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7th Annual

WOLFE HALL (Room 3246) Saturday, April 13, 2002

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Chart Abstracts 7th URS 7th URS Photos  UT Biological Sciences


(click on the presenter name for abstract)

Time

Speaker

Presentation Title

9:30

Dr. Patricia Komuniecki

Welcome

9:35

Plenary Lecture- "Diabetes: Past, present and future" Dr. Sonja Najjar, Medical College of Ohio

Session I - Elizabeth Rex, Moderator

10:30

Kevin E. Aston

"Genetic screening to identify regulators of IF function in Caenorhabditis elegans"  Advisor: Dr. John D. Plenefisch, Department of Biological Sciences, The University of Toledo

10:50

Rebecca M. Hassel

"The genetic engineering of soybean"  Advisor: Dr. S. Goldman and Dr. R.V. Sairam, Department of Earth, Ecological and Environmental Sciences, The University of Toledo

11:10

Kara M. Schafer

"Production of indigazine for fullernophile synthesis" Advisor: Dr. Gary Miracle, Department of Chemistry, The University of Toledo

11:30

Andrew J. Hosken

"Effect of environmental stress on soil nematode life history characteristics" Advisor: Dr. Deborah A. Neher, Department of Earth, Ecological and Environmental Sciences, The University of Toledo

12:00

Pizza lunch

Session II - Katy Smith, Moderator

12:30

Abraham K. Lee

"The localization and expression of the sarcoglycan complex in Caenorhabditis elegans"  Advisor: Dr. John D. Plenefisch, Department of Biological Sciences, The University of Toledo

12:50

Florencia Pauli

"Analysis of 5' untranslated region (UTR) of the Hsp70-2 heat shock transcript of Plasmodium falciparum" Advisors(s): Dr. Dyann Wirth and Dr. Kevin Militello, Department of Immunology and Infectious Diseases, Harvard School of Public Health

1:10

Megan N. Henderson

"Characterization of Caenorhabditis elegans octopamine/ tyramine receptors" Advisors(s): Elizabeth Rex and Dr. Patricia Komuniecki, Department of Biological Sciences, The University of Toledo

1:30

Ann Steck

"Intracellular magnesium and Parkinson's disease" Advisor: Dr. Lawrence Elmer, Department of Neurology, Medical College of Ohio

1:50

Jeniffer Liette

"Analysis of the Acd-1 (accelerated cell death 1) gene promoter in Arabadopsis" Advisor(s): Yang Manli and Dr. John Gray, Department of Biological Sciences, The University of Toledo

2:00

Awards ceremony


Abstract 7th URS


Kevin E. Aston
“Genetic Screening to Identify Regulators of IF Function in Caenorhabditis elegans” Advisor:  Dr. John D. Plenefisch-Department of Biological Sciences, The University of Toledo

Tissue fragility diseases like epidermolysis bullosa are a direct result of the malformation of physical linkages between cells and extracellular matrices known as intermediate filaments (IFs).  Mua-6 is a gene in the nematode C. elegans which codes for IFA2, an IF required for epidermal integrity.  One allele of mua-6, known as rh85, produces an animal which invariably suffers a type of muscular dystrophy.  It is hypothesized that second site mutations that are able to suppress the rh85 phenotype will be able to be observed by performing a genetic screen.  The mutated genes identified are likely to interact directly with mua-6 in its function of cell adhesion or involved in its transcription or translation.  The rh85 strain will be exposed to the random mutation-inducing chemical ethylmethanesulfonate and subsequently screened for suppression of the rh85 phenotype.   The suppressor characteristics will be examined by outcrossing, genetic mapping, and genetic complementation.  Since C. elegans shares 70% of its genes in common with mankind and the C. elegans muscular dystrophy is similar in structure to human tissue fragility diseases, discovering which genes affect muscular dystrophy in C. elegans could lead to an understanding of the occurrence of epidermolysis bullosa in humans. 

Rebecca M. Hassel
“The Genetic Engineering of Soybean”
Advisor:  Dr. S. Goldman and Dr. R.V. Sairam-Department of Earth, Ecological and Environmental Sciences, The University of Toledo

Soybean [Glycine max (L.) Merr.] is one of the most important economic crops in the United States with nearly 70 million acres planted per year. In the present study, we report a simple, high frequency callus, somatic embryogenesis, and genotype independent plant regeneration protocol for soybean. Five different medias were tested for callus induction and plant regeneration. Genotypes Loda and Newton performed best on modified MS medium with NAA (10 mg/l) and 2,4-D (5 mg/l) for callus induction. The modified MS medium with 2 mg/l BA was optimal for plant regeneration in both the genotypes. The callus induction and plant regeneration frequencies in Loda were 100% and 17% respectively, and in Newton they were 100% and 50%. Among the 4 different carbohydrate sources tested, sorbitol was found to be most favorable for both callus induction and plant regeneration. Finally, the rate of T-DNA transfer to soybean cotyledonary nodes was high and was independent of whether super-virulent strains of Agrobacterium were used or not.

Megan N. Henderson
“Characterization of Caenorhabditis elegans  Octopamine/Tyramine Receptors”
Advisor(s): Elizabeth Rex and Dr. Patricia R. Komuniecki-Department of Biological Sciences, The University of Toledo

Octopamine, OA, plays an important role in processes essential for nematode survival, including pharyngeal pumping, locomotion and egg-laying.  Our research has identified a putative Caenorhabditis elegans tyramine (TA)/OA receptor (CO2D4.2) in the C. elegans database.  To date, a cDNA encoding this receptor, OARCe has been cloned, functionally expressed, and pharmacologically characterized.  The OARCe sequence differs from that predicted by Genefinder, in that it lacks exon 10 and 42 basepairs from exon 2.  To identify potential OARCe splice variants, primers were designed to amplify regions encoding exons 2 and 10 from C. elegans cDNA pools.  Interestingly, no fragments were generated that corresponded to either of these regions; however, a short splice variant of OARCe, OARCes, was isolated that lacked 69 nucleotides of exon 7, which corresponded to a 23 amino acid deletion within the third intracellular loop.  The pharmacology and potential coupling of OARCes are currently being characterized and compared to that of OARCe.

Andrew J. Hosken
“Effect of Environmental Stress on Soil Nematode Life History Characteristics”
Advisor:  Dr. Deborah A. Neher-Department of Earth, Ecological and Environmental Sciences, The University of Toledo

Nematodes are found in ecosystems ranging from tropical rainforest to arctic tundra.  Indices of nematode community structure and composition such as species richness, trophic structure, and successional status are useful in monitoring soil health.  Index values decrease in soils contaminated with heavy metals.  The goal was to confirm life history traits of Rhabditidae, a group of nematodes generally believed to tolerate heavy metal contamination.  This family is assigned a rank of 1 in Bonger’s maturity index, on a scale of 1 to 5, with 1 being most tolerant and 5 most sensitive to environmental stress.  Acrobeloides and Rhabditis, two genera of Rhabditidae, were extracted from two soil sites, cultivated in petri dishes containing Nematode Growth Medium, and fed Escherichia coli OP50. The nematodes were then subjected to different metals that were mixed into the medium.  Their survivorship and reproduction were measured.  Both genera are closely related but respond very differently to stress, and this sensitivity increased with duration of time in culture.    Those taken from contaminated soil lost their tolerance to the same soil contaminants; those taken from uncontaminated soil also exhibited lower tolerances to stress.  This suggests that current techniques used for ecotoxicological testing of nematodes should consider the possibility of rapid loss of tolerance in culture.

 

Abraham K. Lee
“The Localization and Expression of the Sarcoglycan Complex in Caenorhabditis elegans”
Advisor: Dr. John D. Plenefisch-Department of Biological Sciences, The University of Toledo

Muscular dystrophy is a collection of inherited disorders characterized by progressive muscle wasting and weakness.  Today there are over twenty different types of muscular dystrophy that have been identified in humans.  Scientists have discovered that a mutation or deficiency in any of the proteins of the dystrophin-glycoprotein complex (DGC), which is located in the cell membrane and bridges the intracellular cytoskeleton to the extracellular matrix, leads to signs of muscular dystrophy or even death.  Within the DGC complex is another set of proteins that make up the sarcoglycan complex (SGC).  The disruption of the SGC leads to the phenotype characterized as limb-girdle muscular dystrophy or LGMD.  In order to aid studies in finding a cure for muscular dystrophy, researchers require the use of model organisms to conduct tests and future studies.  One such versatile organism is the invertebrate Caenorhabditis elegans.  The use of C. elegans can be a practical tool in understanding muscular dystrophy due to its ease of propagation, manipulation, and possession of homologous genes.  Specifically, C. elegans has three genes that are homologous to the alpha-, beta-, and gamma-sarcoglycan genes present in humans.  In the current study, we look to identify whether and in which C. elegans tissues the genes are transcribed.  The expression of the sarcoglycan genes will be confirmed through the use of a green-fluorescence protein (GFP) reporter gene.

Jennifer Liette
“Analysis of the of Acd-1 (accelerated cell death 1) Gene Promoter in Arabadopsis”
Advisor(s): Yang Manli and Dr. John Gray-Department of Biological Sciences, The University of Toledo

Certain genes in plants have been suspected of inhibiting cell death to spread through a plant in response to physical wounding.  Two such genes are the lls1 (lethal leaf spot) gene in maize and the acd-1 (accelerated cell death1 ) gene in Arabadopsis thaliana which are orthologs of one another.  Their orthology was predicted due to the high rate of amino acid similarity as well as the common intron position and protein size.  Genetic analysis of the mutants has confirmed this prediction.  The acd-1 gene was first sequenced from a cDNA clone and then we used PCR to amplify the upstream promoter sequence.  Using a recombinant DNA approach, the gene promoter was then fused to a GUS reporter gene.  Activation of this reporter construct was followed following transformation into plants using Agrobacterium tumefaciens.  The analysis of the reporter gene expression in leaves is a means of identifying where and when the cell death inhibition function of ACD1 is normally present.  We generated transgenic plants containing this promoter fusion but found that it was insufficient to drive reporter gene expression in vivo.  This result indicates that other regions upstream, downstream, or within the acd1 gene are required for normal gene expression.

Florencia Pauli
“Analysis of 5’ Untranslated Region (UTR) of the Hsp70-2 Heat Shock Transcript of Plasmodium falciparum”
Advisor(s):  Dr. Dyann Wirth and Dr. Kevin Militello-Department of Immunology and Infectious Diseases, Harvard School of Public Health

Malaria, caused by Plasmodium falciparum, is the world’s most important parasitic disease.  Many genes in P. falciparum are regulated at the level of transcription, but little is known about the organism’s promoter elements or transcriptional apparatus. It is hypothesized that P. falciparum has a novel mechanism, unlike that of bacteria or other eukaryotes, that could be used as a drug target.  In this study, the 5’ upstream region of the hsp70-2 gene was analyzed by reverse transcriptase PCR mapping to locate an approximate start site for transcription.  Five primer pairs were designed with the forward primers in the 5’ upstream region and the reverse primer in the first exon.  PCR was completed with the five primer pairs on gDNA and cDNA.  All five pairs amplified products of expected size on the gDNA, but only the first three pairs amplified products from cDNA.  This strongly suggests that the transcript begins somewhere between the third and fourth upstream primer which is 662-809 nucleotides upstream of the ATG start codon.  The identities of the PCR products for primer pairs 1-3 were confirmed by a restriction enzyme digest analysis.  The results of this study could eventually be used to identify the organism’s transcriptional apparatus.

 Kara M. Schafer
“Production of Indigazine for Fullerenophile Synthesis”
Advisor:  Dr. Gary Miracle-Department of Chemistry, The University of Toledo

Carbon nanotubes are cylindrical graphene sheets of the nanoscopic scale. Typically isolated in long fibrils, these novel molecules are theorized to have many useful industrial, electrical, and biological applications. However, a practical way to isolate nanotubes has not yet been developed, which greatly hinders the research on nanotubes. A new class of molecules, termed fullerenophiles, has been designed to dissolve carbon nanotubes in perfluorinated solvents. The fullerenophiles are predicted to experience an attractive non-covalent bonding force of over 5 kcal/mol between their rims of electron-deficient N-H and C-H bonds and the relatively electron-rich nanotube surface. This attraction is sufficient to disrupt intermolecular forces between the carbon nanotubes, and to draw them into solution. Synthesis of the fullerenophiles will involve conversion of indigo into an indigazine backbone, and preliminary attempts at this conversion are discussed. These efforts include synthesis of oxalyl indigo, and the attempted synthesis of three disilyl indigo derivatives. The subsequent conversion of oxalyl indigo into oxalyl indigazine was not successful, and the synthesis of one disilyl indigo derivative was not successful. Syntheses of the disilane precursors to the disilyl indigo derivatives are also discussed.

Ann Steck
"Intracellular Magnesium and Parkinson's Disease”
Advisor:  Dr. Lawrence Elmer-Department of Neurology, Medical College of Ohio

Parkinson’s Disease (PD) is a progressive neurodegenerative disorder of unknown etiology.  Numerous potential causes of PD have been proposed, including glutamate excitotoxicity.  Glutamate, when bound to NMDA receptors, facilitates the influx of calcium ions into neurons.  Magnesium is a divalent cation which regulates this calcium influx via specific blockade of the NMDA receptor.  Therefore, sub-cellular deficiencies in magnesium could result in excessive calcium influx, leading to accelerated apoptosis.  Postmortem examinations of PD patients have confirmed lowered magnesium concentrations in multiple regions of the brain.  A prior study of tissue magnesium in buccal cells demonstrated elevated levels of magnesium in PD patients when compared to age-matched controls.  This study examines a separate tissue survey in PD patients and controls.  Participants were recruited at the Medical College of Ohio (MCO) PD clinic, the MCO phlebotomy lab, and support groups throughout Northwest Ohio.  Blood samples were collected from 55 participants, including 20 PD patients, 25 age-matched controls.  The concentrations of intracellular magnesium were obtained using atomic absorption spectrophotometry and normalized to total cellular protein using parallel protein analysis.  Lymphocytes and platelets from age-matched controls showed 81.1 and 83.6 mm/mg protein.  In contrast, lymphocytes from PD patients showed significantly higher amounts of intracellular magnesium, 98.6 mm/mg protein, while platelet concentrations were markedly diminished at 73.5 mm/mg protein compared with age-matched controls.  These data support the hypothesis that intracellular magnesium homeostasis is disturbed in patients with PD.  Future studies will determine the sub-cellular compartment in which magnesium is accumulating in the nucleated cells of PD patients.

7th URS Photos

7th Participants
7th Winners


last updated 29 Dec 2005
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