Microbiology Commons^™

A Three-Component Regulatory System Regulates Biofilm Maturation And Type Iii Secretion In Pseudomonas Aeruginosa, Sherry L. Kuchma, John P. Connolly, George A. O'Toole

Dartmouth Scholarship

Biofilms are structured communities found associated with a wide range of surfaces. Here we report the identification of a three-component regulatory system required for biofilm maturation by Pseudomonas aeruginosa strain PA14. A transposon mutation that altered biofilm formation in a 96-well dish assay originally defined this locus, which is comprised of genes for a putative sensor histidine kinase and two response regulators and has been designated sadARS. Nonpolar mutations in any of the sadARS genes result in biofilms with an altered mature structure but do not confer defects in growth or early biofilm formation, swimming, or twitching motility. After …

Modulation Of Human Herpesvirus 8/Kaposi’S Sarcoma-Associated Herpesvirus Replication And Transcription Activator Transactivation By Interferon Regulatory Factor 7, Jinzhang Wang, Jun Zhang, William Harrington Jr., John T. West, Charles Wood

Nebraska Center for Virology: Faculty Publications

Human herpesvirus 8 (HHV-8)/Kaposi’s sarcoma-associated herpesvirus infection goes through lytic and latent phases that are regulated by viral gene products, but very little is known about the involvement of host proteins. The replication and transcription activator (RTA) is a viral protein sufficient to initiate lytic replication by activating downstream genes, including the viral early gene open reading frame 57 (ORF 57), which codes for a posttranscriptional activator. In this study, we demonstrate that cellular interferon regulatory factor 7 (IRF-7) negatively regulates this process by competing with RTA for binding to the RTA response element in the ORF 57 promoter to …

Human Dendritic Cells Transduced With Herpes Simplex Virus Amplicons Encoding Human Immunodeficiency Virus Type 1 (Hiv-1) Gp120 Elicit Adaptive Immune Responses From Human Cells Engrafted Into Nod/Scid Mice And Confer Partial Protection Against Hiv-1 Challenge, Santhi Gorantla, Kathlyn Santos, Vakara Meyer, Stephen Dewhurst, William J. Bowers, Howard J. Federoff, Howard Gendelman, Larisa Poluektova

Nebraska Center for Virology: Faculty Publications

Small-animal models are needed to test human immunodeficiency virus (HIV) vaccine efficacy following viral challenge. To this end, we examined HIV-1-specific immune responses following immunization of nonobese diabetic-severe combined immunodeficient mice that were repopulated with human peripheral blood lymphocytes (hu-PBL-NOD/SCID mice). Autologous dendritic cells (DC) were transduced ex vivo with replicationdefective, helper virus-free, herpes simplex virus type 1 (HSV-1) amplicons that expressed HIV-1 gp120 and were then injected into the hu-PBL-NOD/SCID mice. This resulted in primary HIV-1-specific humoral and cellular immune responses. Serum samples from vaccinated animals contained human immunoglobulin G that reacted with HIV-1 Env proteins by enzyme-linked immunosorbent …

The Nuclear Localization Of The Arabidopsis Transcription Factor Tip Is Blocked By Its Interaction With The Coat Protein Of Turnip Crinkle Virus, Tao Ren, Feng Qu, Thomas Jack Morris

Nebraska Center for Virology: Faculty Publications

We have previously reported that TIP, an Arabidopsis protein, interacts with the coat protein (CP) of Turnip crinkle virus (TCV) in yeast cells and that this interaction correlated with the resistance response in the TCV-resistant Arabidopsis ecotype Dijon-17. TIP was also able to activate transcription of reporter genes in yeast cells, suggesting that it is likely a transcription factor. We have now verified the physical interaction between TIP and TCV CP in vitro and showed that CP mutants unable to interact with TIP in yeast cells bind TIP with much lower affinity in vitro. Secondly, we have performed gel shift …

In Vitro And In Vivo Translational Efficiencies Of The 5′ Untranslated Region From Eight Genotype 2 Bovine Viral Diarrhea Virus Field Isolates, Christina L. Topliff, Seung K. Chon, Ruben O. Donis, Kent M. Eskridge, Clayton L. Kelling

Nebraska Center for Virology: Faculty Publications

We determined the in vitro and in vivo translational efficiency mediated by the internal ribosomal entry site (IRES) from eight BVDV2 field isolates varying in virulence using a bicistronic reporter vector in rabbit reticulocyte lysates (RRL), and in primate and bovine cell lines. Using a T7-promoter system, the high virulence isolates had greater translational efficiencies in bovine lymphocytes (BL-3 cells), than did the low virulence isolates. The low virulence isolates translated with greater efficiencies than the high virulence isolates in RRL, African green monkey kidney (CV-1) and bovine turbinate (BT) cells. Our results demonstrate that despite a high degree of …

Characterization Of Microbial Activity, Mark P. Buttner, Patricia Cruz, Klaus J. Stetzenbach, Abe Van Luik, Thomas Williams, Amy J. Smiecinski

Publications (YM)

The overall goal of this study is to investigate the phenomena that affect the fate and transport of radionuclides in the environment. The objective of this task, “Characterization of Microbial Activity”, is to develop a molecular biological method for the characterization of the microbial population indigenous to the Yucca Mountain Project site, with emphasis in detection and measurement of species or groups of microorganisms that could be involved in actinide and/or metal reduction, and subsurface transport. Subtasks consist of QA planning and preparation, and literature review. This task is part of a cooperative agreement between the UNLV Research Foundation and …

Novel Protein-Protein Interactions Regulate The Proteolytic Activity Of The Pro- Apoptotic Serine Protease, Omi/Htra2, Supriya Singh

Electronic Theses and Dissertations

Omi/HtrA2 is a mitochondrial serine protease with high homology to the bacterial HtrA proteins. Omi promotes caspase-dependent apoptosis by binding and degrading IAPs-inhibitor of apoptosis proteins. Omi can also induce caspase-independent apoptosis but the actual mechanism is still unknown. IAP's are not the only substrates cleaved by Omi. There are at least two more known substrates of Omi, the HAX-1 and the ped/pea-15 proteins. HS1-associated protein X-1 (HAX-1) is a mitochondrial protein, degraded by Omi after induction of caspase-dependent apoptosis. Ped/pea-15 is also an anti-apoptotic protein and is cleaved by Omi after induction of caspase-independent apoptosis. The proteolytic activity of …

The Effects Of Arsenic On Selenoprotein Biosythesis, Fanta Konate

Electronic Theses and Dissertations

Arsenic contamination of drinking water is a real public health problem in certain areas of South-East Asia where chronic exposure has been correlated to higher rates of lung, skin, bladder, kidney, and liver cancer. Although arsenic carcinogenicity is well established, the mechanism by which it induces cancer is poorly understood. Recent evidence suggests that oxidative stress could be a possible mechanism for the carcinogenic effects of arsenic. Selenium, in the form of selenocysteine, is necessary for the activity of several enzymes with a role in the defense against reactive oxygen species (ROS), primarily thioredoxin reductases (TrxR) and glutathione peroxidases (Gpx). …

Real Time Rt-Pcr For Direct Detection Of Viable Mycobacterium Avium Subspecies Paratuberculosis In Chron's Disease Patients And Association Of Map Infection With Downregulation In Interferon-Gamma Receptor (Infg1) Gene In Crohn's Disease Patients, Mounir Chehtane

Electronic Theses and Dissertations

Association of Mycobacterium avium subspecies paratuberculosis (MAP) with Crohn's disease (CD) and not with ulcerative colitis (UC), two forms of inflammatory bowel disease (IBD), has been vigorously debated in recent years. This theory has been strengthened by recent culture of MAP from breast milk, intestinal tissue and Blood from patients with active Crohn's disease. Culture of MAP from clinical samples remained challenging due to the fastidious nature of MAP including its lack of cell wall in infected patients. The advent of real time PCR has proven to be significant in infectious disease diagnostics. In this study, real time reverse transcriptase …

Expression Of Hepatitis C Viral Non-Structural 3 Antigen In Transgenic Chloroplasts, Anubhuti Bhati

Electronic Theses and Dissertations

Hepatitis C viral infection is the major cause of acute hepatitis and chronic liver disease and remains the leading cause of liver transplants (NIH). An estimated 180 million people are infected globally (WHO). There is no vaccine available to prevent hepatitis C. The treatment with antiviral drugs is expensive, accompanied with various side effects and is limited only to those at risk of developing advanced liver disease. The treatment is also effective in only about 30% to 50% of treated patients and still a high percentage of patients are resistant to therapy. Therefore, there is an urgent need for the …

Expression Of Cholera Toxin B Subunit-Rotavirus Nsp4 Enterotoxin Fusion Protein In Transgenic Chloroplasts, Anila Kalluri

Electronic Theses and Dissertations

Rotavirus, the major cause of life-threatening infantile gastroenteritis, is a member of the Reoviridae family and is considered to be the single most important cause of virus-based severe diarrheal illness in infants and young children particularly 6 months to 2 years of age in industrialized and developing countries. Infection in infants and young children is often accompanied by severe life threatening diarrhea, most commonly following primary infection. Diarrhea is the major cause of death among children around the world. Responsible for 4 to 6 million deaths per year according to the World Health Organization (WHO), diarrhea is especially dangerous for …

Receptor Mediated Oral Delivery Of Bioencapsulated Green Fluorescent Protein Expressed In Transgenic Chloroplasts, Arati Limaye

Electronic Theses and Dissertations

The skyrocketing costs of prescription medicine in developed countries and their lack of availability in developing countries are the most challenging problems of human health. Primary reasons for such high cost are fermentation-based production, expensive purification methods, the need for low temperature storage and transportation and the delivery through sterile injections. Most of these expenses could be minimized or eliminated when therapeutic proteins are expressed and orally delivered via plant cells. Chloroplasts have the machinery to fold complex and biologically active eukaryotic proteins in the soluble chloroplast stromal compartment. Protein expression through chloroplast transformation system offers a number of advantages …

Selenotrisulfide Derivative Of Alpha-Lipoic Acid: Evaluation In A Cell Culture Model For Potential Use As A Topical Antioxidant, Melenie Lee Alonis

Electronic Theses and Dissertations

Selenium is a required micronutrient in mammalian cells. It is incorporated in the form of selenocysteine into selenoenzymes such as glutathione peroxidase and thioredoxin reductase, and is absolutely required for activity. Thioredoxin reductase is necessary for reduction of oxidized thioredoxin and therefore plays a major role in maintaining the redox status of the cell. Glutathione peroxidase is responsible for reducing peroxides into their corresponding alcohols and water. Together, these selenoenzymes constitute a significant part of the cell's arsenal to defend itself against oxidative stress. Exogenous sources of oxidative stress, such as UV radiation, are capable of generating reactive oxygen species …

Efficacy Of Innovative Anti-Microbial Decontamination Of Minimally Processed Vegetables, Christina Busquets I Tortosa

Masters

Chlorine has widespread use commercially as a disinfectant wash for raw vegetables. However, it is an extremely corrosive gas and it may have severe health and environmental implications. The aim of this thesis was to find safe, alternative decontamination treatments for ready-to-eat Iceberg lettuce. The efficacy of 1% acetic acid, 2% citric acid; calcium lactate concentrations of 0.5%, 1.5%, 2.5% and 3% at wash temperature of 4,25 and 50°C, ozone alone (1 mg 1-1) and in combination with calcium lactate (2.5%) were compared with chlorine as decontaminating treatments. Microbiological analyses were performed following decontamination treatments using the pour plate method …

The Effects Of Dams In The Big Sandy Watershed Using A Novel Bacteria-Based Bioindicator Of Water Quality, Kathleen Riha Loughman

Theses, Dissertations and Capstones

During 2003, water samples from the Big Sandy watershed were collected in conjunction with the United States Army Corps of Engineers (USACE). Objectives were to determine the effects of dams on bacterial populations and to test a bioindicator of water quality based on antibiotic-resistant and fecal indicator bacteria. Thirty-five samples were taken each season within the Big Sandy Watershed, which includes six USACE dams. Total cultivable, ciprofloxacin-resistant, erythromycin-resistant, tetracycline- resistant, total coliform, and fecal coliform bacteria were enumerated. Data on water chemistry and physical parameters were collected by the USACE in the spring and summer seasons. Antibiotic-resistant bacteria and fecal …

The Uptake Of Amino Acids By Microbes And Trees In Three Cold-Temperate Forests, Adrien C. Finzi, Sean T. Berthrong

Scholarship and Professional Work - LAS

Amino acids are emerging as a critical component of the terrestrial N cycle, yet there is little understanding of amino acid cycling in temperate forests. This research studied the uptake and turnover of amino acid N by soil microbes and the capacity of forest trees to take up the amino acid glycine in comparison to NH₄⁺ and NO₃⁻. This research was conducted in three temperate forests located in northwest Connecticut, USA. The three forests differed in soil parent material and canopy tree species composition. At all three sites, amino acids were released from soil organic …

Proteomic Fingerprints Distinguish Microglia, Bone Marrow, And Spleen Macrophage Populations, Yoshimi Enose, Christopher J. Destache, Andrea L. Mack, James R. Anderson, Fred Ullrich, Pawel Ciborowski, Howard E. Gendelman

Nebraska Center for Virology: Faculty Publications

Mononuclear phagocytes (MP; dendritic cells, monocytes, tissue macrophages, and microglia) maintain tissue homeostasis and provide a first line of defense against invading pathogens. In specific circumstances, MPs also induce inflammatory responses and as such affect disease onset and progression. Despite intensive research into MP biology, little is known of the functional and molecular properties of individual MP subtypes. Using a novel proteomics platform, unique protein patterns and protein identities were observed among populations of spleen and bone marrow macrophages and microglia. Cells were obtained from C57BL/6 mice and were cultivated in macrophage colony-stimulating factor. MP subtypes were indistinguishable by morphological …

Locatization Of Human Herpesvirus 8 In Human Sperms By In Situ Pcr, Omar Bagasra, Deepa Patel, Lisa Bobroski, Jamil A. Abbasi, Alex U. Bagasra, Hasna Baidouri, Twaina Harris, Albert El-Roeiy, Zsolt Lengvarszky, Homayoon Farzadegan, Charles Wood

Nebraska Center for Virology: Faculty Publications

Objectives: Defining the mechanism of infection with human herpesvirus-8 (HHV-8) or Kaposi’s sarcoma associated herpesvirus (KSHV) is an important clinical issue. HHV-8 has been linked to Kaposi’s sarcoma (KS) development in HIV-1-infected individuals, and KS develops in 40% of those infected with both viruses. A series of epidemiological data suggest that sexual transmission is one of the routes of transmission for HHV-8. In our studies, we sought to assess the cellular reservoirs of HHV-8 DNA in the semen of HIV-1-infected men and the potential role of HHV-8 infected spermatozoa in horizontal transmission. Design and methods: A nested polymerase chain reaction …

Guidelines For The Management Of Hiv Infection In Pregnant Women And The Prevention Of Mother-To-Child Transmission Of Hiv, David Hawkins, M. Blott, P. Clayden, A, De Ruiter, G. Foster, C. Gilling-Smith, B. Gosrani, H. Lyall, D. Mercey, M.-L. Newell, S. O'Shea, R. Smith, J. Sunderland, Charles Wood, G. Taylor

Nebraska Center for Virology: Faculty Publications

The prevalence of HIV infection amongst women giving birth in England and Wales has increased every year since 1990. Results from the Unlinked Anonymous Surveys of infection in pregnancy, show that in 2003, the prevalence reached one in 180 (0.56%) in inner London, one in 271 in outer London (0.37%) and one in 1,282 (0.08%) in the rest of England [1]. The majority of these women are from sub-Saharan Africa. The Department of Health policy of recommending an HIV test to every pregnant woman [2] has resulted in an increase in the proportion of these women who are aware of …

Interferon Resistance And Efficiency Of Hcv Replicon Establishment In Huh7 Cells Expressing Simian Virus 5 V Protein, Nicole E. Wagner

Seton Hall University Dissertations and Theses (ETDs)

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Humboldt Journal Of Microbiology: Volume 7, Number 2- December 2005, Press Humboldt

Humboldt Journal of Microbiology

No abstract provided.

Humboldt Journal Of Microbiology: Volume 7, Number 1- May 2005, Press Humboldt

Humboldt Journal of Microbiology

No abstract provided.

Validation Of An Immunomagnetic-Electrochemiluminescence Detection System For The Detection Of Escherichia Coli 0157 In Food And Comparison With An Iso Standard Method, Eimear Carney

Theses

Rapid and sensitive methods for detecting Escherichia coli 0157 in contaminated food, water and other biological samples are important in limiting and preventing the spread of this pathogen. Traditional culture based methods for bacterial identification and detection can be time consuming and labour intensive. Modem alternative methods should be faster whilst maintaining sensitivity. Immunomagnetic separation (IMS) methodology has been investigated and appears promising for rapid bacterial detection in unrelated food and environmental samples. In this study, a commercial sensor which combined IMS with electrochemiluminescence (ECL) was evaluated for the detection of E. coli 0157 in unrelated food matrices. Emitted signals …

Effect Of Humic Substance Photodegradation On Bacterial Growth And Respiration In Lake Water, Alexandre M. Anesio, William Granéli, George R. Aiken, David J. Kieber, Kenneth Mopper

Chemistry & Biochemistry Faculty Publications

This study addresses how humic substance (HS) chemical composition and photoreactivity affect bacterial growth, respiration, and growth efficiency (BGE) in lake water. Aqueous solutions of HSs from diverse aquatic environments representing different dissolved organic matter sources (autochthonous and allochthonous) were exposed to artificial solar UV radiation. These solutions were added to lake water passed through a 0.7-μm pore-size filter (containing grazer-free lake bacteria) followed by dark incubation for 5, 43, and 65 h. For the 5-h incubation, several irradiated HSs inhibited bacterial carbon production (BCP) and this inhibition was highly correlated with H₂O₂ photoproduction. The H_{2 …}

A Novel Binding Interaction For The Paxillin Ld3 Motif: Paxillin Ld3 Mediates Merlin-Paxillin Binding At Paxillin Binding Domain 1, Sandra E. Beattie Geden

Electronic Theses and Dissertations

Neurofibromatosis type 2, an autosomal dominant genetic disorder, causes predisposed individuals to develop various benign central and peripheral nervous system tumors. The characteristic tumors of this disease are schwannomas, which are tumors of the Schwann cells, typically on the vestibular nerve. These and the other associated tumors slowly compress nervous system structures causing deafness and loss of balance, resulting in an average life-span of less than 40 years. The product of the Nf2 gene is the protein named merlin or schwannomin. In individuals diagnosed with NF2, merlin is either absent or mutated to the point of inactivation. As such, merlin …

Etiology Of Oral Disease In View Of Microbial Complexity, Anne C. R. Tanner, Jacques Izard

Department of Food Science and Technology: Faculty Publications

The number of different microorganisms recognized in the oral cavity using molecular methods has more than doubled compared with the number isolated using cultural techniques. This finding necessitates a reevaluation of which species may be pathogens in dental infections. Molecular methods used to determine microbial diversity include broad range target Polymerase Chain Reaction with ‘universal primers,’ and cloning amplicons or denaturing gradient gel electrophoresis to separate DNA fragments before sequencing. These molecular methods have clarified and expanded the taxonomy of oral microbial species. Discrepancies between comprehensive molecular and cultural methods suggest that neither method alone can adequately evaluate associations of …

Replication And Encapsidation Of Papillomaviruses In Saccharomyces Cerevisiae, Peter C. Angeletti

Nebraska Center for Virology: Faculty Publications

Improvements in methodologies to recapitulate and study particular biological functions of the

papillomavirus life cycle have led to great advances in our knowledge of these viruses. Described in

this chapter are techniques that allow low-copy and high-copy replication of full-length human

papillomavirus (HPV) genomes, as well as assembly of virus-like particles, in Saccharomyces

cerevisiae (yeast). This system has several distinct advantages that make it an attractive complement

to the well-established raft-culturing system. First, yeast are inexpensive, rapid, and simple to culture

in the lab. Second, they provide an ever-widening array of genetic tools to analyze HPV functions

—most recently notable, …

Antigenicity And Immunogenicity Of A Synthetic Human Immunodeficiency Virus Type 1 Group M Consensus Envelope Glycoprotein, Feng Gao, Eric A. Weaver, Zhongjing Lu, Yingying Li, Hua-Xin Liao, Benjiang Ma, S. Munir Alam, Richard M. Scearce, Laura L. Sutherland, Jae-Sung Yu, Julie M. Decker, George M. Shaw, David C. Montefiori, Bette T. Korber, Beatrice H. Hahn, Barton F. Haynes

Nebraska Center for Virology: Faculty Publications

Genetic variation of human immunodeficiency virus (HIV-1) represents a major obstacle for AIDS vaccine development. To decrease the genetic distances between candidate immunogens and field virus strains, we have designed and synthesized an artificial group M consensus env gene (CON6 gene) to be equidistant from contemporary HIV-1 subtypes and recombinants. This novel envelope gene expresses a glycoprotein that binds soluble CD4, utilizes CCR5 but not CXCR4 as a coreceptor, and mediates HIV-1 entry. Key linear, conformational, and glycan-dependent monoclonal antibody epitopes are preserved in CON6, and the glycoprotein is recognized equally well by sera from individuals infected with different HIV-1 …

Antigenicity And Immunogenicity Of A Synthetic Human Immunodeficiency Virus Type 1 Group M Consensus Envelope Glycoprotein, Feng Gao, Eric A. Weaver, Zhongjing Lu, Yingying Li, Hua-Xin Liao, Benjiang Ma, S. Munir Alam, Richard M. Scearce, Laura L. Sutherland, Jae-Sung Yu, Julie M. Decker, George M. Shaw, David C. Montefiori, Bette T. Korber, Beatrice H. Hahn, Barton F. Haynes

Nebraska Center for Virology: Faculty Publications

Genetic variation of human immunodeficiency virus (HIV-1) represents a major obstacle for AIDS vaccine development. To decrease the genetic distances between candidate immunogens and field virus strains, we have designed and synthesized an artificial group M consensus env gene (CON6 gene) to be equidistant from contemporary HIV-1 subtypes and recombinants. This novel envelope gene expresses a glycoprotein that binds soluble CD4, utilizes CCR5 but not CXCR4 as a coreceptor, and mediates HIV-1 entry. Key linear, conformational, and glycan-dependent monoclonal antibody epitopes are preserved in CON6, and the glycoprotein is recognized equally well by sera from individuals infected with different HIV-1 …

Genome Of Deerpox Virus, C. L. Afonso, Gustavo A. Delhon, E. R. Tuhlman, Z. Lu, A. Zsak, V. M. Becerra, L. Zsak, G. F. Kutish, D. L. Rock

Nebraska Center for Virology: Faculty Publications

Deerpox virus (DPV), an uncharacterized and unclassified member of the Poxviridae, has been isolated from North American free-ranging mule deer (Odocoileus hemionus) exhibiting mucocutaneous disease. Here we report the genomic sequence and comparative analysis of two pathogenic DPV isolates, W-848-83 (W83) and W-1170-84 (W84). The W83 and W84 genomes are 166 and 170 kbp, containing 169 and 170 putative genes, respectively. Nucleotide identity between DPVs is 95% over the central 157 kbp. W83 and W84 share similar gene orders and code for similar replicative, structural, virulence, and host range functions. DPV open reading frames (ORFs) with putative …