Microbiology Commons^™

Understanding The Molecular Mechanisms Underlying Rsv1 Mediated Resistance To Smv In Soybean, Mohammed Ali Ahmed Eid

Theses and Dissertations--Plant Pathology

Like humans, viral diseases also affect plants. Of these, viruses belonging to the potyvirus genus are the most prolific. The potyvirus soybean mosaic virus (SMV) is an important pathogen of the crop plant soybean. SMV causes mosaic symptoms (yellow areas alternate with dark green areas on the leaves of the plant) and can affect yield by reducing seed quality. Few cultivars from soybean can resist different SMV strains. To understand soybean defense mechanisms to SMV, I identified soybean proteins that interact with the helper component protease (HC-Pro) of SMV, which also functions as the suppressor of host RNA silencing and …

Transcriptomic Analysis Of Staphylococcal Srnas: Insights Into Species-Specific Adaption And The Evolution Of Pathogenesis, William H. Broach, Andy Weiss, Lindsey N. Shaw

Molecular Biosciences Faculty Publications

Next-generation sequencing technologies have dramatically increased the rate at which new genomes are sequenced. Accordingly, automated annotation programs have become adept at identifying and annotating protein coding regions, as well as common and conserved RNAs. Additionally, RNAseq techniques have advanced our ability to identify and annotate regulatory RNAs (sRNAs), which remain significantly understudied. Recently, our group catalogued and annotated all previously known and newly identified sRNAs in several Staphylococcus aureus strains. These complete annotation files now serve as tools to compare the sRNA content of S. aureus with other bacterial strains to investigate the conservation of their sRNomes. Accordingly, in …

Potential Role Of Pctaire-2, Pctaire-3 And P-Histone H4 In Amyloid Precursor Protein-Dependent Alzheimer Pathology, Dale Chaput, Lisa Kirouac, Stanley M. Stevens Jr., Jaya Padmanabhan

Molecular Biosciences Faculty Publications

Amyloid Precursor Protein (APP) is regulated in a mitosis-specific manner and plays a role in proliferative signaling in cells. Though APP-derived Aβ generation has a well-established role in neurodegeneration, the mechanistic role of APP in this process is not fully understood. Here, we performed an unbiased, comprehensive analysis of the phosphoproteome signature in APP-null neuroblastoma cells (B103) compared to those expressing APP-695 isoform (B103-695) to determine if APP expression affects protein phosphorylation. Stable isotope labeling by amino acids in cell culture (SILAC) followed by mass spectrometry-based phosphoproteomic analysis with PolyMAC identified a total of 2,478 phosphopeptides in the B103 and …

Antibodies Against A Secreted Product Of Staphylococcus Aureus Trigger Phagocytic Killing, Lena Thomer, Carla Emolo, Vilasack Thammavongsa, Hwan Keun Kim, Molly E. Mcadow, Wenqi Yu, Matthew Kieffer, Olaf Schneewind, Dominique Missiakas

Molecular Biosciences Faculty Publications

Host immunity against bacteria typically involves antibodies that recognize the microbial surface and promote phagocytic killing. Methicillin-resistant Staphylococcus aureus (MRSA) is a frequent cause of lethal bloodstream infection; however, vaccines and antibody therapeutics targeting staphylococcal surface molecules have thus far failed to achieve clinical efficacy. S. aureus secretes coagulase (Coa), which activates host prothrombin and generates fibrin fibrils that protect the pathogen against phagocytosis by immune cells. Because of negative selection, the coding sequence for the prothrombin-binding D1-D2 domain is highly variable and does not elicit cross-protective immune responses. The R domain, tandem repeats of a 27-residue peptide that bind …

Isolation And Characterization Of Microbial Community Associated With Diadumene Lineata, The Orange-Striped Sea Anemone, Tiffany Lin, Andrew Weir

Student and Faculty Research Days

The orange-striped sea anemone, Diadumene lineata, is a marine invertebrate that can be found in the Northern Hemisphere. Distribution of Diadumene lineata ranges from Japan to the Gulf of Argentina. This invasive anemone originated from the Pacific Coast of Asia and serves as a host for a diverse microbial community including a photosynthetic alga. The epiphytic bacteria adhere to the outer surface as well as to the gastrovascular cavity of the host. The microbial community associated with Diadumene lineata is relatively unknown. This study attempted to isolate and identify bacteria commonly associated with this invasive marine invertebrate. Diadumene lineata was …

Effect Of Maternal Diet On Select Fecal Bacteria In Mares And Their Foals, Morgan Brewer Pyles

Theses and Dissertations--Animal and Food Sciences

The process of microbial colonization in the gastrointestinal tract of foals and the factors influencing this process are not well understood. The objectives were to determine the effect of starch source in pelleted concentrates in the maternal diet on select fecal bacterial groups in mares and their foals and to evaluate changes in these bacterial groups over time. Thoroughbred foals (n=19) were from dams fed one of two concentrates: an oat-based (OB) or corn and wheat middling-based (CWB) pelleted concentrate. Fecal samples were collected from mares at 324 d of gestation, pre-foaling, 1 d, 14 d, and 28 d after …

Giant Chloroviruses: Five Easy Questions, James L. Van Etten, David Dunigan

Nebraska Center for Virology: Faculty Publications

Chloroviruses are large, icosahedral, dsDNA-containing viruses that replicate in certain unicellular, chlorella-like green algae [1,2]. They exist in freshwater throughout the world with titers as high as thousands of plaque-forming units (PFU) per ml of indigenous water although titers are typically 1–100 PFU/ml. Titers fluctuate during the year with the highest titers typically occurring in the spring and late fall. Known chlorovirus hosts, which are normally symbionts and are often referred to as zoochlorellae, are associated with either the protozoan Paramecium bursaria (Fig 1A), the coelenterate Hydra viridis, or the heliozoan Acanthocystis turfacea. Zoochlorellae are resistant to viruses …

Genetic Barrier To Direct Acting Antivirals In Hcv Sequences Deposited In The European Databank, Dimas Alexandre Kliemann, Cristiane Valle Tovo, Ana Beatriz Gorini Da Veiga, André Luiz Machado, John T. West

Nebraska Center for Virology: Faculty Publications

Background & Aims: Development of resistance results from mutations in the viral genome, and the presence of selective drug pressure leads to the emergence of a resistant virus population. The aim of this study was to analyze the impact of genetic variability on the genetic barrier to drug resistance to DAAs.

Methods: The genetic barrier was quantified based on the number and type of nucleotide mutations required to impart resistance, considering full-length HCV NS3, NS5A and NS5B regions segregated by genotype into subtypes 1a, 1b, 2a, 2b and 3a. This study analyzed 789 NS3 sequences, 708 sequences and 536 NS5B …

Kaposi’S Sarcoma-Associated Herpesvirus Reduces Cellular Myeloid Differentiation Primary-Response Gene 88 (Myd88) Expression Via Modulation Of Its Rna, Amy Lingel, Erica Ehlers, Qianli Wang, Mingxia Cao, Charles Wood, Rongtuan Lin, Luwen Zhang

Nebraska Center for Virology: Faculty Publications

Kaposi’s sarcoma (KS)-associated herpesvirus (KSHV) is a human gammaherpesvirus associated with several human malignancies. The replication and transcription activator (RTA) is necessary and sufficient for the switch from KSHV latency to lytic replication. Interleukin 1 (IL-1) is a major mediator for inflammation and plays an important role in both innate and adaptive immunity. Myeloid differentiation primary response gene 88 (MyD88) is an essential adaptor molecule for IL-1 as well as most Toll-like receptor signaling. In this study, we identified a novel mechanism by which KSHV interferes with host inflammation and immunity. KSHV RTA specifically reduces the steady-state protein levels of …

Nf45 And Nf90 Bind Hiv-1 Rna And Modulate Hiv Gene Expression, Yan Li, Michael Belshan

Nebraska Center for Virology: Faculty Publications

A previous proteomic screen in our laboratory identified nuclear factor 45 (NF45) and nuclear factor 90 (NF90) as potential cellular factors involved in human immunodeficiency virus type 1 (HIV-1) replication. Both are RNA binding proteins that regulate gene expression; and NF90 has been shown to regulate the expression of cyclin T1 which is required for Tat-dependent trans-activation of viral gene expression. In this study the roles of NF45 and NF90 in HIV replication were investigated through overexpression studies. Ectopic expression of either factor potentiated HIV infection, gene expression, and virus production. Deletion of the RNA binding domains of NF45 …

Mucosal Vaccination By Adenoviruses Displaying Reovirus Sigma 1, Eric A. Weaver, Zenaido T. Camacho, Matthew L. Hillestad, Catherine M. Crosby, Mallory A. Turner, Adam J. Guenzel, Hind J. Fadel, George T. Mercier, Michael A. Barry

Nebraska Center for Virology: Faculty Publications

We previously developed adenovirus serotype 5 (Ad5) vectors displaying the sigma 1 protein from reovirus as mucosal vaccines. Ad5-sigma retargets to JAM-1 and sialic acid, but had 40-fold reduced gene delivery when compared to Ad5. While weaker at transduction, Ad5-sigma generated stronger T cell responses than Ad5 when used for mucosal immunization. New Ad5- fiber-sigma vectors were generated here by varying the number of fiber β-spiral shaft repeats (R) fused between fiber tail and the sigma. Ad5 virions encoding R3, R14, and R20 chimeras were rescued. Increasing chimera length led to their decreasing encapsidation of these proteins in the virions. …

Towards The Complete Small Rnome Of Acinetobacter Baumannii, Andy Weiss, William H. Broach, Mackenzie C. Lee, Lindsey N. Shaw

Molecular Biosciences Faculty Publications

In recent years, the Gram-negative bacterium Acinetobacter baumannii has garnered considerable attention for its unprecedented capacity to rapidly develop resistance to antibacterial therapeutics. This is coupled with the seemingly epidemic emergence of new hyper-virulent strains. Although strain-specific differences for A. baumannii isolates have been well described, these studies have primarily focused on proteinaceous factors. At present, only limited publications have investigated the presence and role of small regulatory RNA (sRNA) transcripts. Herein, we perform such an analysis, describing the RNA-seq-based identification of 78 A. baumannii sRNAs in the AB5075 background. Together with six previously identified elements, we include each of …

Peptidoglycan Recycling In Gram-Positive Bacteria Is Crucial For Survival In Stationary Phase, Marina Borisova, Rosmarie Gaupp, Amanda Duckworth, Alexander Schneider, Désirée Dalügge, Maraike Mühleck, Denise Deubel, Sandra Unsleber, Wenqi Yu, Günther Muth, Markus Bischoff, Friedrich Götz, Christoph Mayer

Molecular Biosciences Faculty Publications

Peptidoglycan recycling is a metabolic process by which Gram-negative bacteria reutilize up to half of their cell wall within one generation during vegetative growth. Whether peptidoglycan recycling also occurs in Gram-positive bacteria has so far remained unclear. We show here that three Gram-positive model organisms, Staphylococcus aureus, Bacillus subtilis, and Streptomyces coelicolor, all recycle the sugar N-acetylmuramic acid (MurNAc) of their peptidoglycan during growth in rich medium. They possess MurNAc-6-phosphate (MurNAc-6P) etherase (MurQ in E. coli) enzymes, which are responsible for the intracellular conversion of MurNAc-6P to N-acetylglucosamine-6-phosphate and d-lactate. By applying mass spectrometry, we observed accumulation of MurNAc-6P in …

Self-Regulation Of Functional Pathways By Motifs Inside The Disordered Tails Of Beta-Catenin, Bi Zhao, Bin Xue

Molecular Biosciences Faculty Publications

Background: Beta-catenin has two major functions: coordinating cell-cell adhesion by interacting with cadherin in cadherin junction formation pathway; and regulating gene expression through Wnt signaling pathway. Accomplishing these two functions requires synergistic action of various sequential regions of the same beta-Catenin molecule, including the N-terminal tail, the middle armadillo domain, and the C-terminal tail. Although the middle armadillo domain is the major functional unit of beta-Catenin, the involvement of tails in the regulation of interacti on between beta-Catenin and its partners has been well observed. Nonetheless, the regulatory processes of both tails are still elusive. In addition, it is interesting …

Mir146a-Mediated Targeting Of Fancm During Inflammation Compromises Genome Integrity, Devakumar Sundaravinayagam, Hye R. Kim, Ting T. Wu, Hyun H. Kim, Hyun-Seo Lee, Semo Jun, Jeong-Heon Cha, Younghoon Kee, Ho J. You, Jung-Hee Lee

Molecular Biosciences Faculty Publications

Inflammation is a potent inducer of tumorigenesis. Increased DNA damage or loss of genome integrity is thought to be one of the mechanisms linking inflammation and cancer development. It has been suggested that NF-κB-induced microRNA-146 (miR146a) may be a mediator of the inflammatory response. Based on our initial observation that miR146a overexpression strongly increases DNA damage, we investigated its potential role as a modulator of DNA repair. Here, we demonstrate that FANCM, a component in the Fanconi Anemia pathway, is a novel target of miR146a. miR146a suppressed FANCM expression by directly binding to the 3’ untranslated region of the gene. …

Adeno-Associated Virus-Vegf-165 Mediated Modification Of Adipose Derived Stem Cells For Cell Therapy, Upasana Niyogi

Browse all Theses and Dissertations

Chronic wounds have become a major clinical and economic burden in our society. New approaches that accelerate wound healing are desperately needed. Angiogenesis and vascularization play a critical role in healing. One of the essential angiogenic factors that promote the formation of vascular beds is vascular endothelial growth factor (VEGF). Moreover, adipose-derived stem cells (ASC), through their regeneration and differentiation properties, may promote healing when transplanted into a wound bed. I have proposed to develop a novel organotypic wound model to facilitate the study of wound healing process. I hypothesize that administration of genetically-modified ASC secreting VEGF via an adeno-associated …

An Investigation Of The Use Of Natural Agents To Control Mycobacteria, Olga Lesniak

Theses

Economic losses that are caused by Mycobacterium avium paratuberculosis (MAP) infections represent a major obstacle for farmers and also for the dairy industry. In order to control the population of MAP, alternative control agents must be investigated. One of the studies described in this thesis focused on using a bacteriophage therapy based approach to assess its efficacy on simulated industrial environments and food models. Another study described in this thesis investigated the antimicrobial effects of parsley and garlic on a range of mycobacteria species. Both studies used conventional microbiological methods such as plaque assays, AlamarBlue assay (MABA) and growth curves, …

The Endophytes Of Pediomelum Esculentum: A Unique Case In Legume Evolution, Tyrel Ryan Deutscher

Electronic Theses and Dissertations

Pediomelum esculentum (commonly prairie turnip) is a perennial legume of the Great Plains, consisting of a deep taproot and large edible tuber, and has served as a nutritious staple in Native American diets. The tuber is capable of storing up to 20 percent protein by weight. P. esculentum is a legume, but not a prominent nodule former; instead, it grows in nitrogen-limited soils and produces large amounts of protein. This suggests the involvement of biological nitrogen fixation. We have investigated the presence of diazotrophic endophytes in P. esculentum. Bacteria were isolated from wild plants on nitrogen free media, identified …

Synthetic Biology For Autotrophic And Heterotrophic Production Of Ethanol, Nathanael Braselton

Electronic Theses and Dissertations

Growing energy demand and rising levels of greenhouse gases has put massive strain on the global environment. Alternatives to fossil fuels are being developed in an attempt to curb climate change. Biotechnology has made large strides in order to create a completely renewable energy source by genetically modifying microbes to produce biofuels and other “green” high value compounds. In this thesis project, (1) E. coli ATCC9637 (E. coli W) was genetically modified to produce bioethanol from beet juice which contains mainly sucrose. The ethanol productivity by engineered E. coli W was 18.8 mg/L/H/OD₆₀₀.
(2) Cyanobacterium Anabaena sp. PCC7120 …

Development And Characterization Of A Recombinant Orf Virus Vector Expressing The Spike Protein Of Porcine Epidemic Diarrhea Virus, Kyle Hain

Electronic Theses and Dissertations

Orf virus (ORFV), the type member of the genus Parapoxvirus of the family Poxviridae, causes orf or cutaneous pustular dermatitis in sheep and goats. ORFV is a ubiquitous virus capable of re-infecting its hosts multiple times over time. ORFV causes a non-systemic, self-limiting disease which is usually restricted to the skin surrounding the virus entry sites. ORFV has evolved several immunomodulatory proteins (IMPs) that evade and/or modulate host immune responses to infection and contribute to virus virulence and disease pathogenesis. Given biological properties and unique immunomodula tor y properties, ORFV has gained significant attention in recent years for its …

Cooperation And Punishment In The Arbuscular Mycorrhizal Symbiosis: Implications For Resource Exchange & Biological Market Dynamics, Jerry A. Mensah

Electronic Theses and Dissertations

The arbuscular mycorrhizal (AM) symbiosis is arguably the world’s most abundant and important mutualism, and brings together the roots of the majority of land plants and AM fungi to great mutual advantage. The AM symbiosis can increase the uptake of nutrients, such as phosphorus (P) and nitrogen (N), and improves the abiotic and biotic stress resistance of the host plant. AM fungi have the potential to act as biofertilizers and bioprotectors in sustainable agriculture. However, despite its significance, the mechanisms that control the resource exchange between both partners in the arbuscular mycorrhizal symbiosis are largely unknown. The main aim of …

The Search For Modifiers Of The Maze Gametophyte Factor Ga1-S And Quantitative Trait Polymorphisms Emerging From Doubled-Haploid Maize Lines, Vivek Shrestha

Electronic Theses and Dissertations

The project was designed to conduct two independent projects. The first project was to identify the genomic localization of the modifiers of the maize gametophyte factor (Ga1-s) and the second project was to establish and identify heritable polymorphic lines that have descended from a single doubled-haploid B73 plant. The objectives were (1) to search for modifier genes and loci on the maize chromosomes for the trait and determine genetic effects of them using QTL mapping; (2) to demonstrate the heritable polymorphism of the quantitative traits emerged from the descendants of a single doubled-haploid maize plant. Regarding the QTL …

Enhancing The Industrial Potential Of Filamentous Cyanobacteria, Tylor J. Johnson

Electronic Theses and Dissertations

The objectives of this project were to improve the industrial potential of filamentous N2-fixing cyanobacteria by increasing its biofuel tolerance, and to evaluate the economic feasibility and environmental impacts of a theoretical, cyanobacteria-based biofuel production facility. To develop a method to quantify filamentous cyanobacteria in dilute culture media, a dual-stained fluorescence assay was evaluated. While the viable cell stain (SYTO® 9) was accurate, the non-viable cell stain (propidium iodide) also bound to viable cells. Additional non-viable cell stains were evaluated, but none were accurate at quantifying viability. Thus we concluded that the viable cell stain SYTO® 9 is a reliable …

Diversity Of Free-Living Nitrogen Fixing Bacteria In The Badlands Of South Dakota, Bibha Dahal

Electronic Theses and Dissertations

Biological Nitrogen Fixation is carried out by a highly diverse group of members of the domains Bacteria and Archaea which express a nitrogenase enzyme complex and are termed diazotrophs. The best known diazotrophs form symbiotic associations with plants, but free-living nitrogen fixing bacteria also contribute a substantial amount of nitrogen to ecosystems, including arid lands. The aim of this study was to isolate free-living diazotrophs from the soil crusts of sod tables of South Dakota Badlands. Samples were obtained from sod tables and the surrounding base (matrix) in spring and fall of 2014. A culture dependent approach was used to …

Inactivation Of Acanthamoeba Spp. And Other Ocular Pathogens By Application Of Cold Atmospheric Gas Plasma, Wayne Heaselgrave, Gilbert Shama, Peter W. Andrew, Michael G. Kong

Bioelectrics Publications

Currently there are estimated to be approximately 3.7 million contact lens wearers in the United Kingdom and 39.2 million in North America. Contact lens wear is a major risk factor for developing an infection of the cornea known as keratitis due to poor lens hygiene practices. While there is an international standard for testing disinfection methods against bacteria and fungi (ISO 14729), no such guidelines exist for the protozoan Acanthamoeba, which causes a potentially blinding keratitis most commonly seen in contact lens wearers, and as a result, many commercially available disinfecting solutions show incomplete disinfection after 6 and 24 …

The Contribution Of Marine Aggregate-Associated Bacteria To The Accumulation Of Pathogenic Bacteria In Oysters: An Agent-Based Model, Andrew M. Kramer, J. Evan Ward, Fred C. Dobbs, Melissa L. Pierce

OES Faculty Publications

Bivalves process large volumes of water, leading to their accumulation of bacteria, including potential human pathogens (e.g., vibrios). These bacteria are captured at low efficiencies when freely suspended in the water column, but they also attach to marine aggregates, which are captured with near 100% efficiency. For this reason, and because they are often enriched with heterotrophic bacteria, marine aggregates have been hypothesized to function as important transporters of bacteria into bivalves. The relative contribution of aggregates and unattached bacteria to the accumulation of these cells, however, is unknown. We developed an agent-based model to simulate accumulation of vibrio-type bacteria …

Investigation Of Anaplasma Phagocytophilum And Anaplasma Marginale Adhesin-Host Cell Interactions, Kathryn S. Hebert

Theses and Dissertations

Anaplasma phagocytophilum and A. marginale are the etiologic agents of bovine anaplasmosis and human granulocytic anaplasmosis, respectively. As obligate intracellular pathogens, binding and entry of host cells is a prerequisite for survival. The molecular events associated with these processes are poorly understood. Identifying the adhesins mediating binding, delineating their key functional domains, and determining the molecular determinants to which they bind not only benefits better understanding of Anaplasma spp. pathobiology, but could also benefit the development of novel approaches for protecting against infection. We previously demonstrated that A. phagocytophilum outer membrane protein A (ApOmpA) is critical for bacterial binding and …

Role Of Silencing Rna Fgsir34 In Fusarium Graminearum's Pathogenicity To Wheat, Subha Dahal

Electronic Theses and Dissertations

Fusarium graminearum is an ascomycetous fungal pathogen that causes Fusarium head blight (FHB) disease in wheat and other cereal grains. Mycotoxin produced by the fungus, predominantly deoxynivalenol (DON), is considered as an important virulence factor for the spread of disease. Our previous study of a Dicer-like 2 knockdown mutant has led to our hypothesis that a silencing RNA, fgsiR34, might play a key role in regulating DON biosynthesis and some other virulent factors. To test this hypothesis, we generated an fgsiR34 over-expressing mutant (ΔfgsiR34+) using Inverse Repeat Transgene method and studied the pathogenicity of the mutant in …

Senescence Of Native Perennial Warm Season Grasses Senescence Associated Switchgrass Transcriptome, Michaellong Tran

Electronic Theses and Dissertations

Senescence of perennial crops enable continuous harvests after one sowing event. Perennials senesce at adapted rates of their native environments; however, early senescencing crops do not maximize the growing season as nutrient reallocation takes precedence. Chlorophyll degradation and nitrogen reallocation was observed to occur rapidly between mid to late September. Transcriptome analysis on early and late senescencing switchgrass cultivars reveals upregulation of starch metabolism, light reactions, Calvin-Benson Cycle, and anthocyanin synthesis in late senescencing switchgrass. Morphological variations between the two germplasms prolong the growing season of late senescencing switchgrass, maximizing yield. Expression of mRNA as senescence progresses and between the …

Colony Stimulating Factor-1 Receptor (Csf-1r) Expression, Degradation, And Signaling In Lipid-Laden Macrophages, Gregory Thompson

Electronic Theses and Dissertations

Atherosclerosis is a serious disease affecting a large portion of the world’s population. In atherosclerosis, macrophages become filled with lipid droplets and form fatty plaques in artery walls. The formation of these atherosclerotic plaques is dependent on the macrophage growth factor colony stimulating factor-1 (CSF-1); however, understanding the changes in CSF-1 signaling and related pathways in foam cells remain poorly investigated. Lipid-laden macrophages were formed through exposure of murine bone marrow derived macrophages to 100 μg/ml acetylated LDL for 24 h. Non-LDL treated macrophages were grown in parallel for each experiment and used for comparison. Immunofluorescent staining was used to …