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Deletion Of The Human Cytomegalovirus Us17 Gene Increases The Ratio Of Genomes Per Infectious Unit And Alters Regulation Of Immune And Endoplasmic Reticulum Stress Response Genes At Early And Late Times After Infection, Stephen James Gurczynski, Subhendu Das, Philip E. Pellett

Immunology and Microbiology Faculty Research Publications

Human cytomegalovirus (HCMV) employs numerous strategies to combat, subvert, or co-opt host immunity. One evolutionary strategy for this involves capture of a host gene and then its successive duplication and divergence, forming a family of genes, many of which have immunomodulatory activities. The HCMV US12 family consists of 10 tandemly arranged sequence-related genes in the unique short (US) region of the HCMV genome (US12 to US21). Each gene encodes a protein possessing seven predicted transmembrane domains, patches of sequence similarity with cellular G-protein-coupled receptors, and the Bax inhibitor 1 family of antiapoptotic proteins. We show that one member, US17, plays …

Role Of Peripheral Immune Response In Microglia Activation And Regulation Of Brain Chemokine And Proinflammatory Cytokine Responses Induced During Vsv Encephalitis, Christina D. Steel, Kimberly Breving, Susan Tavakoli, Woong-Ki Kim, Larry D. Sanford, Richard P. Ciavarra

Biological Sciences Faculty Publications

We report herein that neuroinvasion by vesicular stomatitis virus (VSV) activates microglia and induces a peripheral dendritic cell (DC)-dependent inflammatory response in the central nervous system (CNS). VSV neuroinvasion rapidly induces multiple brain chemokine and proinflammatory cytokine mRNAs that display bimodal kinetics. Peripheral DC ablation or T cell depletion suppresses the second wave of this response demonstrating that infiltrating T cells are primarily responsible for the bimodal characteristics of this response. The robust infiltrate associated with VSV encephalitis likely depends on sustained production of brain CCL19 and CCR7 expression on infiltrating inflammatory cells. © 2013 Elsevier B.V. All rights reserved.

Candida Albicans Als5p Amyloid In Host-Microbe Interactions: A Ceanorhabditis Elegans Study, Michael Bois

Dissertations, Theses, and Capstone Projects

Candida albicans, a dimorphic fungus and an opportunistic pathogen, possesses a myriad of adherence factors including members of the agglutinin-like sequence (Als) family of mannoproteins. The adhesin Als5p mediates adhesion to many substrates, and is upregulated during commensal interactions, but is downregulated during active C. albicans infections[1]. An amyloid forming core sequence at residues 325-331 has been shown to be important for Als5p function, because a single amino acid substitution at position 326 (V326N) greatly reduces Als5p-mediated adherence[2]. We evaluated the role of Als5p in host-microbe interactions, using Caenorhabditis elegans as a host model and feeding them Saccharomyces cerevisiae expressing …

Acanthamoeba And Bacteria Produce Antimicrobials To Target Their Counterpart, Junaid Iqbal, Ruqaiyyah Siddiqui, Naveed Ahmed Khan

Department of Biological & Biomedical Sciences

Background

In the microbial ecosystem, microbes compete for space and nutrients. Consequently, some have developed the ability to kill or inhibit the growth of other competing microbes by producing antimicrobial substances. As the ‘producer’ species are generally immune to these substances, their compounds act on the competing microbial species and give the producer more space and access to nutrients for growth. Many currently used antibiotics were developed by exploiting this potential of certain microbes.

Findings

Here, the free-living amoeba, Acanthamoeba castellanii, was investigated for its antibacterial activity against representative Gram positive and Gram negative bacteria, while bacterial isolates were …

Preventing Salmonella And Other Bacterial Contamination In Poultry Using A High-Adhering Strain Of Escherichia Coli, Jeffrey P. North, Dr. Ron Leavitt

Journal of Undergraduate Research

In 1928 Alexander Fleming revolutionized the world with the discovery of penicillin. Penicillin and the antibiotics discovered thereafter revolutionized the treatment of infectious diseases. Equipped with these miracle drugs, scientists thought the battle against pathogenic bacteria was won. However, bacteria began to evolve to meet the antibiotic challenge. As the 20th century progressed, more and more bacteria developed resistance to one or more antibiotics. This resistance is achieved through mutation or through horizontal gene transfer, when one bacterium transfers the gene to another bacterium.

Hiv In The Immune System: The Role Of Specialized Follicular Dendritic Cells In Promoting Viral Disease, Jacob D. Estes, Dr. Gregory F. Burton

Journal of Undergraduate Research

Human Immunodeficiency Virus (HIV) has become a world wild epidemic affecting millions of people across the globe. Even in the face of potent antiviral drugs which are only available in first world countries, HIV continues to evade intervention strategies and eventually destroy the body’s immune system. During the longest stage of HIV disease, clinical latency, much of active infection is confined to the germinal centers of secondary lymphoid tissue (1). Here HIV is trapped on the surface of follicular dendritic cells (FDC) which reside in this region of all secondary lymphoid organs (2). Large amounts of HIV localize on FDC …

Phages Of Non-Diary Lactococci: Isolation And Characterization Of Phi L47, A Phage Infecting The Grass Isolate Lactococcus Lactis Ssp Cremoris Dpc6860, Daniel Cavanagh, Caitríona M. Guinane, Horst Neve, Aidan Coffey, R. Paul Ross, Gerald F. Fitzgerald, Olivia Mcauliffe

Department of Biological Sciences Publications

Lactococci isolated from non-dairy sources have been found to possess enhanced metabolic activity when compared to dairy strains. These capabilities may be harnessed through the use of these strains as starter or adjunct cultures to produce more diverse flavor profiles in cheese and other dairy products. To understand the interactions between these organisms and the phages that infect them, a number of phages were isolated against lactococcal strains of non-dairy origin. One such phage, ΦL47, was isolated from a sewage sample using the grass isolate L. lactis ssp. cremoris DPC6860 as a host. Visualization of phage virions by transmission electron …

Immunologic Special Forces: Anti-Pathogen Cytotoxic T-Lymphocyte Immunotherapy Following Hematopoietic Stem Cell Transplantation, Michael Keller, Catherine M. Bollard

Pediatrics Faculty Publications

Anti-pathogen adoptive T-cell immunotherapy has been proven to be highly effective in preventing or controlling viral infections following hematopoietic stem cell transplantation. Recent advances in manufacturing protocols allow an increased number of targeted pathogens, eliminate the need for viral transduction, broaden the potential donor pool to include pathogen-naïve sources, and reduce the time requirement for production. Early studies suggest that anti-fungal immunotherapy may also have clinical benefit. Future advances include further broadening of the pathogens that can be targeted and development of T-cells with resistance to pharmacologic immunosuppression.

Airborne Staphylococcus Aureus In The Biology Labs At Southern Adventist University, Harrison Filler

Capstone Research Projects

Staphylococcus aureus has been of growing interest to the scientific community in recent years. It is not only easily transferred from one person to another, but its rapidly evolving antibiotic resistance has made it increasingly difficult to treat. Once thought to be a hospital-acquired disease, S. aureus is now often found in the community. Studies indicate that people and clothing can act as carriers of S. aureus not only directly, but through airborne particles. For these reasons a research experiment was designed to determine the prevalence of S. aureus in the microbiology lab and anatomy and physiology lab at Southern …

High Prevalence Of Early Childhood Infection By Kshv In A Minority Population In China, Yifei Cao, Veenu Minhas, Xiaohua Tan, Jin Huang, Baodong Wang, Meiyi Zhu, Yuan Gao, Tingting Zhao, Lei Yang, Charles Wood

Nebraska Center for Virology: Faculty Publications

In China, KSHV seroprevalence varies considerably among different regions and ethnicities. But in Xinjiang province, located in the northwestern China, there is a very high seroprevalence of KSHV in adults of Kazak and Ughur ethnicities. However, KSHV prevalence in children and the risk factors associated with the acquisition of infection are currently not known. The aim of this study was to investigate the prevalence of KSHV infection, identify associated socio-economic or behavioral risk factors and the humoral immune response among children in this population. This is a cross-sectional study (N=178) to screen children and their caregivers from Xinjiang for total …

Global Analysis Of Chlorella Variabilis Nc64a Mrna Profiles During The Early Phase Of Paramecium Bursaria Chlorella Virus-1 Infection, Janet M. Rowe, Adrien Jeanniard, James R. Gurnon, Yuannan Xia, David D. Dunigan, James L. Van Etten, Guillaume Blanc

Nebraska Center for Virology: Faculty Publications

The PBCV-1/Chlorella variabilis NC64A system is a model for studies on interactions between viruses and algae. Here we present the first global analyses of algal host transcripts during the early stages of infection, prior to virus replication. During the course of the experiment stretching over 1 hour, about a third of the host genes displayed significant changes in normalized mRNA abundance that either increased or decreased compared to uninfected levels. The population of genes with significant transcriptional changes gradually increased until stabilizing at 40 minutes post infection. Functional categories including cytoplasmic ribosomal proteins, jasmonic acid biosynthesis and anaphase promoting …

Dynamic Attachment Of Chlorovirus Pbcv-1 To Chlorella Variabilis, Irina Agarkova, Brigitte Hertel, Xinzheng Zhang, Les Lane, Alexander Tchourbanov, David D. Dunigan, Gerhard Thiel, Michael G. Rossmann, James L. Van Etten

Nebraska Center for Virology: Faculty Publications

Chloroviruses infect their hosts by specifically binding to and degrading the cell wall of their algal hosts at the site of attachment, using an intrinsic digesting enzyme(s). Chlorovirus PBCV-1 stored as a lysate survived longer than virus alone, suggesting virus attachment to cellular debris may be reversible. Ghost cells (algal cells extracted with methanol) were used as a model to study reversibility of PBCV-1 attachment because ghost cells are as susceptible to attachment and wall digestion as are live cells. Reversibility of attachment to ghost cells was examined by releasing attached virions with a cell wall degrading enzyme extract. The …

Genetic Variation And Possible Mechanisms Driving The Evolution Of Worldwide Fig Mosaic Virus Isolates, Jeewan Jyot Walia, Anouk Willemsen, Eminur Elci, Kadriye Caglayan, Bryce W. Falk, Luis Rubio

Nebraska Center for Virology: Faculty Publications

Fig mosaic virus (FMV) is a multipartite negative-sense RNA virus infecting fig trees worldwide. FMV is transmitted by vegetative propagation and grafting of plant materials, and by the eriophyid mite Aceria ficus. In this work, the genetic variation and evolutionary mechanisms shaping FMV populations were characterized. Nucleotide sequences from four genomic regions (each within the genomic RNAs 1, 2, 3, and 4) from FMV isolates from different countries were determined and analyzed. FMV genetic variation was low, as is seen for many other plant viruses. Phylogenetic analysis showed some geographically distant FMV isolates which clustered together, suggesting long-distance migration. …

Risk Factors For Early Childhood Infection Of Human Herpesvirus-8 In Zambian Children: The Role Of Early Childhood Feeding Practices, Kay L. Crabtree, Janet M. Wojcicki, Veenu Minhas, David R. Smith, Chipepo Kankasa, Charles D. Mitchell, Charles Wood

Nebraska Center for Virology: Faculty Publications

Background—Human herpesvirus-8 (HHV-8) infection in early childhood is common throughout sub-Saharan Africa with prevalence increasing throughout childhood. Specific routes of transmission have not been clearly delineated, though HHV-8 is present in high concentrations in saliva.

Methods—To understand the horizontal transmission of HHV-8 within households to children we enrolled for cross-sectional analysis, 251 households including 254 children, age two and under, in Lusaka, Zambia. For all children, plasma was screened for HHV-8 and HIV-1 and health and behavioral questionnaires were completed. Multi-level logistic regression analysis was conducted to assess independent factors for HHV-8 infection in children.

Results—Risk factors for HHV-8 infection …

Turnip Crinkle Virus Coat Protein Inhibits The Basal Immune Response To Virus Invasion In Arabidopsis By Binding To The Nac Transcription Factor Tip, Teresa Donze, Feng Qu, Paul Twigg, T. Jack Morris

Nebraska Center for Virology: Faculty Publications

Turnip crinkle virus (TCV) has been shown to interact with a NAC transcription factor, TIP, of Arabidopsis thaliana, via its coat protein (CP). This interaction correlates with the resistance response manifested in TCV-resistant Arabidopsis ecotype Di-17. We report that failure of a mutated CP to interact with TIP triggered the corresponding TCV mutant (R6A) to cause more severe symptoms in the TCV-susceptible ecotype Col-0. We hypothesized that TCV regulates antiviral basal immunity through TIP-CP interaction. Consistent with this hypothesis, we found that the rate of accumulation of R6A was measurably slower than wild-type TCV over the course of an …

Characterization Of Bacteriophages From Environmental Water Samples And The Potential Of Bacteriophages Tailspike Proteins (Tsp) In Bacteria Detection, Gayathri Upeksha Gunathilaka

Wayne State University Theses

ABSTRACT

CHARACTERIZATION OF BACTERIOPHAGES FROM ENVIRONMENTAL WATER SAMPLES AND THE POTENTIAL OF BACTERIOPHAGES TAILSPIKE PROTEINS (TSP) IN BACTERIA DETECTION

by

GAYATHRI UPEKSHA GUNATHILAKA

May 2014

Advisor: Dr. Yifan Zhang

Major: Nutrition and Food Science

Degree: Master of Science

The high abundance of phage in the environment and their specificity with the bacteria host make them an excellent tool for bacteria detection. To characterize phage specific to Salmonella and E. coli from water samples in the Southeastern Michigan and to develop a novel bacterial detection method using phage, environmental water samples were collected from rivers and waste water treatment plant (WWTP) …

Novel Partner Notification Approaches In New York State, And Antibiotic Resistance In The Management Of Chlamydia Trachomatis Infections, Christopher Frank Davis

Legacy Theses & Dissertations (2009 - 2024)

Chlamydia trachomatis (chlamydia) is the most frequently reported notifiable disease in the United States. This dissertation provides an evaluation of chlamydia treatment and control through three different lenses: 1) patient views on a novel strategy to facilitate treatment, 2) student perceptions about materials to promote treatment of sexual partners, and 3) evidence regarding recurrent infection etiology.

Screening Of Natural Antimicrobial Agents And Antimicrobial-Resistant Bacteria Using A Soleris System, Mingyang Huang

Wayne State University Theses

ABSTRACT

SCREENING OF NATURAL ANTIMICROBIAL AGENTS AND ANTIMICROBIAL - RESISTANT BACTERIA USING A SOLERIS SYSTEM

by

MINGYANG HUANG

May 2014

Advisor: Dr. Yifan Zhang

Major: Food Science and Nutrition

Degree: Master of Science

Traditional broth micro-dilution method is a common assay of measuring Minimum Inhibitory Concentration (MIC) to determine the antimicrobial activity of an antimicrobial agent. However, this method is generally time-consuming and labor intensive. Alternatively, an automated optical method using the Soleris system was applied in this study. The system was compared to the traditional broth micro-dilution 96-well assay to test the antimicrobial activity of 11 essential oils and …

Investigating The Interactions Between Cyanobacteria And Vibrio Parahaemolyticus, Caroline E. Ward

Honors Theses and Capstones

One well-known pathogen that has been the topic of many recent studies is Vibrio parahaemolyticus, which causes thousands of foodborne illnesses a year, mostly from the ingestion of raw or undercooked oysters. It has been shown cyanobacteria can act as a long-term reservoir of Vibrio cholerae, another pathogenic Vibrio, by encasing the cells within mucilaginous sheaths during which Vibrios enter a viable but non-culturable state. In this study we investigated the interaction of V. parahaemolyticus with cyanobacteria to determine whether cyanobacteria aid in the longevity and survival of V. parahaemolyticus. We found that non-pathogenic V. parahaemolyticus …

Differentiation Of U-937 Monocytes To Macrophage-Like Cells Polarized Into M1 Or M2 Phenotypes According To Their Specific Environment: A Study Of Morphology, Cell Viability, And Cd Markers Of An In Vitro Model Of Human Macrophages, Fatma Husien S. Abdulhadi

Browse all Theses and Dissertations

In this study, the human leukemic monocyte lymphoma cell line U-937 was used as an in vitro model for monitoring monocyte/macrophage differentiation. Phorbol 12- myristate (13) (PMA) was used to activate U-937 cells into macrophage-like cells (M0). After 24 hours of PMA treatment, non- adherent U-937 cells became tightly adherent to the culture plates forming M0 cells. M0 cells were then polarized into the M1 macrophage phenotype by treatment with LPS and IFN-γ for another 24 hours. Each of the cytokines IL-4, IL-13, or IL-10 was applied separately to three M0 cultures for 24 hours to induce the M2 macrophage …

Different Meal, Same Flavor: Cospeciation And Host Switching Of Haemosporidian Parasites In Some Non-Passerine Birds, Diego Santiago-Alarcon, Adriana Rodríguez-Ferraro, Patricia Parker, Robert Ricklefs

Biology Department Faculty Works

BackgroundPrevious studies have shown that haemosporidian parasites (Haemoproteus(Parahaemoproteus) and Plasmodium) infecting passerine birds have an evolutionary history of host switching with little cospeciation, in particular at low taxonomic levels (e.g., below the family level), which is suggested as the main speciation mechanism of this group of parasites. Recent studies have characterized diverse clades of haemosporidian parasites (H. (Haemoproteus) and H. (Parahaemoproteus)) infecting non-passerine birds (e.g., Columbiformes, Pelecaniiformes). Here, we explore the cospeciation history of H. (Haemoproteus) and H. (Parahaemoproteus) parasites with their non-passerine hosts.MethodsWe sequenced the mtDNA cyt b gene of both haemosporidian parasites and their avian non-passerine hosts. We …

Role Of Viral And Host Factors In Influenza Virus Mediated Inhibition Of Interleukin-23, Ashish Tiwari

Theses and Dissertations--Veterinary Science

Influenza virus is one of the major respiratory pathogens of humans as well as animals, including equines. There is an increasing evidence that bacterial infections are the most common cause of the death during influenza. In horses also, secondary bacterial pneumonia can lead to death, and surviving horses may take up to six months for the complete recovery resulting in heavy economic loss to the equine industry. Interleukin (IL)-23 mediated innate immune response has been shown to protect the host from various respiratory bacterial infections. However, studies to investigate the role of host and viral factors in the regulation of …

Identification And Ss-Lactam Resistance In Aquatic Isolates Of Enterobacter Cloacae And Their Status In Microbiota Of Domica Cave In Slovak Karst (Slovakia), Barbora Gaalova, Alena Donauerova, Milan Seman, Helena Bujdakova

International Journal of Speleology

Domica Cave is located in the Slovak Karst National Park in southern Slovakia. Heterotrophic cultivable psychrophilic and mesophilic microbiota were confirmed to be prevalent in this cave. Escherichia coli was the most abundant bacterium, followed by Enterobacter species and Serrratia species. Enterobacter cloacae isolates belong to the group of faecal contaminants (coliforms) of concern in water. Their status in cave ecosystem is questionable, but we observed E. cloacae in water samples all years round suggesting an autochthonous origin. We were concerned with possible contamination of the cave water with resistant E. cloacae from animal farms located 2-3 km from the …

Virus Purification, Detection And Removal, Khrupa Saagar Vijayaragavan

Dissertations, Master's Theses and Master's Reports - Open

The biopharmaceutical industry has a growing demand and an increasing need to improve the current virus purification technologies, especially as more and more vaccines are produced from cell-culture derived virus particles. Downstream purification strategies can be expensive and account for 70% of the overall manufacturing costs. The economic pressure and purification processes can be particularly challenging when the virus to be purified is small, as in our model virus, porcine parvovirus (PPV). Our efforts are focused on designing an easy, economical, scalable and efficient system for virus purification, and we focused on aqueous two-phase systems. Industry acceptable standards for virus …

Role Of The Pkna And Pknb Kinases In Mycobacterium Tuberculosis, Tripti Anandan

Wayne State University Dissertations

To respond to environmental changes, M. tuberculosis possesses eleven "eukaryotic-type" Ser/Thr protein kinases. The aim of the study described in this dissertation was to identify role of two of these kinases; PknA and PknB that are essential in M. tuberculosis. Two approaches are described to screen for potential in vivo substrates of PknA/PknB. First approach is based on proteomic search by over-expressing PknA/PknB in M. tuberculosis. Proteomic search led to identification of proteasome to be a substrate of PknA and PknB in M. tuberculosis. Furthermore, I demonstrate that the phosphorylation of PrcA and PrcB by PknA regulates processing of Pre-PrcB. …

Characterisation Of The Enzybiotic Potential Of Selected Staphylococcus Aureus Bacteriophages, Ruth Keary

Theses

The work presented in this thesis focuses on harnessing the bactericidal potential of bacteriophage (phage) encoded peptidoglycan hydrolase enzymes to combat the significant pathogen Staphylococcus aureus. In order to identify novel peptidoglycan hydrolases lytic against S. aureus, the complete genomes of two staphylococcal temperate Siphoviridae bacteriophages, DW2 and CSl, were sequenced and annotated. Screening of these genomes identified genes for several putative peptidoglycan hydrolases including two endolysins and three virion-associated peptidoglycan hydrolases. The identification of putative genes relating to virulence and host fitness lends to the understanding of the role played by such temperate phages in the evolution of this …

Socs1/Socs3 Expression And Virus Replication Of Denv2 And Hsv-1 In Cytokine-Polarized Subsets Of Raw 264.7 Macrophages, Nagarjuna Reddy Cheemarla

Browse all Theses and Dissertations

Cytokine activation of macrophages leads to "macrophage polarizatio" The M1 subset is polarized by exposure to lipopolysaccharide (LPS) and interferon-gamma (IFN-γ) and M2 polarized cells develop after exposure to interleukin-4 (IL-4) or IL-13. In this study, the RAW 264.7 murine macrophage cell line was used to study the effect herpes simplex virus-Type I (HSV-1) and Dengue virus serotype-2 (DENV2) infections have on cultures of unpolarized (M0) or polarized subsets. The macrophage subsets were characterized using cluster of differentiation markers CD14 and CD86. Uninfected M1 macrophages showed up regulation in expression of the co-stimulatory factor B7.2 (CD86) compared to expression by …

Development Of A Co-Culture System To Mimic The Transfection Of Hsv-1 From Keratinocytes To Neuronal Cells, David A. Dixon

Browse all Theses and Dissertations

To more closely mimic the in vivo progression of HSV-1 a novel in vitro method was created. In this study HEL-30 keratinocytes were infected with a 0.1 MOI of HSV-1 and plated unto a membrane with 8μm diameter pores. The membrane insert was then placed into culture wells that contained Neuro-2a cells (N2A). The neurites from the neuronal cells made cell-to-cell contact with the infected HEL-30 cells and the virus was transmitted into the neuronal cells. To determine if infection occurred in the N2A cells the cells were lysed and the lysate incubated with Vero cells to titrate virus plaque …

White Blood Cell Counts, Parasite Prevalence, And Plasma Cortisol Levels Of Dogs In A County Animal Shelter: Changes Over Days And Impact Of A Program Of Repeated Human Interaction, Emily S. Dudley

Browse all Theses and Dissertations

Animal shelter housing is highly stressful for a dog, compromising welfare and leading to undesirable behaviors and unknown health consequences. We documented the changes in circulating numbers of white blood cells, plasma cortisol, and fecal parasite shedding of dogs housed for 10 days at a county animal shelter. White blood cell changes were most prominent on Day 10 after arrival to the shelter. Changes included increased total leukocytes, mature neutrophils, and lymphocytes, with less consistent increases in monocytes and neutrophil to lymphocyte ratio (N:L). Fecal parasite shedding was elevated and not affected by day. Cortisol levels of shelter dogs declined …

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 …