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Chart Abstracts 8th URS 8th URS Photos  UT Biological Sciences


(click on the presenter name for abstract)

Time

Speaker

Presentation Title

9:00

Dr. Patricia Komuniecki

Welcome

9:05 Plenary Lecture- "From Bench to Clinics: Translating basic science into human therapies"- Dr. Lawrence Elmer, Medical College of Ohio

9:50

Coffee Break

Session I - Sarah Miller, Moderator

10:00

Allison Cihla

"The mechanism of NF-kappaB Regulation in Pancreatic Cancer"  Advisor: Dr. Brian Ashburner, Department of Biological Sciences, The University of Toledo

10:20

Dia Smiley

"Development of Activation Tagging for Gene Discovery in Maize"  Advisor: Dr. John Gray, Department of Biological Sciences, The University of Toledo

10:40

Jeff Henke

"Preparation of Candidate Genes for Microinjection Analysis of mua-5" Advisor: Dr. John Plenefisch, Department of Biological Sciences, The University of Toledo

11:00

Meredith Snapp

"Attentional Blink Comparison of Central Versus Lateral Presentations" Advisor: Dr. V. Kavcic and Dr. R. Doty, Department of Neurobiology and Anatomy, University of Rochester

11:20

Shaddy Swade

"Uneven Magnesium Distribution in Parkinson's Disease Patients" Advisor: Dr. Lawrence Elmer, Department of Neurology, Medical College of Ohio

11:40

Margaret Uhrich

"NF-KB: Midnight in the Garden of Good and Evil" Advisor: Dr. Brian Ashburner, Department of Biological Sciences, The University of Toledo

12:00

Pizza lunch

Session II - Rhea Busick, Moderator

12:30

Lyndsey Emery

"Gene Knock-Out Study of Oxygenases in Cyanobacteria"  Advisor: Dr. John Gray, Department of Biological Sciences, The University of Toledo

12:50

Emily Davis

"ANovel Protocol for Regeneration of Soybean from Mature Immature Cotyledon " Advisors(s): Dr. G. Franklin and Dr. R.V. Sairam, Department of Earth, Ecological and Environental Sciences, The University of Toledo

1:10

Sean Ehinger

"Hemidesmosome Localization in Caenorhabditis elegans Cell-matrix Adhesion Mutants" Advisor): Dr. John Plenefisch, Department of Biological Sciences, The University of Toledo

1:30

Steve Driver

"The Potential Use of Tr1 Type Regulatory T-Cells in Antigen-Specific Multiple Sclerosis Immunotherapies" Advisor: Dr. B. Bielekova and Dr. R. Martin, Cellular Immunology Section NIH, NINDS

1:50

Paul Nolan

"The Effects of IL-13 on CD38/cAPDR Mediated Calcium Signaling in Human Airway Smooth Muscle Cells" Advisor: Dr. Mathus Kannan, University of Minnesota

2:10

Courtney Maunz

"Identification of Viral Protein Inducing Peptides" Advisor: Dr. Scott Leisner, Department of Biological Sciences, The University of Toledo

2:30

Nancy Parquet

"The Role of Transcriptional Coactivator: Proteins on the Regulation of NF-kB Activity" Advisor: Dr. Brian Ashburner, Department of Biological Sciences, The University of Toledo

2:50

Break

3:00

Awards ceremony




Abstract 8th URS


Allison Cihla
“The mechanism of NF-KappaB Regulation in Pancreatic Cancer”
Advisor:  Dr. Brian P. Ashburner-Department of Biological Sciences, The University of Toledo

NF-kappaB is a transcription factor important in mediating immune and inflammatory responses, and in protecting cells from programmed cell death.  Because of this, NF-kappaB is dysregulated in many cancers but the mechanism for this dysregulation is not known.  Ras is one of the most frequently mutated oncogenes in all human cancers, and greater than 90% of pancreatic tumors displayed on activating mutation in the K-Ras gene.  In addition, many pancreatic tumors also display constitutively activated NF-kappaB.  Ras activates the p38 MAP kinase pathway, which contributes to NF-kappaB activation.  Although several reports indicate that p38 is antagonistic to Ras-mediated transformation, kappaB is required for Ras-mediated oncogenesis, understanding how Ras regulates NF-kappaB activity is important.  We are using pancreatic cancer cell lines that express onocegenic K-Ras to determine if Ras activation of -38 contributes to NF-kappaB activity.  Our results indicate that inhibition of p38 partially blocks NF-kappaB activity in these pancreatic cell lines.  Future experiments will look at the roll of p38 on expression of NF-kappaB regulated genes in pancreatic tumor cells to determine whether p38 plays an important role in the dysregulation of NF-kappaB in pancreatic cancer cells.


Emily Davis
“A novel protocol for regeneration of soybean from mature and immature cotyledon”
Advisors:  Dr. S. Goldman, Dr. G. Franklin and Dr. R. V. Sairam-Department of Earth, Ecological and Environmental Sciences, The University of Toledo

Soybean (Glycine max L.) plantlets were efficiently regenerated from mature and immature cotyledons.  Deep greenish organogenic nodules were induced which subsequently differentiated into shoot buds from the proximal end on modified Murashige and Skoog (1962) medium.  Among the various combinations of plant growth regulators tested, the medium containing two cytokinins (6-benzlaminopurine and thidiazuron) was found to be superior.  The regenerated shoot buds elongated and rooted on MS medium containing 0.1 mgl-1 gibberellic acid and 0.5 mgl-1 a-naphthaleneacticacid respectively.  Rooted plants transferred to greenhouse with 87% success produced viable seeds.  Preliminary studies with Agrobacterium show great promise for transforming soybean using this protocol.


Steve Driver
“The potential Use of Tr1 Type Regulatory T-Cells Antigen-Specific Multiple Sclerosis Immunotherapies”
Advisors:  Dr. B. Bielekova and Dr. R. Martin-cellular Immunology Section, NIH, NINDS

Multiple sclerosis (MS) is a chronic inflammatory demyelinating disease of the central nervous system, and is second only to trauma as the leading cause of neurological disability among young and middle aged adults.   Approved treatments are antigen-non-sepcific and unable to cure mose patients.  It is widely accepted that auroreactive T lymphocytes specific for some antigenic constituent of myelin play a crucial role in the disease’s pathogenesis.  Evidence now exists suggesting that, in healthy individuals, deleterious activation of autoreactive T cells is regulated by several active mechanisms including regulatory T cells.  Type 1 Regulatory Cells (Tr1 cells), primed from human peripheral blood by polyclonal stimuli, are potent inhibitors of immune responses.  Therefore using MS patient samples, we asked whether Tr1 cells specific for myelin antigens may be isolated from memory T cells pools, and/or primed from naïve T cells under selected in-vitro conditions.  In order to define optimal priming conditions and to characterize the phenotypes of the immunoregulatory cells, we used proliferation assays, functional supernatant mediated suppressions assays, and ELISA from cultured supernatants.  Preliminary data suggest that such cells can be isolated from some MS patients, and to some degree, cells with moderate immunosuppressive activity (~30%) can also be primed.


Sean Ehinger
“Hemidesmosome Localization in Caenorhabditis elegans Cell-matrix Adhesion Mutatnts”
Advisor:  Dr. John D. Plenefisch-Department of Biological Sciences, The University of Toledo

The purpose of this study is to understand how adhesion between tissues in maintained.  Adhesion genes are connected to disorders such as muscular dystrophy and cancers which involve problems in cellular adhesion.  A simple model used in this research is the nematode Caenorhabditis elegans.  Genetice mutations in the muscle attachment genes are used in the experimentation.  These mutations knows as MUA mutations, exhibit abnormalities in the cellular junctions of the animals.  A protein marker simply called Green Fluorescent Protein (GFP) is used to mark specific junctional proteins involved in cellular adhesion.  A strain of C. elegans (EE86) contains a GFP marked MUP-4 protein that is connective protein in the hemidesmosomes (a type of cellular junction).  Mutant strains mua-2, mua-6, and mua-10 are crossbred with EE86 strain.  Through the crossbreeding the MUP-4::GFP is incorporated into the mutant strains.  Animals are selected from the mutant strains that exhibit the mutant phenotype and fluorescent MUP-4::GFP.  These animals are examined using fluorescent microscopy to detect the MUP-4::GFP and determine the location of the hemidesmosomes.  Expected results include disruptions of hemidesmosome positions and determination of cellular detachment areas.


Lyndsey Emery
“Gene Knock-Out Study of Oxygenases in Cyanobacteria”
Advisor:  Dr. John Gray-Department of Biological Sciences, The University of Toledo

The LLS1 protein provides an important protection against light-dependent cell death in higher plants.  Several lines of evidence, including localizations studies, show that LLS1 functions inside plant chloroplasts.  The biochemical function of the LLS1 protein has not been elucidated but the predicted amino acids sequence exhibits two non-heme iron binding motifs that are conserved amongst aromatic right-hydroxylating (ARH) oxygenases in bacteria.  Phylogenetic analysis has revealed the presence of several genes homologous to LLS1 in photosynthetic bacteria.  In an attempt to functionally analyze these oxygenases in Synechocystis sp. PCC 6803, the technique of gene knock-out via homologous double recombination was employed.  Theree genes of interests, sir7447, still 1297, and sir1869 were targeted for gene knockout.  The Polymerase Chain Reaction was used with specifically designed primers to amplify the flanking chromosomal regions of each of the genes of interest.  Then, these amplified products were cloned into the TA vector and sequenced to verify that no mutations had occurred.  Verified clones were subcloned into the knockout vector pKCm and checked for proper orientation.  These deletion constructs are currently being transformed into Synechocystis sp. PCC 6803.  During this transformation process, the cyanbacteria should incorporate this plasmid into their chromosome by the process of homologous double recombination, and a cointegrate intermediate should be formed.  With successful recombination, slr1747, sll1297, and slr1869 will be deleted from the Synechocystis genome, respectfully.  These mutated strains can then be compared with the wild type strain to help determine the functions of these genes and hopefully lead to further insight into the function of lls1 in plants.

 

Jeff Henke
“Preparation of candidate genes for microinjection analysis of mua-5”
Advisor:  Dr. John D. Plenefisch-Department of Biological Sciences, The University of Toledo

Locomotion in Caenorhabditis elegans is dependent upon the transmission of contractile force from skeletal muscles through a basal lamina and epidermis to external cuticle.  Various cytoskeletal proteins and matrix components are crucial to formation of this mechanical pathway.  The mua (muscle attachment) class of genes in caenorhabditis elegons results in fragility of these structural linkages when mutated.  This study focuses on mua-5.  Mua-5 mutants typically have normal muscle assembly and differentiation, and their movement in unaffected as L1 larvae.  However, as they grow, muscles detach from the body wall resulting in animals that are paralyzed. typically lying in a S-shaped coil position.  Using three-factor mapping, mua-5 was positioned between the genes unc-44 and lin-45.  Within this region, ten cosmids containing genomic DNA spanning this region were obtained and prepared for rescue analysis.  Preparation of these cosmids for microinjection was the main goal of this research.  The cosmids C11D2, CU9D4, and CO1LBIO, were grown and harvested according to conventional plasmid miniprep methodology.  Bacterial cells were lysed and the contents run on agarose gel to determine if an appropriately sized cosmid (40 kb) was present.  Then, cleanup was done using either a combination of PEG-800 and phenol/chloroform extraction of CTAB.  Subsequent gels revealed a loss of desired cosmids.  Testing of reagents found no contamination.  Further digestions, which utilized the same reagents, with know concentrated DNA resulted in no loss of the DNA.  It was determined at this point in the research, that the focus should be placed on the seven remaining cosmids.  Of great interest is the cosmid F58F9, which encodes a trombospondin type 1 repeat protein.  After isolation, microinjection analysis will be carried out on this candidate gene.

 

Courney Maunz
“Identification of Viral Protein Interacting Peptides”
Advisor:  Dr. Scott Leisner-Department of Biological Sciences, The University of Toledo

Plant viruses are a major economic problem for many crops worldwide.  Therefore, it is important to develop new strategies for dealing with plant viruses.  Since these pathogens are related to animal viruses, such research may also have implications for human disease.  Our laboratory studies Cauliflower mosaic virus (CaMV).  This virus is relatively well-characterized and the function of most of its genes is known.  One of the viral genes, VI, encodes a protein (P6) that is essential for viral infection.  The P6 protein is multifunctional regulating: viral host range, translation of viral proteins, and the formation of large intracellular structures called inclusion bodies (IBs).  IBs are sites where viral proteins synthesis, genome replication, and virus particle assembly occur.  Hence P6 is a key protein for viral infection and  as such, it is an attractive target for antiviral therapy.  To identify potential antiviral agents, we initially sought peptides that interacted with P6.  This was accomplished using a modified form of the yeast two-hybrid assay.  After testing a random peptide library, we identified at least 2 peptides that appeared to interact with P6.  We are now in the process of characterizing these peptides.  We will be isolating the DNA encoding the peptides from yeast and cloning it is Escherichia coli.  The DNA encoding the peptides will then be sequenced.  Finally, the portion of P6 to which the peptides bind will be determined.


Paul Nolan
“The Effects of IL-13 on CD38/cADPR Mediated Calcium Signaling in Human Airway Smooth Muscle Cells”
Advisor:  Dr. Mathur Kannan from the University of Minnesota

Bronchial inflammatory disease is a serious problem affecting a large population, and bronchoconstriction is a major affliction associated with airway diseases.  The exact mechanism of bronchoconstriction is not known, however, increased intracellular calcium response in smooth muscle has been shown to be responsible for this process.  Studies have demonstrated the role of cADPR as a second messenger to mobilize calcium, an essential component for smooth muscle contraction, through ryanidine receptor channels.  cADRP is produced and hydrolyzed by a single bifunctional membrane protein called CD38.  Furthermore, TH2 cytokines have been shown to be the primary mediators of changes in the contractile properties of human airway smooth muscle (HASM).  The present study was designed to determine the effect of IL-13 on CD38/cADPR mediated calcium mobilization in HASM cells.  The effect of IL-13 on CD38 expression was measured using RT-PCR, and real-time PCR.  Calcium responses to acetylcholine, bradykinin, and thrombin were measured in Fura-2 loaded cells by fluorescence microscopy.  The treatment of HASM cells with IL-13 caused augmentation of CD38 expression, and calcium response to the agonist.  In the presence of cADPR antagonist the net calcium response was attenuated indicating a significant contribution of cADPR mediated calcium mobilization during airway hyperresponsiveness.  The finding of this study demonstrates the underlying mechanism to cytokine induced HASM hyperresponsiveness. 


Nancy Parquet
“The Role of Transcriptional Coactivator:  Proteins on the Regulation of NF-kB Activity”
Advisor:  Dr. Brian P. Ashburner-Department of Biological Sciences, The University of Toledo

NF-kB is a transcription factor found in essentially all cell types, and controls the expression of numerous genes involved in immune response, inflammation, and cell growth and differentiations.  NF-kB-mediated gene expression requires transcriptional coactivator proteins.  Many coactivator proteins possess histone acetyltransferase (HAT) activity, and it is well established that histone acetylation is important for transcriptional activation.  Recently histone methylation has been shown to be important for transcriptional regulation.  Depending upon the residue methylated, histone methylation can contribute to transcriptional activation or repression.  Methylation of arginine residues on aminoterminal histone tails correlates with transcriptional activation.  We are currently trying to determine whether the family of protein arginine methyltransferases (PRMTs) acts as coactivators with histone acetyltransferases for transcriptional activation by NF-kB.  Our results indicate that PRMT1 and PRMT4, in conjuction with the steroid receptor coactivators SRC-1 and SRC-3, play an important role in NF-kB-mediated gene expression.


Dia Smiley
“Development of Activation Tagging for Gene Discovery in Maize”
Advisor:  Dr. John Gray-Department of Biological Sciences, The University of Toledo

The study of plant gene function has been classically carried out by investigating the effect of loss of function mutations in particular genes, either using forward or reverse genetics.  Although this approach has proven to be very powerful, often no phenotypes result when a particular gene is knocked-out.  The “functional genomics” strategy named “Activation Tagging” allows the generation of novel overexpression mutatnts (dominant) and recovery of mutant genes of interest in a short time with minimal effort.  The recovered genes can then be assessed for their agronomic utility and contributions to basic plant science.  In this project the aim was to construct and test genetically engineered maize transposons for use as activation tagging elements in maize.  Four Trigger DS reporter constructs were engineered to test the ability of 4X35S enhancer repeats to activate a nearby luciferase gene with a functional maize promoter (A1), in the context of the right and left Terminal Inverted Repeats (TIR) of the Ac/Ds transporson.  The activity of each of the four constructs was shown by bombarding each plasmid into maize BMS cells.  These constructs are also being further modified to contain a selectable marker (bar gene) in order to avoid the need for co-transformation.  The activation tag will be engineered to contain a plasmid to allow the quick recovery (“rescue”) of the flanking DNA into which the element has inserted.  The founder lines will be generated using Agrobacterium-mediated transformation.

 

Meredith Snapp
“Attentional Blink Comparison of Central Versus Lateralized Presentations”
Advisors: Dr. V. Kavcic and Dr. R. Doty, Department of Neurobiology and Anatomy, University of Rochester

Each half of the visual field projects to the contralateral hemisphere.  Thus by presenting visual stimuli to one or the other side, one can address stimuli to either half of the brain; or for stimuli in the middle of the field, to both hemispheres together.  The “attentional blink” is a momentary deficit in visual processing of a second target occurring shortly after allocation of attention to a first target within a series of stimuli.  The effect of lateralizing these stimuli was studied on different runs by presenting stimuli for the “blink” to right, left or center of the visual field.  The “blink” is a deeper and lasts longer following presentation in left and right visual fields as compared to central visual field.  There is no significant difference with stimulus presentations in the left or right field.  The “blink” is deeper and lasts longer following presentation in left and right visual fields as compared to the central visual field.  There is no significant difference with stimulus presentations in the left or right fields.  Poor performance of lateral presentations (20 left or right of center) is not likely to be cause by diminished acuity, as demonstrated by similar performance for presentations 40 from the fovea.  It is suggested that processing of stimuli initially received by only one hemisphere requires more interhemispheric communication, and is therefore more complex, than processing of central stimuli that engages both hemispheres concurrently.


Shaddy Swade
“Uneven Magnesium Distribution in Parkinson’s Disease Patients”
Advisor:  Dr. Lawrence Elmer, Department of Neurology, Medical College of Ohio

It is believed that the gating mechanism which binds the NMDA receptor in the human brain to a neurotransmitter called glutamate is changed by the presence of the divalent cation magnesium.  Blood was drawn from patients previously diagnosed with Parkinson’s Disease as well as from age-matched controls and fractionated in a method copyrighted by the OptiPreptm corporation.  Blood was fractionated into whole serum, platelets, leucocytes, and red blood cells by means of gravity and by density centrifugation.  These elements were then analyzed by atomic absorption spectrophotmetry and protein analysis for magnesium and protein content respectively, and compared to standard levels.  Epithelial cheeks cells were also collected with soft nylon brushes and placed into test tubes containing double distilled water and analyzed in a matter similar to the blood components.  Results indicate that there is an abnormal distribution of magnesium in patients with Parkinson’s as compared to those individuals can be recognized and understood, then some type of magnesium therapy may be plausible as an alternative treatment for Parkinson’s Disease that can be put into place at a stage in the illness before the symptoms become readily apparent.


Margaret Uhrich
“NF-xB:  Midnight in the Garden of Good and Evil”
Advisor:  Dr. Brian P. Ashburner-Department of Biological Sciences, The University of Toledo

Nuclear factor – kappa B (NF-kB) is a member of the Rel family of DNA-binding transcription factors that is important in regulating expression of the genes involved immune and inflammatory responses, and pro-survival signals.  NF-kB is activated by many stimuli that lead to a variety of responses.  Many oncogenes, including ras, activated NF-0B, which is thought to be important for proficing pro-survival signals to tumor cells.  Ras activates many signal transduction pathways including the p38 MAP kinase pathway.  Although p38 is known to play a role in activating NF-kB the exact mechanism is not understood.  Preliminary evidence indicates that p38 interacts with CREB binding protein (CBP), a transcriptional coactivator protein, to activate the p65 subuit of NF-kB.  This interaction is important for NF-kB stimulated transcription.  Results from in vitro binding assays show that p38 interacts with amino terminal region of CBP. Moreover, in vitro kinase assay using activated p38 show that p38 can phosphorylate CBP within the binding region.  Currently, we are generating a series of CBP carboxyl-terminal deletions to further map the p38 interacting regions and aid in identifying the phosphorylations sites.

8th URS Photos

 

Presenters
8th Presenters
Left to right: Dr. Emilio Duran, Dr. Lawrence Elmer (plenary speaker), Dia Smiley, Sean Ehinger, Shaddy Swade, Emily Davis, Nancy Parquet, Meredith Snapp, Margaret Uhrich, Allison Cihla, Lyndsey Emery, Courtney Maunz Derrer, Jeffrey Henke, Steven Driver, Paul Nolan, and Dr. Patricia Komuniecki.

 

Winners
8th Winners
First Place - Meredith Snapp, Second Place - Allison Cihla, Second Place - Nancy Parquet, Third Place - Steven Driver


last updated 29 Dec 2005
please contact brenda.leady@utoledo.edu with comments or concerns
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Last Updated: 3/22/15