Cell and Developmental Biology Commons^™

Increases In Cytosolic Calcium Ion Levels In Human Natural Killer Cells In Response To Butyltin Exposure, Rhonda Lane, Sabah O. Ghazi, Margaret M. Whalen

Chemistry Faculty Research

This study investigated whether exposures to butyltins (BTs), tributylin (TBT), and dibutyltin (DBT) were able to alter cytosolic calcium levels in human natural killer (NK) cells. Additionally, the effects of cytosolic calcium ion increases on the activation state of mitogen-activated protein kinases (MAPKs) in NK cells were also investigated. NK cells are an intital immune defense against the development of tumors or viral infections. TBT and DBT are widespread environmental contaminants, due to their various industrial applications. Both TBT and DBT have been shown to decrease the ability of NK cells to lyse tumor cells (lytic function). TBT has also …

Modeling Brain Injury: Expression Of Glial Fibrillary Acidic Protein In Stretch-Injured Astrocytes, Lindsey Cooke

Undergraduate Theses and Capstone Projects

Traumatic brain injury (TBI) is the leading cause of death and disability in individuals under 45. Individuals who survive TBI may be disabled for the rest of their lives and suffer from cognitive, physical, social and financial problems. Following TBI it has been shown that astrocytic gene expression of many proteins, including glial fibrillary acidic protein (GFAP), increases. These “reactive” astrocytes are thought to have both beneficial and detrimental effects on neuronal survival and function. The current study uses an in vitro model of injury which grows astrocytes on deformable silastic membranes to allow for dynamic stretch of cultured cells …

Study Of The Dna Damage Complexes Within The Htlv-1 Tax Oncoprotein Interactome, Sidi Mehdi Belgnaoui

Theses and Dissertations in Biomedical Sciences

Human T-cell leukemia virus type 1 (HTLV-1) is a transforming retrovirus that can give rise to adult T-cell leukemia (ATL) and HTLV-1 associated myelopathy/tropical spastic paraparesis (HAM/TSP). Tax is a virally encoded oncoprotein that is involved in HTLV-1 mediated cellular transformation. It has been hypothesized that Tax induces genomic instability via repression of the cellular DNA damage repair response. Our laboratory has previously shown that the interaction between Tax and various proteins involved in the DNA-damage response pathway impairs the ability of these proteins to mount an efficient repair response. As part of these observations, we proposed that Tax induces …

Altered Regulation Of Aquaporin Gene Expression In Allergen And Il-13-Induced Mouse Models Of Asthma, Carissa M. Krane, Bijia Deng, Venkateshwar Mutyam, Casey A. Mcdonald, Stephen Pazdziorko, Lawrence Mason, Samuel Goldman, Marion Kasaian, Divya Chaudhary, Cara Williams, Melisa W.Y. Ho

Biology Faculty Publications

IL-13 is known to affect many processes that contribute to an asthmatic phenotype, including inflammation, fibrosis, and mucus production. Members of the aquaporin (AQP) family of transmembrane water channels are targets of regulation in models of lung injury and inflammation. Therefore, we examined AQP mRNA and protein expression in allergen and IL-13-induced mouse models of asthma. Lungs from ovalbumin sensitized and ovalbumin challenged (OVA/OVA) and IL-13 treated mice showed airway thickening, increased mucus production, and pulmonary eosinophilia. Pulmonary function tests showed a significant increase in methacholine-induced airway hyperreactivity in OVA/OVA and IL-13-treated mice as compared with controls. Quantitative PCR analysis …

Comparison Of Replication-Competent, First Generation, And Helper-Dependent Adenoviral Vaccines, Eric A. Weaver, Pramod N. Nehete, Stephanie S. Buchl, Julien S. Senac, Donna Palmer, Philip Ng, K. Jagannadha Sastry, Michael A. Barry

Nebraska Center for Virology: Faculty Publications

All studies using human serotype 5 Adenovirus (Ad) vectors must address two major obstacles: safety and the presence of pre-existing neutralizing antibodies. Helper-Dependent (HD) Ads have been proposed as alternative vectors for gene therapy and vaccine development because they have an improved safety profile. To evaluate the potential of HD-Ad vaccines, we compared replication-competent (RC), first-generation (FG) and HD vectors for their ability to induce immune responses in mice. We show that RC-Ad5 and HD-Ad5 vectors generate stronger immune responses than FG-Ad5 vectors. HD-Ad5 vectors gave lower side effects than RC or FG-Ad, producing lower levels of tissue damage and …

Comparison Of Gene Ontology Term Annotations Between E.Coli K12 Databases, Reddysailaja Marpuri

Student Research Conference Select Presentations

The objective of this project was to get up-to-date functional information on all genes of E.coli K12 strains based on Genome Ontology terms. Gene Ontology is described by a defined library of terms related the biological process, cellular components and molecular functions of a gene in an organism. The genome sequence of an organism gains its value when it is annotated with gene ontology terms, which bridges the gap from the sequence to the biology of an organism. Since we use annotated gene database in the prediction of the function of newly sequenced genes, it is important to have databases …

Food Entrainment Of Circadian Gene Expression Altered In Pparα-/- Brown Fat And Heart, Brian Chun Kim Goh

Honors Capstones

No abstract provided.

Biarsenical Labeling Of Vesicular Stomatitis Virus Encoding Tetracysteine-Tagged M Protein Allows Dynamic Imaging Of M Protein And Virus Uncoating In Infected Cells, Subash C. Das, Debasis Panda, Debasis Nayak, Asit K. Pattnaik

School of Veterinary and Biomedical Sciences: Faculty Publications

A recombinant vesicular stomatitis virus (VSV-PeGFP-M-MmRFP) encoding enhanced green fluorescent protein fused in frame with P (PeGFP) in place of P and a fusion matrix protein (monomeric red fluorescent protein fused in frame at the carboxy terminus of M [MmRFP]) at the G-L gene junction, in addition to wild-type (wt) M protein in its normal location, was recovered, but the MmRFP was not incorporated into the virions. Subsequently, we generated recombinant viruses (VSV-PeGFP-∆M-Mtc and VSV-∆M-Mtc) encoding Mprotein with a carboxy-terminal tetracysteine tag (Mtc) in place of the M protein. These recombinant viruses incorporated Mtc at levels similar to M in …

Naloxone's Pentapeptide Binding Site On Filamin A Blocks Mu Opioid Receptor–Gs Coupling And Creb Activation Of Acute Morphine, Hoau-Yan Wang, Lindsay H. Burns

Publications and Research

Chronic morphine causes the mu opioid receptor (MOR) to switch its coupling from Gi/o to Gs, resulting in excitatory signaling via both Gαs and its Gβγ dimer. Ultra-low-dose naloxone (NLX) prevents this switch and attenuates opioid tolerance and dependence. This protective effect is mediated via a high-affinity interaction of NLX to a pentapeptide region in c-terminal filamin A (FLNA), a scaffolding protein interacting with MOR. In organotypic striatal slice cultures, we now show that acute morphine induces a dose-dependent Go-to-Gs coupling switch at 5 and 15 min that resolves by 1 hr. The acute Gs coupling induced by 100 µM …

Glutathione Diminishes Tributyltin- And Dibutyltin-Induced Loss Of Lytic Function In Human Natural Killer Cells, Jeralyn J. Powell, Mclisa V. Davis, Margaret M. Whalen

Chemistry Faculty Research

This study investigated whether reduced glutathione (GSH) was able to alter the negative effects of tributyltin (TBT) or dibutyltin (DBT) on the lytic function of human natural killer (NK) cells. NK cells are an initial immune defense against the development of tumors or viral infections. TBT and DBT are widespread environmental contaminants, due to their various industrial applications. Both TBT and DBT have been shown to decrease the ability of NK cells to lyse tumor cells (lytic function). The results indicated that the presence of GSH during the exposure of NK cells to TBT or DBT diminished the negative effect …

In Vitro Guidance Of Retinal Axons By A Tectal Lamina-Specific Glycoprotein Nel, Yulan Jiang, Hiroya Obama, Soh-Leh Kuan, Ritsuko Nakamura, Chizu Nakamoto, Zhufeng Ouyang, Masaru Nakamoto

Biology Faculty Publications

Nel is a glycoprotein containing five chordin-like and six epidermal growth factor-like domains and is strongly expressed in the nervous system. In this study, we have examined expression patterns and in vitro functions of Nel in the chicken retinotectal system. We have found that in the developing tectum, expression of Nel is localized in specific laminae that retinal axons normally do not enter, including the border between the retinorecipient and non-retinorecipient laminae. Nel-binding activity is detected on retinal axons both in vivo and in vitro, suggesting that retinal axons express a receptor for Nel. In vitro, Nel inhibits retinal axon …

Characterization And Differentiation Of Peripheral Blood Derived Multipotent Adult Progenitor Cells, Hari Satya Shankar Addagarla

Theses, Dissertations and Capstones

Stem cells are populations of undifferentiated cells that are found in most tissues and act as precursors for regeneration and maintenance. In the future, they could provide promising therapies for diseases which are to date incurable. Our lab developed a novel cell line from the peripheral blood of adult transgenic green fluorescent protein swine and designated them as Peripheral Blood Derived Multipotent Adult Progenitor Cells (PBD-MAPCs). In this study we characterized the mRNA and protein expression profiles of PBD-MAPCs before and after neural differentiation and investigated the potential of PBD-MAPCs to differentiate into myocardial or neural lineages in vitro. We …

Loss Of Ski Expression In Testicular Cancer Leads To An Enhanced Invasive Phenotype Through Both Bmp-Dependent And Bmp-Independent Pathways, Amy N. Nash

Theses, Dissertations and Capstones

The proto-oncogene SKI is a transcription factor and a co-repressor of the TGFβ superfamily, including TGF

β and BMP. However, additional data suggests that SKI may function as a tumor suppressor in some cell types. The TGFβ superfamily has been implicated in cancer progression and germ cell migration. Testicular cancer afflicts men during their peak reproductive years and is the most common cancer among men of this age group. Cisplatin-based chemotherapy is the standard treatment for testicular cancer. This treatment can lead to undesirable side effects, including infertility. We have shown that SKI expression is decreased in testicular germ cell …

Enumeration Of Endothelial Progenitor Cells In Hind Limb Suspended Rats And The Mechanics Of Endothelial Wound Healing, Jarrod Matthew Pennington

Theses, Dissertations and Capstones

The endothelium repairs itself through two methods. One is by the activity of circulating endothelial progenitor cells (EPCs). EPCs are immature endothelial cells that circulate the bloodstream that are capable of proliferation and differentiation into mature endothelium. It is thought that EPCs contribute to the repair and replacement of damaged endothelial cells in the process of reendothelialization. Physical inactivity and bed rest are known to be deleterious to the endothelium. It is possible that this inactivity is also deleterious to the number of viable EPCs, which would reduce the ability of the endothelium to repair itself. We used hind limb …

Effects Of Aging On Regulators Of Muscle Apoptosis In The Female F344bn Rat, Murali K. Gadde

Theses, Dissertations and Capstones

Age-related muscle atrophy is a consequence of normal aging characterized by decreases in muscle mass and strength. The mechanism(s) underlying the loss of muscle mass with increasing age is not fully understood, however recent data has suggested that muscle cell apoptosis may be involved. Here we investigate how aging affects the regulation of muscle apoptosis in the extensor digitorum longus (EDL) and soleus muscles of young (6-month), aged (26-month), and very aged (30-month) female Fischer 344/NNiaHSD X Brown Norway / BiNia (F344BN) rats. EDL and soleus muscle mass/body weight ratios were lower in aged animals but not different between 26- …

Evolution Of Gene Structure In Multicellular Eukaryotes, Maria Hester

Inquiry: The University of Arkansas Undergraduate Research Journal

We investigated the patterns of intron conservation in eukaryotes for five different genes. The genes examined were ribosomal proteins L8, S14 and S17, along with elongation factor 2B and triose phosphate isomerase. Intron conservation for S14, S17, and triose phosphate isomerase was determined for 32 species representing the major branches of multicellular eukaryotes. For 25 conserved introns 16 were phase 0, five were phase 1, and four were phase 2. Triose phosphate isomerase had five of nine conserved introns shared between plants and animals, where S14 had one of nine and S17 had one of seven. However, there were two …

Light-Induced Fos Expression In Intrinsically Photosensitive Retinal Ganglion Cells In Melanopsin Knockout (Opn4-/-) Mice, Gary E. Pickard, Scott B. Baver, Malcolm D. Ogilvie, Patricia J. Sollars

School of Veterinary and Biomedical Sciences: Faculty Publications

Retinal ganglion cells that express the photopigment melanopsin are intrinsically photosensitive (ipRGCs) and exhibit robust synaptically driven ON-responses to light, yet they will continue to depolarize in response to light when all synaptic input from rod and cone photoreceptors is removed. The light-evoked increase in firing of classical ganglion cells is determined by synaptic input from ON-bipolar cells in the proximal sublamina of the inner plexiform layer. OFF-bipolar cells synapse with ganglion cell dendrites in the distal sublamina of the inner plexiform layer. Of the several types of ipRGC that have been described, M1 ipRGCs send dendrites exclusively into the …

A Microrna Imparts Robustness Against Environmental Fluctuation During Development, Xin Li, Justin J. Cassidy, Catherine A. Reinke, Stephen Fischboeck, Richard W. Carthew

Faculty Publications

The microRNA miR-7 is perfectly conserved from annelids to humans, and yet some of the genes that it regulates in Drosophila are not regulated in mammals. We have explored the role of lineage restricted targets, using Drosophila , in order to better understand the evolutionary significance of microRNA-target relationships. From studies of two well characterized developmental regulatory networks, we find that miR-7 functions in several interlocking feedback and feedforward loops, and propose that its role in these networks is to buffer them against perturbation. To directly demonstrate this function for miR-7, we subjected the networks to temperature fluctuation and found …

X4 Human Immunodeficiency Virus Type 1 Gp120 Down-Modulates Expression And Immunogenicity Of Codelivered Antigens, Avi-Hai Hovav, Michael Santosuosso, Maytal Bivas-Benita, Andre Plair, Alex Cheng, Mazal Elnekave, Elda Righi, Tao Chen, Satoshi Kashiwagi, Michael W. Panas, Shi-Hua Xiang, Karina Furmanov, Norman L. Letvin, Mark C. Poznansky

Nebraska Center for Virology: Faculty Publications

In order to increase the immune breadth of human immunodeficiency virus (HIV) vaccines, strategies such

as immunization with several HIV antigens or centralized immunogens have been examined. HIV-1 gp120

protein is a major immunogen of HIV and has been routinely considered for inclusion in both present and

future AIDS vaccines. However, recent studies proposed that gp120 interferes with the generation of immune

response to codelivered antigens. Here, we investigate whether coimmunization with plasmid-encoded gp120

alters the immune response to other coadministered plasmid encoded antigens such as luciferase or ovalbumin

in a mouse model. We found that the presence of gp120 …

Identification And Characterization Of A New Kaposi’S Sarcoma-Associated Herpesvirus Replication And Transcription Activator (Rta)- Responsive Element Involved In Rta-Mediated Transactivation, Hui-Ju Wen, Veenu Minhas, Charles Wood

Nebraska Center for Virology: Faculty Publications

Kaposi’s sarcoma-associated herpesvirus (KSHV) replication and transcription activator (RTA) is well established as a key transcriptional activator that regulates the KSHV life cycle from latency to lytic replication. It is expressed immediately after infection and activates a number of viral genes leading to virus replication. The RTA-responsive element (RRE) in the RTA target gene promoters is critical for RTA to mediate this transactivation. A number of non-conserved RREs have been identified in various RTA-responsive promoters, and AT-rich sequences have been proposed to serve as RTA targets, but no consensus RRE sequence has been identified so far. Two nonconserved RREs (RRE1 …

Use Of Average Mutual Information For Studying Changes In Hiv Populations, Khalid Sayood, Federico Hoffman, Charles Wood

Nebraska Center for Virology: Faculty Publications

Average mutual information (AMI) has been used in a number of applications in bioinformatics. In this paper we present its use to study genetic changes in populations; in particular populations of HIV viruses. Disease progression of HIV-1 infection in infants can be rapid resulting in death within the the first year, or slow, allowing the infant to survive beyond the first year. We study the development of rapid and slow progressing HIV population using AMI charts based on average mutual information among amino acids in the env gene from a population of 1142 clones derived from seven infants with slow …

Comparison Of Replication-Competent, First Generation, And Helper-Dependent Adenoviral Vaccines, Eric A. Weaver, Pramod N. Nehete, Stephanie S. Buchl, Julien S. Senac, Donna Palmer, Philip Ng, K. Jagannadha Sastry, Michael A. Barry

Nebraska Center for Virology: Faculty Publications

All studies using human serotype 5 Adenovirus (Ad) vectors must address two major obstacles: safety and the presence of pre-existing neutralizing antibodies. Helper-Dependent (HD) Ads have been proposed as alternative vectors for gene therapy and vaccine development because they have an improved safety profile. To evaluate the potential of HD-Ad vaccines, we compared replication-competent (RC), first-generation (FG) and HD vectors for their ability to induce immune responses in mice. We show that RC-Ad5 and HD-Ad5 vectors generate stronger immune responses than FG-Ad5 vectors. HD-Ad5 vectors gave lower side effects than RC or FG-Ad, producing lower levels of tissue damage and …

The Guinea Pig Ileum Lacks The Direct, High-Potency, M2-Muscarinic, Contractile Mechanism Of The Mouse Ileum, Michael T. Griffin, Minoru Matsui, Rennolds S. Ostrom, Frederick J. Ehlert

Pharmacy Faculty Articles and Research

We explored whether the M2 muscarinic receptor in the guinea pig ileum elicits a highly potent, direct-contractile response, like that from the M3 muscarinic receptor knockout mouse. First, we characterized the irreversible receptor-blocking activity of 4-DAMP mustard in ileum from muscarinic receptor knockout mice to verify its M3 selectivity. Then, we used 4-DAMP mustard to inactivate M3 responses in the guinea pig ileum to attempt to reveal direct, M2 receptor-mediated contractions. The muscarinic agonist, oxotremorine-M, elicited potent contractions in ileum from wild-type, M2 receptor knockout, and M3 receptor knockout mice characterized by negative log EC50 (pEC50) values ± SEM of …

Overexpression Of Truncated Erg From Tmprss2-Erg Fusion And Prostate Cancer Development , Melanie Leong, Wenfeng Shi, Jun Tian, Ellen Cho, Abbas Raza, Saquib A. Siddiqi, Abdulhafez Selim, Hanchun Chen, Dianzheng Zhang

PCOM Scholarly Works

No abstract provided.

Repressive Effects Of Resveratrol On Androgen Receptor Transcriptional Activity, Wenfeng Shi, Melanie Leong, Ellen Cho, Joseph Farrell, Hanchun Chen, Jun Tian, Dianzheng Zhang

PCOM Scholarly Works

Background: The chemopreventive effects of resveratrol (RSV) on prostate cancer have been well established; the androgen receptor (AR) plays pivotal roles in prostatic tumorigenesis. However, the exact underlying molecular mechanisms about the effects of RSV on AR have not been fully elucidated. A model system is needed to determine whether and how RSV represses AR transcriptional activity. Methodology: The AR cDNA was first cloned into the retroviral vector pOZ-N and then integrated into the genome of ARnegative HeLa cells to generate the AR(+) cells. The constitutively expressed AR was characterized by monitoring hormonestimulated nuclear translocation, DNA binding, and transcriptional activation, …

Live-Cell Studies On Mitotic Slippage In Humans, Daniela A. Brito

Legacy Theses & Dissertations (2009 - 2024)

Checkpoints are regulatory pathways that control the order and timing of specific cell-cycle events. In the presence of unattached/weakly-attached kinetochores, the mitotic checkpoint (MC) arrests cells in mitosis by inhibiting the degradation of cyclin B, the regulatory subunit of Cdk1 (cyclin dependent kinase 1). Checkpoints do not arrest cells permanently, and escaping mitosis with an unsatisfied MC requires cyclin B/Cdk1 inactivation. In yeast, this occurs through an “adaptation” mechanism involving inhibitory phosphorylations and/or Cdk1-inhibitors. To determine how vertebrate cells escape mitosis when the MC cannot be satisfied I conducted live-cell imaging and immunofluorescence studies on nocodazole-treated rat kangaroo (PtK) and …

An Rnai Screen Targeting The Protein Tyrosine Kinases Identifies Bruton's Tyrosine Kinase (Btk) As A Breast Cancer Cell Survival Factor, Cheryl Lynne Eifert

Legacy Theses & Dissertations (2009 - 2024)

The receptor protein tyrosine kinases (RPTKs) and the non- receptor protein tyrosine kinases (PTKs) are among the most commonly up-regulated genes found in all types of cancers. Although, a large body of data implicates a majority of tyrosine kinases (TKs) in cancer, few have been extensively evaluated for any potential therapeutic benefit in any of the many subtypes of breast cancer. We have used RNA interference (RNAi) to perform a large-scale loss-of-function analysis to facilitate the identification of individual factors necessary for the survival of an ErbB2 positive breast cancer cell line. We have found that 30% of the TKs …

The C1 And C2 Domains Target Human Type 6 Adenylyl Cyclase To Lipid Rafts And Caveolae, Muthusamy Thangavel, Xiaoqiu Liu, Shu Qiang Sun, Joseph Kaminsky, Rennolds S. Ostrom

Pharmacy Faculty Articles and Research

Previous data has shown that adenylyl cyclase type 6 (AC6) is expressed principally in lipid rafts or caveolae of cardiac myocytes and other cell types while certain other isoforms of AC are excluded from these microdomains. The mechanism by which AC6 is localized to lipid rafts or caveolae is unknown. In this study, we show AC6 is localized in lipid rafts of COS-7 cells (expressing caveolin-1) and in HEK-293 cells or cardiac fibroblasts isolated from caveolin-1 knock-out mice (both of which lack prototypical caveolins). To determine the region of AC6 that confers raft localization, we independently expressed each of the …

Hmgb1 Mediates Endogenous Tlr2 Activation And Brain Tumor Regression, James Curtin, Naiyou Liu, Marianela Candolfi, Weidong Xiong, Hikmat Assi, Kader Yagiz, Matthew Edwards, Kathrin Michelsen, Kurt Kroeger, Chunyan Liu, Akm Ghulam Muhammad, Mary Clark, Moshe Arditi, Begonya Comin-Anduix, Antoni Ribas, Pedro Lowenstein, Maria Castro

Articles

BACKGROUND: Glioblastoma multiforme (GBM) is the most aggressive primary brain tumor that carries a 5-y survival rate of 5%. Attempts at eliciting a clinically relevant anti-GBM immune response in brain tumor patients have met with limited success, which is due to brain immune privilege, tumor immune evasion, and a paucity of dendritic cells (DCs) within the central nervous system. Herein we uncovered a novel pathway for the activation of an effective anti-GBM immune response mediated by high-mobility-group box 1 (HMGB1), an alarmin protein released from dying tumor cells, which acts as an endogenous ligand for Toll-like receptor 2 (TLR2) signaling …

Patterning Defects In Silkworm Embryos Analysed Through Cuticle Preparations, Amit Singh, Madhuri Kango-Singh, K. P. Gopinathan

Biology Faculty Publications

The mulberry silkworm, Bombyx mori, a holometabolous lepidopteran insect, has a metameric body plan. Due to its functional adaptation, B. mori presents some unique deviations in its pal/ern from the evolutionarily advanced dipteran insect, Drosophila melanogaster. Previous studies on mutant phenotype analysis in B. mori have been carried out in late stages of larval development. Here we employ, the cuticle preparation approach during embryonic development to study morphological landmarks associated with B. mori, Eri, another race a/silkworm, and pattern defects associated with Ekp mutant of B.mori. The homeotic mutant Ekp, generates ectopic abdominallegs, a feature …