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Through the FUTURE in Biomedicine℠ program, the University of Iowa Carver College of Medicine is committed to:
Consistent with these commitments, the goal of the Better Futures for Iowans program was to make state-of-the-art core reseach facilities at the University of Iowa available to academic classes and research projects conducted primarily by undergraduates at 2-year and 4-year institutions in Iowa.
Faculty throughout the state were eligible to apply for small grants to support laboratory-intensive projects involving undergraduate students in "hands-on" inquiry. Students took responsibility for the preparation of samples and analysis of data obtained. All participants were invited to present a poster about their work at the FUTURE in Biomedicine℠ Research Symposium. This activity, which was supported primarily by the Office of the Provost, extended University resources to Iowans and addressed an important goal of the University Strategic Plan - to provide better futures for Iowans.
"We have engineered the expression of the four major protein components of a contractile fabric associated with the cortical membrane systems of the ciliated protozoan Tetrahymena (Tcb2, Epc1, Epk1, and Mrn1). We have GFPtagged versions of each of the proteins to study their distribution in living cells. We would use this facility grant to do the following: (1) We have used fluorescence microscopy to look at the distribution of three of these proteins using the confocal laser scanning microscope in the Central Microscopy Research Facility, but we have not yet imaged the 3D distributions of the Mrn1 protein and two closely related homologs (Mrn2 and Mrn3). All three of these MORN-domain proteins are hypothesized to function as adaptors that link the fibrous portion of the ciliate membrane skeleton to cortical membrane systems (the plasma membrane and closely apposed alveolar sacs). We propose to use one 3-4 hour session at the Central Microscopy Research Facility to complete these studies. (2) In addition, we had carried out electron microscopic studies of negative stained filaments of the Tcb2 protein this summer. We propose to use the electron microscope in the Central Microscopy Research Facility to image at high resolution the assembled structures formed by Tcb2 and Epc1 proteins. We suspect that two of these proteins (Epc1 and Tcb2) constitute distinct filamentous systems that bind to each other to form a contractile fabric, and that this interaction may be regulated by calcium ions. We propose to use one 4-hour session at the Central Microscopy facility to image negatively stained protein filaments formed by the Tcb2 and Epc1 proteins, both separately, and in combination. If time permits, we would also look at the effect of Epk1-mediated phosphorylation of the assemblies formed by these proteins. We have purified the kinase domain of Epk1, a Ser/Thr-specific protein kinase that co-purifies with these proteins when they are extracted from Tetrahymena cells. Phophoproteomics studies indicate that Epc1 and Tcb2 proteins are each phosphorylated at several sites by a Ser/Thr-specific protein kinase."
Maria Bohorquez, PhD and Jerry Honts, PhDBFFI Awardees
"In summer 2013, our lab began a transgenerational study to investigate the effects of naturally administered endocrine disrupting chemicals (EDCs) on growth and reproduction in multiple generations of zebra finch offspring. To do so, we administered drinking water to paired zebra finches via either glass bottle (negative control), glass bottle supplemented with water-soluble estrogen (E2+ positive control), plastic bottle suspected of containing BPA (baby bottle, BPA+), or a plastic bottle claiming to be BPA-free (FIJI water bottle). Bottles were hand-washed daily with regular dish washing liquid and hot water. Nearly all food- and beverage-safe plastics tested have been found to leach EDCs into the contents, so we wish to determine the extent to which that has occurred in our study. While we have collected and continue to collect measurements on the offspring, we also collected water samples from each of these delivery vessels twice per week. We wish to use the High Resolution Mass Spectrometry Core facility to determine if our bottles leached EDCs into the drinking water and, if so, the identity and quantity of each of the compounds that leached into the water. Drake students have already been extensively involved in this project. Matt Wright has been working with chemistry and pharmacy departments at Drake and have determined that equipment available on campus is not sensitive enough for our purposes. Matt and I will work together to communicate with core facility staff and travel to the University of Iowa to perform water analysis."
Debora Christensen, PhDBFFI Awardee
"The cytoskeleton of the ciliated protozoan Tetrahymena thermophila is remarkably complex, but little is known about the molecular and structural bases of its organization and regulation. Several proteins have been recently identified during proteomic analysis of alciumdependent contractile fibers isolated from this cytoskeleton. These include a putative calcium-binding protein (Tcb2), a filament-forming structural protein (Epc1), and a protein kinase (Epk1), among others. The function of these proteins is unclear, but it has been proposed that they play regulatory and structural roles in the calcium-dependent regulation of ciliary movement. The goal of this project is to investigate the structure of the calcium sensor Tcb2. The full-length protein forms filamentous structures and becomes insoluble in the presence of calcium and/or upon concentration, but its C-terminal domain is highly soluble and amenable to structural characterization. We propose to use NMR spectroscopy to probe the calcium-binding properties of this domain, and to investigate its structure and dynamics. We will express isotopically labeled protein in bacterial cells, and purify it using standard chromatography methods. NMR titrations will enable us to quantify its interaction with calcium. We will also acquire three-dimensional NMR experiments in order to achieve the sequential assignment of all backbone and sidechain resonances of the labeled polypeptide chain. Structure determination will be performed with the program CSRosetta or by using additional NMR distance constraints. This research will enable us to explore the structure-function relationship in Tcb2, and provide insights into the molecular basis of calcium sensing during ciliary movement in Tetrahymena."
Adina Kilpatrick, PhDBFFI Awardee
"The phase behavior of amphiphilic lipids and surfactants at various relative humidity levels and water content is of great interest for many technical and pharmaceutical applications. Lipid matrices containing mixtures of glyceryl monooleate (GMO) and glyceryl monostearate (GMS) have been evaluated for their potential application as magnetically induced thermo-responsive local drug delivery systems. However, the presence of excess moisture or hydrophilic additives such as ethanol is reported to influence the phase behavior of monoglycerides. Aims: The aims of this study are to investigate the thermal properties of GMO and GMS, determine the phase behavior of various monoglycerides blends, and investigate the effect of moisture on the crystallization of GMO:GMS matrices at 25, 37, and 42 degrees Celsius. Various compositions of GMO:GMS matrices will be prepared by a fusion method and characterized using differential scanning calorimetry (DSC) and scanning electron microcopy (SEM). We will determine the phase behavior of the monoglyceride blends and study the crystallization behaviors such as melting point, glass transition and heat of energy. Those properties are directly influenced by polymorphism and influenced by several external factors such as temperature, rate of crystallization and impurities. The DSC and SEM results will provide the best composition of GMO:GMS matrix for the monoglycerides-based thermal responsive drug delivery systems and evaluate the effect of moisture on the crystalline arrangement of the system."
Abebe Mengesha, PhDBFFI Awardee
"Historically, Iowa had more than 12.5 million hectares of prairie. Currently less than a tenth of 1% remains—the majority of that land has been converted to row crops. With the advent of Round-up ready technology, Glyphosate, the active ingredient in the product, is now being sprayed over a large majority of the land area in Iowa. The commonly accepted mechanism of action of glyphosate is the interruption of the Shikimate pathway through competitive inhibition of 5-enolpyruvylshikimate-3-phosphate (EPSP) synthase. This pathway is lacking in mammals but used by plants in the synthesis of aromatic amino acids. This seems to make it an ideal herbicide. However, bacteria and fungi also utilize the shikimate pathway. Field assessments have been conducted investigating the effect of glyphosate application on soil microbial community composition and overall microbial activity but the ability of specific soil bacteria to develop resistance to glyphosate has not been assessed. As a part of the advanced Cell and Molecular biology class at Dordt College, we would like to screen soil microbes from local fields for resistance to glyphosate, and sequence the EPSP synthase gene to look for patterns of variation in resistant strains. Organisms known to be inherently resistant will be excluded."
Jeffrey Ploegstra, PhDBFFI Awardee
"Funds are requested to support an undergraduate project to clone a putative catalase from the bacterium Deinococcus maricopensis. D. maricopensis has potential as a bioremediation agent due to its natural resistance to environmental stressors including oxidative
damage, dessication and ionizing radiation. The genome of the bacterium has recently been sequenced as part of the GEBA (Genomic Enclyclopedia of Bacteria and Archaea) project headed by the Joint Genome Institute of the Department of Energy. A goal of GEBA
is to provide undergraduates an opportunity to contriubte to genomics research, through the MGAN (Microbial Genome Annoation Network, supported by the National Science Foundation) network. Grand View is part of the MGAN network, and as part of the this
group, has access to bioinformatiics tools geared at genome annotations. I am currently working with undergraduate mentees to more closely examine the genomic content of D. maricopensis. In particular, we are collecting and annotating genes believed to be related to
stress tolerance phenotypes observed for D. maricopensis. The funding requested here will help support these efforts, by providing a means to validate our in silico findings through “wet-lab” work."
Michael LaGier, PhDBFFI Awardee
"I am working with a student, Allie Byrne, on a research project to determine whether perisynaptic Schwann cells
(PSCs), the glial cells at the neuromuscular junction (NMJ), modulate neurotransmitter release. Although there is growing consensus that glial cells play an essential role in synaptic plasticity in the CNS, it is not yet clear whether PSCs play an analogous role at the NMJ. Previous work has shown that synaptic activity increases calcium levels in the PSCs in a frequency dependent manner. This calcium increase depends on the binding of acetylcholine (ACh) or ATP to receptors on the PSCs, suggesting that a feedback mechanism exists by which ACh and/or ATP released from motor terminals activates PSCs. The activated PSCs may then signal back to the nerve terminal, reducing the subsequent release of neurotransmitter. To test this idea Allie and I are measuring high frequency synaptic depression in muscles in which the PSCs have been immunologically ablated. In parallel with these experiments, we are attempting to confirm that the ablation has abolished calcium signals in the PSCs. Surprisingly, we still observe “PSC-like” calcium signals in muscles with ablated PSCs. Since the calcium indicator we are using, Fluo-4 AM, loads into both the PSCs and the nerve terminal, these anomalous calcium signals may be originating in the nerve terminals. Unfortunately, our spinning disk confocal microscope lacks sufficient sensitivity and resolution to distinguish fluorescence originating from PSCs vs. nerve terminals. We propose using a confocal microscope at the University of Iowa to resolve Fluo-4 signals originating from these two components and determine the extent to which calcium signals have been lost or altered at NMJs with ablated PSCs."
Clark Lindgren, PhDBFFI Awardee
"The purpose of the project is to optimize preparation conditions for live samples for transmission electron microscopy. Standard procedures have been established to prepare tissue culture samples for transmission electron microscopy sample preparation. However, standardization of procedures for live samples is more difficult because live samples are not as concentrated and require different chemicals for processing. We are focused on optimizing preparation conditions for feather follicles infected with Marek’s disease virus (MDV), which is an excellent pathogen-host model. MDV is an excellent host because the early and late stages of infection of chickens with MDV largely mimic those of varicella-zoster virus, which commonly causes chicken pox in children and adults. It is essential to prepare these live samples for TEM because it will help us confirm and identify that a live host can be infected with multiple herpesviruses. The infection of a host with multiple herpesviruses, known as superinfection is rare due to the phenomenon known as superinfection inhibition. However, we hypothesize the superinfection is the cause of the increased virulence to MDV in chickens. Thus, confirmation of the dual infection of cells can help us better understand pathogenic principles of human herpesvirus disease."
Joseph Nguyen, PhDBFFI Awardee
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