Microbiology Commons^™

Identification Of The Sites Of Action Of Inhibitors Of Mammalian Phospholipase D2 (Pld2) And The Role Of Interacting Protein Partners, Ramya Ganesan

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Phospholipase D (PLD) is a key enzyme for the remodeling of phospholipids in the cell membrane. PLD has been implicated in many physiological functions such as chemotaxis and phagocytosis as well as pathological functions such as ischemia/reperfusion and cancer metastasis. Several small molecule inhibitors of PLD have been recently developed to overcome these pathological effects. We have concentrated on 5-Fluoro-2-Indolyl des-Chlorohalopemide (FIPI) which is an indole derivative of halopemide that inhibits the two mammalian isoforms PLD1 and PLD2. We also concentrate on N-[2-[1-(3-Fluorophenyl)-4-oxo-1,3,-8-triazaspiro[4.5]dec-8-yl]ethyl]-2-naphthalenecarboxamide (NFOT) and N-[2-(4-oxo-1-phenyl-1,3,8-triazaspiro[4,5]dec-8-yl)ethyl]-2-naphthalenecarboxamide (NOPT) that are specific inhibitors of the PLD2 isoform. In spite of knowing their …

Comparison Between Flow Cytometry And Bead Method In Counting Cd4 And Cd8 T Lymphocytes In Mouse Spleen Cells Suspension, Abdulrahman Abdalla Allabidi

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The flow cytometry is gold standard method to count CD4, CD8 human T lymphocytes, and calculate CD8 to CD4 ratio in HIV patients peripheral blood to stage them and monitor their response to treatment. In this study, a comparison was made by counting CD4and CD8 mouse spleen T lymphocytes by flow cytometry, and a method involving antibody coated latex beads. The bead method yielded results comparable to those obtained by flow cytometry. These results indicate that antibody coated beads are suitable to determine CD4 and CD8 + T lymphocytes counts in situation where flow cytometry is not readily available.

Effect Of Herpes Simplex Virus-1 On Macrophage Cd Marker Expression, Hind Obaid Albeshri

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Macrophages are specialized phagocytic cells derived from the peripheral blood mononuclear cells. Macrophages can assume M1 or M2 cellular states upon activation with specific cytokines. Upon activation, macrophages produce either pro- or anti- inflammatory cytokines and thereby perform their functions of antigen engulfment and debris clearance. Macrophages have several cell surface cluster differentiation (CD) markers including CD80, CD163 and CD200R that differentiate M1 macrophages from M2. Downstream signaling and cytokine production varies based on the interaction between the specific CD marker and antigen.

The present study was aimed at analyzing and comparing the effects of HSV-1 infection on un-polarized and …

Elucidating The Effects Of Tcdd On The Polymorphic Human Hs1,2 Enhancer, Abdullah Freiwan

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2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a potent environmental toxin known to inhibit immunoglobulin (Ig) gene expression in various animal studies. We have identified the mouse 3'Ig heavy chain regulatory region (3'IgH RR) as a sensitive transcriptional target of TCDD, which may mediate the inhibitory effect of TCDD on Ig expression. Interestingly, the human hs1,2 enhancer is polymorphic and has been associated with a number of autoimmune diseases.

Suggesting a species difference, TCDD inhibited mouse hs1,2 enhancer activation and activated basal human hs1,2 (hs-hs1,2) enhancer activity in the mouse B-cell line. The objective of this study was to elucidate the effects of TCDD …

Adeno-Associated Virus (Aav) Transduction Of Primary Human Cd4+T Lymphocytes, Sahar Hussein Kamel

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Development of latent HIV-infected CD4+ T lymphocytes is the major cause of HIV treatment failure. Adeno-associated virus (AAV) is an attractive vector for anti-HIV gene therapy due to its lack of pathogenicity, low immunogenicity, and persistent transgene expression. However, a major limitation for AAV gene transfer is the cell-specific tropism of each serotype. Only AAV serotypes 2 and 5 have been investigated in hematopoietic cells, both of these serotypes have shown low transduction efficiency. Since each of the currently described AAV serotypes demonstrate distinct tissue tropism, I hypothesized that other serotypes such as AAV1, 4, 8, or 9 may demonstrate …

Expression Of The Alpha, Beta, And Gamma Subunits Of The Interleukin-2 Receptor By Human Vascular Smooth Muscle Cells, Sultan Mohammed Alhayyani

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Interleukin 2 (IL-2) is a member of the cytokine family and contributes to the proliferation, survival, and death of lymphocytes [1]. The interleukin-2 receptor (IL-2) is a tripartite receptor commonly expressed on the surfaces of many lymphoid cells and is composed of three non-covalently associated subunits, alpha (CD25), beta (CD122), and gamma (CD132) [2]. Our laboratory has previously described IL-2 receptor Beta expression by vasculature smooth muscle cells (VSMC) in mice and humans [3]. The current work expands our previous observations by assessing the expression of the alpha and gamma subunits of the IL-2R by VSMCs. Analysis of IL-2R expression …

Construction Of A Live-Attenuated Hiv-1 Vaccine Through Genetic Code Expansion, Nanxi Wang, Yue Li, Wei Niu, Ming Sun, Ronald Cerny, Qingsheng Li, Jiantao Guo

Qingsheng Li Publications

A safe and effective vaccine against human immunodeficiency virus type 1 (HIV-1) is urgently needed to combat the worldwide AIDS pandemic, but still remains elusive. The fact that uncontrolled replication of an attenuated vaccine can lead to regaining of its virulence creates safety concerns precluding many vaccines from clinical application. We introduce a novel approach to control HIV-1 replication, which entails the manipulation of essential HIV-1 protein biosynthesis through unnatural amino acid (UAA*)-mediated suppression of genome-encoded blank codon. We successfully demonstrate that HIV-1 replication can be precisely turned on and off in vitro.

Includes supporting information.

Glut 4 And Insulin Resistance, Salmin Ali

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Five types of glucose transporters stimulate the transport of glucose from the bloodstream into cells such as muscle cells and adipocytes in response to insulin signaling or exercise. Skeletal muscle and adipose tissue are the main sites for glucose uptake and dispose in insulin action dependent pathway. In non-insulin stimulated condition, the GLUT4 storage vesicles stay away from the plasma membrane, so there is no GLUT4 translocation or GLUT4 mediated glucose uptake. However, in insulin stimulated state, insulin initiates insulin signaling cascade which results in rapid GLUT4 translocation and increased glucose uptake at the cell surface. Failure of the translocation …

Ticks And Spotted Fever Group Rickettsiae Of Southeastern Virginia, Robyn M. Nadolny, Chelsea L. Wright, Daniel E. Sonenshine, Wayne L. Hynes, Holly D. Gaff

Biological Sciences Faculty Publications

The incidence of tick-borne rickettsial disease in the southeastern United States has been rising steadily through the past decade, and the range expansions of tick species and tick-borne infectious agents, new and old, has resulted in an unprecedented mix of vectors and pathogens. The results of an ongoing 4-year surveillance project describe the relative abundance of questing tick populations in southeastern Virginia. Since 2009, more than 66,000 questing ticks of 7 species have been collected from vegetation in a variety of habitats, with Amblyomma americanum constituting over 95% of ticks collected. Other species represented included Ixodes scapularis, Dermacentor variabilis …

Single-Tube Real-Time Pcr Assay For Differentiation Of Ixodes Affinis And Ixodes Scapularis, Chelsea L. Wright, Wayne L. Hynes, Breanna T. White, Mindy N. Marshall, Holly D. Gaff, David T. Gauthier

Biological Sciences Faculty Publications

Ixodes affinis Neumann (1899) and Ixodes scapularis Say (1821) are tick vectors of the etiologic agent of Lyme disease, Borrelia burgdorferi sensu stricto. Ixodes affinis and I. scapularis are morphologically very similar, and as they are sympatric in the mid- and south-Atlantic U.S. coastal states, their accurate identification is crucial to studies of disease and vector ecology in this area. This work describes a rapid, single-tube SYBR® Green-based real-time PCR assay for differentiation of I. affinis and I. scapularis at all life stages. The assay employs 2 pairs of species specific primers directed against the internal transcribed spacer 2 (ITS2) …

Prevalence Of Ehrlichia Chaffeensis And Ehrlichia Ewingii In Amblyomma Americanum And Dermacentor Variabilis Collected From Southeastern Virginia, 2010-2011, Chelsea L. Wright, Holly D. Gaff, Wayne L. Hynes

Biological Sciences Faculty Publications

Amblyomma americanum is the most commonly-encountered tick species in southeastern Virginia, representing approximately 95% of the human-biting tick population in this area. Here we investigated the prevalence of Ehrlichia chaffeensis and Ehrlichia ewingii in questing Amblyomma americanum and Dermacentor variabilis ticks collected from multiple sites in southeastern Virginia from 2010–2011. Although both Ehrlichia species were detected in Amblyomma americanum, no evidence of either pathogen was found in Dermacentor variabilis. Prevalence of E. chaffeensis varied by location, ranging from 0 – 5.08% among Amblyomma americanum populations. Ehrlichia ewingii prevalence was slightly higher, ranging from 0 – 8.20% among A. …

Characterizing Dsrna Production In Virus-Infected Fish Cells, Amal Brek Aloufi

Theses and Dissertations (Comprehensive)

Viral dsRNA is produced by almost all viruses sometime during their replicative cycle. These viral nucleic acids are potent inducers of both innate and adaptive immune responses, and are therefore considered important immuno-modulators. Previous studies have shown that viruses produce dsRNA when replicating in mammalian cells; however, to date no one has demonstrated viral dsRNA production in virus infected fish cells. Therefore, the goal of this study is to investigate dsRNA production by fish viruses in fish cells, verifying production and performing initial characterization of the dsRNA molecules being produced. Three different rainbow trout cell lines were used in this …

C-Jun N-Terminal Kinases Regulate Adenovirus-Mediated Autophagy And Antigen Presentation, Sarah R. Klein

Dissertations and Theses (Open Access)

Targeted immunotherapy with recombinant, oncolytic adenoviruses is under investigation for the treatment of cancer. Evidence indicates adenoviruses induce autophagy that is required for oncolysis, but the molecular regulation of autophagy in infected cells remains under investigation. Our data suggested the canonical pathway regulating starvation-induced autophagy was not implemented in adenovirus-induced autophagy; however, adenovirus infection triggered phosphorylation of c-Jun N-terminal kinases (JNK) that was essential for autophagy. Adenoviral replication within the host cell elicited JNK pathway activation leading to B cell lymphoma-2 (Bcl-2) phosphorylation. JNK-dependent Bcl-2 phosphorylation stimulated the dissociation of Bcl-2/beclin 1 heterodimers, enabling beclin 1 to initiate autophagy. Moreover, …

Barrier To Autointegration Factor (Baf) Inhibits Vaccinia Virus Intermediate Transcription In The Absence Of The Viral B1 Kinase, Nouhou Ibrahim, April Wicklund, Augusta Jamin, Matthew S. Wiebe

Nebraska Center for Virology: Faculty Publications

Barrier to autointegration factor (BAF/BANF1) is a cellular DNA-binding protein found in the nucleus and cytoplasm. Cytoplasmic BAF binds to foreign DNA and can act as a defense against vaccinia DNA replication. To evade BAF, vaccinia expresses the B1 kinase, which phosphorylates BAF and blocks its ability to bind DNA. Interestingly, B1 is also needed for viral intermediate gene expression via an unknown mechanism. Therefore, we evaluated the impact of B1-BAF signaling on vaccinia transcription. Strikingly, the decrease in vaccinia transcription caused by loss of B1 can be rescued by depletion of BAF. The repressive action of BAF is greatest …

A Supramolecular Strategy To Assemble Multifunctional Viral Nanoparticles, Limin Chen, Xia Zhao, Yuan Lin, Yubin Huang, Qian Wang

Faculty Publications

Using a one-pot approach driven by the supramolecular interaction between β-cyclodextrin and adamantyl moieties, multifunctional viral nanoparticles can be facilely formulated for biomedical applications.

Coccidia Of Gerbils From Mongolia, Ethan Thomas Jensen

School of Biological Sciences: Dissertations, Theses, and Student Research

In this study, gerbils collected in the Mongolia over the summers of 2009-2012 were examined for coccidia. In total, 171 gerbils of three species from 22 localities were examined for coccidia. Coccidian oocysts were identified from 21 gerbils, but those found in 1 of those gerbils were probably pseudoparasites of the host from which they were recovered. From the remaining 20 gerbils, 7 morphotypes of Eimeria and 1 morphotype of Isospora were identified. Four of the 7 morphotypes of Eimeria were attributed to new species which were described in this study. In addition, 10 previously described species of Eimeria were …

Characterization Of The Gut Microbiota And Colitogenic Bacterial Species In Core 1 O-Glycans Deficient Mice, Maria E. Perez-Munoz

Department of Food Science and Technology: Dissertations, Theses, and Student Research

Mucus is one important component of the intestinal mucosal barrier, and loss of its functionality is associated with colitis in humans and mice. Mice deficient in core 1 O-glycans (TM-IEC C1galt1^-/-) display a thinner mucus layer upon mutation of the glycosyltransferase enzyme (C1GALT1) responsible for core 1 O-glycans attachment, thus constituting an experimental model for human Ulcerative Colitis (UC). Under conventional conditions, these mice experience spontaneous inflammation of the colon. However, the exact role of the microbiota in colitis development in these mice has not been systematically investigated.

Aimed to gain insights into the role of …

Neuropathogenic Escherichia Coli K1 Does Not Exhibit Proteolytic Activities To Exert Its Pathogenicity, Junaid Iqbal, Mehak Rajan, Ruqaiyyah Siddiqu, Naveed Ahmed Khan

Department of Biological & Biomedical Sciences

Background: Proteases are well-known virulence factors that promote survival, pathogenesis and immune evasion of many pathogens. Several lines of evidence suggest that the blood-brain barrier permeability is a prerequisite in microbial invasion of the central nervous system. Because proteases are frequently associated with vascular permeability by targeting junctional proteins, here it is hypothesized that neuropathogenic Escherichia coli K1 exhibit proteolytic activities to exert its pathogenicity.
Methods: Zymographic assays were performed using collagen and gelatin as substrates. The lysates of whole E. coli K1 strain E44, or E. coli K-12 strain HB101 were tested for proteolytic activities. The conditioned media were …

Analysis Of The Chondroitinase Operon Of Flavobacterium Columnare, Erin L. Sorlien

Senior Honors Projects

Analysis of the chondroitinase operon of Flavobacterium columnare

Erin Sorlien

Major

Cell and Molecular Biology, Chemistry

Advisor

Dr. David R. Nelson

Date

May 2, 2013

Keywords

Flavobacterium columnare, columnaris disease, chondroitin AC lyase, complementation, csl operon

Abstract
Flavobacterium columnare, an opportunistic bacterial pathogen of fish, is the causative agent of columnaris disease (CD). The bacterium is a Gram-negative rod that exhibits gliding motility and avidly forms biofilms. CD affects both wild and cultured freshwater fish, and continues to cause large economic losses to the fish farming industry. According to an investigation conducted by the National Animal Health Monitoring System, CD …

Interaction Of Bacillus Anthracis Exosporium Protein Bcla With Complement Factor H And Spore Persistence In The Lung, Sarah A. Jenkins

Dissertations and Theses (Open Access)

Anthrax outbreaks in the United States and Europe and its potential use as a bioweapon have made Bacillus anthracis an interest of study. Anthrax infections are caused by the entry of B. anthracis spores into the host via the respiratory system, the gastrointestinal tract, cuts or wounds in the skin, and injection. Among these four forms, inhalational anthrax has the highest lethality rate and persistence of spores in the lungs of animals following pulmonary exposure has been noted for decades. However, details or mechanisms of spore persistence were not known. In this study, we investigated spore persistence in a mouse …

Enrollment In Yfv Vaccine Trial: An Evaluation Of Recruitment Outcomes Associated With A Randomized Controlled Double-Blind Trial Of A Live Attenuated Yellow Fever Vaccine, Paula M. Frew, Eve T. Shapiro, Lu Lu, Srilatha Edupuganti, Harry L. Keyserling, Mark J. Mulligan

Environmental & Global Health Faculty Research

This investigation evaluated several factors associated with diverse participant enrollment of a clinical trial assessing safety, immunogenicity, and comparative viremia associated with administration of 17-D live, attenuated yellow fever vaccine given alone or in combination with human immune globulin. We obtained baseline participant information (e.g., sociodemographic, medical) and followed recruitment outcomes from 2005 to 2007. Of 355 potential Yellow Fever vaccine study participants, 231 cases were analyzed. Strong interest in study participation was observed among racial and ethnically diverse persons with 36.34% eligible following initial study screening, resulting in 18.75% enrollment. The percentage of white participants increased from 63.66% (prescreened …

Novel Neutralizing Antibody Assays For Recombinant Human Hookworm Na-Gst-1 Vaccine, Xi Chen, Brian Keegan, Peter J. Hotez, Jeffrey M. Bethony, Amar R. Jariwala

GW Research Days 2013

BACKGROUND

Necator Americanus, a human hookworm causes approximately 85% of the global hookworm infections. Hookworm ingest hemoglobin containing erythrocytes. Hemoglobin is further digested to Heme and Globin by hookworm's gut enzymes. Iron-containing Heme is a potent enzyme inhibitor and generates toxic reactive oxygen species which is toxic to hookworms. Hookworm's gut enzyme Na-GST-1 (Necator Americanus Glutathione S-Transferase-1) has been hypothesized to detoxify Heme. Na-GST-1 adjuvanted with Alhydrogel® is a new vaccine which is currently under clinical development. Na-GST-1 has two active sites, the ligand binding or Heme detoxification site (H-site) and the catalytic active glutathione binding …

Phi X 174, David J. Hall

Infectious Pathogens

Phi X 174 database number # 1CD3. Phi X 174 is a virus that infects bacteria and was the first DNA-based genome to be sequenced in 1977. This bacteriophage has a [+] circular single-stranded DNA genome. The assembly of the capsid has been successfully done in vitro and is the focus of many studies on self-assembly, drug delivery and nanotechnology.

Dengue Virus, David J. Hall

Infectious Pathogens

Dengue virus, database # 1THD Dengue fever, is an infectious tropical disease caused by the dengue virus and transmitted by mosquito. Symptoms include fever, headache, muscle and joint pains, and a characteristic skin rash that is similar to measles. In a small proportion of cases the disease develops into the life-threatening dengue hemorrhagic fever. Dengue is a positive strand RNA virus that is part of the flavivirus family. Dengue is an enveloped virus meaning it has a nucleocapsid protecting the RNA and the nucleocapsid is surrounded by lipids (hence the name enveloped).

Human Papilloma Virus 16, David J. Hall

Infectious Pathogens

Human Papilloma virus 16, database# 1DZL Infection by most papillomaviruses is either asymptomatic or causes small benign tumors, known as papillomas or warts (e.g. human papillomavirus HPV6 or HPV11). Papillomas caused by some types, however, such as human papillomaviruses 16 and 18, carry a risk of becoming cancerous. Human papilloma virus is a non-enveloped, double stranded, circular DNA virus.

Hepatitis B, David J. Hall

Infectious Pathogens

Hepatitis B, database # 2g33. Hepatitis B is an infectious inflammatory illness of the liver caused by the hepatitis B virus (HBV). Hepatitis B virus (HBV) is a member of the Hepadnavirus family.The virus particle, (virion) consists of an outer lipid envelope and an icosahedral nucleocapsid core composed of protein. These virions are 42 nM in diameter. The nucleocapsid encloses the viral DNA and a DNA polymerase that has reverse transcriptase activity.

Equine Serum Antibody Responses To Streptococcus Equi And Streptococcus Zooepidemicus, Rafaela De Negri

Theses and Dissertations--Veterinary Science

Streptococcus zooepidemicus (Sz) and Streptococcus equi (Se) share 98% DNA sequence homology, but display different pathogenic properties. Infection by one organism does not cross-protect against the other. To better understand pathogenic differences between these organisms and gain information about which proteins are expressed in horses infected experimentally with Se, intrauterine Sz or naturally with respiratory Sz we compared antibody specificities of convalescent sera using ELISA. These comparisons were based on sets of 8 and 14 immunoreactive recombinant proteins of Se strain CF32 and Sz strain NC78, respectively. Sera from donkeys that were previously naturally affected with strangles and later developed …

Hypoxia Inducible Factor 1 Alpha (Hif-1a): A Major Regulator Of Placental Development, Renee E. Albers

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Hypoxia-inducible factor-1 alpha (HIF-1a) is a critical component of the cellular oxygen-sensing machinery and is essential for placental formation and embryonic survival. In this study, we promoted prolonged expression of HIF-1a, by using a form that is insensitive to oxygen, denoted as CA-HIF-1a. In order to have continual placental specific expression of the CA-HIF-1a, lentiviral infection was performed on embryos at the blastocyst stage of development and transferred into pseudo-pregnant mothers. Placental analysis was performed via in situ hybridization on embryonic day 14.5 (E14.5) to determine the effects of CA-HIF-1a prolonged expression. Data indicate that prolonged activity of CA-HIF-1a restricted …

Validation Of N-Myristoyltransferase As Potential Chemotherapeutic Target In Chagas Disease, Linda Herrera

Open Access Theses & Dissertations

The protozoan parasite Trypanosoma cruzi is the etiological agent of Chagas disease, a problem endemic to Central and South America. In recent years, this neglected infectious disease has become a global health concern. The only clinically available drugs for the chemotherapy of Chagas disease have important disadvantages such as, high toxicity, strain resistance and variable efficacy. Therefore, there is an urgent need for the development of new antichagasic agents. The enzyme N-myristoyltransferase (NMT) has been characterized in a range of eukaryotes, from Saccharomyces cerevisae to Homo sapiens. Moreover, NMT has been shown to be essential in protozoan parasites, including Leishmania …

Type I Interferon Activation Of Natural Killer (Nk) Cells By Cytomegalovirus (Cmv) And Their Interaction With Dendritic (Dc) And Nkt Cells, Howaida A. Oulad Abdelati

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In our current study the roles of natural killer (NK) cells in regulation of the acute phase of murine cytomegalovirus (MCMV) infection were demonstrated. NK cells utilize perforin and gamma interferon to kill MCMV infected cells. Activation of NK cells is controlled by the balance between inhibitory and activation receptors and modulated by cytokines produced from the infected cells. Type I interferons are produced mainly by plasmacytoid dendritic cells (PDCs). Type I interferons have a substantial role in enhancing NK cells cytotoxic activity and NK cells proliferation and maintain cell survival through STAT mediated signaling pathway. IL-12 produced by dendritic …