North Carolina State University
ACCIAC Fellows in Creativity and Innovation Program
September 20, 2010
Campus Director: George T. Barthalmus
Executive Summary: The Office of Undergraduate Research at North Carolina State University
is pleased to submit this final report on our ACCIAC Undergraduate Fellows. These awards,
especially during difficult financial times, are most appreciated. Note that faculty and
administrators are pleased to see this effort of support coming from the academic side of the
ACC. Please know that our faculty would embrace the expansion of this program.
A campus-wide call for proposals was distributed in spring 2010. A faculty review committee
ranked the candidates and identified two students within the College of Agriculture and Life
Sciences: Ms. Mary Pat Bulfin (mentored by Dr. Chris Ashwell in the Department of Poultry
Science) and Mr. David Higgins (mentored by Dr. Wendy Boss with Dr. Yang Ju Im in the
Department of Plant Biology). The selection committee considered the scholarship and
completeness of the application as well as the reputation of the faculty as both past mentors and
scholars. Photographs of the students, alone and with their mentors, are attached and we expect
to showcase them on the homepages of the Undergraduate Research website and the NC State
University’s website.
Student Biographies, Career Goals, and Abstracts
Mary Pat Bulfin is a sophomore majoring in both Animal Science and Biological Sciences, the
latter with a concentration in integrative physiology and neurobiology. Born in Hillsborough,
North Carolina she attended Cedar Ridge high school. Fascinated by the vast, integrative nature
of the sciences, she participated in the Reaching Incoming Students Enrichment (RISE) research
program at NC State before beginning her freshman year. She investigated methane production
in respiring plants. Funded by the Howard Hughes Medical Institute (HHMI), the RISE
experience introduced her to undergraduate research. She is grateful to continue in HHMIsupported research that focuses on the isolation and characterization of a novel
mycobacteriophage, culminating in genome annotation. Interested in learning more about
genomics, she was delighted to have the opportunity to pursue genomic research with the
ACCIAC fellowship under the guidance of Dr. Chris Ashwell and research technician Shelly
Nolin in the Department of Poultry Science Department. “It has been a privilege beyond
measure!” Through this experience, she has learned how dietary manipulation influences gene
expression and can affect disease susceptibility. The fellowship has enhanced her appreciation of
the interconnectedness of living systems and has reinforced the applicability of her coursework in
biology, thus developing an exciting new context in which to interpret her academic studies.
Realizing genomic similarity across many diverse organisms, the fellowship has helped confirm
her belief that research is a humbling form of service in which time, effort, and knowledge can
improve the lives of many. Ms. Bulfin is grateful for the generous support and expertise that her
mentors shared with her each day. They exemplify how quality research is the process of
collective diligence and joyful collaboration in pursuit of new knowledge. After graduation from
NC State, Ms. Bulfin will look forward to life-long service involving research and the pursuit of
M.S. and Ph.D. degrees in neurobiology or genetics.
Career Goals: Fascinated by the intricacy of the brain and its central role in both human and
animal function and behavior, Ms. Bulfin is interested in pursuing a career in neurobiology.
Brain circuitry and the linkage of neural pathways to potential gene expression patterns amaze
her, and she is motivated to learn more. While she enjoys the sciences, her career goals are
ultimately dictated by the desire to serve people in a meaningful way with both her mind and
heart through relationships and collaboration. As a hobby, she is intrigued by the remarkable
capacity of the canine-human bond to surmount challenges and empower others as demonstrated
by search and rescue (SAR) and guide and assistance dog partnerships, and she is excited to serve
in such a partnership in the future.
Abstract: Effect of exogenous enzyme and DFM supplementation on growth and disease vulnerability in
broiler chickens based on ileal IL-10, and IFN-γ RNA expression patterns. Due to rising feed costs and the
growing global demand for meat products, commercial poultry producers are challenged to identify
methods for raising healthy birds resistant to illness, especially Clostridium-related infection, through
dietary manipulation. Two common approaches that consider both financial pressures and biological
metabolism are exogenous protease supplementation and supplementation of a direct-fed microbial.
Utilizing focused microarray analysis and real-time PCR quantification of interleukin (IL)-10 and
interferongamma (INF-γ) mRNA expression patterns in broiler chickens, we observed the contrasting
effects on ileal gene expression between control birds (T2) and those given supplemental exogenous dietary
enzymes (T4 and T5) and/or a DFM (Avicorr) (T8) in the presence of C. perfringens (CP) disease
challenge. We hypothesized that protease supplementation would result in decreased body weight and
protease supplementation would negatively impact immunity and inflammation response, and a direct-fed
microbial (DFM) would have positive effects. Our results supported our hypotheses, and body weights of
the birds followed the following pattern T2 < T4 < T5 < T8 with higher body weight achieved with the
DFM. The lesion scores detected in the intestine followed the following pattern T4 > T2 > T5 = T8 across
the treatment groups. Relative gene expression of IFN-γ was statistically different between T2 and T8,
while relative IFN-gamma gene expression was not statistically different between T4 and T8. Relative gene
expression of IL-10 was overall higher across all treatment groups than IFN-gamma. Using a mixed-model
ANOVA, 8 differentially expressed genes in addition to IL-10 and IFN-γ were identified. MetaCore
pathway analysis of the significant genes identified many known processes involved in signal transduction,
carbohydrate metabolism, and cell death. We can deduce that protease enzyme supplementation increases
body weight. The negative inflammatory and immunological effect increases with multiple enzymes, and
the severity of negative effects is lessened by a supplemental DFM.
_________
David Higgins is a junior hoping to major in both the new undergraduate Genetics degree and in
Plant Biology with a minor in Biotechnology. He is a member of the University Honors Program
and the College of Agriculture and Life Sciences’ Honors Program. Mr. Higgins attended high
school in Winston-Salem, NC at West Forsyth High School. There he found interest in science
by attending the University of NC system-sponsored Summer Ventures in Science and
Mathematics program at Western Carolina University. His first college research was conducted
through an NSF REU program at the Donald Danforth Plant Science Center in St. Louis,
Missouri, one of the top independent plant research centers in the world, where he studied the
biofuel potential of locally collected algae strains. He used that experience to formulate a project
with his mentor, Dr. Wendy Boss. That research was based on material learned through taking
her Plant Physiology course. The fellowship provided Mr. Higgins with an opportunity to expand
his learning beyond the classroom. He discovered that laboratory research is much more than
putting together facts learned in class, but also requires an understanding of techniques, concepts,
and the complexities of living systems as a whole.
Career Goals: Mr. Higgins plans to attend graduate school and pursue a MS and/or a PhD in
Plant Cell Biology or Genetics. Then, his plan includes a career in biological research at a
university. He hopes to study fields with high-impact implications in helping society while
imparting his love of biology to future students.
Abstract: Optimizing the transformation of Arabidopsis thaliana with Tap-tagged Phosphatidylinositol
phosphate kinase. The phosphoinositide (PI) pathway is involved in sensing changes in gravity, osmotic
stress, and light-mediated signaling in plants. The nature of the regulation of the pathway is currently
unknown, but is hypothesized to involve the binding of proteins to the key enzyme phosphatidylinositol 4
phosphate 5-kinase (PIPK). To test this hypothesis, I have been attempting to generate plant cells
expressing tandem affinity purification (TAP)-tagged forms of the PIPK in order to recover interacting
proteins for further analysis. Three approaches were used to achieve this goal: vector-mediated
transformation of Arabidopsis cell liquid cultures through co-cultivation with the bacterium,
Agrobacterium tumefaciens; a similar method involving callus growth of co-cultivated cell cultures; and a
floral dip method involving the submersion of whole Arabidopsis flowers into a solution of A. tumefaciens.
Conditions were optimized for the liquid culture transformation; however, this potentially rapid
transformation method has yet to yield transformants. Callus growth experiments have produced putative
transformed calli and the plants used in the floral dip procedure have begun producing seeds. In the future,
I will check for the presence of the TAP-tag mRNA and tagged protein. Once the presence of the protein is
confirmed, I will purify the TAP-tagged PIPK and optimize conditions for the recovery of interacting
proteins.
_______
Emailed attachment:
The first attachment is the above 2.5 page document. The required brief executive summary is at
the top of page one. A biography for each student participant with her/his completed research
abstract follows the summary.
The second attachment is of Ms. Mary Pat Bulfin (ACCIAC Fellow) with her mentor, Dr. Chris
Ashwell in the mentor's laboratory in the Department of Poultry Science.
The third attachment is of mentor Dr. Wendy Boss (left) with Mr. David Higgins (ACCIAC
Fellow), and Dr. Yang Ju Im (postdoc and assisting mentor) in the Plant Biology Laboratory at
the NCSU Centennial Campus.
The fourth is the detailed manuscript of Ms. Mary Pat Bulfin.
The fifth is the detailed manuscript of Mr. David Higgins.