Bio 351: Group Investigations in Epithelial Cell Biology
Independent Investigations of the Effects of Pollution on Lung Epithelial Cell
Biology
Epithelial cells are the body’s first line of defense against airborne pollutants. We are
interested in understanding the effects of pollution on the function of lung epithelial
cells-- the ‘cellular side’ of environmental biology. To study this question we have
designed and built a cell culture, ozone exposure chamber and are working to define
the parameters of the system and determine methods for measuring necrosis,
apoptosis and changes in stress-related genes’ expression.
Current work includes:
Characterizing growth characteristic of rat alveolar cells (L2 cell culture) grown
+/- polarity, +/- ozone in assays measuring relative levels of apoptosis and
necrosis
Developing increase metabolism and increased toxin L2 culture systems to
model additional cellular stress (thyroid hormone and EtOH exposure based,
respectively) and extend ozone exposure studies.
Future work will continue in these directions and:
Examine contributions of ozone’s interactions with media
Determine whether/how manipulating cellular surfactant layer composition affects
cells’ ability to survive stressors
Use above systems to examine the effect of ozone levels found in Davidson on L2
cultures (collaboration with site monitoring project with Dr Hauser)
The group investigation includes library research and bench research. The ‘two parts’
will be carried out concurrently. In the course of this research you will gain
experience in identification and critical reading of scientific literature,
experimental design, execution and troubleshooting, data collection and
analysis, and scientific writing. Experimental procedures will include
maintenance of mammalian cell culture, harvesting proteins and nucleic
acids and assays using colorimetry, fluorimetry, fluorescence microscopy to
measure cell viability and metabolism.
Expectations:
I expect that you are dedicated and excited about this research
opportunity and this research area. If you are not—don’t sign up.
As part of the Group Investigation you will:
++ Dedicate at least 10- 15hr every week to your research project.
++ Use all scientific equipment and facilities conscientiously, obeying all safety
regulations, reporting problems with equipment, maintaining supplies and cleaning
lab and glassware.
++ Keep a detailed notebook that records your activities. Notebook entries will
include the date, ‘question’, plan/approach, and detailed notes describing what was
actually done, side observations or notes about equipment, the data that were
collected, where that data is stored (and backed up), analysis of the data and
conclusions drawn.
+++Be proactive in making sure that your understanding extends beyond the
mechanics of the protocols and includes the focus of the research project and how it
fits and adds to the scientific field.
Group Meetings: (Monday 1:30- 4:20)
During the first hour of this meeting each person will
a) Present a summary of their data from the previous week and plans for the
current week. The group will help troubleshoot, provide feedback
b) Turn in an annotation of a paper relevant to your research. These will
become
part of your annotated bibliography. A copy of the paper should be
emailed to me
beforehand.
The rest of the group meeting time will be used for experimentation (mandatory lab
time)
THE FIRST DAY OF CLASSES we will MEET to DISCUSS PROPOSAL DRAFTS
and LUNG PAPERS
Individual research: In addition to Mondays, you will schedule another 3hr
weekday block of time when you will be in lab. Additional research hours are needed
and will be determined according to cell growth/ assay time requirements.
Assignments:
1) Consistent effort in laboratory research
2) Weekly meetings—Monday 1:30-4:30 (in Wat 261), individual 3hr research block
Written assignments: directions on next page
3) Lung and Ozone Background paper—Due Monday of the week BEFORE classes
start.
4) Research Proposal—Due Friday of the first week of classes. (noon)
5) Annotated Bibliography—Due in an accessible Refworks folder seventh week of
semester (Wed 10/10 9:30am)
6) Final Article--- expands upon your initial proposal and includes your results.
Experimental Procedures Draft Due 11/7/07)
Results Figures due
11/14/07 (data collection stops by 11/11)
Introduction Draft due
11/19/07
Final article submitted for editorial comments 11/28/07
Final version due
12/10/07
7) Protocols—Protocols are more detailed than Exp. Procedures sections. You will
prepare formal copies of all protocols/ set ups that you develop. Hard copy in the
protocol notebook. Electronic Copy saved on Louise; Public in the laboratory folder.
8) Laboratory Notebook—Due December 12th before 5pm
(optional) I strongly encourage you to plan to present your research at the
NC Academy of Sci. (3/14) and/or Assoc for Southeastern Biologist (4/16)
annual meetings in the spring.
I will be attending the American Society for Cell Biology meeting in Washington DC in
December. I welcome and encourage you to attend with me. (Unfortunately
abstracts are due in Aug so we will not be able to submit fall research if it is a new
project.) You are part of a long-term project and I encourage long-term
commitment to research. During the semester we can discuss options for
Independent Research in the spring.
The best thing to do would be to start with the basics. By that I mean— lung
anatomy.
How does a lung work? What are its parts?
What is an alveolae? (Type I cells? Type II cells? Any other cells in there? How does
it (the whole alveolae) work?
What is epithelia? What is polarized epithelia? How does it work (blocking and getting
across it)? Why is it important to have this sort of barrier in the lung?
What is surfactant? Why do we have it?
What is apoptosis vs necrosis? How can you tell them apart?
What is ozone? Why is it ‘good’ in the upper atmosphere but not so good to breathe
all the time?
What are the ‘acceptable’ levels of exposure to ozone according to the EPA— both in
amt (ppb) and amt of time you can be exposed to that amt?
What happens to ozone in the lungs (the basics of its oxidant properties) -- if lungs
are exposer to the allowable amounts or more than the allowable amounts what
happens? And If you have interest— what happens to people that have asthma?
People that have emphysema? Or some other respiratory problem OR what happens
if a ‘normal’ person is exercising in high ozone (increase metabolic rate)
For all of these things I do mean the basics— upper level textbooks, reputable
websites (CDC, EPA— not wikipedia), review articles rather than the nitty gritty of
research articles and analyzing mass Specs or microscopy. It is very important that
you have a broad understanding of the research area and its importance before you
dive in to particular sub-areas.
Once you have a handle on this then you can move on to defining an area that is
interesting to you— within constraints of the lab and equipment, of course, so having
a general area or two is good that we can then define further (later) according to the
parameters we already have.
This proposal is an ~5pg (1.5space) description of your research project. It should
include an introduction to the area of research and its importance and your specific
area within it (and its relevance) and describe your research goals for the semester.
It should have a projected timeline and a budget. I will help you revise the budget
before we submit the proposal for departmental support.
—a bibliography containing brief descriptions of the journal articles, their findings
and how this research connects with your project. It should contain at least 15
articles that are pertinent to your research focus. This may include papers
researching lung epithelia (Type 1, Type 2, tracheal, bronchial, alveoli), surfactant,
ozone research/ environmental chemistry and biochemistry, methods of cell culture
maintenance, methods of increasing metabolic activity, previously I strongly suggest
that you maintain a bibliography in Refworks (available through library homepage)