Work at the University of MD

From 2000 to 2005, I served as a Research Associate/Lab manager for a lab in the Radiation Oncology Research department at the University of Maryland. Our laboratory was investigating the role of genomic instability (1) in Cancer using radiation-induced unstable clones of Chinese Hamster Ovary cells having a single Human Chromosome 4 (CHOC4). As lab manager, I was the direct supervisor for six graduate students and two laboratory technicians.  My job was to troubleshoot and solve problems that arose during the graduate student’s research work in addition to guiding their research projects. I was the “go-to” person for scientific advice. If a student had an exceptional large tissue culture experiment, I would help in plating of cells into flasks or 96 well plates. I also reviewed their laboratory notebooks. One of my primary responsibilities was to train graduate students to write “scientifically’. ‘ I edited and proofread their articles prior to submission to scientific journals. I was responsible for organizing, running and participating in all research and journal club meetings. I ordered and monitored the inventory for all laboratory supplies and was responsible for maintenance of basic laboratory instruments including Coulter Counters, laboratory incubators, sterile hoods, liquid nitrogen storage and -80oC freezers, refrigerators, fluorescence and light microscopes, and sterile water still. I was responsible for making sure that they obtained the proper training for use of radioactive materials and had the appropriate monitoring equipment from the Environmental Health and Safety Department.

I introduced a number of new technologies into lab including RNase protection assays (2) and Affymatrix macro arrays (3) to determine increased or decreased expression of proteins due to genomic instability. I introduced TUNEL assays (4) to determine apoptosis in our unstable clones. I optimized and wrote standard operating procedures for hybridization of Human Chromosome painting probes (4). I maintained a bacterial repository for all of the Human chromosome painting probes as well as a repository for all of the unstable clones. A loss of a -80^oC freezer or liquid nitrogen storage unit could be severely detrimental to our research aims. I was one of the contact people for emergencies in the laboratory. I developed optimized protocols for preparing metaphase and interphase spreads for the humid Maryland climate. I also introduced PCR in situ (PRINS) (6) to identify small size repetitive elements in the human and hamster genome in metaphase (7) and interphase (8) spreads. We moved from a chromosomal scale to protein scale during my tenure in the lab. I was responsible for ordering SDS-Page and immunoblotting equipment and determining which HRP (9) conjugated antibodies to order for chemiluminescent detection (10).

In addition to the above responsibilities, I had my own projects. I identified the mechanism of action in one of our unstable clones. I also had my own projects investigating the possible mechanisms involved in genomic instability of clones exposed to radiation or accelerated heavy metal ions. I determined the instability in one clone. The genomic instability in this particular clone was due to the lack of a caspase crucial to apoptosis (11). I noticed that the fluorescently labeled DNA in this clone appeared punctuate in appearance in interphase spreads suggesting that it was undergoing initial but not complete caspase (12) cleavage.  Immunoblotting using HRP-conjugated antibodies demonstrated that this particular clone had initiator caspases eight (13) and nine (14) but was completely lacking in the primary effector Caspase 3 (15). Incomplete cleavage appeared to accounted for duplication and translocation of chromosomal material observed in this unstable clone.

During my time in the lab, I was a primary author of a review of quantity and quality of radiation involved in genomic instability. entitled, ‘ Radiation-Induced Genomic instability:

Radiation Quality and Dose Response’ (16) which appeared in the Health Physics Journal in 2003. I was a coauthor on five papers published by my students in journals such as Mutagenesis and Cancer Research. My position was a non-tenured faculty position and in this capacity, I taught sections of Molecular Biology for Graduate students. I also team taught Radiation Biology for Oncology interns studying for their Oncology boards. My portion of the course was DNA structure, DNA damage and repair. I judged the annual Molecular Biology Poster sessions and served as the coordinator for review of abstracts and the award of travel fellowships for the Molecular Mutagenesis Society.

I have to say that out off all the positions that I have held in my research career. This position of Research Associate /Lab manager was the most fulfilling.  I truly enjoyed the interactions that I had with the students and students from other labs working in the field of Radiation/Cancer Research. Our lab really functioned as a team even though my students were investigating in a wide variety of aspects of genomic instability from programmed cell death to bioinformatics.  I enjoyed being head of that team. All of the students under my supervision obtained their PhD’s and the two laboratory technicians went on to obtain higher degrees in science

 

 

Links and citations:

 

1.       http://www.wikigenes.org/e/mesh/e/22690.html

2.       http://en.wikipedia.org/wiki/Nuclease_protection_assay

3.       http://www.everythingbio.com/glos/definition.php?word=macroarray

4.       http://en.wikipedia.org/wiki/TUNEL_assay

5.       http://hmg.oxfordjournals.org/content/7/10/1619.full

6.       http://www.pcrstation.com/in-situ-pcr/

7.       http://en.wikipedia.org/wiki/Metaphase

8.       http://en.wikipedia.org/wiki/Interphase

9.       http://en.wikipedia.org/wiki/Horseradish_peroxidase

10.   http://en.wikipedia.org/wiki/Chemiluminescence

11.   http://en.wikipedia.org/wiki/Apoptosis

12.   http://en.wikipedia.org/wiki/Caspase

13.   http://en.wikipedia.org/wiki/Caspase_8

14.   http://en.wikipedia.org/wiki/Caspase-9

15.   [http://en.wikipedia.org/wiki/Caspase_3]

16.   http://www.ncbi.nlm.nih.gov/pubmed?term=Smith%20LE%2C%20Healthphysics%202003