Midterm Review Flashcards
Order the following metric system units in order from largest to smallest:
Picometer, Milliliter, Nanoliter, Microliter
- Milliliter
- Microliter
- Nanoliter
- Picoliter
In the Collins paper, you learned about a cell line derived from a patient with acute promyelocytic leukemia. What is the name of this cell line?
HL-60
Describe the steps of the scientific method in order.
- Observation
- Hypothesis
- Experimental Design
- Data Collection
- Data Analysis
- Conclusion
A _______ article is a scientific literature source that is written by people who actually performed the experiments and are presenting their own data.
Primary
A ______ article summarizes and analyzes the work of several different groups of scientists.
Review
You are viewing a stained blood cell with a compound light microscope under the 40X objective lens and the 10X ocular lens. What is the total magnification of the stained cell when viewed at these parameters?
400x
In the Xie paper, the authors described a certain collagenase/gelatinase enzyme called _________, which is found to be highly expressed in HL-60 cells that have differentiated into monocytes.
MMP-9
Our microscopes are _______ and _______ (the first means the microscope stays relatively in focus between objectives, while the second means the sample will stay relatively in the center of the field of view).
- Parfocal
2. Paracentral
What are HL-60 cells, and why do they make a good model system for stem cell research?
HL-60 cells are cells derived from a woman with acute promyelocytic leukemia that have been made into a cell line (they are immortal). They are a good model system for stem cell research because they are multipotent cells, able to differentiate into a variety of different cell types.
PMA induces differentiation of HL-60 cells, and activation of the production of a protein called MMP-9. It does this by binding to a receptor on the surface of the cell, and activating a second messenger called ________.
PKC-B
The May-Grunwald/Giemsa staining procedure was used to view changes in the HL-60 cells’ ________.
Morphology
List the 2 types of apoptotic pathways
- Extrinsic
2. Intrinsic
The Central Dogma of Molecular Biology is:
DNA is transcribed into mRNA, which is then translated into protein
Name the 2 chemicals we have used to induce HL-60 cells to differentiate, and the cell types they were expected to differentiate primarily into based on the treatment
- PMA = Monocytes
2. DMSO = Granulocytes
Enzymes called _______ are special proteases that cleave DNA and cellular components into fragments during apoptosis.
Caspases (or executioner caspases)
Like stem cells, HL-60 cells are capable of becoming a different cell type upon the proper inductive stimulus. HL-60 cells are ________ and can be induced to differentiate into cells of the white blood cell lineage.
Multipotent
MMP-9 is a member of a family of enzymes called matrix metalloproteinases, released by ________, that digest the ______ in order to improve cell motility during immune response.
Monocytes, extracellular matrix
________ is a housekeeping gene, and therefore very abundant in all cell types. For this reason, we used it as our control for PCR.
B-actin
In order to visualize our PCR results, we used _______, a method for separating molecules based on size and charge.
Gel electrophoresis
What does PCR stand for?
Polymerase Chain Reaction
What do we used PCR for?
To amplify a certain segment of DNA that is of interest
Name the PCR step:
Addition of nucleic acid bases complementary to a template strand, to the end of an oligonucleotide primer by Taq polymerase
Extending the primers to generate a PCR product
Name the PCR step:
Pairing of short strands of DNA by hydrogen bonding to form complementary double-stranded sequences targeting the sequence of interest
Annealing of Primers
Name the PCR step:
Splitting the hydrogen bonds between complementary strands of DNA to form single strands
Denaturing the DNA
We typically make cDNA in order to view changes in:
Transcription
An agarose gel is used to separate (DNA, proteins) by size
DNA
Put the following signaling pathway components in order from start to finish
PKC-B MMP-9 PMA Transcription Factor Membrane Receptor
PMA Membrane Receptor PKC-B Transcription Factor MMP-9
We used RT-PCR to view changes in:
Transcription
For statistical reason, it is necessary to state a ________ and an _______.
Null hypothesis, alternate hypothesis
What is a null hypothesis?
A statement of no change
What is an alternate hypothesis?
A statement that describes any possible outcome in addition to your prediction
What is the function of a positive control?
Ensures the experimental method is functional
What is the function of a negative control?
Eliminates the potential for a false positive
A ______ is a special slide used to determine the density of cells
Hemacytometer
The _____________ is used to determine the number of viable cells present in a cell suspension
Dye exclusion test
The HL-60 cell line has been used as an (in vitro, in vivo) model system
In vitro
Cells of the white blood cell lineage are known as ________.
Hematopoietic
Define signal transduction
Intermolecular reactions involved in transferring and amplifying extracellular signals detected by the cell
What are 3 kinds of granulocytes?
Basophils, neutrophils, and eosinophils
Apoptosis is programmed cell death and differs from the other 2 types of cell death, __________ and _______.
Autophagy and necrosis
What is autophagy?
Self-ingestion of the cell, may occur when apoptosis is blocked
What is necrosis?
Cell death by which a cell swells, bursts, and decomposes
Apoptosis begins with nuclear fragmentation followed by the formation of cellular fragments known as __________.
Apoptotic bodies
Apoptotic bodies are engulfed by:
Phagocytosis
Describe the general apoptotic pathway, including where it takes place
- Adaptor proteins bind with 2 identical initiator caspases to form an active enzyme
- The initiator caspases cleave and activate the executioner caspases
- The executioner caspases cleave hundreds of substrates resulting in cellular apoptosis, fragmenting the cell into small apoptotic bodies
- Phagocytosis of apoptotic bodies.
Takes place in cytoplasm
When cells are treated with PMA or DMSO, apoptosis takes place via the _______ pathway.
Extrinsic
Describe the steps in extrinsic apoptosis
- Death ligands attach to
- Death receptors attracting
- Adaptor proteins form death-inducing complex (DISC) resulting in
- Caspase activation: Initiator cascade and executioner caspase
- Substrate cleavage follows and leads to
- Cell death
Why is RNA so unstable?
It is single-stranded (making it susceptible to ribonuclease, RNases) with a free hydroxyl group on the 2’ carbon (making the RNA molecule susceptible to hydrolysis)
What is an oligonucleotide?
A short single-stranded nucleic acid of fewer than 20 bases
Why is a forward and reverse primer necessary?
So that a region of interest will be specifically amplified
Why is Taq polymerase commonly used in RT-PCR?
It is heat-stable, and resists denaturation from the heat used in the denaturation step
Why does DNA have an overall negative charge?
Due to the phosphate backbone of DNA
To separate proteins, ____________ gels are used rather than agarose
Polyacrylamide
How did we know how large the proteins were when the zymogram gel was run?
Regions where MMPs had degraded the gelatin appeared white on the blue gel.
