Sample
Problems
Choosing between paired and unpaired t tests
Sometimes the choice of which type of t test to
run is obvious, and sometimes the choice requires
some careful reflection. In all of the cases below
the numbers of data points are low enough that
the statistical analysis should be based upon a
t distribution rather than a normal distribution.
Assume that the variances are equal for all data
sets.
Problem #1
As a biochemist working for a pharmaceutical company,
your job is to test new drugs for possible side
effects, both deterimental and beneficial. The
chemistry of agent TFK-05W suggests that it may
have the side effect of reducing the tendency toward
obesity. The Zucker rat is an established genetic
model for both obesity and hypertension. As rats
of the obese strain age they gain weight much more
rapidly than do the so-called "lean" Zucker
strain. You plan to treat a group of animals
over a period of one year and compare their average
weight with that of a group of untreated animals.
The odds of the drug actually showing this side
effect are small and maintenance of rats for a
year is expensive, so you limited the scope of
the study to twelve animals in each group. The
null hypothesis is that treated animals
will show an average weight gain over one year
that is no different from the average weight gain
of untreated animals. What statistical test will
you use to compare the two means?
Reference TW Kurtz, RC Morris and HA Pershadsingh,
The Zucker fatty rat as a genetic model of obesity
and hypertension, Hypertension, Vol 13, 896-901,
1989
Problem #2
This study follows from problem #1. Not only did
your study suggest that the agent TFK-05W indeed
does affect weight gain, but it also proved effective
and safe (so far) and it is in clinical trials.
Because the drug was designed to treat symptoms
that have nothing to do with obesity, the clinical
trials do not focus on that problem and won't answer
the question of whether or not the agent is a potential
weight loss drug. The company, however, has permitted
you to test the agent on a group of 12 people with
morbid obesity, who have signed the appropriate
consent forms.
This time the plan is to treat the 12 obese individuals
for a year, having measured their weights on the
day treatment was started. The paid participants
will be monitoring their weight regularly, taking
the drug, and are required to keep a daily log
of activity and eating habits so that the experiment
can be properly controlled. Nevertheless, the simplest
initial test of the hypothesis that obese individuals
treated with TFK-05W for a year will show an average
weight loss is to compare average weight at the
beginning and end of the experiment. Thus, as with
problem #1, you will have two sets of 12 data points
each to compare. What statistical test will you
use to compare the two means?
Problem #3
Embryonic cells (stem cells) from a single human
blastula are genetically equivalent. Any of them
has the potential to form any kind of tissue that
is normally found in an adult human body. Exploitation
of stem cells for therapeutic purposes has the
potential to revolutionize medicine and expand
the average human lifespan considerably. Stem cells
have very complex cultural requirements. So far
your stem cell lines require the addition of fetal
bovine serum to the culture medium, and the
exact composition (and efficacy) of animal sera
varies from lot to lot, To exercise the greatest
control over your experiments, it would be valuable
to be able to culture your cells in a synthetic
medium that includes only those components that
are essential to support survival and growth of
your cultures.
You have developed a synthetic medium that keeps
your cells going for several days, but not indefinitely.
You think that you can extend the life of your
cultures by adding an expensive hormone to the
medium. To test the hypothesis that stem cells
cultured with medium 2 will survive longer than
stem cells cultured with medium 1, you will set
up twenty cultures from ten original embryos, growing
them in complex medium to the point at which each
culture contains about 100 cells. You will then
remove the original medium and feed the cultures
from now on with synthetic medium. One culture
from each original embryo will receive medium 1
and the other medium 2. For your data you will
record the time at which each culture declines
to the point of having only 50% of its original
viable cells remaining. The null hypothesis is
that this average "survival time" will be the same
for cultures treated with either synthetic medium.
Alternative hypotheses, of course, are that feeding
with one or the other medium will enhance survival
time by comparison.
What statistical test will you run on the two
sets of 10 data points each?
Problem #4
You suspect that a cause of decline of your stem
cell cultures is a failure to produce sufficient
superoxide dismutase to rid the cells of oxygen
free radicals. You have an assay for the enzyme,
but to conduct the assay you must destroy the culture.
From a single source of stem cells you can prepare
about thirty cultures that remain healthy about
10 days in your synthetic medium after growing
to a sufficient number of cells to permit you to
run your assay. From then on, they
decline rapidly.
You prepared thirty cultures from the same source
and sampled half of them when alll thirty cultures
had reached the point at which the assay was feasible.
You then sampled the other half 10 days later.
Your null hypothesis is that superoxide dismutase
activity will not be significantly different between
the two sets of cultures. What statistical test
will you run to determine whether or not the difference
is significant?
Normally, this would be a rather poor experimental
design, because all of the cultures are identical.
Why conduct replicate sampling on the exact same
culture? The issue is that enzyme assays are notoriously
inaccurate. The chances of mulitple comparisons
yielding dubious results are much smaller than
for a single comparison. The p value that
we obtain will give us a fairly accurate estimate
of the level of confidence with which we can interpret
the result.
Interpreting t test results
Problem #5
Referring to problem #1, one of your rats died
of natural causes during the study, leaving 11
animals in one group while you still had 12 animals
in the other. Does this turn of events ruin the
experiment? If you do conduct the analysis, how
will you modify it, if at all?
Problem #6
Referring to problem #2, one of your human subjects
died of a massive heart attack halfway through
the study. The death was clearly not related to
the drug treatments. Please anwer the same questions
as for problem #5.
Problem #7
In problem #2 it was stated that Zucker
rats treated with the agent TFK-05W were significantly
less obese than untreated rats. Does this mean
that when the t test was run it returned a probability
(p) value of > 0.05 or a p value of < 0.05?
Problem #8
For the third problem the difference between means
was 12 hours, that is, the average half life of
one culture was 12 hours longer than the average
half life of the other one. The t test returned
a p value of 0.33. What is your conclusion regarding
the original hypothesis?
Would you be correct in stating that the result
is significant or insignificant?
Problem #9
In problem #4, average superoxide dismutase activity
was 30% lower after 10 days than it was in the
beginning. The t test gave a p value of 0.07. What
result do you report? It turns out that this study
is very important, and if you indeed find that
a decline in superoxide dismutase activity is a
primary cause of cell death then good things will
happen to your career. How will you proceed from
this point?
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