Molecular and

Cellular Biology

Faculty and Student

Retreat

November 1, 2014

Nelson Commons

Ohio University

Translational Research in

Cell and Molecular Biology Nelson Commons, Ohio University

Athens Ohio

8:30 am - 9:00 am Welcome coffee/tea

9:00 am - 9:30 am Dr. Ramiro Malgor, "Wnt5a in T1 translational research

process”

9:30 am - 10:00 am Dr. Jaime Costales, "A case of human infection with

Trypanosoma cruzi lineage l in the U.S.A."

10:00 am - 10:30 am Dr. Darlene Berryman, "Growth hormone's effect on

adipose tissue: too much of a good thing?"

10:30 am - 11:00 am Dr. Tomo Sugiyama, "Biochemical Nature of Damage-

Induced Mutagenesis"

Panel Discussion

11:00 am - 12:00 pm NIH/NSF funding, how to get funded, stay funded

and current trends in funding.

Dr. Sarah Wyatt, Environmental and Plant Biology

Dr. Monica Burdick, Chemical and Biomolecular Engineering

Dr. Robert Colvin, Biological Science

12:00 pm - 1:30 pm Lunch Break (lunch provided) / View Posters

1:30 pm - 2:30 pm Keynote Address

Keynote

Defects in Growth Hormone

Signaling and Beyond: Impacts on

Human Growth, Immunity, Insulin

Sensitivity

Dr. Vivian Hwa, Associate Professor of

Pediatrics, Division of Endocrinology

Cincinnati Center for Growth Disorders

Cincinnati Children’s Hospital Medical Center

Panel Discussion

2:30 pm - 3:30 pm What is Translational Research, Past, Present, and

Future?

Dr. Sonsoles de Lacalle, HCOM

Dr. Michael Boyle, Senior Scientist, V.P. for Research

Dr. Shiyong Wu, Director Edison Biotechnology Institute

3:30 pm - 4:00 pm Dr. Felicia Nowak, "Establishing the Role of Pro-apoptotic

Porf-2 in Cognitive Development and Decline"

4:00 pm - 4:30 pm Dr. Ronan Carroll , "The Role of PPlases in Staphylococcus

aureus"

4:30 pm - 5:00 pm Dr. Erin Murphy, "RNA & Shigella virulence"

5:00 pm - 5:30 pm Dr. Zhihua Hua, "Understanding Plant Ubiquitylation Tar

geted by the F-box Gene Superfamily"

5:30 pm - 5:45 pm Debarati Basu, PhD , "Two Hydroxyproline Galactosyl

transferases, GALT5 and GALT2, Function in Arabinogalactan- Protein

Glycosylation, Growth and Development in Arabidopsis"

Award Ceremony

6:30 pm - 7:30 pm Social / View Posters

7:30 pm - 9:00 pm Banquet and Kopchick Fellowship Awards

9:00 pm - 11:00 pm Music and open bar

John J. Kopchick FellowshipProvided by a generous gift from John J. Kopchick, PhD, Goll-Ohio Eminent

Scholar and Distinguished Professor of Molecular Biology and Char Kopchick, Assistant Dean of Students as part of the Promise Campaign 2014.

Faculty John J. Kopchick Molecular and Cellular Biology (MCB)/Translational

Biomedical Sciences (TBS) Faculty Support Fund

Funding up to $10,000 for MCB/TBS faculty for research activities and confer-

ence travel.

Dr. Shiyong Wu, Director Edison Biotechnology Institute

Title of Project: Identifying Antioxidants with Optimum Properties.

Graduate Student John J. Kopchick Molecular and Cellular Biology (MCB)/Translational Bio-

medical Sciences (TBS) Research Fellowship Award

Funding of up to $10,000 for PhD and DO/PhD students in the MCB or TBS

programs to support translational biomedical research, with an additional

$5,000 for an off-campus internship (min. 4 weeks , up to one semester).

Amrita Basu, 2015, Biomedical Sciences, Biological Sciences, MCB program

Mentor: Dr. John Kopchick

Title of Project: Influence of GH/IGF-! Signaling on Development and Progres-

sion of Amyloid-beta Toxicity in Relation with Alzheimer’s disease.

Xuan Wang, 2016, Biological Sciences, MCB program

Mentor: Dr. Xiaozhuo Chen

Title of Project: Anti-cancer mechanism study of a novel small molecule glu-

cose transporter 1 inhibitor.

Mark Slayton, 2018, Biological Sciences, MCB program

Mentor: Dr. Bonita Biegalke

Title of Project: Mutational Analysis of UL34 Protein-DNA Binding Sites in the

Manindra Singh, 2016, Biological Sciences, MCB program

Mentor: Dr. Fabian Benencia

Title of Project: Investigation of Endothelial Cell-Leukocyte interaction regu-

lated by the Adipokine Visfatin.

Qiping Lu, 2017, Biological Sciences, MCB program

Mentor: Dr. Yang Li

Title of Project: The relationship among Zinc, ROS and mitochondria under

hypoxia and re-oxygenation.

Undergraduate Student John J. Kopchick Molecular and Cellular Biology (MCB)/Translational Bio-

medical Sciences (TBS) Undergraduate Student Support Fund

Funding up to $1,500 for undergraduate students working MCB/TBS faculty

and conducting translational medical research and scholarly activities.

Nicole Brooks, 2016, HTC, Biological Sciences

Mentor: Dr. Darlene Berryman

Title of Project: FGF21 Expression in Growth Hormone Modified Mice: Links

to Obesity, Type 2 Diabetes Mellitus, and Increased Longevity.

Phillip Craigmile, 2016, Biological Sciences

Mentor: Dr. Shiyong Wu

Title of Project: The effect of p53 mutation on ionizing radiation-induced can-

cer cell invasion.

Kara Finley, 2015, Biological Sciences

Mentor: Dr.Sochi Tanda

Title of Project: Genetic and histochemical analysis of Moesin phosphoryla-

tion in light-sensing rhabdomere differentiation in Drosophila melanogaster.

Grant Gase, 2016, Biological Sciences

Mentor: Dr. Darlene Berryman

Title of Project: Using Isolated Mitochondria from Adipose Tissue to Assess

the Impact of Depot and Aging on Cellular Metabolism.

Jacob Sieg, 2017, Chemistry and Biochemistry

Mentor: Dr. Jennifer Hines

Title of Project: Development of a fluorescence anisotropy assay for the

Poster Presentations (1) Marilyn Hayden, MS Graduate Student

MCB / Department of Environmental and Plant Biology

Advisor: Dr. Sarah Wyatt

Title: Creating a Foundational Gene Network: a Systems Approach for

Understanding the Plant Response to Spaceflight.

Abstract: Advances in technology for mRNA analysis and continuous devel-

opment of the plant biology research facilities on the International Space Sta-

tion have resulted in an explosion of gene expression data. Initial examina-

tion of gene expression data from plants subjected to spaceflight show that

plants do respond but with modified gene expression patterns when com-

pared to ground controls. Currently, considerable gene expression data are

available, but the treatments and parameters are varied in each approach.

The sheer quantity of the genes expressed in response to spaceflight and the

variability between the gene expression data has limited further analysis.

Thus, to leverage these data sets for a better understanding of how plants

respond to spaceflight, we undertook a global examination of all the current

spaceflight gene expression data through creation of a gene network using

Cytoscape. Publicly available Arabidopsis Protein-Protein Interaction data sets

were extracted and imported into Cytoscape. The PPIs were merged into a

single network, and network analysis performed using publicly available gene

expression data sets to filter out genes not correlated or co-regulated and

genes unrelated to spaceflight. Optimization of the filtered network was facil-

itated through annotation of genes and their potential interactions using

Gene Ontology, Aracyc and text mining. The optimized network resulted in a

directed acyclic network, consisting of nodes (genes) and directed edges

(interactions) involved in the plant response to spaceflight. Ultimately, this

network will help identify hub and bottleneck genes involved in the plant re-

sponse to spaceflight. This network will also allow users to simulate the

effects of subtle changes of a single gene on the rest of the network. Both

identification of hub and bottleneck genes and simulation of the subtle

changes within the network will allow the network to serve as a foundation

for future gene expression analysis, as well as provide insight to the physio-

logical effects of spaceflight on plants.

This work was partially funded by NASA.

(2) Proma Basu, MS Graduate Student

MCB / Department of Environmental and Plant Biology

Advisor: Dr. Sarah Wyatt

Title: A Study of Posttranslational Modification of Proteins Expressed During

Gravitropic Response in Plants.

Abstract: Proteins are the work horses of the cell. Changes in the genome of

an organism manifest itself through the differential expression of proteins.

This is true for plants too. Plant response to gravity also takes effect through

changes in its proteome. Although gene expression studies provide infor-

mation on the genes expressed during gravity response, they do not provide

the full picture. After transcription the RNA generated may get translated to

proteins or it may be degraded. Thus studying the transcriptome can only

give a transient picture of the response, but not the final outcome of the re-

sponse. This final outcome can be visualized by studying the proteome. The

aim of this study is to identify the function of the proteins involved in the sig-

nal transduction events that occur in Arabidopsis prior to the redistribution of

auxin in response to gravity. The Gravity Persistent Signal (GPS) treatment

allows the plants to perceive gravity but provides a cold treatment that inhib-

its auxin transport thus isolating the events of signal transduction. Tissue was

collected from the plants subjected to the GPS treatment at 2min and 4 min

after the cold treatment to study the proteins expressed during signal trans-

duction. Previous proteomic analysis identified 82 proteins to be differentially

expressed between treatment and control. Post translational modifications of

proteins determine the function and sub cellular localization of proteins. A

SysPTM search of the differentially expressed proteins shows that 26 out of

the 82 differentially expressed proteins have post translational modifications

and 24 of the 26 proteins show phosphorylation at multiple sites. Segregation

of these proteins into functional groups may reveal evidence of phosphoregu-

lation in signal transduction pathways.

This work has been partially funded by NASA.

(3) Yanrong Qian, PhD Graduate Student

MCB / Department of Chemistry and Biochemistry

Advisor: Dr. Xiaozhuo Chen

Title: Extracellular ATP-induced intracellular ATP concentration elevation

mediated by macropinocytosis promotes growth, survival, and drug re-

sistance of cancer cells.

Abstract: Despite progress made in research of the Warburg effect, the bio-

logical reasons for ATP synthesis by aerobic glycolysis in cancer cells are only

partially understood. Intriguingly, intratumoral (extracellular) ATP levels are

103 to 104 times higher than those in normal tissues. We recently showed

that although extracellular ATP is not known to cross the plasma membrane

by itself, extracellular ATP in the range of the intratumoral ATP levels induced

large intracellular ATP concentration increase in A549 human lung cancer

cells. Based on these results, we hypothesized that extracellular ATP is taken

up by cancer cells and promotes cancer cell survival. Here we report that, a

nonhydrolyzable fluorescent ATP was internalized by A549 human lung cancer

cells through macropinocytosis, a process known as non-specific large fluid

drinking. The induced ATP increase, which involved neither transcription nor

translation, was reduced by the macropinocytosis inhibitor EIPA but persisted

even when mitochondrial oxidative phosphorylation and glycolysis were inhib-

ited. The increases were also observed in several other cancer cell lines, but

not in noncancerous cells. Furthermore, extracellular ATP enhanced cancer

cell survival under various stress conditions and promoted drug resistance to

tyrosine kinase inhibitors that compete with ATP for their anticancer action.

Together, these results provide the first evidence that extracellular ATP is in-

ternalized by cancer cells via macropinocytosis, which significantly contributes

to their growth and drug resistance. These findings potentially change our

understanding of ATP supply and sharing among cancer cells, expanding the

current interpretation of the Warburg effect and highlighting a novel anti-

cancer target.

(4) Qiping Lu, PhD Graduate Student

MCB / Department of Biological Sciences / Department of Biomed Sciences

Advisor: Dr. Shiyong Wu

Title: Intracellular Zinc distribution in mitochondria, ER and the Golgi

apparatus.

Qiping Lu1,2, Hariprakash Haragopal3, Yang V Li1,3

1. Molecular and Cellular Biology Interdisciplinary Program 2. Department of

Biological Sciences 3. Department of Biomedical Sciences

Abstract: Zinc is an essential trace element in human body involved in various

physiological processes. Zinc functions within cells in both protein-bound

form and free/labile state. The majority of Zinc is bound to proteins and en-

zymes, contributing to protein structures and enzymatic activities. The labile

Zinc, in a small but measurable pool, may play an equally important role in

modulating intracellular activities through its transient action on effector pro-

teins. An interesting phenomenon is that, the level of labile zinc in cells is

strictly regulated, which is called zinc homeostasis. Disruptions of zinc homeo-

stasis cause diseases and cell death. Intracellular zinc distribution and storage,

is a critical step toward understanding zinc homeostasis. However, there is

much debate on the exact distribution of free Zinc within a live cell. The data,

currently available, are still inconsistent in showing which organelles act as

Zinc stores. Clear understanding of the intracellular Zinc stores is an immedi-

ate necessity for successful mechanistic modeling of various Zinc-involving

pathologies like diabetes, stroke, epilepsy and Alzheimer’s disease. To this

end, we undertook the present study with a comprehensive approach to

screen for organelles, although qualitatively, that represent potential intracel-

lular Zinc stores across various cell types: HeLa, HIT-T15 and cultured rat pri-

mary cortical neurons. The cells were co-stained with Zinpyr-1, a Zinc fluores-

cent indicator, and organelle-specific fluorescent dyes (Mitochondrion: Mito-

Fluor Red 589; Endoplasmic Reticulum: ER Tracker red; the Golgi apparatus:

BODIPY TR Ceramide; Nucleus: SYTO Red 64) to locate free intracellular Zinc

within the cells. The co-localization was analyzed by overlaying fluorescence

signals from Zinpyr-1 and organelle-specific dyes. Here, we identify Zinpyr-1

signals co-localizing with MitoFluor Red 589, ER Tracker Red, and BODIPY TR

(5) Yunyi Feng, PhD Graduate Student

Department of Environmental and Plant Biology

Advisor: Dr. Ahmed Faik

Title: In silico modeling of a synthase complex involved in the biosynthesis

of plant cell wall hemicellulose.

Abstract: Plant cell wall is the most abundant source of biomass on earth and

thus is of high interest for industrial applications. Cell wall is also important

for plant development. Hemicellulose polymers are one of important compo-

nents of cell walls. They are critical for cell wall integrity, as they cross-link

cellulose microfibrils. In grasses glucuronoarabinoxylan (GAX) is the most im-

portant hemicellulose. Previous study from our lab purified a GAX synthase

from wheat and found that at least six proteins form a complex and work in a

cooperative manner to synthesize this polymer. However, the exact biochemi-

cal function of individual proteins and their interactions are still elusive. This

study uses computational tools to model these six proteins in conjunction

with molecular dynamics simulation. The aim of the study is to provide useful

information for further experimental studies on biosynthetic mechanism, and

protein-protein interactions that regulate the assembly of this complex. Ulti-

mately, the outcome of this study should facilitate biochemical engineering of

xylan synthesis.

(6) Nan Jiang, PhD Graduate Student

Department of Environmental and Plant Biology

Advisor: Dr. Ahmed Faik

Title: Characterization of glucuronoarabinoxylan synthase complex from

bread wheat.

Nan Jiang, Rebekah Whitley, Prameela Jyothi, Michael Held, and Ahmed Faik

Abstract: Xylan represents a major constituent of grass cell walls. Its struc-

ture consists of a ß-(1,4)-linked D-xylose residues substituted with either a-

(1,2 or 1,3)-L-arabinofuranose (Araf), as in arabinoxylan, a-(1,2)-D-glucuronic

acid residues (GlcA), as in glucuronoxylan, or both Araf and GlcA residues as in

glucuronoarabinoxylan (GAX). Proteomics and transcriptomics analyses spe-

cifically associated three wheat glycosyltransferases, namely TaGT43-4, TaG-

T47-13, and TaGT75-4, to purified GAX synthase activity. Co-

immunoprecipitation experiments indicated these proteins physically interact

to form a GAX synthase complex. In addition, we demonstrated that GAX syn-

thase activity reqiures cooperation between xylosyltransferase, arabinosyl-

transferase, and glucuronosyltransferase activities. Bimolecular fluorescence

complementation (BiFC) methods demonstrated that TaGT75-4 and TaGT43-4

physically interact to form homodimers as well as heterodimers. The interac-

tion between TaGT43-4 and TaGT47-13 was weak but reproducible. Alterna-

tively, biochemical approach is used to reconstitute a functional GAX synthase

complex in vitro. GAX synthase activity can be partly reconstituted from het-

erologous expression of both TaGT43-4 and TaGT47-13 proteins in Pichia pas-

toris cells. Interestingly, much higher GAX synthase activity was recovered as

co-expression of TaGT43-4, TaGT47-13, and TaGT75 members, which possess

UDP-Arap mutase activity. The results of product analysis and GAX biosynthe-

sis mechanism will be presented. To demonstrate that wheat proteins are

functionally similar to Arabidopsis xylan synthesizing GTs such as IRREGULAR

XYLEM (IRX) 10 and IRX14, genetic complementation experiments of Ara-

bidopsis irx mutants (irx10, irx10-L, irx14, and irx14-L) were carried out using

TaGT43-4 and TaGT47-13 genes. Our results showed that these mutants can

be functional complemented by their homologous wheat proteins. Characteri-

zation of complemented mutants will be discussed in the meeting. This work

(7) Aditi Vyas, PhD Graduate Student

MCB / Department of Biological Sciences

Advisor: Dr. Soichi Tanda

Title: Identification of novel STAT92E target genes in hematopoiesis using in

silico methods.

Abstract: The Jak/Stat pathway is one of the pathways controlling hematopoi-

esis in Drosophila. A dominant allele of the Drosophila Jak gene

called hopTum-l causes a tenfold increase in the total hemocyte count. Re-

moval of a negative regulator of the Jak/Stat pathway increases the pathway

activity beyond the hopTum-l level and surprisingly results in reduction of the

total cell count. These results suggest that Stat92E controls expression of two

sets of target genes; the low threshold genes, expressed at moderate levels of

the pathway and the high threshold genes, activated at much higher levels of

pathway activity. We hypothesize that high threshold genes are possibly in-

volved in limiting cell proliferation, and their expression is negatively regulat-

ed by transcriptional repressors. To test this, we screened for effect of co-

repressor and repressor mutants on hematopoiesis. We found that loss of

function of C-terminal Binding Protein (CtBP) as well as its DNA binding part-

ner Suppressor of Hairless [Su(H)], dramatically decreased the hemocyte

count. We then aimed to identify the high threshold genes. We used two in

silico approaches to discover possible regions in the Drosophila melano-

gaster genome, that have Stat92E and repressor binding sites. In one ap-

proach, we searched modENCODE's CtBP ChIP-Chip dataset for presence of

Stat92E and repressor binding sites using the bioinformatics tool Target Ex-

plorer. This method gave us a list of 12 candidate genes. In a second ap-

proach, we scanned the entire Drosophila melanogaster genome for Stat92E,

Su(H) and insulator motif occurrences, using the publicly available FIMO tool,

and performed post processing using Perl. This in silico scan yielded a list of 18

genes. We performed RNAi screening against these candidates in the hopTum

-l background, to see their effect on larval hematopoiesis. RNAi against genes

that caused significantly higher increase in the hemocyte count, have been

selected as potential high threshold genes.

(8) Xuan Wang, PhD Graduate Student

MCB / Department of Biological Sciences

Advisor: Dr. Xiaozhuo Chen

Title: Extracellular ATP promotes cancer cell growth, survival and drug re-

sistance.

Xuan Wang1,4,5, Yanrong Qian2,4,5, Yunsheng Li5, Xiaozhuo Chen1-5.

1 Department of Biological Sciences, 2 Department of Chemistry and Bio-

chemistry, 3 Department of Biomedical Sciences, 4 Interdisciplinary Graduate

Program in Molecular and Cellular Biology, 5 Edison Biotechnology Institute,

Ohio University, Athens, OH 45701 USA

Abstract: ATP is the most direct energy form in cells including cancer cells.

The fact that cancer cells undergo rapid proliferation suggests that they must

possess mechanisms to meet their high energy need. Previous studies report-

ed that extracellular ATP levels of various cancer types are 103 to 104 times

higher than those in corresponding normal tissues and that ATP can be taken

up directly by cancer cells. Based on these results, we hypothesized that the

internalized extracellular ATP contributes to intracellular ATP pool and pro-

motes cancer cells growth, survival and drug resistance. In this study, human

lung cancer A549 and breast cancer MCF7 cells were used to determine the

effects of extracellular ATP in the presence or absence of anticancer drugs.

Cancer cell proliferation rate, viability and intracellular ATP concentration

were determined. The results suggest that the proliferation rate of cancer

cells was increased in the presence of extracellular ATP. Furthermore, extra-

cellular ATP promoted cancer drug resistance to tyrosine kinase inhibitors by

increasing intracellular ATP levels. Moreover, the drug resistance was reduced

by ATP, but not its derived molecules ADP, AMP or adenosine. The findings of

the study may expand our understanding on the roles of the high extracellular

ATP in tumors and lead to novel potent anti-cancer strategies.

(9) Debra Walter, PhD Graduate Student

MCB / Biological Sciences / HCOM Biomedical Sciences

Advisors: Dr. Kelly McCall, Dr. Karen Coschigano

Title: Kidney Damage Associated with Viral-Induced Diabetes.

Walter D.L., McCall K.D. and Coschigano K.T.

Abstract: Over twenty-five million people in the United States are affected by

diabetes, requiring a lifetime of treatment for the disease and resulting com-

plications. Diabetes is the leading cause of end stage renal disease and kidney

failure. We are interested in identifying reasons for the effects of diabetes on

the kidneys since the development of kidney damage is likely to occur in these

patients. Unfortunately, it is difficult to study kidney damage using human

subjects. Our goal is to develop a new experimental method that makes the

use of mice more time and cost effective to study diabetic kidney disease pro-

gression. We are utilizing the non-obese diabetic (NOD) mouse that spontane-

ously develops diabetes, mimicking the disease seen in humans and making it

a potentially useful mouse in which to study diabetic complications. When

infected with coxsackievirus, the NOD mouse develops diabetes nearly twice

as fast as uninfected mice. We hypothesize that infection of NOD mice with

coxsackievirus will also accelerate the onset of diabetic kidney damage. In

order to successfully use the NOD mouse for a model of diabetic kidney dam-

age, we need to distinguish kidney damage caused by coxsackievirus infection

and kidney damage caused by hyperglycemia. Thus far we have demonstrated

that coxsackievirus can infect the kidneys of NOD mice. Coxsackievirus infec-

tion also transiently but significantly alters kidney weight and albuminuria in

NOD mice, indicators of kidney damage. We are also measuring levels of mo-

lecular markers known to be found in the kidneys, urine and serum of mice

with kidney damage. Demonstration of viral-induced acceleration of diabetic

kidney damage in this study would provide a new tool for the development of

beneficial treatment strategies for diabetic kidney disease and possibly other

complications associated with diabetes.

This work has been partially funded by an Ohio University Student Enhance-

ment Award, an Original Works Grant from the Ohio University Graduate Stu-

(10) Chunxi Zeng, PhD Graduate Student

MCB / Department of Chemistry and Biochemistry

Advisor: Dr. Jennifer Hines

Title: Molecular modulation of T box riboswitch function.

Chunxi Zeng,1,2 George Acquaah-Harrison,2 Crina Orac,

2 Stephen C. Bergmeier,1,2 Jennifer V. Hines1,2*

1Molecular and Cellular Biology Program, Ohio University; 2Department of

Chemistry and Biochemistry, Ohio University

Abstract: The T box riboswitch is a transcription regulation mechanism that

controls expression of essential genes in many bacteria. Uncharged cognate

tRNA induces read-through (antitermination) of the T box riboswitch and al-

lows expression of downstream genes. Two fluorescently labeled probes were

designed to study the riboswitch in real time during multi-round in vitro tran-

scription of the leader region of the glyQS gene. The read-through probe

showed good photostability, integrity and target sensitivity. Specific ri-

boswitch response to uncharged cognate tRNAgly was detected with a low

nanomolar EC50. Using this validated, moderate throughput riboswitch func-

tional assay, the effect of molecular modulators of T box riboswitch function

was then determined.

(11) Sofia Ocana-Mayorga, PhD

Center for Infectious Disease Research Catholic University of Ecuador

Title: Role of vectors and reservoirs of Trypanosoma cruzi, causal agent of

Chagas disease, in parasite population dispersal in Southern Ecuador.

Sofia Ocana-Mayorga1,2, Mario J. Grijalva1,2

1 Ohio University, Department of Biomedical Sciences, Heritage College of

Osteopathic Medicine, Tropical Disease Institute, Athens, Ohio.

2 Pontifical Catholic University of Ecuador, School of Biological Sciences, Cen-

ter for Infectious and Chronic Disease Research, Quito, Ecuador.

Abstract: Chagas disease is a parasitic infection caused by the parasite Trypa-

nosoma cruzi. This disease affects an estimated 200,000 people in Ecuador,

and the presence of this parasite has been demonstrated in humans, vectors

and reservoirs. The biological complexity of this disease greatly hampers

transmission control efforts. Currently, diagnosis and treatment are unsatis-

factory. Moreover, control strategies applied to control vector populations are

partially effective. A former study has unveiled important genetic characteris-

tics about the parasite population dynamics which demonstrates a genetic

subdivision of populations of T. cruzi and a dispersal process related with the

geographic distance. However, the impact of parasite dispersal in areas where

control strategies have been applied, the role of the different species of hosts

in parasite dispersal and its influence on the genetic characteristics of T. cruzi

population have not yet determined. The current research propose to use

genetic typing techniques as the analysis of microsatellites (MLMT) and se-

quencing (MLST) to establish the role of hosts in dispersal of parasite popula-

tions, based on 1) the influence of reservoirs species (mammals) in genetic

flow and dispersal of parasites along geographic areas and 2) the impact of

vectors (triatomines) in dispersal of parasite populations in areas where the

application of insecticide spraying for vector control has been carried out.

Through the analysis of multi locus microsatellite (MLMT) typing and se-

quence typing (MLST) of T. cruzi isolates from vertebrate and invertebrate

hosts circulating in domestic, peridomestic and sylvatic environments we aim

to gather important data to determine how parasites are moving among

transmission areas and the origin of the parasites found in human dwellings

(12) Manindra Singh, PhD Graduate Student

MCB / Biological Sciences, Biomedical Sciences

Advisor: Dr. Fabian Benencia

Title: Investigation of Adipokine-Induced Activation Responses in Endotheli-

al Cells.

Abstract: Adipokines are fat tissue-derived signaling molecules that control a

variety of physiological functions, including metabolism, insulin resistance,

energy balance, angiogenesis and inflammation. Obesity-induced dysregula-

tion in the secretion of adipokines has been reported as a major contributive

factor for cardiovascular pathogenesis and metabolic syndrome. Previous

studies have shown that various chemotactic and inflammatory signals origi-

nating from the visceral adipose tissue may cause the recruitment of leuko-

cytes from the circulation towards the adipose tissue. But the mechanisms

that integrate abnormal adipokine function and adipose tissue inflammation

are poorly understood. In the present study, the function of adipokines:

visfatin, leptin and vaspin was assessed for their capability to induce inflam-

matory responses in vascular endothelial cells. A large-scale antibody-based

protein micro array was used to detect the chemokines secreted by the adi-

pokine-treated cells. Specifically, visfatin treatment upregulated the produc-

tion of CXCL1, CCL11 and VEGF. Further analysis using enzyme-linked im-

munosorbant assays revealed that visfatin also upregulated the production of

inflammatory molecules: MCP-1 and GM-CSF, by the endothelial cells. Quanti-

tative real-time PCR analysis showed that visfatin induced the expression of

inflammatory genes: IL-6, MCP-1 and MMP-9 at the transcript level. Addition-

ally, a colorimetric cell-proliferation assay revealed that visfatin decreased cell

viability over the treatment period. Overall, these results indicate that the

adipokine visfatin might be involved in the inflammatory activation of endo-

thelial cells and may contribute to inflammation of adipose tissue and patho-

genesis.

(13) Rami Al-Ouran PhD Graduate Student

Computer Sciences (Bioinformatics)

Advisor: Dr. Lonnie Welch

Title: Identification of functional regulatory elements in Brugia malayi for

Lymphatic Filariasis (LF) disease .

Abstract: Gene transcription initiation and gene regulation are complex bio-

logical mechanisms that involve several molecular components working in a

precise manner. Transcription factors (TFs) and the transcription factor bind-

ing sites (TFBSs) are functional elements that control gene regulation. Identi-

fying TFBSs will assist in deciphering regulation of transcription and represent

potential target sites for disease prevention. Lymphatic filariasis (LF), also

known as elephantiasis, is a neglected tropical disease that affects over 120

million people worldwide. Brugia malayi is one of the nematode

(roundworm) parasites that cause LF. Each stage of the B. malayi lifecycle has

a unique gene transcriptional signature and third-stage filarial larvae (L3) are

of particular interest as they represent the infective stage. In this study we

aim to discover the putative TFBSs that are unique to genes over expressed in

the L3 phase of the B. malayi lifecycle. Identifying the B. malayi stage specific

regulatory elements could help in developing intervention strategies for the

control of LF.