α-fetoprotein (AFP) is elevated?
α-fetoprotein (AFP) is determined by Which
method?
What is the other name of Carcinoembryonic
Antigen (CEA)?
Which is the origin of Cytokeratins?
What happened if Tissue polypeptide antigen
(TPA) is elevated?
What is the use of Tissue polypeptide antigen
(TPA)?
What is tissue Polypeptide-Specific Antigen ?
-Elevated concentrations are seen in
pregnancy, trophoblast disease and germ cell
tumors
-Also elevated in biliary cancer, pancreatic cancer
and other cancers including bladder, renal, prostate,
liver, colorectal, non-small cell lung, breast and
head and neck cancers
GC together with α-fetoprotein (AFP) is useful
in detecting nonseminomatous testicular tumor
measured in the laboratory by a immunometric
(sandwich) technique
Oncofetal antigens
are proteins produced during fetal life
-present in high concentration in the sera of
fetuses and decrease to low concentrations or
disappear after birth
-This proteins often reappear in cancer
patients, indicating the reactivation of certain
genes as a result of the malignant
transformation of cells
α-fetoprotein (AFP)
Carcinoembryonic Antigen (CEA)
Elevated AFP and serum bilirubin
concentrations (>2 mg/dL) are associated with
a decreased survival time
Levels are determined by immunometric assay
"Gold" antigen
Cytokeratins group of proteins that make up
the cytoskeletal intermediate filaments of
epithelial cells and cells of epithelial origin.
Tissue polypeptide antigen (TPA)
Elevated serum concentrations are related to
the proliferative activity and turnover of cells,
allowing it to be used as a proliferation marker.
In monitoring of metastatic disease, TPA is
useful when comb (ined with CEA and CA 153 in breast cancer, with CEA and CA 19-9 in
colon cancer, and with CA 125 in ovarian
cancer.
TPA may be helpful in the differentiation of
cholangiocarcinoma (increased TPA) from
hepatocellular carcinoma (in which TPA is not
elevated)
an antigenic site on cytokeratin 18 that is
specifically recognized by the M3 monoclonal
antibody
64
Chem PPT Flashcards Unit 2
CYTOKERATINS AS TUMOR MARKERS
have been proposed as a specific marker for
cell proliferation and is detectable in serum
with the use of a _____ _____?
CYFRA 21-1 is elevated in all types of _____
_____, although it is most sensitive for nonsmall cell ____ _____.
Squamous Cell Carcinoma Antigen (SCCA) is
a _______ referred to as tumor associated
antigen 4.
Squamous Cell Carcinoma Antigen (SCCA) is
useful in detecting recurrence of ______ and in
monitoring treatment and disease progression
CA 15-3 is detected by a _____ ______ _____
against a membrane-enriched extract of the
human breast cancer metastatic to liver
CA 15-3 elevated concentrations are found in
______ including pancreatic, lung, breast,
ovarian, colorectal and liver cancers.
CA 27.29 is used for detecting recurrent _____
_____ in patients with stage II or stage III
disease and for monitoring response to therapy
in patients with stage IV (metastatic) disease
CA 125 is a marker for monitoring what?
CA 125 is also used to detect ______ or _____
disease in patients who have undergone firstline therapy and would be considered for
second-look procedures
CA 125 is also elevated in non-_____
carcinoma including pancreatic, lung, breast,
colorectal, gastrointestinal tumors and useful in
determining the prognosis of patients with
endometrial carinoma
CA 125 is determined by what?
CA 19-9 (sialylated Lexa) a marker for
gastrointestinal cancer and is used primarily in
patients with _____ _____?
CA 19-9 has been approved by the FDA for
quantitative measurement in serum and as an
aid in monitoring _____ _____ patient
Monoclonal immunoglobulin has been used as
a marker for multiple ______?
Monoclonal paraproteins appear as sharp bands
in the _____ region of serum electrophoretic
patterns.
specific immunoassay
lung cancer, lung cancer
glycoprotein
cancer
murine monoclonal antibody
malignancies
breast cancer
epithelial ovarian cancer
residual or recurrent
ovarian
determined by immunoradiometric essay
pancreatic carcinoma
pancreatic cancer
myeloma
Globulin
65
Chem PPT Flashcards Unit 2
Bence Jones proteins is what?
What type of cancer is detected through
cytoscopy or cytology of shed cells using
tumor markers like NMP22, complement
factor-H (CFH), fibronectin, telomerase,
cytokeratins and survivin.
Bladder cancer is measured what type of
technique?
What type of proteins have functions that have
been associated with regulating key reactions
in the nucleus, such as DNA replication and
RNA synthesis
What detects the antigen human complement
factor-H related protein, which is a variant of
human complement factor-H
Bladder Tumor Associated analytes
functions in the alternative complement
pathway by interacting with complement factor
C3b to prevent what?
_______ is a cell surface glycoprotein express
on mesothelial cells, and mesothelin fragments,
found in the circulation of patients with
mesothelial tumor.
Mesothelioma is a rare cancer of the
mesothelial surfaces of the pleural and
peritoneal cavities or the pericardium that is
linked to what?
What type of tumor markers are used as
Calcium binding proteins for diagnostic
histological marker of melanoma and
melanoma metastases
What type of protein is a family of proteins
that are major components of the secretory
granules of most neuroendocrine cells
Chromogranins is useful in detecting _____
______ like carcinoid tumor,
pheochromocytoma and neuroblastoma
Thyroglobulin and antibodies
produced by the _____ gland as the precursor
to ____ hormone
Thyroglobulin and antibodies are used for
monitoring patients with a diagnosis of
differentiated ____ cancer after thyroid gland
ablation.
a free monoclonal immunoglobulin light chain
in the urine
Bladder Cancer
fluorescence in situ hybridization (FISH)
Nuclear Matrix Protein (NMP22)
Bladder Tumor Associated analytes
cell lysis
Mesothelin
asbestos exposure
S-100 Proteins
Chromogranins
neuroendocrine tumors
thyroid
Thyroid
66
Chem PPT Flashcards Unit 2
What antibodies are used to monitor residual
disease and or recurrence?
Thyroglobulin and antibodies
are measured by what?
Estrogen and progesterone receptors are
members of the nuclear ____ hormone receptor
family and are involved in hormone directed
transcriptional activation
Estrogen and progesterone receptors are used
in tumor markers as indicators of what type of
cancer for hormonal therapy?
Immunohistochemistry assay is used to
measure steroid hormone receptors in what
type of tumor tissue specimens?
Epidermal Growth Factor Receptor (EGFR) is
a prototype of what type of family receptors?
What is used as a companion diagnostic to aid
in identifying colorectal cancer patients?
Prostate Cancer Gene or Antigen3 (PCA3) a
molecular test for what?
Single-Nucleotide Polymorphisms
genes are associated with what?
Cell-Free Nucleic Acids are markers for what 2
types of cancer?
Creatinine is a diagnostic indicator of what
function?
What three organs is Creatinine synthesized in?
True or False: Creatinine is present in all body
fluids and secretions.
What is Creatinine a marker of?
What reaction is used in Creatinine?
The Jaffe reaction of Creatinine has a reaction
with what?
Creatinine analytical methods tend to lack
what?
Name two enzymes of Creatinine
Antithyroglobulin
Immunometric assays and RIA
steroid
Breast cancer
breast
tyrosine kinase
Epidermal Growth Factor Receptor
prostate cancer
human disease
lung cancer and colorectal cancer
Kidney




Kidneys
Liver
Pancreas
True
 Glomerular Filtration Rate (GFR)
 Jaffe reaction
 Picrate ion
 Specificity


What are four analytical methodology factors

that are used in Creatinine?

Enzymatic methods (Fig. 21-1)


What type of state is Urea?
•
Urea is an underestimation of what?
•
Urea is an outdated indicator of what function? •
Creatininase
creatinase
Hydroxide
Wavelength
Temperature
“Concentration”
Azotemic state
Glomerular filtration rate
Kidney
67
Chem PPT Flashcards Unit 2
What is a chemical method of Urea?
What is an enzymatic method of Urea based
on?
What three types of Urea enzymatic methods
approached is used?
What does a spectrophotometric approach
consist of?
What does the biochemistry and physiology of
Uric Acid focus on?
A clinical significance of Uric Acid that
involves, 20-plus inherited disorders of what
metabolism?
Uric Acid is used as a measurement in
management of what?
What does Uric Acid help determine on during
Hypouricemia?
Name two analytical methodology of Uric
Acid?
Which is more specific, PTA or Uricase?
What does HPLC-based methods focus on
during Uric Acid?
What does reference values of Uric Acid
indicate?
What are Carbohydrates ?
What is a Monosaccharides ?
What is an example of a Monosaccharide?
What are Oligosaccharides ? Give an example
•
•
Fearon reaction
Preliminary hydrolysis of urea with urease
 Spectrophotometric
 Dry Chemistry Systems
• Specificity
Berthelot method and enzymatic assay
 Urate pool
Renal handling of uric acid
 Purine metabolism
 Gout
Link to underlying conditions
 Other causes
 Phosphotungstic acid (PTA)
 Uricase
Uricase
Ion-exchange or reversed-phase
 Gout
 Purine-containing and purine-free diets
•Aldehyde or ketone derivatives of
polyhydroxy (more than one -OH group)
alcohols composed of carbon, hydrogen, and
oxygen in the ratio of 1:2:1
•they serve as structural components in RNA
and DNA sugars
•considered to be the main source of energy
Simple sugars that contain 4-8 carbons and
only 1 aldehyde or ketone group.
They are reducing sugars with the general
formula of Cn(H2O) n that cannot be broken
down to simpler substances by acid hydrolysis.
Glucose
Sometimes referred to as disaccharides which
are formed by the interaction of two
monosaccharides with the loss of water
molecule.
Examples are: Maltose → two molecules of
glucose
;Lactose → glucose and galactose; Sucrose →
glucose and fructose
68
Chem PPT Flashcards Unit 2
Describe Polysaccharides?
What is considered the primary energy source
for the human body?
What are the results of Glucose metabolism?
What is Glycogenesis and Glycolysis?
What is Hypoglycemia and its symptoms?
Describe neuroglycopenia and its
characteristics
How is a 72-hour test for Hypoglycemia
conducted?
How to determine a true Hypoglycemia?
How does Ethanol produces hypoglycemia?
One way to determine an insulin producing
pancreatic islet cell tumor is?
Carbohydrates formed by the interaction
between many units of simple sugars.
Examples include starch and glycogen
Glucose
•energy production by conversion to carbon dioxide
and water
•storage as glycogen in the liver or as triglycerides in
adipose tissue
•conversion to keto acids, amino acids or
protein
Glycogenesis is the conversion of glucose to
glycogen and Glycolysis is the conversion of
glucose or other hexoses into lactate or
pyruvate.
Hypoglycemia is a blood glucose concentration
below the fasting value (<50 mg/dL or 2.75
mmol/L). Symptoms include trembling,
sweating, nausea, rapid pulse, lightheadedness,
hunger and upper abdominal discomfort
Neuroglycopenia is caused by the lack of
glucose supply in the brain. It is characterized
by headache, confusion, blurred vision,
dizziness, seizures, loss of consciousness and
even death.
•Fasting hypoglycemia in adults can be
monitored by the 72 hour fasting test
•symptoms usually begin at plasma glucose
concentrations below 55 mg/dL, and
impairment of cerebral function begins when
glucose is less than 50 mg/dL
•the 72 hour fast should be conducted in the
hospital
•the patient should be allowed a liberal intake
of calorie free and caffeine free fluids
True hypoglycemia will show an abnormally
low value within 12 hours of beginning a fast
Ethanol produces hypoglycemia by inhibiting
gluconeogenesis, and this inhibition is
aggravated by malnutrition (low glycogen
stores) in individuals with chronic alcoholism.
The demonstration of a low plasma glucose
concentration in the presence of a high plasma
insulin value
69
Chem PPT Flashcards Unit 2
Name possible causes of Postprandial
Hypoglycemia
What are some criteria of Whipple's triad
overflow blood glucose concentration?
What are the Pathophysiological mechanisms
that contribute to hypoglycemia in patients
with diabetes?
Where does the Lactic acid predominantly
derived from?
What is the Cori cycle ?
How can the measurement of pyruvate be
useful?
What can a lactate-to-pyruvate ratio indicate?
Describe Lactic acidosis
When testing for carbohydrate metabolism, a
deficiency or absence of an enzyme that
participates in carbohydrate metabolism may
result in what inborn error?
Name the 2 sugars of clinical interest that are
found in the urine?
•drugs
•antibodies to insulin or insulin receptor
•inborn errors (fructose-1,6- diphosphatase
deficiency)
•reactive hypoglycemia (functional
hypoglycemia)
•symptoms known or likely to be caused by
hypoglycemia
•low glucose when symptoms occur
•relief of symptoms when glucose is increased
the normal
Defective glucose counter regulation (ability to
increase glucose, counter to the effect of
insulin)
unaware of hypoglycemia
white skeletal muscle, brain, skin, renal
medulla and erythrocytes, and are metabolized
in the liver and kidneys
The Cori cycle is the conversion of glucose to
lactate in the periphery and reconversion of
lactate to glucose in the liver
Measurement of pyruvate is useful in the
evaluation of patients with inborn error of
metabolism who have increase serum lactate
concentration.
Pyruvate is also measured in clinical studies
evaluating reperfusion after myocardial
ischemia
A lactate-to-pyruvate ratio of less than 25
suggests a defect in gluconeogenesis, whereas
an increase ratio (<35) indicates reduced
intercellular conditions found in hypoxia
Lactic acidosis is a lactate concentrations
exceeding 5mmol/L with pH less than 7.25.
Occurs in 2 clinical settings:
•Type A (hypoxic) associated with decreased
tissue oxygenation
•Type B (metabolic) associated with diabetes
mellitus, neoplasia, liver disease, drugs and
inborn error of metabolism
Accumulation of monosaccharides, which are
measured in the urine
Glucose and Galactose
70
Chem PPT Flashcards Unit 2
Glucose can be checked by what?
Why should urine from infants and children be
tested by both the glucose oxidase and copper
reduction test?
Name the 5 techniques used to separate and
identify sugars?
Where in the body is glycogen predominantly
stored?
What is the cause of Glycogen Storage
Disease?
Lipids are a class of compounds that are
soluble in
but insoluble
in
?
Which organic compound can serve as
hormones, energy source, assist with ingestion
and act as structural components in cell
membranes?
Name the 6 forms of lipids:
What form of lipid is an unsaturated steroid
alcohol whose structure is based on the CPPP
nucleus?
CPPP stands for?
Where does absorption of cholesterol mostly
occur?
Absorption of cholesterol is mediated by what
enterocyte protein?
In the intestinal mucosal cells, cholesterol is
packaged with triglycerides, phospholipids and
a large protein, called apolipoprotein (apo) B48, into large lipoprotein particles called?
What is the function of the chylomicrons?
Cholesterol is synthesized by all cells in the
body, but particularly by what organs?
Glucose oxidase strips
To identify individuals with inborn error of
metabolism



fermentation
optical rotation
osazone formation with
phenylhydrazine
 specific chemical test
 paper or thin layer chromatography
Liver and Skeletal muscles
Deficiency of a specific enzyme in glycogen
metabolism
Organic solvents; water
Lipids
 Cholesterol
 Fatty acids
 Acylglycerols (Glycerol esters)
 Sphingolipids
 Prostaglandins
 Terpenes
Cholesterol
Cyclopentanoperhydrophenanthrene
In the middle jejunum and the terminal ileum of the
small intestines
NPC1L1 (Niemann-Pick C1-like1).
Chylomicrons
Chylomicrons are secreted into the lymph and
enter the circulation, where they deliver the
absorbed dietary lipid to the liver and
peripheral tissues
Liver and intestines
71
Chem PPT Flashcards Unit 2
What are the three stages to cholesterol
biosynthesis?
Cholesterol is esterified to a fatty acid to form
a cholesteryl ester by what two enzymes?
What happens to cholesterol when it is
esterfied?
Specialized endocrine cells can use cholesterol
for the synthesis of what?
How is most cholesterol catabolized?
What is referred by the term, essential fatty
acids?
Name the only 2 fatty acids known to be
essential to humans:
What is the term that is used for other fatty
acids that are sometimes essential under some
developmental or disease conditions?
What 2 fatty acids are conditionally essential?
Where does the fatty acid get metabolized and
produced energy by a series of reactions
known as β-oxidation?
When fatty acid is metabolized it produces
energy by a series of reactions, this reaction is
called what?
What is Krebs cycle?
What is the main energy storage molecule in
the body?
True or False
Ketone formation develops from excessive
production of acetyl-CoA (ketosis)

involvement of acetyl-coenzyme A
(CoA)
 involvement of the microsomal enzyme
HMG-CoA ( hydroxy-methyl-glutaryl
CoA) reductase
 a series of oxidation-decarboxylation
reactions in the endoplasmic reticulum
 acylcholesterol acyltransferase (ACAT)
 lecithin cholestrol acyltransferase
(LCAT)
It loses its free hydroxyl group and becomes
more hydrophobic
Steroid hormones
Most cholesterol is converted in the liver into
bile acids
The term "essential fatty acid" refers to fatty
acids required for biological processes but does
not include the fats that only act as fuel.
 alpha-linolenic acid (an omega-3 fatty
acid)
 linoleic acid (an omega-6 fatty acid)
Conditionally essential
 docosahexaenoic acid
 gamma-linolenic acid
Mitochondria
β-oxidation
A common pathway for the final oxidation of
nearly all metabolic fuels (carbohydrate, fat or
protein) and ultimately results in the
production of ATP (adenosine triphosphate)
ATP (adenosine triphosphate),
True
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Chem PPT Flashcards Unit 2
True or False
In ketone formation, as the body attempts to
obtain necessary energy from stored fat, in the
absence of an adequate supply of carbohydrate
metabolites as in cases of prolonged starvation
or when carbohydrate metabolism is impaired
(as in diabetes mellitus)
What are the forms of ketone bodies?
What do you call a three-carbon alcohol that
contains a hydroxyl group on each of its
carbon and is classified by the number fatty
acyl groups present?
What is acylglycerol?
How many carbons does ACYLGLYCEROLS
contain?
What do you call an ACYLGLYCEROL that
has one fatty acid?
What do you call an ACYLGLYCEROL that
has two fatty acids?
What do you call an ACYLGLYCEROL that
has three fatty acids?
What type of lipid that constitutes 95% of
tissue storage fat and are the predominant form
of glyceryl esters found in plasma?
Triglycerides from plant sources (corn,
sunflower, safflower) are liquid oils at room
temperature tend to be enriched with what kind
of fatty acid?
Triglycerides from animals (ruminants) that are
solid at room temperature and tend to have this
kind of fatty acid, what is this fatty acid called?
Where does dietary triglyceride get digested?
After being digested in the duodenum, dietary
triglycerides get absorbed where?
True
acetoacetic acid
Β-hydroxybutyric acid
acetone
ACYLGLYCEROLS
(GLYCEROL ESTERS)
A three-carbon alcohol that contains a
hydroxyl group on each of its carbon and is
classified by the number fatty acyl groups
present: one fatty acid - monoacylglycerols
(monoglycerides); two fatty acids diacylglycerols (diglycerides); and three fatty
acids - triacylglycerols (triglycerides)?
3
monoacylglycerols
diacylglycerols (diglycerides);
triacylglycerols (triglycerides)
Triglycerides
Unsaturated fatty acids
saturated fatty acids
Dietary triglycerides are digested in the
duodenum
Absorbed in the proximal ileum.
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Chem PPT Flashcards Unit 2
Dietary triglycerides through the action of
pancreatic and intestinal lipases and in the
presence of bile acids, they are first hydrolyzed
to what?
Through the action of and intestinal lipases and
in the presence of bile acids, they are first
hydrolyzed to ____(1)___, ___(2)___,and
___(3)_. After absorption, they are
reassembled as triglycerides in the intestinal
epithelial cells and then are package with
cholesterol and apoB-48 to form ___(4)_____
What do you call the lipoprotein that carries
triglycerides to the liver and peripheral cells?
______________are the main metabolic fuel
carried by chylomicrons to the liver and
peripheral cells.
A ________ is a dihydric 18-carbon alcohol,
derived from the amino alcohol sphingosine.
What form of lipids that have a more general
role in cellular interactions and serves as a
source of blood group and tumor antigens?
What are prostaglandins?
What group of physiologically active lipid
compounds, which have diverse hormone-like
effects in animals and found in almost every
tissue in humans and other animals?
Prostaglandins are derived enzymatically
from?
True or False
The structural differences between
prostaglandins account for their different
biological activities.
True or False
A given prostaglandin may have different and
even opposite effects in different tissues.
The ability of the same prostaglandin to
stimulate a reaction in one tissue and inhibit
the same reaction in another tissue is
determined by?
With their target cells present in the immediate
vicinity of the site of their secretion,
Prostaglandin acts as what?
To glycerol, monoglycerides, and fatty acids.
1) glycerol
2) monoglycerides
3) Fatty acids
4)chylomicrons
chylomicrons
Triglycerides
Sphingolipids
Sphingolipids
Any of the group of compounds derived from
unsaturated 20-carbon fatty acids (primarily
arachidonic acid) via the cyclo-oxygenase
pathway
prostaglandins
fatty acids
True
True
The type of receptor which the prostaglandins
bind.
autocrine or paracrine factors
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Chem PPT Flashcards Unit 2
How does Prostaglandins differ from endocrine Prostaglandins differ from endocrine hormones
hormones?
in that they are not produced at a specific site
but in many places throughout the human
body.
What are the two derivatives of prostaglandin? prostacyclins and thromboxanes
What derivative of prostaglandin that is
Prostacyclins
powerful locally acting vasodilators and
inhibits the aggregation of blood platelets?
Through their role of this prostaglandin
Prostacyclins
derivative in vasodilation, they are also
involved in inflammation. What is this
derivative called?
Where does Prostacyclins get synthesized?
They are synthesized in the walls of blood
vessels and serve the physiological function of
preventing needless clot formation, as well as
regulating the contraction of smooth muscle
tissue
What derivative of prostaglandins produced by Thromboxanes
platelet cells?
What Prostaglandin derivatives are
vasoconstrictors and facilitate platelet
aggregation?
What is this polymer consisting of the fivecarbon isoprene unit and include vitamins A,
E, and K and the dolichols, which play
important roles in protein glycation?
_____(1)____ and _____(2)______ are the
primary constituents of the essential oils of
many types of plants and flowers. Essential
oils are used widely as fragrances in
perfumery, and in medicine and alternative
medicines such as aromatherapy.
What are these macromolecular complexes that
serve as transport vehicle of insoluble lipids in
the plasma?
What is the role of the lipoprotein in insoluble
lipids in the plasma?
What is the appearance of a lipoprotein?
What do you call this major protein component
that is found on the surface of a lipoprotein?
Thromboxanes
Terpenes
1)Terpenes
2)terpenoids
Lipoprotein
Serves as transport vehicle of insoluble lipids
in the plasma.
They are typically spherical particles with
nonpolar neutral lipids (triglycerides and
cholesterol esters) in their core and more
amphipathic lipids (phospholipids and free
cholesterol) at their surface.
Apolipoproteins
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Chem PPT Flashcards Unit 2
What are the categories of lipoprotein?
True of False.
The larger lipoproteins contain more core
lipids, triglycerides and cholesterol esters and
are lighter in density and contain a small
percentage of protein.
True or False
In the fasting state, most plasma triglycerides
are present in VLDL, but 10 to 12 hours after a
meal, most triglycerides are transported on
chylomicrons.
How much total of plasma cholesterol is
carried by LDL?
What Lipoprotein contains about 20% to 30%
of plasma cholesterol?
True or False
Lower density corresponds to larger particle
and lower protein-to-lipid ratio, while higher
density are smaller and have higher proportions
of protein
In lipoprotein, density is inversely proportional
to the protein content and directly proportional
to the lipid content and size of the molecule
Complete the chart:
_(1)_ density = _(2)_ protein content
↑ density
= _(3)_ lipid content
_(4)_ density =
↓ size of the
molecule (smaller)
Are the Apolipoproteins; major components of
lipoproteins?
True or False
A. True
B. False
1)Chylomicrons
2)Very Low Density Lipoprotein (VLDL) or
pre-β-Lipoprotein
3)Intermediate Density Lipoprotein (IDL)
4)Low Density Lipoprotein (LDL) or βLipoprotein
5)High Density Lipoprotein (HDL) or αLipoprotein
6)Lipoprotein (a) [Lp(a)]
True
False (2 – 6 hours after a meal)
carries about 70% of total plasma cholesterol
HDL
True
False (directly proportional to the protein
content and inversely proportional to the lipid
content and size of the molecule)
1) ↑
2) ↑
3) ↓
4) ↑
A
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Chem PPT Flashcards Unit 2
Apolilipoproteins function to:
A. modulate the activity of enzymes that
act on lipoproteins
B. maintain the structural integrity of the
lipoprotein complex
C. facilitate the uptake of lipoprotein by
acting as ligands for a specific cellsurface receptors
D. All of the above
Apo A-1 is not the major protein in HDL. True
or False
A. True
B. False
Chylomicrons have what type of
Apolipoprotein?
A. Apo A-1
B. Apo B-100
C. Apo B-48
D. none of the above
Lipoproteins have several pathways. Identify
them.
A. Exogenous Pathway and the
Endogenous Pathway
B. Intercellular-Cholesterol Transport
Pathway and the Reverse-Cholesterol
Transport Pathway
C. A and B
D. None of the Above
The transport of dietary lipids from the
intestines to the liver and peripheral cells
mediated by:
A. Chylomicrons
B. RBC’s
C. WBC’s
D. Adipose Tissues
The endogenous pathway’s is transfer _______
derived________ .
A. Hepatically, Lipids
B. Splenically, Triglycerides
C. Kidney, Monoglycerides
D. Stomach, Diglycerides
D
B
C
C
A
A
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Chem PPT Flashcards Unit 2
The endogenous pathway has which mediator
lipoprotein?
A. Apo A-1
B. Apo B-100
C. Apo B-48
D. all of the above
Cells use the cholesterol for several roles.
Identify the appropriate role.
A. Membrane Biogenesis
B. Vesicular Transport
C. Osmosis
D. Exocytosis
Do cells have a surface receptor for
cholesterol?
A. Yes
B. No
The Reverse Cholesterol Transport Pathway’s
function is to remove excess cellular
cholesterol from peripheral cells and return it
to the liver for excretion. What is the mediator?
A. HDL
B. VLDL
C. LDL
D. chylomicrons
Are Coronary Heart Disease, Type V
Hyperlipoproteinemia, and Familial
hypercholesterolemia clinically significant?
A. Yes
B. No
The Lab has been asked to conduct a total lipid
panel. What tests must be run?
A. Total Cholesterol
B. Triglyceride
C. HDL-C
D. LDL-C
E. All of the above
B
A
A
A
A
E
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