Laboratory
Notebook
AAPS-PSPG Science Day
• 2009 •
Name:
Age:
Tour Guide:
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1/10/2009
MAP
Nephron
Lab
Liver
Gut
Lung
Brain

Tour starts here
• Registration
• Missing persons 
2
• Food
• ice cream/liquid nitrogen
demonstration
1/10/2009
Table of Contents
Introduction
Lecture by Dr. Steven Hamilton (LUNG room)
Absorption
Inhaler lung model (LUNG room)
Dissolution experiment (GUT room)
Distribution
Diffusion experiment (GUT room)
Metabolism
Enzymatic activity (LIVER room)
DNA extraction (LIVER room)
Excretion
Exit human body Drug Tour via the NEPHRON (hallway)
Other activities include:
• Build-a-Drug in the LAB
• See if you can tell the difference between candy vs.
medication in the LAB
• Learn about the blood-brain barrier in the BRAIN room
• Watch how scientists can make ice cream with liquid
nitrogen in the main FOOD area
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Introduction
One thing I learned about drugs from Dr. Hamilton
is…
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Intro of tour.
Ask kids – why do your parents give you medicine (cough, cold, fever,
stomach ache, etc.)? What are some common ways that you take medicine
(liquid, pill – swallow, shots)?
The first thing a drug must do after you take it before it can go do what it is
supposed to do is be ABSORBED into your body. Does anyone know what
absorbed means? (use example like water in a sponge – definition: to take in
or soak up by chemical or physical action, typically gradually)
The most common way we take medicine at home is by swallowing a pill or
liquid. So where do you think these drugs would be absorbed? (gut)
But some people also take medicine in a special way to help get the drug to
the place it needs to work. For example, in asthma – which affects how well
your lungs work to help you breathe – people use inhalers to get the
medicine to their lungs. The medicine is delivered in a fine powder that
helps coat all the alveoli of the lungs so that it can have an effect. The drug
can then be ABSORBED into the lungs.
We are going to build a model of a lung and simulate how someone would
breathe in a fine mist of medicine and how that medicine would be absorbed
into the lungs. In this example, lemon juice will represent the drug and
cotton balls will represent the lung’s alveoli.
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Absorption
Inhaler lung model
Lung room
Introduction
In order to work, medication must first be ABSORBED into your body. Absorption is
the process of taking in or soaking up something.
In our experiment, we will build a model of a lung and simulate how someone would
breathe in a fine mist of medication so it can be absorbed into the lungs to help the person
breathe better. In this example, lemon juice will represent the drug and cotton balls will
represent the lung’s alveoli. We will test whether the ‘drug’ (lemon juice) reaches the
lung using a colored pH indicator. Pink represents absorption and blue represents no
absorption.
Reagents
1.
2.
3.
4.
5.
6.
40 cotton balls
1 plastic bottle
Tweezers
Red Cabbage (pH indicator)
Lemon juice
Goggles
Procedure
1.
Fill plastic bottle with 40 cotton balls. Keep 1 ball out and set aside on a paper
towel & label as “Control.”
2.
Simulate breathing by squeezing sides of bottle to suck air into “lung” (make
sure the lid is OFF).
3.
Put on your safety goggles.
4.
When it is your turn, “breathe in” the lemon juice mist (student volunteer)
with your “lung” bottle to simulate inhaling asthma medication. Cotton balls
should absorb the lemon juice.
5.
Use tweezers to pull out 1 cotton ball from the top of your bottle. Put this on
your paper towel and label as “Top.”
6.
Use tweezers to pull out 10 more cotton balls and set these aside. Then pull
out 1 cotton ball from the middle of your bottle. Put this on the towel and
label as “Middle.”
7.
Pull out the rest of the cotton balls and set aside. Keep the last cotton ball and
place on towel, labeling it as “Bottom.”
8.
Test your cotton balls for absorption by applying the pH indicator (student
volunteer) on the cotton balls you pulled out of different parts of the bottle.
9.
Observe differences in color. Compare to untreated “Control” ball.
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Questions
1.
What color is the top cotton ball
Pink or purple
2.
What color is the middle cotton ball?
Pink/purple or blue
3.
What color is the bottom cotton ball?
blue
4.
Which part of the lungs absorbed the most medication?
Top
5.
Why is taking too much medication bad for absorption?
Your body can’t absorb that much all at once
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Dissolution
Gut Room
Introduction
Medicine has to dissolve in order to get where it needs to work. When you swallow a
pill, it will dissolve in your stomach before it can move to its target.
Dissolution is the process of dissolving a solid substance into a solvent to yield a
solution. In our experiment we will use water as our solvent and we will attempt to
dissolve 3 different substances in it. A substance that has dissolved becomes invisible
to your eye.
Reagents
1. Three cups
2. Three spoons
3. Citric acid
4. Alka Seltzer
5. Sand
6. Water (solvent)
7. Pipette
Procedure
1. Label 3 cups as follows:
a. Citric acid
b. Alka Seltzer
c. Sand
2. Add one spoonful of citric acid or sand, or one tablet of alka seltzer, to the
corresponding cup.
3. Add water in 1 mL increments until the solute has dissolved. Stir with spoon as
needed to help dissolve the material.
4. Determine the amount of water needed to dissolve each substance.
5. Save your solutions for the next experiment: Diffusion.
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Questions
1. Which substance(s) dissolved?
Citric acid and alka seltzer
2. How much water did it take to dissolve each solute?
a. Citric acid? 14 pipette squeezes
b. Alka Seltzer? 16 pipette squeezes
c. Sand? 20 pipette squeezes - DID NOT DISSOLVE
3. Based on your results which substance would make the best medicine?
Citric Acid
4. Do you think any of the solutes would dissolve better or faster in warm water?
Yes
5. Why is taking too much medication bad for dissolution?
If there is too much it can’t all dissolve
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Distribution
Diffusion
Gut Room
Introduction
Simple diffusion is the net movement of dissolved particles (dissolution experiment)
down their concentration gradient, from a region of high concentration to a region of low
concentration. This is a passive process. In order for medication to reach its target, it
must diffuse across barriers.
This experiment will utilize the color change associated with a pH indicator to
determine the extent of diffusion. A blue solution indicates no diffusion while a pink
or purple solution signifies diffusion.
Reagents
1. Four cups
2. The dissolved solutions from the dissolution experiment:
a. Citric acid (pH = 2)
b. Alka Seltzer (pH = 5)
c. Sand (pH = 7)
3. Three pieces of 5 inch dialysis tubing
4. String
5. Red Cabbage (pH indicator)
Procedure
1. Fill all four cups halfway with water
2. Add pH indicator (student volunteer) to the three cups until it turns blue
3. Wet each piece of dialysis tubing in the fourth non blue cup of water
4. Rub the wet tubing until it opens up
5. Twist one end and tie off that end of the dialysis tubing with a string
6. Add each of the following solutions from the previous dissolution experiment to a
separate piece of dialysis tubing:
a. Citric Acid
b. Alka Seltzer
c. Sand
7. Tie off the other end of the tubing with string
8. Drop the dialysis tubing into the blue water
9. Observe the color change
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Questions
1. What was the color of the water when you dropped the citric acid/dialysis tubing
into the blue water?
Pink
2. What was the color of the water when you dropped the alka seltzer/dialysis tubing
into the blue water?
Purple
3. What was the color of the water when you dropped the sand/dialysis tubing into
the blue water
Blue (No change)
If there is a color change, possible reasons may include contamination by one of
the other items on dirty hands used to tie the string/touch the outside of the
dialysis tubing. Can test this hypothesis by testing pH of just sand solution and
compare color to diffusion expt.
4. Based on your results, which substance would make the best medicine?
Citric Acid or alka seltzer
5. Why is taking too much medication bad for diffusion?
There is a limit to the amount that can diffuse into the body
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Metabolism
Metabolism
Liver Room
Introduction
Metabolism is the conversion of one molecule into another molecule. All foreign
substances, including food and medication, are metabolized in the body.
Starchy substances make up a major part of the human diet. They are made naturally in
a variety of plants. Some plants with high starch content are corn, potatoes, rice, and
wheat. Starch molecules are made up of a number of sugar molecules linked together. In
order to make use of the energy stored in starch, the body uses an enzyme known as
amylase to break down the starch into sugars. Starch metabolism begins in the mouth
where the enzyme salivary amylase in saliva hydrolyzes starches into simple sugars.
This experiment will utilize alpha amylase and salivary amylase in spit to metabolize
starches into sugars. The extent of metabolism will be determined by using iodine as an
indicator of starch. If all of the starch remains unconverted, then the solution will be
dark blue. Partially degraded starch solution will appear brown-red. The solution
will be clear when all of the starch is metabolized into simple sugars.
Reagents
1. Alpha amylase (Enzyme 1)
2. Human salivary amylase (spit)
3. Starch solution (1 g/L)
4. Iodine
5. Water
6. Three cups
7. Water bath
8. Plastic pipette
Procedure
1. Label three cups accordingly:
a. Control
b. Enzyme 1
c. Spit
2. Add 1ml of starch solution to all three cups
3. Add 1ml of water to all three cups, mix
4. To the appropriate cup, add an additional:
a. Control1 ml of water
b. Enzyme 11 ml of fungal amylase (student volunteer)
c. Spitspit into cup (~ 1 ml)
5. Put in water bath for at least 10 min
6. Add three drops of iodine (student volunteer) to each cup, mix
7. Observe the color in each cup
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Questions
1. What was the color of the Control solution?
Blue
2. What was the color of the Enzyme 1 solution?
Green
3. What was the color of the Spit solution?
Yellow (may be green depending upon how much they spit into the cup)
4. What happens when you put your cups in the water bath for longer than 10
minutes?
Control- no change (blue)
Enzyme 1-clear (may take an hour)
Spit-clear (may take an hour)
5. Why is taking too much medication bad for metabolism?
Your body’s enzymes won’t be able to metabolize (break down) all of the drug
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Strawberry DNA extraction
Liver Room
Introduction
Deoxyribonucleic acid (DNA) is a nucleic acid that contains the genetic instructions
used in the development and functioning of all living organisms and some viruses. The
main role of DNA molecules is the long-term storage of information. DNA is often
compared to a set of blueprints, a recipe, or a code, since it contains the instructions
needed to construct other components of cells, such as proteins and RNA molecules.
Differences in our DNA sequences help make each of us unique and different from any
other person. Because of these differences, each person may also respond to medications
in slightly different ways. In this experiment, we will extract DNA from strawberries.
Reagents
1. coffee filter
2. plastic cup
3. rubber band
4. strawberry
5. plastic bag
6. Lysis Buffer (Tube 1)
7. wooden stick
8. isopropanol (Tube 2)
9. Eppendorf tube
Procedure
1. Put coffee filter over top of plastic cup and secure with a rubber band.
2. Put a single strawberry in a Ziploc bag, remove as much air as you can and close
the bag.
3. Smash up the strawberry with your hand and fingers for 2 minutes. Be careful not
to break your bag. The best way to mix it is to massage the mixture at the bottom
of the bag.
4. Add 12ml of Lysis Buffer (Tube 1) to the bag and zip it closed. Mash again for 1
minute.
5. Tilt the bag so that the mush collects in one bottom corner of the bag, and open
the bag. Carefully pour the mush into the coffee filter. Let the liquid drip through
into the cup. You can gently stir with the flat wooden stick.
6. After most of the reddish liquid has dripped into the cup, carefully remove the
filter paper with the strawberry mush and throw it in the trash. Tilt the cup a little
and gently pour the 10ml of Isopropanol (Tube 2) in the cup - letting it slowly
pour down the side of the cup. DO NOT MIX!
7. Observe and wait a little. You'll see the DNA start to collect as a goopy glob, and
you can "spool it out" on the tip of the wooden stick.
8. Transfer the spooled DNA into an Eppendorff tube with some isopropanol. You
can take the DNA home with you, but keep it tightly closed to avoid evaporation
of the alcohol. The DNA is stable in this form for many years.
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Questions
1. What does DNA look like?
Like snot
2. Besides strawberries, what else contains DNA?
Pretty much anything living…
3. How might differences in a person’s DNA sequence affect how they respond to
medication?
Genetic differences may affect how drugs are absorbed, distributed throughout the
body, metabolized and/or excreted
Can reference back to Steve’s talk on pharmacogenetics (i.e. genotype-dependent
alterations in dosing regimes etc.)
FYI - the lysis buffer is composed of dish-washing soap and salt in water.
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Genetics Fun Facts
o It takes about eight hours for one of your cells to completely copy its DNA.
o The human DNA code is made up of about three thousand million A,T, C, and Gs
on each side of the DNA strand.
o If you were to start reciting the order of the ATCGs in your DNA tomorrow
morning, at a rate of 100 each minute, 57 years would pass before you reached the
end (provided that you did not stop to eat, drink, sleep, use the bathroom etc.)
o If you were to stretch out the DNA from those 46 chromosomes in one cell and
lay it end to end, it would be over 2 yards in length.
o If the total DNA in one person were laid in a straight line, it would stretch to the
sun and back over 30 times (it’s 93 million miles from here to the sun).
o You could fit one thousand nuclei across the period at the end of this sentence.
o You could fit one million threads of DNA across the period at the end of this
sentence.
o If the genome was a book, it would be the equivalent of 800 dictionaries. It would
take a person typing 60 words per minute, eight hours a day, around 50 years to
type the human genome. You would need 3 gigabytes of storage space on a
computer to hold all of this
o Humans are 99.9% genetically identical – only 0.1% of our genetic make-up
differs
o Our genes are remarkably similar to those of other life forms. For example, we
share 98% of our genes with chimpanzees, 90% with mice, 85% with zebra fish,
21% with worms, and 7% with a simple bacterium such as E. coli.
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Excretion
Nephron - Hallway
Introduction
Excretion is the process of eliminating or expelling waste matter from the body. At
some point, the medication that enters the body is broken down into smaller
molecules called metabolites. Many metabolites are then filtered out of the blood by
the kidneys so they can be excreted from the body. In this activity, we will be exiting
the Drug Tour body via the nephron of the kidney (represented by the crawl tunnel).
Procedure
Follow instructions given by tour guide.
You can have the kids crawl through one at a time. Then demonstrate how different
sized items might be harder to pass by having kids take 1, 3 or a lot of balloons
through the tube with them.
Questions
1. What was one challenge of excretion through the nephron?
How fast you can go, what size item can fit through the tube, etc.
2. Why is taking too much medication bad for excretion?
There is a limit to the amount and speed that things can be excreted from the body
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Build-a-Drug
Lab
Introduction
Many drugs work by targeting the “active site” of a certain receptor. A drug fits into a
receptor like a key fits into a lock.
Some drugs only fit into 1 type of receptor. Other drugs fit many different receptors.
Sometimes drugs that fit the right receptor may also fit the wrong (“off-target”) receptor.
This can lead to bad side effects. That’s why it is important that drugs are specific to the
target receptor.
Reagents
1. Modeling clay
2. 3 types of receptors (on posters in hallway):
a. Basic (blue)
b. Intermediate (yellow)
c. Advanced (orange)
Procedure - Please build your drugs in the LAB!
Basic activity
1.
Build a drug with your drug modeling compounds that fits into 1 receptor.
Advanced activities
2.
Build 1 drug that will fit many different receptors without remodeling the drug
(hint: pick receptors with the same shape active site).
3.
Build a drug that will activate 1 receptor and not any other off-target receptors
(hint: make your drug so that it won’t fit into any other active sites).
Notes
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Blood-Brain Barrier
Brain Room
Introduction
The brain is one of the most important organs in the body. It controls how we move our
arms and legs, the pace of our heartbeat, and how fast we breathe in and out. Since it is
such an important organ, it has a special layer of protection to keep poisons and toxins
that may get into the blood from getting to the brain. This protection is called the bloodbrain barrier.
Procedure
Follow the instructions of the blood-brain activity leader and see if you can answer the
following questions afterwards.
Questions
1. What types of molecules does the brain need to get from the blood?
oxygen, glucose, proteins, ethanol
2. What types of molecules can diffuse across the blood-brain barrier without any help?
oxygen, carbon dioxide, sedatives, alcohol
3. If a molecule can’t diffuse across the blood-brain barrier, how does it get across?
Using transporters, in a process called transcytosis
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