Cell and Developmental Biology Commons^™

Mechanisms Of Adenovirus-Mediated Autophagy, Erin White

Dissertations and Theses (Open Access)

A patient diagnosed with a glioma, generally, has an average of 14 months year to live after implementation of conventional therapies such as surgery, chemotherapy, and radiation. Glioblastomas are highly lethal because of their aggressive nature and resistance to conventional therapies and apoptosis. Thus other avenues of cell death urgently need to be explored. Autophagy, which is also known as programmed cell death type II, has recently been identified as an alternative mechanism to kill apoptosis- resistant cancer cells. Traditionally, researchers have studied how cells undergo autophagy during viral infection as an immune response mechanism, but recently researchers have discovered …

Mechanisms Of Adenovirus-Mediated Autophagy, Erin White

Dissertations and Theses (Open Access)

A patient diagnosed with a glioma, generally, has an average of 14 months year to live after implementation of conventional therapies such as surgery, chemotherapy, and radiation. Glioblastomas are highly lethal because of their aggressive nature and resistance to conventional therapies and apoptosis. Thus other avenues of cell death urgently need to be explored. Autophagy, which is also known as programmed cell death type II, has recently been identified as an alternative mechanism to kill apoptosis- resistant cancer cells. Traditionally, researchers have studied how cells undergo autophagy during viral infection as an immune response mechanism, but recently researchers have discovered …

Mmp9 Cleavage Of The Β4 Integrin Ectodomain Leads To Recurrent Epithelial Erosions In Mice, Sonali Pal-Ghosh, Tomas Blanco, Gauri Tadvalkar, Ahdeah Pajoohesh-Ganji, Arpitha Parthasarathy, James D Zieske, Mary Ann Stepp

Anatomy and Regenerative Biology Faculty Publications

Integrin α6β4 is an integral membrane protein within hemidesmosomes and it mediates adhesion of epithelial cells to their underlying basement membrane. During wound healing, disassembly of hemidesmosomes must occur for sheet movement-mediated cell migration. The mechanisms of disassembly and reassembly of hemidesmosomes are not fully understood. The current study was initiated to understand the underlying cause of recurrent corneal erosions in the mouse. Here, we show that in vivo: (1) MMP9 levels are elevated and β4 integrin is partially cleaved in epithelial cell extracts derived from debridement wounded corneas; (2) the β4 ectodomain is missing from sites where erosions develop; …

Embryonic Development Of The Grass Shrimp, Palaemonetes Pugio, And The Influence Of Salinity On Cardiac Physiology, Amie Lynn Romney

UNLV Theses, Dissertations, Professional Papers, and Capstones

The grass shrimp, Palaemonetes pugio, inhabit the brackish waters along the Atlantic coast of North America and are a key component of the estuarine ecosystem. These decapod shrimp can tolerate large fluctuations in environmental parameters including daily and seasonal fluctuations in salinity between 0 to 55 parts per thousand (seawater at 30-32 ppt). Any observed distribution patterns of adult P. pugio in relation to salinity may be accounted for by their relative ability to tolerate a range in salinity and their ability to maintain internal water volumes and cardiac performance in the earliest life stage, the embryonic period. This thesis …

Combating Bioterrorism: Purification And Structural Analysis Of The E3 Proteins Of The Venezuelan Equine Encephalitis Virus And The Chikungunya Virus, Tyshia Gwin

The Journal of Purdue Undergraduate Research

Outbreaks of the Venezuelan Equine Encephalitis Virus (VEEV) and the Chikungunya Virus (CHIKV) continue to emerge in Central and South America, Africa, and Asia, but there are currently no vaccines or anti-virals for these viruses. Given their ease of transmission, debilitating symptoms, and genetic alterability, VEEV and CHIKV have great potential for development into biological weapons. Therefore, there is an urgent need to determine possible methods of treatment or prevention. One possibility of prevention lies in determining the structure and biological function of the E3 protein that plays a key role in the infectivity of the viruses. In this research, …

Na/K-Atpase Mimetic Pnaktide Peptide Inhibits The Growth Of Human Cancer Cells, Zhichuan Li, Zhongbing Zhang, Joe X. Xie, Xin Li, Jiang Tian, Ting Cai, Hongaun Cui, Hanfei Ding, Joseph I. Shapiro Md, Zijian Xie

Biochemistry and Microbiology

Cells contain a large pool of non-pumping Na/K-ATPase that participates in signal transduction. Here, we show that the expression of α1 Na/K-ATPase is significantly reduced in human prostate carcinoma as well as in several human cancer cell lines. This down-regulation impairs the ability of Na/K-ATPase to regulate Src-related signaling processes. Supplement of pNaKtide, a peptide derived from α1 Na/K-ATPase, reduces activities of Src and Src effectors. Consequently, these treatments stimulate apoptosis and inhibit growth in cultures of human cancer cells. Moreover, administration of pNaKtide inhibits angiogenesis and growth of tumor xenograft. Thus, the new findings demonstrate the in vivo effectiveness …

Comparison Of Adenoviruses As Oncolytics And Cancer Vaccines In An Immunocompetent B Cell Lymphoma Model, Eric A. Weaver, Christopher Y. Chen, Shannon M. May, Mary E. Barry, Michael A. Barry

Nebraska Center for Virology: Faculty Publications

We have screened human adenoviruses (Ads) for oncolytic activity against a variety of mouse and hamster cell lines and have found a number that are susceptible to a variety of Ad serotypes. A20 lymphoma is derived from BALB/c mice and is susceptible to infection and killing by a variety of human Ads. A20 is also a suitable cancer vaccine model, because these cells express a unique immunoglobulin variable region that can be targeted by vaccination. To compare Ads as cancer vaccines versus Ads as oncolytics, A20 tumors were initiated in immunocompetent BALB/c mice. Mice immunized with first-generation Ad5 expressing the …

Regulation Of Cytosolic Phospholipase A2 (Cpla2) And Its Association With Cell Proliferation In Human Lens Epithelial Cells, Yin Wang, Kui-Yi Xing, Marjorie F. Lou

School of Veterinary and Biomedical Sciences: Faculty Publications

PURPOSE. To investigate the molecular mechanism for cytosolic phospholipase A2 (cPLA2) regulation and its association to platelet-derived growth factor (PDGF)-induced cell proliferation.

METHODS. cPLA2 was examined using human lens epithelial (HLE) B3 cells. Reactive oxygen species (ROS) generation induced by PDGF was analyzed by luminescence assay. Cell proliferation was measured by cell counting and by BrdU assay. Human cPLA2 gene was cloned via RT-PCR followed by sitedirected mutagenesis to construct HLE B3 cells expressing either inactive cPLA2 enzyme with S228A mutation (S228A), or cPLA2 truncated at the calcium-binding C2 domain (C2D). Activity of cPLA2 was measured by arachidonic acid (AA) …

Calcineurin Is Required For Trpv1-Induced Ltd Of Ca1 Stratum Radiatum Interneurons, Tyron Deray Jensen

Theses and Dissertations

Learning and memory in the brain are thought to be dependent on synaptic plasticity. In response to sensory input, synapses can be strengthened or weakened, known as long-term potentiation or long-term depression (LTD), respectively. Transient receptor potential vanilloid 1 (TRPV1) has been shown to mediate a novel form of presynaptic LTD in hippocampal interneurons. TRPV1 is currently being heavily studied in the PNS and being targeted by pharmaceuticals for its anti-nociceptive and anti-inflammatory properties. However, much less is known regarding TRPV1 function in the CNS, including the signal mechanism mediating hippocampal LTD despite its obvious importance. Here we performed whole-cell …

The Effects Of Β-Amyloid On Α7 Nicotinic Acetylcholine Receptors Expressed In Xenopus Oocytes, Malia L. Anderson

Theses and Dissertations

The exact mechanism and progression of Alzheimer's disease (AD) at present is not fully understood. In patients suffering from AD, damage to the hippocampal region and impairment of learning and memory is present. It is also known that a buildup of β-amyloid plaques occur in AD patients and that β-amyloid interacts with some subtypes of neuronal nicotinic acetylcholine receptors (neuronal nAChRs). These receptors are composed of five subunits. The most prevalent nAChR subunit composition through the brain as a whole is α7. Previous data produced from our lab suggests that α7 nAChRs are also one of the most prevalent subunits …

Mechanical Test Methods For Assessing Porcine Carotid And Uterine Artery Burst Pressure Following Ex Vivo Ultrasonic Ligature Seal And Transection, Carissa M. Krane, Margaret Pinnell, Courtney Gardner, Mercedes Thompson, James Coleman, Robert Wilkens

Biology Faculty Publications

A test method was developed to identify those variables important for assessing the performance of ultrasonic surgical devices in ex vivo ligature sealing of porcine carotid and uterine arteries. Ruggedness testing using a small sample size in pilot experiments was conducted using a newly developed test method in an effort to assess the usefulness of this methodology and to identify test variables that might warrant further testing. The development of this test method included the use of a custom-designed prototypic tension device for load-controlled ex vivo vessel stretching during saline perfusion and subsequent seal and transection of porcine arteries with …

Generation Of A Kupffer Cell-Evading Adenovirus For Systemic And Liver-Directed Gene Transfer, Reeti Khare, Shannon M. May, Francesco Vetrini, Eric A. Weaver, Donna Palmer, Amanda Rosewell, Nathan Grove, Philip Ng, Michael A. Barry

Nebraska Center for Virology: Faculty Publications

As much as 90% of an intravenously (i.v.) injected dose of adenovirus serotype 5 (Ad5) is absorbed and destroyed by liver Kupffer cells. Viruses that escape these cells can then transduce hepatocytes after binding factor X (FX). Given that interactions with FX and Kupffer cells are thought to occur on the Ad5 hexon protein, we replaced its exposed hypervariable regions (HVR) with those from Ad6. When tested in vivo in BALB/c mice and in hamsters, the Ad5/6 chimera mediated >10 times higher transduction in the liver. This effect was not due to changes in FX binding. Rather, Ad5/6 appeared to …

Age-Related Hamster Mitochondrial Changes And Oocyte Changes Following Autologous Platelet Mitochondrial Microinjection, Fang Li

Theses and Dissertations in Biomedical Sciences

This study's objective was to verify age-related mitochondrial changes in oocytes from old hamsters compared with those from young hamsters. We used autologous platelet mitochondrial microinjection to improve the mitochondrial quality and quantity of aged oocytes to improve their pregnancy potential. Metaphase II oocytes were collected from super-ovulated female hamsters and prepared for biochemical and morphological analysis. Adenosine triphosphate (ATP) levels, mitochondrial DNA (mtDNA) number, reactive oxygen species (ROS) level and mitochondrial membrane potential (MMP) were determined in individual oocytes from young and old hamsters. Transmission electron microscopy (TEM) allowed evaluation of oocyte ultra-structure. In oocytes from old hamsters, ATP …

Nanosecond Pulsed Electric Field Induction Of Programmed Cell Death Is Cell Type Dependent: An In Vitro Study, Wei Ren

Theses and Dissertations in Biomedical Sciences

Nanosecond pulsed electric fields (nsPEFs) present a novel and effective method for cancer ablation by eradicating the ubiquitous cancer hallmark of apoptosis evasion and enforcing cancer programmed cell death. To develop nsPEFs as an anticancer method, a comprehensive understanding of cell death mechanisms is required. The overall objective of this dissertation is to elucidate molecular mechanisms underlying effects of nsPEFs on E4 murine squamous cell carcinoma and human T-cell Jurkat clones that are wildtype, deficient in FADD (ΔFADD) and deficient in caspase-8 (ACas-8). The overall hypothesis is that nsPEFs eliminate cancer cells through activating caspase-dependent and caspase-independent programmed cell death …

Altering Nitric Oxide Bioavailability And Lipid Profiles In Endothelial Cells By Polycyclic Aromatic Hydrocarbons In Particulate Matter, Liang Yu

Theses and Dissertations in Biomedical Sciences

Epidemiologic studies have demonstrated a significant association between exposure to particulate matter (PM) and atherosclerosis. Polycyclic aromatic hydrocarbons (PAHs) present in particulate matter, are well known to induce oxidative stress and lipid peroxidation via generation of reactive oxygen species (ROS). Lipid peroxidation involves regulating endothelial nitric oxide synthase via inhibition of its activity, and as a result, mediates dilation of coronary arterioles is involved with the pathogenesis of atherosclerosis. However, data on assessment of oxidized lipid formation is limited due to low resolution of mass spectrometer methods. Taking the advantage of a Fourier Transform Ion Cyclotron Resonance Mass Spectrometer (FT-ICR-MS) …

Transcription And Translation Of Human F11r Gene Are Required For An Initial Step Of Atherogenesis Induced By Inflammatory Cytokines, Bani M. Azari, Jonathan D. Marmur, Moro O. Salifu, Yigal H. Ehrlich, Elizabeth Kornecki, Anna Babinska

Publications and Research

Background - The F11 Receptor (F11R; aka JAM-A, JAM-1) is a cell adhesion protein present constitutively on the membrane surface of circulating platelets and within tight junctions of endothelial cells (ECs). Previous reports demonstrated that exposure of ECs to pro-inflammatory cytokines causes insertion of F11R molecules into the luminal surface of ECs, ensuing with homologous interactions between F11R molecules of platelets and ECs, and a resultant adhesion of platelets to the inflamed ECs. The main new finding of the present report is that the first step in this chain of events is the de-novo transcription and translation of F11R molecules, …

Sonic Hedgehog Dependent Phosphorylation By Ck1Α And Grk2 Is Required For Ciliary Accumulation And Activation Of Smoothened, Yongbin Chen, Noriaki Sasai, Guoqiang Ma, Tao Yue, Jianhang Jia, James Briscoe, Jin Jiang

Markey Cancer Center Faculty Publications

Hedgehog (Hh) signaling regulates embryonic development and adult tissue homeostasis through the GPCR-like protein Smoothened (Smo), but how vertebrate Smo is activated remains poorly understood. In Drosophila, Hh dependent phosphorylation activates Smo. Whether this is also the case in vertebrates is unclear, owing to the marked sequence divergence between vertebrate and Drosophila Smo (dSmo) and the involvement of primary cilia in vertebrate Hh signaling. Here we demonstrate that mammalian Smo (mSmo) is activated through multi-site phosphorylation of its carboxyl-terminal tail by CK1α and GRK2. Phosphorylation of mSmo induces its active conformation and simultaneously promotes its ciliary accumulation. We demonstrate that …

Imaging Mass Spectrometry Detection Of Gangliosides Species Within The Mouse Brain Following Transient Focal Cerebral Ischemia, Shawn N. Whitehead, Kenneth H. N. Chan, Sandhya Gangaraju, Jacqueline Slinn, Jianjun Li, Sheng T. Hou

Anatomy and Cell Biology Publications

Gangliosides, a member of the glycosphingolipid family, are heterogeneously expressed in biological membranes and are particularly enriched within the central nervous system. Gangliosides consist of mono- or poly-sialylated oligosaccharide chains of variable lengths attached to a ceramide unit and are found to be intimately involved in brain disease development. The purpose of this study is to examine the spatial profile of ganglioside species using matrix-assisted laser desorption/ionization (MALDI) imaging (IMS) following middle cerebral artery occlusion (MCAO) reperfusion injury in the mouse. IMS is a powerful method to not only discriminate gangliosides by their oligosaccharide components, but also by their carbon …

Use Of Imaging Biomarkers To Assess Perfusion And Glucose Metabolism In The Skeletal Muscle Of Dystrophic Mice, Nabeel Ahmad, Ian Welch, Robert Grange, Jennifer Hadway, Savita Dhanvantari, David Hill, Ting-Yim Lee, Lisa M Hoffman

Robarts Imaging Publications

BACKGROUND: Duchenne muscular dystrophy (DMD) is a severe neuromuscular disease that affects 1 in 3500 boys. The disease is characterized by progressive muscle degeneration that results from mutations in or loss of the cytoskeletal protein, dystrophin, from the glycoprotein membrane complex, thus increasing the susceptibility of contractile muscle to injury. To date, disease progression is typically assessed using invasive techniques such as muscle biopsies, and while there are recent reports of the use of magnetic resonance, ultrasound and optical imaging technologies to address the issue of disease progression and monitoring therapeutic intervention in dystrophic mice, our study aims to validate …

Vacht Knock-Down Mice Show Normal Prepulse Inhibition But Disrupted Long-Term Habituation, S. Schmid, E. Azzopardi, X. De Jaeger, Marco Prado, V. F. Prado

Anatomy and Cell Biology Publications

The neurotransmitter acetylcholine (ACh) plays a crucial role in both the central and peripheral nervous system. Central cholinergic transmission is important for cognitive functions and cholinergic disruptions have been associated with different neural disorders. We here tested the role of cholinergic transmission in basic cognitive functions, i.e. in prepulse inhibition (PPI) and short-term habituation (STH) as well as long-term habituation (LTH) of startle using mice with a 65% knockdown (KD) of the vesicular ACh transporter (VAChT). These mice are slow in refilling cholinergic synaptic transmitter vesicles, leading to a reduced cholinergic tone. Prepulse inhibition has been assumed to be mediated …

Multimodal Spectral Imaging Of Cells Using A Transmission Diffraction Grating On A Light Microscope, Dragan Isailovic, Yang Xu, Tyler Copus, Suraj Saraswat, Surya M. Nauli

Pharmacy Faculty Articles and Research

A multimodal methodology for spectral imaging of cells is presented. The spectral imaging setup uses a transmission diffraction grating on a light microscope to concurrently record spectral images of cells and cellular organelles by fluorescence, darkfield, brightfield, and differential interference contrast (DIC) spectral microscopy. Initially, the setup was applied for fluorescence spectral imaging of yeast and mammalian cells labeled with multiple fluorophores. Fluorescence signals originating from fluorescently labeled biomolecules in cells were collected through triple or single filter cubes, separated by the grating, and imaged using a charge-coupled device (CCD) camera. Cellular components such as nuclei, cytoskeleton, and mitochondria were …

Immune Evasion Of Porcine Reproductive And Respiratory Syndrome Virus Through Glycan Shielding Involves Both Glycoprotein 5 As Well As Glycoprotein 3, Hiep Vu, Byungjoon Kwon, Kyoung-Jin Yoon, William W. Laegreid, Asit K. Pattnaik, Fernando A. Osorio

School of Veterinary and Biomedical Sciences: Faculty Publications

Passive administration of porcine reproductive and respiratory syndrome virus (PRRSV) neutralizing antibodies (NAbs) can effectively protect pigs against PRRSV infection. However, after PRRSV infection, pigs typically develop a weak and deferred NAb response. One major reason for such a meager NAb response is the phenomenon of glycan shielding involving GP5, a major glycoprotein carrying one major neutralizing epitope. We describe here a type II PRRSV field isolate (PRRSV-01) that is highly susceptible to neutralization and induces an atypically rapid, robust NAb response in vivo. Sequence analysis shows that PRRSV-01 lacks two N-glycosylation sites, normally present in wild-type (wt) PRRSV strains, …

Tetrabromobisphenol A Decreases Cell-Surface Proteins Involved In Human Natural Killer (Nk) Cell–Dependent Target Cell Lysis, Tasia Hurd, Margaret M. Whalen

Chemistry Faculty Research

Human natural killer (NK) lymphocytes are able to destroy tumor cells and virally-infected cells. Interference with their function can leave an individual with increased susceptibility to cancer development and/or viral infection. We have shown that the tumor-destroying (lytic) function of NK cells can be dramatically decreased by exposure to the environmental contaminant tetrabromobisphenol A (TBBPA). TBBPA is a flame retardant used in a variety of materials including circuit boards, carpeting, and upholstery and has been found in human blood samples. TBBPA interferes with NK cell lytic function, in part, by decreasing the ability of NK cells to bind to target …

Variations In Mre11/Rad50/Nbs1 Status And Dna Damage-Induced S-Phase Arrest In The Cell Lines Of The Nci60 Panel, Kristen M. K. Garner, Alan Eastman

Dartmouth Scholarship

The Mre11/Rad50/Nbs1 (MRN) complex is a regulator of cell cycle checkpoints and DNA repair. Defects in MRN can lead to defective S-phase arrest when cells are damaged. Such defects may elicit sensitivity to selected drugs providing a chemical synthetic lethal interaction that could be used to target therapy to tumors with these defects. The goal of this study was to identify these defects in the NCI60 panel of cell lines and identify compounds that might elicit selective cytotoxicity.

Homoplastic Microinversions And The Avian Tree Of Life, E. L. Braun, R T. Kimball, K. L. Han, N. R. Luhasz-Velez, A. J. Bonilla, Jena L. Chojnowski, J. V. Smith, R. C. Bowie, M. J. Braun, S. J. Hackett, J. Harshman, C. J. Huddleston, B. D. Marks, W. S. Moore, S. Reddy, F. H. Sheldon, C. C. Witt, T. Yuri

Natural Sciences Faculty Publications

BACKGROUND:

Microinversions are cytologically undetectable inversions of DNA sequences that accumulate slowly in genomes. Like many other rare genomic changes (RGCs), microinversions are thought to be virtually homoplasy-free evolutionary characters, suggesting that they may be very useful for difficult phylogenetic problems such as the avian tree of life. However, few detailed surveys of these genomic rearrangements have been conducted, making it difficult to assess this hypothesis or understand the impact of microinversions upon genome evolution.

RESULTS:

We surveyed non-coding sequence data from a recent avian phylogenetic study and found substantially more microinversions than expected based upon prior information about vertebrate …

Possible Molecular Mechanism To Account For Wavelength Dependence Of Equilibration Rates Of Patman And Laurdan In Phosphatidylcholine Bilayers, Hannabeth A. Franchino

Theses and Dissertations

Patman is a fluorescent membrane probe related to Laurdan. The structural distinctions between the two probes are the lengths of the aliphatic tails (eleven carbons in Laurdan and fifteen in Patman) and the presence of a trimethylammonium group on Patman that produces a positively-charged head. Preliminary studies exploring Patman as a probe to detect membrane properties during apoptosis revealed that the fluorescence intensity of two edges of the emission spectrum (435 and 500 nm) stabilizes at different rates as the probe binds to the cell membrane. To test whether these differences represent dissimilarities in probe binding to ordered and disordered …

Quorum Sensing In Archaea, Charles Mackin

Honors Scholar Theses

Bacteria coordinate cell density dependent behaviors by communicating through chemical intermediaries in a process called quorum sensing. In a bacterial culture, individual cells will constitutively produce signal molecules, termed autoinducers, and export them into the environment. When the concentration of autoinducers reaches a threshold, the cells sense that they are in a specific situation, which requires the upregulation of certain genes. This upregulation causes the bacteria to produce proteins that allow them to take part in a coordinated population-wide behavior.

In bacteria that are naturally competent, or capable of importing DNA from the environment, the expression of competence genes is …

Design And Bio-Production Of A Nanoparticle Avian Influenza Vaccine, Kashif N. Ather

Honors Scholar Theses

Influenza is one of the most common diseases in the world and the cause for numerous deaths every year. The primary method of combating the disease is the influenza vaccine, which is produced by inoculating chicken eggs with inactivated virus. An emerging solution is to use Self-Assembling Polypeptide Nanoparticles (SAPN) to elicit an immune response in the body, rather than using inactivated viruses. This project focuses on the synthesis, purification, and refolding of two peptide constructs, BN5C and S43, which are specific protein sequences that under the right conditions will refold into the 3-dimensional structures necessary for producing an immune …

Characterizing The Role Of The Bacterial Metallothionein, Smta, In Mammalian Infection, Stephanie R. Davis

Honors Scholar Theses

Mammalian metallothioneins (MT) are induced by various immunomodulatory molecules and are involved in a spectrum of immune processes such as essential metal homeostasis, detoxification of certain heavy metals, inflammation, and immune cell trafficking [1-3]. A bacterial metallothionein, SmtA, shares some sequence homology with mammalian MT as well as its metal-binding capabilities [4]. In addition to its ability to sequester heavy metals, eukaryotic MT has also been shown to scavenge free radicals such as reactive oxygen and nitrogen species (ROS, RNS), interfering with their toxic effects on cells and potentially influencing their regulatory roles in cell proliferation and differentiation [5, 6]. …

Biomarker Signature Classification Of Various Stress Forms, Chana L. Rich

Honors Scholar Theses

Various types of stressful conditions can have unique and important effects on immunity and can lead to dramatic consequences to health. For my University Scholar project, the characteristic biomarker signatures produced from a set of diverse stressors (e.g. psychological, biological and chemical) are being investigated. A biomarker signature is a distinctive biological indicator of a specific condition. High-throughput tools for the measurement of different cellular products have the potential to further our understanding of human disease and facilitated the identification of new biomarkers in all areas of medicine. The hypothesis that each form of stress, psychological, chemical and physical, will …