Cell and Developmental Biology Commons^™

Characterization Of The Interaction Between The Fanconi Anemia (Fa) D2 Protein And Proliferating Cell Nuclear Antigen (Pcna), Carolyn Schmiedel

Senior Honors Projects

No abstract provided.

Investigating The Affects Of Cucurbitacin-I On Cellular Motility, Rebecca Lafleur

Honors Scholar Theses

Cellular migration is an integral component of many biological processes including immune function, wound healing and cancer cell metastasis. A complete model illustrating the mechanism by which cells accomplish movement is still lacking. Exploring the affects of various drugs on cell motility may be instrumental in discovering new proteins which mediate cell movement. This project aims ultimately to characterize the molecular target of the drug Cucurbitacin-I, a natural plant product. This drug has been shown to inhibit migration of epithelial sheets and may have anti-tumor activity.

In this paper, we show that Cucurbitacin-I inhibits the migration of MDCK and B16F1 …

Expression Analysis Of Plant Defense Responses During The Establishment Of Biotrophy And Role Of Abiotic Stress In The Infection Of Dyer’S Woad (Isatis Tinctoria) By Puccinia Thlaspeos, Elizabeth Thomas

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

The kinetics and amplitude of the salicylic acid-responsive pathogenesis-related (PR) genes and the cytochrome P450 gene ItCYP79B2 in the compatible interaction between Puccinia thlaspeos and dyer’s woad (Isatis tinctoria) during the first 72 hours of inoculation were examined. Immediately following penetration of the host by the rust pathogen, there was a modest up-regulation of PR genes but a significant down-regulation of ItCYP79B2 expression. During haustoria formation, a significant pathogen-mediated suppression of PR genes was observed with a corresponding up-regulation of ItCYP79B2. This potentially facilitates haustoria formation by P. thlaspeos. After haustoria formation, a more …

Cloning And Biochemical Characterization Of The Recq Helicase And Topoisomerase Iii In Schizosaccharomyces Pombe, Rebecca Wood

All Theses And Dissertations

RecQ and Topoisomerase III proteins are both essential for proper chromosomal maintenance in species ranging from bacteria to humans. Due to their highly conserved nature throughout evolution, understanding their basic biochemical properties as well as their interactions together is important for understanding human health, disease, and even to provide potential evolutionary insight. S. pombe has been chosen as a model organism because little research has been done on the biochemical analysis of its RecQ homolog, Rqhl. This study shows developments in the biochemical characterization of a truncation protein of Rqh1 including AFM (Atomic Force Microscopy) Imaging, DNA binding, and ATP …

Stochastic Calculations For Computation Of Radiation Effects And Cell Survivability Under Voltage Pulsing, Madhuri Ganapathiraju

Electrical & Computer Engineering Theses & Dissertations

Statistical computations are an important tool for the analysis of stochastic phenomena and processes that are characterized by variability. Biological systems (e.g., cells, tissues etc.) are perfect examples wherein response to a given external stimulus can be varied and needs to be adequately considered. The Monte Carlo method of analysis has now been recognized as the most effective way of treating stochastic variability.

This thesis uses Monte Carlo based simulations to probe two problems that require the quantification and modeling of effects caused by energy deposition onto biological matter from external sources. One problem involves the probabilistic study of the …

Treg Depletion Inhibits Efficacy Of Cancer Immunotherapy: Implications For Clinical Trials., James Curtin, Marianela Candolfi, Tamer Fakhouri, Chunyan Liu, Anderson Alden, Matthew Edwards, Pedro Lowenstein, Maria Castro

Articles

BACKGROUND: Regulatory T lymphocytes (Treg) infiltrate human glioblastoma (GBM); are involved in tumor progression and correlate with tumor grade. Transient elimination of Tregs using CD25 depleting antibodies (PC61) has been found to mediate GBM regression in preclinical models of brain tumors. Clinical trials that combine Treg depletion with tumor vaccination are underway to determine whether transient Treg depletion can enhance anti-tumor immune responses and improve long term survival in cancer patients. FINDINGS: Using a syngeneic intracrabial glioblastoma (GBM) mouse model we show that systemic depletion of Tregs 15 days after tumor implantation using PC61 resulted in a decrease in Tregs …

The Effects Of A Resveratrol Derivative On Regulatory Behaviors And Reproductive Health In Male And Female Long-Evans Rats, Kimberly Michelle Fabick

Theses and Dissertations

Phytoestrogens are chemicals produced by plants that act like estrogens and have the ability to bind to the mammalian estrogen receptor system. The purpose of this study is to evaluate a new phytoestrogen analog called 4-acetoxy Resveratrol. Resveratrol is a phytoestrogen that has been found in the skin of grapes. Resveratrol has been shown to be able to bind to the estrogen receptors and has a similar molecular structure as estradiol. Resveratrol has been shown to have many positive health benefits such as improving cardiovascular health, serving as a neuroprotective agent, acting as an anti-inflammatory agent, working as an anti-cancer …

Mechanisms By Which Apoptotic Membranes Become Susceptible To Secretory Phospholipase A2, Rachel Williams Bailey

Theses and Dissertations

During apoptosis, changes occur in T-lymphocyte membranes that render them susceptible to hydrolysis by secretory phospholipase A2 (sPLA2). To study the relevant mechanisms, a simplified model of apoptosis using a calcium ionophore was first applied. Kinetic and flow cytometry experiments provided key observations regarding ionophore treatment: initial hydrolysis rate was elevated, total reaction product was increased four-fold, and adsorption of the enzyme to the membrane surface was unaltered. Analysis of these results suggested that susceptibility during calcium-induced apoptosis is limited by substrate availability rather than enzyme adsorption. Fluorescence experiments identified three membrane alterations that might affect substrate access to the …

The Effects Of Prenatally Administered Phytoestrogens On The Reproductive And Behavioral Development Of Long-Evans Rats, Crystal Blake

Theses and Dissertations

Equol is known to be a selective androgen modulator and has the ability to bind and inhibit 5-alpha dihydrotestosterone (5α-DHT). Equol is also a selective estrogen receptor modulator and is able to bind beta estrogen receptors with high affinity. As estrogen receptors are found in the hypothalamus, pituitary, and gonads, prenatally administered equol could affect the morphological and reproductive development of offspring. To test this hypothesis, during gestational days 14 to 20, forty-two pregnant Long-Evans rats were given one of six treatments: 1) no treatment, 2) injection with dimethyl sulfoxide (DMSO), 3) injection with 10 mg/kg equol, 4) injection with …

Turning The Gene Tap Off; Implications Of Regulating Gene Expression For Cancer Therapeutics, James Curtin, Marianela Candolfi, Weidong Xiong, Pedro Lowenstein, Maria Castro

Articles

Cancer poses a tremendous therapeutic challenge worldwide, highlighting the critical need for developing novel therapeutics. A promising cancer treatment modality is gene therapy, which is a form of molecular medicine designed to introduce into target cells genetic material with therapeutic intent. Anticancer gene therapy strategies currently used in preclinical models, and in some cases in the clinic, include proapoptotic genes, oncolytic/replicative vectors, conditional cytotoxic approaches, inhibition of angiogenesis, inhibition of growth factor signaling, inactivation of oncogenes, inhibition of tumor invasion and stimulation of the immune system. The translation of these novel therapeutic modalities from the preclinical setting to the clinic …

Novel Role Of Antioxidant-1 (Atox1) As A Copper-Dependent Transcription Factor Involved In Cell Proliferation, S. Itoh, H. W. Kim, O. Nakagawa, K. Ozumi, Susan M. Lessner, H. Aoki, K. Akram, R. D. Mckinney, M. Ushio-Fukai, T. Fukai

Faculty Publications

Copper plays a fundamental role in regulating cell growth. Many types of human cancer tissues have higher copper levels than normal tissues. Copper can also induce gene expression. However, transcription factors that mediate copper-induced cell proliferation have not been identified in mammals. Here we show that antioxidant-1 (Atox1), previously appreciated as a copper chaperone, represents a novel copper-dependent transcription factor that mediates copper-induced cell proliferation. Stimulation of mouse embryonic fibroblasts (MEFs) with copper markedly increased cell proliferation, cyclin D1 expression, and entry into S phase, which were completely abolished in Atox1^-/- MEFs. Promoter analysis and EMSA revealed that copper …

Adenomatous Polyposis Coli Is Present Near The Minimal Level Required For Accurate Graded Responses To The Wingless Morphogen, Hassina Benchabane, Edward G. Hughes, Carter M. Takacs, Jason R. Baird, Yashi Ahmed

Dartmouth Scholarship

The mechanisms by which the Wingless (Wg) morphogen modulates the activity of the transcriptional activator Armadillo (Arm) to elicit precise, concentration-dependent cellular responses remain uncertain. Arm is targeted for proteolysis by the Axin/Adenomatous polyposis coli (Apc1 and Apc2)/Zeste-white 3 destruction complex, and Wg-dependent inactivation of destruction complex activity is crucial to trigger Arm signaling. In the prevailing model for Wg transduction, only Axin levels limit destruction complex activity, whereas Apc is present in vast excess. To test this model, we reduced Apc activity to different degrees, and analyzed the effects on three concentration-dependent responses to Arm signaling that specify distinct …

Prostasin Is Expressed In Benign Prostatic Hyperplasia And Regulates Cell Proliferation And Invasion Via Inos, Icam-1, And Cycli, Meghan Hatfield

Electronic Theses and Dissertations

Prostasin is expressed in normal prostate epithelial cells but down-regulated in prostate cancers, while prostasin re-expression in invasive prostate cancer cells reduced invasion. We examined prostasin expression and function in benign prostatic hyperplasia (BPH). We evaluated prostasin expression in 12 BPH specimens by immunohistochemistry, and evaluated the impact of prostasin silencing by siRNA on the expression of the inducible nitric oxide synthase (iNOS), intercellular adhesion molecule-1 (ICAM-1), and cyclin D1, as well as on cell proliferation and invasion, using the BPH-1 human prostate epithelial cell line model. Prostasin expression was localized in the glands of BPH tissues by immunohistochemistry, in …

Role Of Transient Receptor Potential Canonical-6 (Trpc6) Channel In Metastasis Of Glioblastoma Multiforme, Rajarajeshwari Venkataraman

Electronic Theses and Dissertations

Glioblastoma multiforme (GBM) is one of the extremely fatal brain tumors. The main reason that makes it so lethal is its capability to invade and spread to other parts of CNS producing secondary tumors. Among other factors hypoxia, reduced oxygen availability, is linked to higher metastatic potential of cancers. Hypoxia causes numerous changes in genome and proteome of the cell. These changes help a normal cell to adapt to nutritional deficiency, but the same changes can increase the malignancy and metastasis in tumor cells. Extensive research by a number of curious scientists reveal that various pathways involving numerous proteins cross-talk …

Comparing Digit Morphology Of An Arboreal Salamander With Potential Competitors, Eric Hugh Diefenbacher

Theses, Dissertations and Capstones

Green Salamanders (Aneides aeneus) are the only salamanders in West Virginia to exhibit an arboreal lifestyle. The focus of this study was to determine the cellular anatomy of the distal digit structures and how these structures may influence climbing ability. Comparative histology, comparative morphometrics, and comparative osteology of Green Salamanders, Cumberland Plateau Salamanders (Plethodon kentucki), and Slimy Salamanders (Plethodon glutinosus), were also used to determine if Cumberland Plateau Salamanders had the potential to compete with Green Salamanders for arboreal habitats. Histologically, Cumberland Plateau Salamanders had cell layer thickening similar to that of Green Salamanders. Morphometrically, Green Salamanders and Cumberland Plateau …

Molybdate Treatment And Sulfate Starvation Decrease Atp And Dna Levels In Ferroplasma Acidarmanus, Kai Hung

Faculty Research & Creative Activity

Sulfate is a primary source of sulfur for most microbes and in some prokaryotes it is used an electron acceptor. The acidophile Ferroplasma acidarmanus (strain fer1) requires a minimum of 150 mM of a sulfate-containing salt for growth. Sulfate is assimilated by F. acidarmanus into proteins and reduced to form the volatile organic sulfur compounds methanethiol and dimethyldisulfide. In the absence of sulfate, cell death occurs by an unknown mechanism. In this study, cell viability and genomic DNA and ATP contents of F. acidarmanus were monitored in response to the absence of sulfate or the presence of sulfate and the …

Molybdate Treatment And Sulfate Starvation Decrease Atp And Dna Levels In Ferroplasma Acidarmanus, Kai F. Hung

Faculty Research & Creative Activity

Sulfate is a primary source of sulfur for most microbes and in some prokaryotes it is used an electron acceptor. The acidophile Ferroplasma acidarmanus (strain fer1) requires a minimum of 150 mM of a sulfate-containing salt for growth. Sulfate is assimilated by F. acidarmanus into proteins and reduced to form the volatile organic sulfur compounds methanethiol and dimethyldisulfide. In the absence of sulfate, cell death occurs by an unknown mechanism. In this study, cell viability and genomic DNA and ATP contents of F. acidarmanus were monitored in response to the absence of sulfate or the presence of sulfate and the …

Purinergic Receptors In The Circumvallate Papillae Of Rats, Alana Marselle Montoya

Electronic Theses and Dissertations

P2X2 receptors and P2Y4 receptors are ATP-activated cell surface receptors that gate movement of K+, Na+, and Ca2+. We used immunocytochemistry for P2X2 and P2Y4 receptors as well as taste cell type markers to learn if P2X2 immunoreactivity (IR) is present in nerve processes in close apposition to Type II and/or Type III taste cells. We also tested to see if P2Y4 IR is present in Type II and/or Type III taste cells. Our results indicate that P2X2 is present only in intragemmal nerve processes. P2X2-LIR nerve processes form close contacts with Type II and Type III taste cells. P2Y4 …

Annotation Of The Bacteriophage 933w Genome: An In-Class Interactive Web-Based Exercise, Kai F. Hung

Faculty Research & Creative Activity

No abstract provided.

Different Requirements Of The Swi/Snf Complex For Robust Nucleosome Displacement At Promoters Of Heat Shock Factor And Msn2- And Msn4-Regulated Heat Shock Genes, Tamara Y. Erkina, P. A. Tschetter, Alexander M. Erkine

Scholarship and Professional Work – COPHS

The stress response in yeast cells is regulated by at least two classes of transcription activators—HSF and Msn2/4, which differentially affect promoter chromatin remodeling. We demonstrate that the deletion of SNF2, an ATPase activity-containing subunit of the chromatin remodeling SWI/SNF complex, eliminates histone displacement, RNA polymerase II recruitment, and heat shock factor (HSF) binding at the HSP12 promoter while delaying these processes at the HSP82 and SSA4 promoters. Out of the three promoters, the double deletion of MSN2 and MSN4 eliminates both chromatin remodeling and HSF binding only at the HSP12 promoter, suggesting that Msn2/4 activators are primary determinants of …

Cholinergic Neurons Regulate And Utilize Gdnf Secreted By C2c12 Skeletal Muscle Cells In Culture, John-Mary Vianney, John Spitsbergen

Biological Sciences Faculty and Graduate Student Research

Glialcell line-derived neurotrophicfactor (GDNF) has been regarded as a potent survival factor for a subpopulation of neurons. It has been shown that GDNF expression is upregulated in skeletal muscle of patients with early stage of neuromuscular diseases such as amyotrophic lateral sclerosis (ALS). Previous results from our laboratory showed that neural cells regulate GDNF secretion by skeletal muscle; non-innervated skeletal muscle appear to secrete more GDNF compared to innervated skeletal muscle. Two aims were examined in the current study. First, to examine whether neural cells inhibit GDNF through acetylcholine release. Second was to examine whether differentiated NG108-15 neural cells secrete …

Bmp Signaling Goes Posttranscriptional In A Microrna Sort Of Way, Catherine A. Reinke, Richard W. Carthew

Faculty Publications

Aberrant microRNA (miRNA) expression correlates with human diseases such as cardiac disorders and cancer. Treatment of such disorders using miRNA-targeted therapeutics requires a thorough understanding of miRNA regulation in vivo. A recent paper in Nature by Davis et al. expands our understanding of miRNA biogenesis and maturation, elucidating a mechanism by which extracellular signaling directs cell differentiation via posttranscriptional regulation of miRNA expression.

A Versatile Assay For The Identification Of Rna Silencing Suppressors Based On Complementation Of Viral Movement, Jason G. Powers, Tim L. Sit, Feng Qu, T. Jack Morris, Kook-Hyung Kim, Steven A. Lommel

Nebraska Center for Virology: Faculty Publications

The cell-to-cell movement of Turnip crinkle virus (TCV) in Nicotiana benthamiana requires the presence of its coat protein (CP), a known suppressor of RNA silencing. RNA transcripts of a TCV construct containing a reporter gene (green fluorescent protein) (TCV-sGFP) in place of the CP open reading frame generated foci of three to five cells. TCV CP delivered in trans by Agrobacterium tumefaciens infiltration potentiated movement of TCV-sGFP and increased foci diameter, on average, by a factor of four. Deletion of the TCV movement proteins in TCV-sGFP (construct TCVΔ92-sGFP) abolished the movement complementation ability of TCV CP. Other known suppressors of …

Interactions Between Changing Pco2, N2 Fixation, And Fe Limitation In The Marine Unicellular Cyanobacterium Crocosphaera, Fei-Xue Fu, Margaret R. Mulholland, Nathan S. Garcia, Aaron Beck, Mark E. Warner, Sergio A. Sañudo, David A. Hutchins

OES Faculty Publications

We examined the physiological responses of steady-state iron (Fe)-replete and Fe-limited cultures of the biogeochemically critical marine unicellular diazotrophic cyanobacterium Crocosphaera at glacial (19 Pa; 190 ppm), current (39 Pa; 380 ppm), and projected year 2100 (76 Pa; 750 ppm) CO₂ levels. Rates of N₂ and CO₂ fixation and growth increased in step with increasing partial pressure of CO₂ (pCO₂), but only under Fe- replete conditions. N₂) and carbon fixation rates at 75 Pa CO₂ were 1.4-1.8-fold and 1.2-2.0-fold higher, respectively, relative to those at present day and glacial pCO_{2 …}

Adenylyl Cyclase Type 6 Overexpression Selectively Enhances Ss-Adrenergic And Prostacyclin Receptor Mediated Inhibition Of Rat Cardiac Fibroblast Function Due To Co-Localization In Lipid Rafts, Xiaoqiu Liu, Muthusamy Thangavel, Shu Qiang Sun, Joseph Kaminsky, Penden Mahautmr, Jeremiah Stitham, John Hwa, Rennolds S. Ostrom

Pharmacy Faculty Articles and Research

Cardiac fibroblasts produce and degrade extracellular matrix and are critical in regulating cardiac remodeling and hypertrophy. Fibroblasts are activated by factors such as transforming growth factor β and inhibited by agents that elevate 3′,5′-cyclic adenosine monophosphate (cAMP) levels. cAMP signal generation and response is known to be compartmentalized in many cell types in part through the colocalization of receptors and specific adenylyl cyclase isoforms in lipid rafts and caveolae. The present study sought to define the localization of key G protein-coupled receptors with adenylyl cyclase type 6 (AC6) in lipid rafts of rat cardiac fibroblasts and to determine if this …

Genotoxic Stress-Induced Expression Of P53 And Restoration Of Apoptosis In Leukemic Clam Hemocytes With Cytoplasmically Sequestered P53, Stefanie Boettger, Emily Jerszyk, Ben Low, Charles Walker

Biology Faculty Publications

No abstract provided.

Identification Of Lactic Acid Bacteria In Michigan Cherry Wines, Emily Henk, Margaret Dietrich, Terri Weese

Student Summer Scholars Manuscripts

Many factors contribute to the final flavor of wine. One factor is malolactic fermentation, during which lactic acid bacteria (LAB) transform the harsh tasting malic acid into a more drinkable lactic acid in grape wine. The role of LAB in the production of cherry wine is completely unknown. The goal of this study is to identify the species of LAB in cherry wine and compare them to those found in grape wines. Bacteria from cherry wine were grown on general media plates and plates fortified with malic acid, which may provide optimal growing conditions for the LAB. To identify the …

Towards Cloning The Clk-3 Gene In Caenorhabditis Elegans, Suchita Umesh Desai

University of Kentucky Master's Theses

Mutations in the clk-1, clk-2, clk-3 and gro-1 genes in Caenorhabditis elegans show alterations in developmental and behavioral timing and lifespan, collectively termed the Clk phenotype. While the clk-1, clk-2, and gro-1 genes have been cloned, clk-3 gene has not been identified. Gene expression changes in clk-3 mutant worms were determined using microarray expression data. I examined genes in the region to which clk-3 gene maps, for strongly reduced expression in the clk-3 mutants and identified thirteen clk-3 candidate genes. RNAi feeding vectors for all these candidate genes were picked and cultured from the RNAi library. …

The Effects Of Oxidative Stress And Age On Human Spermatozoa, Taryn Mcandrew

Theses : Honours

Male infertility is now recognised as a significant factor in couples having difficulty conceiving. The impact of maternal age has long been known as a limiting factor, however recent research indicates that advancing paternal age can also negatively impact on a couple's chances of conception. One of the major contributing causes of male infertility has now been linked to spermatozoa exposure to reactive oxygen species (ROS). Such exposure induces oxidative stress when coupled with reduced total antioxidant capacity (TAC). Measures of both ROS and TAC are used as tests of oxidative stress status (OSS) which are used together to give …

Aging Predisposes Oocytes To Meiotic Nondisjunction When The Cohesin Subunit Smc1 Is Reduced, Vijayalakshmi V. Subramanian, Sharon E. Bickel

Dartmouth Scholarship

In humans, meiotic chromosome segregation errors increase dramatically as women age, but the molecular defects responsible are largely unknown. Cohesion along the arms of meiotic sister chromatids provides an evolutionarily conserved mechanism to keep recombinant chromosomes associated until anaphase I. One attractive hypothesis to explain age- dependent nondisjunction (NDJ) is that loss of cohesion over time causes recombinant homologues to dissociate prematurely and segregate randomly during the first meiotic division. Using Drosophila as a model system, we have tested this hypothesis and observe a significant increase in meiosis I NDJ in experimentally aged Drosophila oocytes when the cohesin protein SMC1 …