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Binding Of Equine Infectious Anemia Virus Rev To An Exon Splicing Enhancer Mediates Alternative Splicing And Nuclear Export Of Viral Mrnas, Michael Belshan, Gregory S. Park, Patricia Bilodeau, C. Martin Stoltzfus, Susan Carpenter

Nebraska Center for Virology: Faculty Publications

In addition to facilitating the nuclear export of incompletely spliced viral mRNAs, equine infectious anemia virus (EIAV) Rev regulates alternative splicing of the third exon of the tat/rev mRNA. In the presence of Rev, this exon of the bicistronic RNA is skipped in a fraction of the spliced mRNAs. In this report, the cis-acting requirements for exon 3 usage were correlated with sequences necessary for Rev binding and transport of incompletely spliced RNA. The presence of a purine-rich exon splicing enhancer (ESE) was required for exon 3 recognition, and the addition of Rev inhibited exon 3 splicing. Glutathione-S-transferase (GST)-Rev bound …

Unusual Polymorphisms In Human Immunodeficiency Virus Type 1 Associated With Nonprogressive Infection, Louis Alexander, Emma Weiskopf, Thomas C. Greenough, Nathan C. Gaddis, Marcy C. Auerbach, Michael H. Malim, Stephen J. O'Brien, Bruce D. Walker, John L. Sullivan, Ronald C. Desrosiers

Biology Faculty Articles

Factors accounting for long-term nonprogression may include infection with an attenuated strain of human immunodeficiency virus type 1 (HIV-1), genetic polymorphisms in the host, and virus-specific immune responses. In this study, we examined eight individuals with nonprogressing or slowly progressing HIV-1 infection, none of whom were homozygous for host-specific polymorphisms (CCR5-Δ32, CCR2-64I, and SDF-1-3^'A) which have been associated with slower disease progression. HIV-1 was recovered from seven of the eight, and recovered virus was used for sequencing the full-length HIV-1 genome; full-length HIV-1 genome sequences from the eighth were determined following amplification of viral …

Jembrana Disease Virus Tat Can Regulate Human Immunodeficiency Virus (Hiv) Long Terminal Repeat-Directed Gene Expression And Can Substitute For Hiv Tat In Viral Replication, Hexin Chen, Jun He, Steven Fong, Graham Wilcox, Charles Wood

Nebraska Center for Virology: Faculty Publications

Jembrana disease virus (JDV) is a bovine lentivirus genetically similar to bovine immunodeficiency virus; it causes an acute and sometimes fatal disease in infected animals. This virus carries a very potent Tat that can strongly activate not only its own long terminal repeat (LTR) but also the human immunodeficiency virus (HIV) LTR. In contrast, HIV Tat cannot reciprocally activate the JDV LTR (H. Chen, G. E. Wilcox, G. Kertayadnya, and C. Wood, J. Virol. 73:658–666, 1999). This indicates that in transactivation JDV Tat may utilize a mechanism similar to but not the same as that of the HIV Tat. To …

Interferon Regulatory Factor 7 Is Induced By Epstein-Barr Virus Latent Membrane Protein 1, Luwen Zhang, Joseph S. Pagono

Nebraska Center for Virology: Faculty Publications

Infection by Epstein-Barr virus (EBV) generates several types of latency with different profiles of gene expression but with expression of Epstein-Barr nuclear antigen 1 (EBNA-1) in common. The BamHI Q promoter (Qp) is used for the transcription of EBNA-1 mRNA in type I latency, which is an EBV infection state exemplified by Burkitt’s lymphoma (BL). However, Qp is inactive in type III latency, and other promoters (C/Wp) are used for transcription of EBNA-1, which raises the question of how usage of these promoters is governed. Interferon (IFN) regulatory factor 7 (IRF-7) was identified first as a negative regulator of …

Integronlike Structures In Campylobacter Spp. Of Human And Animal Origin, Brigid Lucey, D. Crowley, P. Moloney, B. Cryan, M. Daly, F. O'Halloran, E. J. Threlfall, S. Fanning

Department of Biological Sciences Publications

Resistance to antimicrobial agents used to treat severe Campylobacter spp. gastroenteritis is increasing worldwide. We assessed the antimicrobial resistance patterns of Campylobacter spp. isolates of human and animal origin. More than half (n = 32) were resistant to sulphonamide, a feature known to be associated with the presence of integrons. Analysis of these integrons will further our understanding of Campylobacter spp. epidemiology.

Identification Of The Transactivation Domain Of The Transcription Factor Sox-2 And An Associated Co-Activator, Tamara K. Nowling, Lance R. Johnson, Matthew S. Wiebe, Angie Rizzino

Nebraska Center for Virology: Faculty Publications

The importance of interactions between Sox and POU transcription factors in the regulation of gene expression is becoming increasingly apparent. Recently, many examples of the involvement of Sox-POU partnerships in transcription have been discovered, including a partnership between Sox-2 and Oct-3. Little is known about the mechanisms by which these factors modulate transcription. To better understand the molecular interactions involved, we mapped the location of the transactivation do-main of Sox-2. This was done in the context of its interaction with Oct-3, as well as its ability to transactivate as a fusion protein linked to the DNA-binding domain of Gal4. Both …

Quantitative Image Analysis Of Simian Immunodeficiency Virus Replication In Macrophages Coinfected With Mycobacterium Avium Complex, Qingsheng Li, Keith G. Mansfield, Andrew Lackner, Ashley T. Haase

Qingsheng Li Publications

Mycobacterium avium is the most frequent cause of disseminated bacterial infection in patients with human immunodeficiency virus type 1 infection and in rhesus macaques with simian immunodeficiency virus (SIV) infection. This animal model of AIDS was used to test the hypothesis that this frequent association is the result of reciprocal enhancement of replication of both microorganisms. The replication of M. avium and SIV was analyzed in lymphatic tissues obtained from rhesus macaques experimentally inoculated with SIVmac who developed or remained free of overt M. avium infection. In situ hybridization, quantitative image analysis, and staining of M. avium and of macrophages …

Antigenic Epitope Analysis Of The P101 Nucleocapsid Protein Of Human Herpes Virus 6 (Var.B), Katherine Anne Francis

Student Works (2000-2009)

Human herpes virus 6 (HHV6) is an ubiquitous, lymphoproliferative P-herpesvirus, which infects almost all individuals during infancy and persists lifelong. Two variants of HHV6 (A and B) have been described. The present study is on HHV6B, which causes exanthem subitum, also known as roseola infantum, a common febrile childhood illness. Two methods of detection for anti-HHV6B in body fluids were used, being indirect immunofluorescence assay (IFA) and peptidylated pin-ELISA. For IFA, the target cells were HHV68-infected human cord blood mononuclear cells (HCBMC). Ninety eight infant sera were tested by IFA and all were found to be IgG-HHV6 positive. Seventeen breast …

The Molecular Characterisation Of Selected Epidemic Associated Gram Negative Pathogens In Southern Ireland, Mairéad Daly

Theses

This study was undertaken to evaluate various molecular techniques for the analysis of Gram negative bacterial associated epidemics. The first investigation dealt with bovine mastitis outbreaks, where the causative agent was identified as Pseudomonas aeruginosa. Epidemiological findings, suggested that all herds were infected from teat wipes that were contaminated with this organism. Initial investigations using the polymerase chain reaction (PCR), indicated a possible clonal relationship between all outbreak linked strains, with one exception. This finding was confirmed following pulsed-field gel electrophoresis (PFGE) and ribotype analysis. PCR was again employed to study a rather unusual epidemic, involving Salmonella tel-el-kebir over a …

A Preclinical Model Of Ionizing Radiation And Tnf-Alpha Gene Therapy For Cancer, Jun Li

Loma Linda University Electronic Theses, Dissertations & Projects

Long term control of high-grade brain tumors is rarely achieved with current therapeutic regimens. This study sought to determine if pretreatment with plasmid DNA expressing Tumor Necrosis Factor-α (TNF-α) could enhance the effect of ionizing radiation in an aggressive, rapidly growing glioma tumor (C6) model. Plasmid DNA is an attractive vector system for delivery of foreign genes into eukaryotic cells due to its advantages including low immunogenicity in vivo, lesser toxicity and relatively low cost. We have constructed a plasmid-based mammalian expression vector (pGL1-TNF) to secrete human TNF-α from cancer cells. Plasmid DNA was successfully transfected into C6 cells …

Dna-Dna Checkerboard Hybridization To Identify Bacteria In Human Plaque Samples, Jasan Lee Zimmerman

Loma Linda University Electronic Theses, Dissertations & Projects

Previous research has shown that using DNA probes to identify the presence of specific bacteria in human plaque samples is superior to the commonly used anaerobic cultural procedures. Some periodontal pathogens are either uncultivable or very difficult to grow, such as Prevotella intermedia, P. nigrescens, and Eubacterium brachy. Digoxygenin probes were prepared for 15 oral bacteria. These probes were used with a checkerboard DNA-DNA hybridization scheme to analyze whether human plaque samples contained any of the 15 bacterial species. Our results have shown that this method is successful in identifying bacterial DNA present in the subgingival patient plaque …

Engineering Secreted Proteins For Gene Transfer And Dna Vaccination, Jingxue Liu

Loma Linda University Electronic Theses, Dissertations & Projects

In recent years gene therapy has become a promising way of alleviating incurable human ailments, its concept emerging as the ultimate therapy for many infectious and genetic diseases. Two important aspects of the development of successful gene therapy protocols are the ability to monitor gene transfer readily, and the establishment of new protocols for treating specific diseases. In this work, Renilla luciferase and human glutamic acid decarboxylase (GAD) 65 were engineered for secretion to address some aspects of these issues.

Secreted reporter proteins are promising tools to study gene transfer and expression in a non-destructive manner, and bioluminescent proteins are …

Interferon Regulatory Factor 2 Represses The Epstein-Barr Virus BamHi Q Latency Promoter In Type Iii Latency, Luwen Zhang, Joseph S. Pagano

Nebraska Center for Virology: Faculty Publications

Epstein-Barr virus (EBV) nuclear antigen 1 (EBNA-1) is the essential protein for maintenance of the EBV episome and establishment of latency. The BamHI Q promoter (Qp) is used for the transcription of EBNA-1 mRNA in type I and type II latency, which are EBV infection states exemplified by Burkitt’s lymphoma and nasopharyngeal carcinoma. However, Qp is inactive in type III latency, and other promoters (the BamHI C promoter and/or the BamHI W promoter) are used for EBNA-1. The involvement of interferon regulatory factors (IRFs) in the regulation of Qp is suggested by the presence of an essential interferon-stimulated response element …

Genetic And Transcriptional Analyses Of The Vibrio Cholerae Mannose-Sensitive Hemagglutinin Type 4 Pilus Gene Locus, Jane W. Marsh, Ronald K. Taylor

Dartmouth Scholarship

The mannose-sensitive hemagglutinin (MSHA) of the Vibrio cholerae O1 El Tor biotype is a member of the family of type 4 pili. Type 4 pili are found on the surface of a variety of gram-negative bacteria and have demonstrated importance as host colonization factors, bacteriophage receptors, and mediators of DNA transfer. The gene locus required for the assembly and secretion of the MSHA pilus has been localized to a 16.7-kb region of the V. cholerae chromosome. Sixteen genes required for hemagglutination, including five that encode prepilin or prepilin-like proteins, have been identified. Examination of MSHA-specific cDNAs has localized two promoters …

Microbial Colonization Of The Cloacae Of Nestling Tree Swallows, Tamara K. Mills, Michael P. Lombardo, Patrick A. Thorpe

Peer Reviewed Publications

Microbes have the potential to be important selective forces in many aspects of avian biology. Microbes can affect fitness as a result of either their pathogenic or beneficial effects on host health. Little is known about the chronology of microbial colonization of nestlings or the effects of microbes on fledgling condition. We set out to (1) characterize the time course of microbial colonization of the cloacae of nestling Tree Swallows (Tachycineta bicolor), (2) examine the relationship between cloacal microbes and fledgling condition, and (3) determine if nest mates had similar assemblages of cloacal microbes. We repeatedly measured nestlings …

Characterization Of The Jembrana Disease Virus Tat Gene And The Cis- And Trans-Regulatory Elements In Its Long Terminal Repeats, Hexin Chen, Graham Wilcox, Gde Kertayadnya, Charles Wood

Nebraska Center for Virology: Faculty Publications

Jembrana disease virus (JDV) is a newly identified bovine lentivirus that is closely related to the bovine immunodeficiency virus (BIV). JDV contains a tat gene, encoded by two exons, which has potent transactivation activity. Cotransfection of the JDV tat expression plasmid with the JDV promoter chloramphenicol acetyltransferase (CAT) construct pJDV-U3R resulted in a substantial increase in the level of CAT mRNA transcribed from the JDV long terminal repeat (LTR) and a dramatic increase in the CAT protein level. Deletion analysis of the LTR sequences showed that sequences spanning nucleotides 268 to 153, including the TATA box and the predicted first …

Carmoviruses (Tombusviridae), Feng Qu, Thomas Jack Morris

School of Biological Sciences: Faculty Publications

Carmovirus is one of two officially recognized genera of Tombusviridae. Members of Tombusviridae all have icosahedral virions of about 30 nm in diameter with T = 3 symmetry that consists of 180 coat protein (CP) subunits of about 38- 43 kDa and a single-stranded (ss) RNA genome ranging in size from 4.0 to 4.7 kb. Carmoviruses share recognizable sequence similarity in both the polymerase and structural genes with members of the other genus, Tombusvirus. Sequence comparisons and phylogenetic analysis of these genes support the view that the genera are distinct but closely related. In addition, the genome organization of the …

Biological Characterization Of Rev Variation In Equine Infectious Anemia Virus, Michael Belshan, Matthew E. Harris, Anne E. Shoemaker, Thomas J. Hope, Susan Carpenter

Nebraska Center for Virology: Faculty Publications

Sequence analysis identified significant variation in the second exon of equine infectious anemia virus (EIAV) rev. Functional analysis indicated that limited amino acid variation in Rev significantly altered the export activity of the protein but did not affect Rev-dependent alternative splicing. EIAV Rev can mediate export through two independent cis -acting Rev-responsive elements (RREs), and differences among Rev variants were more pronounced when both RREs were present. Variation in Rev may be an important mechanism for regulation of virus replication in vivo and may contribute to changes in clinical disease.

Human Immunodeficiency Virus Neurotropism: An Analysis Of Viral Replication And Cytopathicity For Divergent Strains In Monocytes And Microglia, Anuja Ghorpade, Adeline Nukuna, Myhanh Che, Sheryl Haggerty, Yuri Persidsky, Eboni Carter, Leeroy Carhart, Laura Shafer, Howard Gendelman

Nebraska Center for Virology: Faculty Publications

Productive replication of human immunodeficiency virus type 1 (HIV-1) in brain macrophages and microglia is a critical component of viral neuropathogenesis. However, how virus-macrophage interactions lead to neurological disease remains incompletely understood. Possibly, a differential ability of virus to replicate in brain tissue macrophages versus macrophages in other tissues underlies HIV-1 neurovirulence. To these ends, we established systems for the isolation and propagation of pure populations of human microglia and then analyzed the viral life cycles of divergent HIV-1 strains in these cells and in cultured monocytes by using identical viral inocula and indicator systems. The HIV-1 isolates included those …

An Analysis Of Fecal Coliform Bacteria As A Water Quality Indicator, Janet Heyl Vail

Dissertations

The focus of this study is to assess the efficacy of fecal coliform bacteria as a microbiological water quality indicator. The scientific and educational context of fecal coliform bacteria is explored through analysis of large sets of water quality data, focused field monitoring projects, and review of commonly used resources for school-based and volunteer water-related studies. Analyzed data sets include long term sampling (10 years), multiple sites, daily samples, storm event samples, and other parameters monitored in parallel with bacteria. Special attention is given to data sets from Kent County, Michigan. A background reference site in Michigan unimpacted by humans …

Human Immunodeficiency Virus Neurotropism: An Analysis Of Viral Replication And Cytopathicity For Divergent Strains In Monocytes And Microglia, Anuja Ghorpade, Adeline Nukuna, Myhanh Che, Sherryl Haggerty, Yuri Persidsky, Eboni Carter, Leeroy Carhart, Laura Shafer, Howard Gendelman

Nebraska Center for Virology: Faculty Publications

Productive replication of human immunodeficiency virus type 1 (HIV-1) in brain macrophages and microglia is a critical component of viral neuropathogenesis. However, how virus-macrophage interactions lead to neurological disease remains incompletely understood. Possibly, a differential ability of virus to replicate in brain tissue macrophages versus macrophages in other tissues underlies HIV-1 neurovirulence. To these ends, we established systems for the isolation and propagation of pure populations of human microglia and then analyzed the viral life cycles of divergent HIV-1 strains in these cells and in cultured monocytes by using identical viral inocula and indicator systems. The HIV-1 isolates included those …

Human Herpesvirus 8 As A Potential Sexually Transmitted Agent In Honduras, C. Sosa, W. Klaskala, B. Chandran, R. Soto, L. Sieczkowski, M. H. Wu, M. Baum, Charles Wood

Nebraska Center for Virology: Faculty Publications

The seroprevalence of human herpesvirus 8 (HHV-8) was studied in 326 human immunodeficiency virus (HIV)–positive and –negative persons from Honduras; women constituted 77% (n = 251) of the subjects. Sera were tested for lytic HHV-8 antibodies by an IFA, and positive samples were confirmed by a radioimmunoprecipitation assay. Of the 326 persons tested, 58 (17.8%) had HHV- 8 antibodies. Among the HIV-infected women, 22.7% were seropositive; 11.3% of the HIV-negative women were seropositive. HHV-8 seroprevalence was almost four times higher in HIV-positive female commercial sex workers (36%) than in HIV-negative female non–commercial sex workers (9.9%; odds ratio = 3.8, 95% …

Seroprevalence Of Human Herpesvirus 8 Among Zambian Women Of Childbearing Age Without Kaposi’S Sarcoma (Ks) And Mother-Child Pairs With Ks, Jun He, Ganapati Bhat, Chipepo Kankasa, Chifumbe Chintu, Charles Mitchell, Wenjing Duan, Charles Wood

Nebraska Center for Virology: Faculty Publications

The seroprevalence of human herpesvirus 8 (HHV-8) among a group of Zambian women of reproductive age and among mother-child pairs in which either one of them has Kaposi’s sarcoma (KS) was determined. A cross-sectional group of 378 pregnant women was randomly recruited into the study, and 183 (48.4%) had HHV-8 antibodies. Among the human immunodeficiency virus (HIV)-1–infected women, 51.1% were HHV-8–seropositive, whereas of HIV- 1–negative women, 47.3% were HHV-8–seropositive. In addition, 21 women index patients with KS and 5 young children index patients with KS were studied. All children with KS had mothers who were HHV-8–seropositive, while not all children …

In Vitro Growth Characteristics Of Two Cryptococcus Neoformans Isolates, Kristy L. Jones, Juneann W. Murphy

Journal of the Arkansas Academy of Science

Cryptococcus neoformans is an opportunistic pathogen which attacks individuals with weakened immune systems. Two C. neoformans isolates, 184A and NU-2, were examined to determine characteristics that contribute to their difference in virulence. Both isolates were grown under tissue culture conditions, and different characteristics were tested at timed intervals. Isolate 184A was found to grow at a faster rate than isolate NU-2, with doubling times of 4 and 12 hours respectively. The polysaccharide capsule of isolate NU-2 doubled in size in four hours, while that of isolate 184A stayed approximately the same. The NU-2 yeast cell diameter without the polysaccharide capsule …

Vh Gene Usage In Vha2 Allotype-Producing B Cells From Mutant Alicia Rabbits, Anusorn Boonthum

Dissertations

No abstract provided.

Porcine Reproductive And Respiratory Syndrome Virus Replicates In Testicular Germ Cells, Alters Spermatogenesis, And Induces Germ Cell Death By Apoptosis, Jung-Hyang Sur, Alan R. Doster, James S. Christian, Judith A. Galeota, Robert W. Wills, Jeff J. Zimmerman, Fernando A. Osorio

Nebraska Center for Virology: Faculty Publications

Like other arteriviruses, porcine reproductive and respiratory syndrome virus (PRRSV) is shed in semen, a feature that is critical for the venereal transmission of this group of viruses. In spite of its epidemiological importance, little is known of the association of PRRSV or other arteriviruses with gonadal tissues. We experimentally infected a group of boars with PRRSV 12068-96, a virulent field strain. By combined use of in situ hybridization and immunohistochemistry, we detected infection by PRRSV in the testes of these boars. The PRRSV testicular replication in testis centers on two types of cells: (i) epithelial germ cells of the …

Isolation And Characterization Of Luxi And Luxr Homologues From Edwardsiella Ictaluri, Edward Todd Jacobs

Morehead State Theses and Dissertations

A thesis presented to the faculty of the College of Science and Technology at Morehead State University in partial fulfillment of the requirements for the Degree of Master of Science by Edward Todd Jacobs on August 1, 1997.

Study Of The Hiv-1 Gene Regulatory Proteins Tat And Rev By The Two-Hybrid System In Yeast, David Allen Elkins

Loma Linda University Electronic Theses, Dissertations & Projects

Tat and Rev, small proteins encoded by the Human Immunodeficiency Virus type 1 (HIV-1), regulate its pattern of gene expression within infected cells. Tat increases transcription from the integrated pro virus by approximately a thousand-fold; Rev effects a shift in the pattern of splicing of the viral mRNA. Both are likely to interact with cellular proteins in executing their respective functions. Tat has additional activities (including neurotoxicity and inhibition of lymphocyte activation) which may be extraneous to its viral functions.

In order to identify cellular proteins interacting with Tat and Rev, the genes for both were cloned into plasmid vectors …

Genomic Diversity In Staphylococcus Aureus, Bret Max Barton

Loma Linda University Electronic Theses, Dissertations & Projects

Staphylococcus aureus chromosomal DNA from 125 methicillin-resistant and 10 methicillin-sensitive clinical isolates was digested with rare-cutting restriction endonucleases and subjected to pulsed-field gel electrophoresis (PFGE). Thirty-nine distinct genomic macrorestriction patterns (GPs) were identified using SmaI; 29 of these patterns are from methicillin-resistant S. aureus (MRSA).

A dendrogram showing percent similarity among the patterns was constructed which revealed the considerable genomic diversity of the collection even though the isolates had been obtained during outbreaks. The majority of the MRSA formed one broad group with two outbreak subclasses. However, 8 of the 29 MRSA GPs (28%) were diverse.

Digestion with Csp …

Human Herpesvirus Type 8 Dna Sequences In Cell-Free Plasma And Mononuclear Cells Of Kaposi's Sarcoma Patients, William J. Harrington, Omar Bagasra, Carlos E. Sosa, Lisa E. Bobroski, Marianna Baum, Xue Lan Wen, Lisa Cabral, Gerald E. Byrne, Roger J. Pomerantz, Charles Wood

Nebraska Center for Virology: Faculty Publications

Human herpesvirus (HHV) type 8 bas been detected in both classical and AIDS-related Kaposi's sarcoma, body-cavity lymphomas, and other types of tumors. HHV-8 has also been detected in DNA from peripheral blood mononuclear cells (PBMC) of some Kaposi's sarcoma patients and more readily in B cell fractions derived from panned cell subpopulations. Two patients were followed using several methods: in situ hybridization, solution-based polymerase chain reaction (PCR), and in situ PCR. HHV-8 was intermittently detected in plasma and detection correlated with detection in PBMC. In situ PCR demonstrated HHV-8 sequences in both peripheral blood B lymphocytes and, to a lesser …