Microbiology Commons^™

The Bradyrhizobium Japonicum Fega Gene Encodes An Iron-Regulated Outer Membrane Protein With Similarity To Hydroxamate-Type Siderophore Receptors., Kristin Levier, Mary Lou Guerinot

Dartmouth Scholarship

Iron is important in the symbiosis between soybean and its nitrogen-fixing endosymbiont Bradyrhizobium japonicum, yet little is known about rhizobial iron acquisition strategies. Analysis of outer membrane proteins (OMPs) from B. japonicum 61A152 identified three iron-regulated OMPs in the size range of several known receptors for Fe(III)-scavenging siderophores. One of the iron-regulated proteins, FegA, was purified and microsequenced, and a reverse genetics approach was used to clone a fegA-containing DNA fragment. Sequencing of this fragment revealed a single open reading frame of 750 amino acids. A putative N-terminal signal sequence of 14 amino acids which would result in a mature …

About The Cellular Tropism Of The Gammaherpesvirus Bovine Herpesvirus Type 4, Fernando A. Osorio, J. A. Galeota-Wheeler, D. E. Reed, O. Lopez

Nebraska Center for Virology: Faculty Publications

We read with interest the recent report by Egyed et al. (4) about the distribution in tissue, in the natural host, of bovine herpesvirus type 4 (BHV-4), a member of the gammaherpesvirus group. Using PCR, those authors were able to trace BHV-4 (Movar strain) in several tissues of experimentally infected cattle (mostly spleen, lung, trachea, and nasal epithelium tissues) at different times, ranging from 3 to 48 days postinfection. The authors also report consistent detection of BHV-4 DNA in what they consider to be a pure population of lymphocytes obtained from peripheral blood of the BHV-4- infected cattle, and then …

Structural And Functional Analysis Of Secondary Structure Block To Mrna Utilization, Juan Lin

Loma Linda University Electronic Theses, Dissertations & Projects

Experimentally introduced secondary structure, formed by a cis-targeted hammerhead ribozyme in exon 2 two nucleotides dowstream of the 3' splice site of a yeast ACT-E. coli lacZ fusion gene, virtually abolishes splicing in vivo and inhibits B-galactosidase production. This is the first demonstration of the splicing inhibitory effect of secondary structures occurring in exons in the absence of splice site sequestration. Our goal is to understand the mechanism by which utilization of this splice site is impeded. Two cis - and one trans-acting mutations that alleviate the splicing blockage were obtained by screening for restoration of B-galactosidase activity. Both cis …

Diseased Thyrocytes Have Aberrant Communication And Tnf Receptor Secretion, Mary Labue

Loma Linda University Electronic Theses, Dissertations & Projects

Autoimmune diseases are a world-wide health concern and are an intense area of investigation. However, one area of research has been largely ignored. This involves the contribution of the target tissue to the disease process. We hypothesized that cell-cell communication is disrupted by lymphocytic infiltration and advancing fibrosis and this disruption results in altered target cell responses to immune components produced at the site of inflammation, including Tumor Necrosis Factor-alpha (TNF- α).

We have used thyroiditis as our model of autoimmunity and have documented the development of spontaneous thyroiditis in MRL-lpr/lpr mice. We have found aberrant connexin expression in …

Effect Of Sediment Particle Size And Temperature On Fecal Bacteria Mortality Rates And The Fecal Coliform/Fecal Streptococci Ratio, J. M. Howell, Mark S. Coyne, P. L. Cornelius

Plant and Soil Sciences Faculty Publications

Extended survival of fecal bacteria in sediment can obscure the source and extent of fecal contamination in agricultural settings. The variability in fecal coliform/fecal streptococci (FC/FS) ratios with time and discrepancies between observable fecal sources and measured FC/FS ratios in shallow surface water from agricultural watersheds may be explained by examining FC and FS mortality rates in response to ambient temperature and sediment particle size. We measured FC and FS mortality rates at three different temperatures and in three feces-amended sediments with different particle size in a laboratory study. In controlled conditions, using physiological saline to reduce cell death by …

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 …

Parotid Gland Lipoma: A Rare Entity, S Muzaffar, N Kayani, S H. Hasan

Department of Pathology and Laboratory Medicine

No abstract provided.

Bacillus Thuringiensis Hd-73 Spores Have Surface-Localized Cry1ac Toxin: Physiological And Pathogenic Consequences, Cheng Du, Kenneth Nickerson

Papers in Microbiology

Spores from Cry⁺ strains of Bacillus thuringiensis bound fluorescein isothiocyanate-labeled antibodies specific for the 65-kDa activated Cry 1Ac toxin, whereas spores from Bacillus cereus and Cry^- strains of B. thuringiensis did not. The Cry⁺ spores could be activated for germination by alkaline conditions (pH 10.3), whereas Cry^- spores could not. Once the surrounding exosporia had been removed or permeabilized, Cry⁺ spores were able to bind the toxin receptor(s) from insect gut brush border membrane vesicle preparations, and their germination rates were increased ca. threefold in the presence of brush border membrane vesicles. A model is …

Genetic And Phylogenetic Divergence Of Feline Immunodeficiency Virus In The Puma (Puma Concolor), Margaret A. Carpenter, Eric W. Brown, Melanie Culver, Warren E. Johnson, Jill Pecon-Slattery, Dulce Brousset, Stephen J. O'Brien

Biology Faculty Articles

Feline immunodeficiency virus (FIV) is a lentivirus which causes an AIDS-like disease in domestic cats (Felis catus). A number of other felid species, including the puma (Puma concolor), carry a virus closely related to domestic cat FIV. Serological testing revealed the presence of antibodies to FIV in 22% of 434 samples from throughout the geographic range of the puma. FIV-Pco pol gene sequences isolated from pumas revealed extensive sequence diversity, greater than has been documented in the domestic cat. The puma sequences formed two highly divergent groups, analogous to the clades which have been defined for …

A Role For A Rab4-Like Gtpase In Endocytosis And In Regulation Of Contractile Vacuole Structure And Function In Dictyostelium Discoideum, John Bush, Lesly A. Temesvari, Juan Rodriguez-Paris, Greg Buczynski, James Cardelli

Publications

The small Mr Rab4-like GTPase, RabD, localizes to the endosomal pathway and the contractile vacuole membrane system in Dictyostelium discoideum. Stably transformed cell lines overexpressing a dominant negative functioning RabD internalized fluid phase marker at 50% of the rate of wild-type cells. Mutant cells were also slower at recycling internalized fluid. Microscopic and biochemical approaches indicated that the transport of fluid to large postlysosome vacuoles was delayed in mutant cells, resulting in an accumulation in acidic smaller vesicles, probably lysosomes. Also, RabD N121I-expressing cell lines missorted a small but significant percentage of newly synthesized lysosomal alpha-mannosidase precursor polypeptides. However, the …

Characterization Of P39, A Borrelia Burgdorferi Specific Protein, Jill M. Troyer

Theses and Dissertations in Biomedical Sciences

Borrelia burgdorferi, the causative agent of Lyme disease, expresses a species specific, 39 kiloDalton protein of unknown function called P39. This protein is highly immunogenic and the presence of anti-P39 antibodies in patient sera is used as an indicator of B. burgdorferi infection. P39 is also a candidate for vaccine developement due to conserved expression of P39 among B. burgdorferi isolates. Since little is presently known about this important molecule, the present study was designed to characterize P39 and was carried out in six separate investigations. First, infectious and noninfectious isolates of B. burgdorferi were established in vitro, and …

Crystal Protein Expression During Vegetative Growth Of Dictyostelium, Kristy K. Taylor, Catherine P. Chia

Papers in Microbiology

We are studying interactions between membrane proteins and the cytoskeleton of Dictyostelium discoideum. Detergent (Triton X-100) -insoluble cytoskeletons from vegetative (AX2) amebae are enriched in a 70 kDa Concanavalin A (Con A)-binding protein (gp70). An enriched fraction of gp70 was prepared by Con A affinity chromatography of cytoskeletons from axenically-grown cells. N-terminal amino acid sequence of gel-purified gp70 established its identity to a 69 kDa 'crystal protein' previously characterized by Bomblies, et al. (1990, J. Cell Biol. 110:669 -679). Enzymatic deglycosylation of gp70 resulted in a 65 kDa product that did not bind Con A, verifying the predicted number …

Activation Of The Human Thymidine Kinase (Tk) Promoter By Simian Virus 40 Large T Antigen Requires Both The T Antigen Prb Family-Binding Domain And Tk Promoter Sequences Resembling E2f-Binding Sites., Michelle M. Anderson, Jun Chen, Charles N. Cole, Susan E. Conrad

Dartmouth Scholarship

Infection of quiescent cells with the DNA tumor virus simian virus 40 induces expression of the cellular thymidine kinase (TK) gene a minimum of 10- to 20-fold, and this induction depends upon the viral protein large T antigen (T-Ag). To define both human TK promoter elements and T-Ag functional domains required for transcriptional induction, we have established a system in which stable Rat-1 transfectants harboring TK promoter-luciferase hybrid genes are infected with recombinant adenoviruses expressing either wild-type or mutant forms of T-Ag and luciferase expression is measured as an indicator of promoter activity. The results show that (i) a 135-bp …

In Vivo Detection Of Porcine Reproductive And Respiratory Syndrome Virus Rna By In Situ Hybridization At Different Times Post-Infection, Jung-Hyang Sur, Vicki Cooper, Judith A. Galeota, Richard A. Hesse, Alan R. Doster, Fernando A. Osorio

Nebraska Center for Virology: Faculty Publications

We studied the distribution of porcine reproductive and respiratory syndrome virus (PRRSV) RNA in tissues by in situ hybridization at different times postinfection (p.i.). The probe used for in situ hybridization was prepared by reverse transcription of PRRSV RNA, followed by PCR amplification of the cDNA. The sequence amplified corresponded to 433 bp from PRRSV open reading frame 7, which is contained in the nucleocapsid protein gene and which is highly conserved in both European and American strains (H. Mardassi, L. Wilson, S. Mounir, and S. Dea, J. Clin. Microbiol. 32:2197–2203, 1994). An immunohistochemical technique was used to detect PRRSV …

The Bacillus Thuringiensis Insecticidal Toxin Binds Biotin-Containing Proteins, Cheng Du, Kenneth Nickerson

Papers in Microbiology

Brush border membrane vesicles from larvae of the tobacco hornworm, Manduca sexta, contain protein bands of 85 and 120 kDa which react directly with streptavidin conjugated to alkaline phosphatase. The binding could be prevented either by including 10 μM biotin in the reaction mixture or by prior incubation of the brush border membrane vesicles with an activated 60- to 65-kDa toxin from Bacillus thuringiensis HD-73. The ability of B. thuringiensis toxins to recognize biotin-containing proteins was confirmed by their binding to pyruvate carboxylase, a biotin-containing enzyme, as well as to biotinylated ovalbumin and biotinylated bovine serum albumin but not …

Vif-Negative Human Immunodeficiency Virus Type 1 Persistently Replicates In Primary Macrophages, Producing Attenuated Progeny Virus, Iqbal Chowdhury, Wei Chao, Mary Jane Potash, Pavel Sova, Howard Gendelman, David J. Volsky

Nebraska Center for Virology: Faculty Publications

The vif gene of human immunodeficiency virus type 1 (HIV-1) is required for efficient infection of primary T lymphocytes. In this study, we investigated in detail the role of vif in productive infection of primary monocyte-derived macrophages (MDM). Viruses carrying missense or deletion mutations in vif were constructed on the background of the monocytotropic recombinant NLHXADA-GP. Using MDM from multiple donors, we found that vif mutants produced in complementing or partially complementing cell lines were approximately 10% as infectious as wild-type virus when assayed for incomplete, complete, and circularized viral DNA molecules by quantitative PCR amplification or for viral core …

Resistance Of Blastocystis Hominis Cysts To Chlorine, M Zaki, V Zaman, N A. Sheikh

Department of Pathology and Laboratory Medicine

No abstract provided.

A Truncated Form Of The Pho80 Cyclin Of Saccharomyces Cerevisiae Induces Expression Of A Small Cytosolic Factor Which Inhibits Vacuole Inheritance., Teresa Nicolson, Barbara Conradt, William Wickner

Dartmouth Scholarship

Vacuoles project streams of vesicles and membranous tubules into the yeast bud where they fuse, founding the daughter cell organelle, vac5-1, which encodes a truncated form of the Pho80 cyclin, inhibits normal vacuole inheritance. An in vitro inheritance assay which measures the fusion of vacuoles serves as a model for several steps of this process. We find that cytosol isolated from the vac5-1 mutant is unable to promote the fusion of wild-type vacuoles in the in vitro assay. Wild-type vacuoles are irreversibly inactivated in a time- and temperature-dependent manner if preincubated with vac5-1 cytosol and ATP, suggesting the presence of …

Oxidative Stress Detection With Escherichia Coli Harboring A Katg'::Lux Fusion, S. Belkin, D. R. Smulski, Amy Cheng Vollmer, T. K. Van Dyk, R. A. Larossa

Biology Faculty Works

A plasmid containing a transcriptional fusion of the Escherichia coli katG promoter to a truncated Vibrio fischeri lux operon (luxCDABE) was constructed. An E. coli strain bearing this plasmid (strain DPD2511) exhibited low basal levels of luminescence, which increased up to 1,000-fold in the presence of hydrogen peroxide, organic peroxides, redox-cycling agents (methyl viologen and menadione), a hydrogen peroxide-producing enzyme system (xanthine and xanthine oxidase), and cigarette smoke. An oxyR deletion abolished hydrogen peroxide-dependent induction, confirming that oxyR controlled katG'::lux luminescence. Light emission was also induced by ethanol by an unexplained mechanism. A marked synergistic response was observed when cells …

A Computer-Simulated Restriction Fragment Length Polymorphism Analysis Of Bacterial Small-Subunit Rrna Genes: Efficacy Of Selected Tetrameric Restriction Enzymes For Studies Of Microbial Diversity In Nature, Craig L. Moyer, James M. Tiedje, Fred C. Dobbs, David M. Karl

OES Faculty Publications

An assessment of 10 tetrameric restriction enzymes (TREs) was conducted by using a computer-simulated restriction fragment length polymorphism (RFLP) analysis for over 100 proximally and distally related bacterial small-subunit (SSU) rRNA gene sequences. Screening SSU rDNA clone libraries with TREs has become an effective strategy because of logistic simplicity, commercial availability, and economy. However, the rationale for selecting the type and number of TREs has not been systematically evaluated. Our objective nas to identify the optimal combination of TREs for RnP screening of cloned SSU rRNA genes from undefined bacterial clone libraries. After computer-simulated TRE digestion, the resultant fragments were …

Subcutaneous Phaeohyphomycosis: Infection With Pigmented Fungi, I N. Soomro, A Kennedy

Department of Pathology and Laboratory Medicine

No abstract provided.

Molecular Studies On Biv Infection And Its Interaction With Other Bovine Viruses, Charles Wood, Harish Minocha, Yunqi Geng

Nebraska Center for Virology: Faculty Publications

Bovine immunodeficiency virus (BIV) is a bovine lentivirus that infects animals world-wide. The authors have cloned the BIV env and gag proteins and several sub-unit peptides spanning the entire coding regions of the two proteins as fusions to the bacterial trpE protein and expressed them in Escherichia coli (Spindler et al. 1984). These recombinant proteins can readily be expressed but the expression levels of these peptides varied. When these proteins were tested against a panel of bovine sera, only sera from BIV-infected animals reacted specifically. One major antigenic determinant was identified in the cartoxyl terminus of the p26 gag protein …

Potential New Method: A Comparability Study Of M-Coliblue24 And M-Endo Les For The Verified Recoveries Of Total Coliforms And E. Coli, Kevin Robert Geotz

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 in Biology by Kevin Robert Goetz on June 10, 1996.

Longitudinal Studies Of Viral Sequence, Viral Phenotype, And Immunologic Parameters Of Human Immunodeficiency Virus Type 1 Infection In Perinatally Infected Twins With Discordant Disease Courses, Cecelia Hutto, Yi Zhou, Jun He, Rebeca Geffin, Martin Hill, Walter Scott, Charles Wood

Nebraska Center for Virology: Faculty Publications

Perinatal human immunodeficiency virus type 1 (HIV-1) infections cause a broad spectrum of clinical disease and are variable in both the age of the patient at onset of serious disease and the progression of the clinical course. Heterozygotic perinatally infected twins with a marked difference in their clinical courses were monitored during the first 2 years of life. Twin B, the second-born twin, developed AIDS by 6 months of age and died at 22 months of age, while twin A remained minimally symptomatic through the first 2 years. Sequential blood specimens were obtained from the twins in order to characterize …

Involvement Of The Vacuolar Proton-Translocating Atpase In Multiple Steps Of The Endo-Lysosomal System And In The Contractile Vacuole System Of Dictyostelium Discoideum, Lesly A. Temesvari, Juan M. Rodriguez-Paris, John M. Bush, Linyi Zhang, James A. Cardelli

Publications

We have investigated the effects of Concanamycin A (CMA), a specific inhibitor of vacuolar type H(+)-ATPases, on acidification and function of the endo-lysosomal and contractile vacuole (CV) systems of D. discoideum. This drug inhibited acidification and increased the pH of endo-lysosomal vesicles both in vivo and in vitro in a dose dependent manner. Treatment also inhibited endocytosis and exocytosis of fluid phase, and phagocytosis of latex beads. This report also confirms our previous conclusions (Cardelli et al. (1989) J. Biol. Chem. 264, 3454–3463) that maintenance of acidic pH in lumenal compartments is required for efficient processing and targeting of a …

Expression Of The Human Milk Protein, Beta-Casein : In Transgenic Potato Plants, Wendy Roberts

Loma Linda University Electronic Theses, Dissertations & Projects

Mother’s milk is recommended by the nutritional and pediatric communities as the best food for suckling infants. For non breast-fed infants, bovine milk-based formulas have been regarded as a suitable substitute. In the early 1900's, physicians found that foreign proteins in cow milk were responsible for gastrointestinal disease and even digestive collapse of the newborn fed a cow milk-based diet¹⁶. The allergenicity of cow milk for the human infant appears to be caused predominantly by p-casein¹⁷. In this research, the cDNA encoding human p-casein was introduced into potato cells under the control of the bidirectional, auxin-inducible …

Tyrosine Kinase-Dependent Release Of An Adenovirus Preterminal Protein Complex From The Nuclear Matrix, Peter C. Angeletti, Jeffrey A. Engler

Nebraska Center for Virology: Faculty Publications

Adenovirus (Ad) replicative complexes form at discrete sites on the nuclear matrix (NM) through the interaction of Ad preterminal protein (pTP). The NM is a highly salt-resistant fibrillar network which is known to anchor transcription, mRNA splicing, and DNA replication complexes. Incubation of rATP with NM to which pTP was bound caused the release of pTP as a pTP-NM complex with a size of 220 to 230 kDa; incubation with 5’ adenylylimidodiphosphate (rAMP-PNP) showed no significant release, indicating that rATP hydrolysis was required. With NM extracts, it was shown that a pTP-NM complex which was capable of binding Ad origin …

Molecular Approaches In Attempts To Identify The Flagellin B-Type Gene In Clone Pkw52, Yolanda F. Kirkpatrick

Masters Theses

Pseudomonas aeruginosa is an important pathogen for susceptible individuals particularly those immunocompromised due to the prolonged use of drugs, noscomial infections, burns, or cancer. In addition, P. aeruginosa infections are high among cystic fibrosis patients. P. aeruginosa presents a unique health concern and challenge for researchers due to its multiple virulence factors and the increasing number of patients developing P. aeruginosa infections. P. aeruginosa produces a variety of toxins and enzymes. The flagellum of P. aeruginosa allows the bacterium to be motile. Motility has enabled the P. aeruginosa bacterium to rapidly colonize the host's body.

The major structural component of …

Report Of Significant Findings--Las Vegas Bay/Boulder Basin Investigations, James F. Labounty, Michael Horn, Bureau Of Reclamation

Publications (WR)

Field sampling was carried out between 0830 and 1500 beginning at the confluence of Las Vegas Wash and the Inner Las Vegas Bay. Ten (10) locations were sampled, each in a similar manner. Locations of sampling stations are in line from the Wash-Bay confluence to a point midway between Saddle and Black Islands. In addition, sampling was done at a location midway between Sentinel Island and the base of Fortification Hill, and at the buoy line in front of Hoover Dam. Sampling stations are labeled from LV01, at Wash-Bay confluence, to LV17 at Hoover Dam. A significant data collection point …

Antibody To The Ligand For Cd40 (Gp39) Inhibits Murine Aids-Associated Splenomegaly, Hypergammaglobulinemia, And Immunodeficiency In Disease-Susceptible C57bl/6 Mice., Kathy A. Green, Karen M. Crassi, Jon D. Laman, Arjan Schoneveld, Rendall R. Strawbridge, Teresa M. Foy, Randolph J. Noelle, William R. Green

Dartmouth Scholarship

Infection of genetically susceptible C57BL/6 mice with the LP-BM5 isolate of murine retroviruses cause profound splenomegaly, hypergammaglobulinemia, lymphadenopathy, and an immunodeficiency syndrome which includes the development of terminal B-cell lymphomas. Because many of these and the other manifestations of LP-BM5 virus-induced disease are similar to those seen in AIDS, this syndrome has been named murine AIDS, or MAIDS. Previous reports have shown that the onset of MAIDS depends on the presence of both CD41 T cells and B cells and have suggested that CD41 T-cell-B-cell interactions are important to disease pathogenesis. Here, we assessed the possibility that interactions between CD40 …