Microbiology Commons^™

Effects Of Conalbumin Bound Iron On The Growth Of Salmonella Paratyphi B And Salmonella Thompson, John Nicholas Mason

Dissertations and Theses

I have investigated the possibility that specific conalbumin (ovotransferrin) iron saturation levels enable less virulent strains of Salmonella to become more virulent. Iron starved cells of two pathogenic Salmonella strains, S. paratyphi B var. java and S. thompson, were cultured in iron limited media at 3 different iron conalbumin saturation levels. Results indicate that strains differ significantly at both low and high iron saturation conalbumin. These differences depict a growth advantage for S. paratyphi B which correlates with reports by the Centers for Disease Control that S. paratyphi B was 3 times more frequent in blood isolates than S. …

The Ability Of Simian Virus 40 Large T Antigen To Immortalize Primary Mouse Embryo Fibroblasts Cosegregates With Its Ability To Bind To P53., Jiyue Y. Zhu, Marina Abate, Philip W. Rice, Charles N. Cole

Dartmouth Scholarship

The large T antigen encoded by simian virus 40 (SV40) plays essential roles in the infection of permissive cells, leading to production of progeny virions, and in the infection of nonpermissive cells, leading to malignant transformation. Primary mouse embryo fibroblasts (MEFs) are nonpermissive for SV40, and infection by wild-type SV40 leads to immortalization and transformation of a small percentage of infected cells. We examined the ability of an extensive set of mutants whose lesions affect SV40 large T antigen to immortalize MEFs. We found that immortalization activity was retained by all mutants whose lesions are located upstream of codon 346. …

Hepatic Dysfunction In Falciparum Malaria, J Khan, J Akhter, H Sheikh, W Jafri

Department of Pathology and Laboratory Medicine

No abstract provided.

Nutritional Complementation Of Oxidative Glucose Metabolism In Escherichia Coli Via Pyrroloquinoline Quinone-Dependent Glucose Dehydrogenase And The Entner-Doudoroff Pathway, Michael Adamowicz, Tyrrell Conway, Kenneth W. Nickerson

Papers in Microbiology

Two glucose-negative Escherichia coli mutants (ZSC113 and DF214) were unable to grow on glucose as the sole carbon source unless supplemented with pyrroloquinoline quinone (PQQ). PQQ is the cofactor for the periplasmic enzyme glucose dehydrogenase, which converts glucose to gluconate. Aerobically, E. coli ZSC113 grew on glucose plus PQQ with a generation time of 65 min, a generation time about the same as that for wildtype E. coli in a defined glucose-salts medium. Thus, for E. coli ZSC113 the Entner-Doudoroff pathway was fully able to replace the Embden-Meyerhof-Parnas pathway. In the presence of 5% sodium dodecyl sulfate, PQQ no longer …

Control Of Salmonella On Beef Tissue Surfaces In A Model System By Pre- And Post-Evisceration Washing And Sanitizing, With And Without Spray Chilling, James S. Dickson, Maynard E. Anderson

James S. Dickson

Beef tissue was inoculated with a nalidixic acid resistant strain of Salmonella California and processed under conditions simulating pre- and post-evisceration carcass washing and sanitizing. These treatments, using distilled water to wash and 2% acetic acid to sanitize, reduced the population of salmonellae by as much as 2-log10 cycles when compared to samples which were washed only in distilled water. Increasing the acid temperature to 55°C reduced the bacterial populations further. Spray chilling, when used in series with the pre- and post-evisceration treatments, apparently resulted in recovery of some injured Salmonella.

The Inability Of Human Immunodeficiency Virus To Infect Chimpanzee Monocytes Can Be Overcome By Serial Viral Passage In Vivo, Howard Gendelman, Garth D. Ehrlich, Lisa M. Baca, Shawn Conley, Jorge Ribas, D. Chester Kalter, Monte S. Meltzer, Bernard J. Poiesz, Peter Nara

Nebraska Center for Virology: Faculty Publications

Studies of lentivirus infection in ruminants, nonhuman primates, and humans suggest that virus infection of macrophages plays a central role in the disease process. To investigate whether human immunodeficiency virus type 1 (HIV-1) can infect chimpanzee macrophages, we recovered monocytes from peripheral blood mononuclear cells of HIV-1-negative animals and inoculated these and control human monocytes with a panel of four human-passaged monocytotropic virus strains and one chimpanzee-passaged isolate. HIV-1-infected human monocytes synthesized proviral DNA, viral mRNA, p24 antigen, and progeny virions. In contrast, except for the chimpanzee-passaged HIV-1 isolate, chimpanzee monocytes failed to support HIV-1 replication when cultured under both …

Transactivation Of The Human Immunodeficiency Virus Promoter By Human Herpesvirus 6 (Hhv-6) Strains Gs And Z-29 In Primary Human T Lymphocytes And Identification Of Transactivating Hhv-6(Gs) Gene Fragments, Rebecca Horvat, Charles Wood, Steven Josephs, N. Balanchandran

Nebraska Center for Virology: Faculty Publications

Human herpesvirus 6 (HHV-6) can activate the human immunodeficiency virus (HIV) promoter and accelerate cytopathic effects in HIV-infected human T cells. This study examines the regions of the HIV promoter required for HHV-6 transactivation in a heterogeneous population of primary human T lymphocytes with or without antigenic stimulation. Two different strains of HHV-6, GS and Z29, transactivated the HIV promoter. The GS strain transactivated the promoter in both stimulated and resting T cells, while the Z29 strain increased HIV promoter activity only in stimulated T cells. Three DNA clones containing HHV-6(GS) genomic fragments transactivated the HIV promoter in cotransfected T …

Mapping The Transcriptional Transactivation Function Of Simian Virus 40 Large T Antigen., Jiyue Y. Zhu, Philip W. Rice, Michele Chamberlain, Charles N. Cole

Dartmouth Scholarship

T antigen is able to transactivate gene expression from the simian virus 40 (SV40) late promoter and from several other viral and cellular promoters. Neither the mechanisms of transactivation by T antigen nor the regions of T antigen required for this activity have been determined. To address the latter point, we have measured the ability of a set of SV40 large T antigen mutants to stimulate gene expression in CV-1 monkey kidney cells from the SV40 late promoter and Rous sarcoma virus (RSV) long terminal repeat (LTR) promoter. Transactivation, although reduced, was retained by an N-terminal 138-amino-acid fragment of T …

Transactivation Of The Human Immunodeficiency Virus Promoter By Human Herpesvirus 6 (Hhv-6) Strains Gs And 2-29 In Primary Human T Lymphocytes And Identification Of Transactivating Hhv-6(Gs) Gene Fragments, Rebecca Horvat, Charles Wood, Steven Josephs, N. Balachandran

Nebraska Center for Virology: Faculty Publications

Human herpesvirus 6 (HHV-6) can activate the human immunodeficiency virus (HIV) promoter and accelerate cytopathic effects in HIV-infected human T cells. This study examines the regions of the HIV promoter required for HHVd transactivation in a heterogeneous population of primary human T lymphocytes with or without antigenic stimulation. Two different strains of HHV-6, GS and 229, transactivated the HIV promoter. The GS strain transactivated the promoter in both stimulated and resting T cells, while the 229 strain increased HIV promoter activity only in stimulated T cells. Three DNA clones containing HHV-6(GS) genomic fragments transactivated the HIV promoter in cotransfected T …

Aerotaxis In Halobacterium Halobium, Jemima Clara Lindbeck

Loma Linda University Electronic Theses, Dissertations & Projects

Aerotaxis, previously studied in eubacteria was investigated in the archaebacterium Halobacterium halobium. A quantitative temporal assay for aerotaxis of H. halobium was developed using computer assisted motion analysis. Aerotaxis was most pronounced in early log phase cultures. Peak aerotaxis was found to coincide with peak respiration consistent with a model in which respiration is required for aerotaxis. In a spatial assay a mutant strain lacking the proton motive force (pmf)-generating pigments, bacteriorhodopsin (bR) and halorhodopsin (hR), exhibited enhanced aerotaxis bands relative to the bR⁺hR⁺ parent. The enhanced aerotaxis in the mutant cells was not due solely to …

A Flow Cytometric Analysis Of Immunocyte Populations In Peripheral Blood And Pleural Effusions Of Patients With Cancer, Mandy L. Bohn

Loma Linda University Electronic Theses, Dissertations & Projects

Two-color flow cytometric analysis was performed on paired samples of peripheral blood (PB) and pleural effusions (PE) of patients with metastatic malignancies. The purpose of these analyses was to test the hypothesis that there are significant differences in the distribution of immunocyte populations in the PE compared to that of the PB.

The majority of the pleural fluid immunocytes were CD3+ cells. The ratio of CD4+ to CD8+ cells was higher in the PE compared to PB. Levels of CD8+, CD19+ and CD14+ cells were not significantly different in the PE compared to the levels in the PB. CD16+CD56+ cells …

Use Of Antisense Rna In The Study Of Essential Cell Division Genes In Escherichia Coli, Quynh V. Nguyen

Honors Theses

No abstract provided.

Ecdysterone Regulatory Elements Function As Both Transcriptional Activators And Repressors., Leonard Dobens, Karen Rudolph, Edward M. Berger

Dartmouth Scholarship

A synthetic, 23-bp ecdysterone regulatory element (EcRE), derived from the upstream region of the Drosophila melanogaster hsp27 gene, was inserted adjacent to the herpes simplex virus thymidine kinase promoter fused to a bacterial gene for chloramphenicol acetyltransferase (CAT). Hybrid constructs were transfected into Drosophila S3 cells and assayed for ecdysterone-inducible CAT expression. In the absence of ecdysterone a tandem pair of EcREs repressed the high constitutive level of CAT activity found after transfection with the parent reporter plasmid alone. After hormone addition very high levels of CAT activity were observed. Insertion of the EcRE pair 3' of the CAT gene …

Strain-Specific Neutralizing Determinant In The Transmembrane Protein Of Simian Immunodeficiency Virus, Toshiaki Kodama, Dawn P. Wooley, Daniel P. Silva, Fulvia Dimarzo Veronese, Ronald C. Desrosiers

Neuroscience, Cell Biology & Physiology Faculty Publications

Monoclonal antibody SF8/5E11, which recognizes the transmembrane protein (TMP) of simian immunodeficiency virus of macaque monkeys (SIVmac), displayed strict strain specificity. It reacted with cloned and uncloned SIVmac251 but not with cloned SIVmac142 and SIVmac239 on immunoblots. This monoclonal antibody neutralized infection by cloned, cell-free SIVmac251 and inhibited formation of syncytia by cloned SIVmac251-infected cells; these activities were specific to cloned SIVmac251 and did not occur with the other viruses. Site-specific mutagenesis was used to show that TMP amino acids 106 to 110 (Asp-Trp-Asn-Asn-Asp) determined the strain specificity of the monoclonal antibody. This strain-specific neutralizing determinant is located within a …

The Study Of Natural Product(S) From Opsanus Tau (Linnaeus), The Oyster Toadfish, Miki Taira

Chemistry & Biochemistry Theses & Dissertations

Opsanus tau (Linnaeus), the Oyster toadfish, commonly inhabits the Chesapeake Bay. The Oyster toadfish has been categorized as a venomous and/or ichthyotoxic fish; however, chemical studies to substantiate these claims have not been forthcoming.

The oyster toadfish has dorsal and opercular spines which are believed to secrete a venom(s). Based on our findings, the oyster toadfish does not secrete venom(s) from these spines.

The oyster toadfish secretes large amounts of a mucus when it is irritated. 3-Octanone was believed to be the major ichthyotoxic component of the mucus from the oyster toadfish. Based on our investigation, 3-octanone is present only …

Selection Of Genetic Variants Of Simian Immunodeficiency Virus In Persistently Infected Rhesus Monkeys, Dawn P. Wooley, Ronald C. Desrosiers

Neuroscience, Cell Biology & Physiology Faculty Publications

Genetic and antigenic variation may be one means by which lentiviruses that cause AIDS avoid elimination by host immune responses. Genetic variation in the envelope gene (env) was studied by comparing the nucleotide sequences of 27 clones obtained from two rhesus monkeys infected with molecularly cloned simian immunodeficiency virus. All 27 clones differed from each other and differed from the input clone in the gp120 (SU) portion of the envelope gene. Nucleotide substitutions were shown to accumulate with time at an average rate of 8.5 per 1,000 per year in SU. Surprisingly, the majority of nucleotide substitutions (81%) resulted in …

Rapid Detection Of Bovine Viral Diarrhea Virus By Polymerase Chain Reaction, O. J. Lopez, Fernando A. Osorio, Ruben O. Donis

Nebraska Center for Virology: Faculty Publications

The polymerase chain reaction was used to detect genomic sequences of the positive-stranded RNA of bovine viral diarrhea virus (BVDV), a member of the family Togaviridae. Using a set of 20-bp primers located within the conserved 3' region of the BVDV genome, we were able to consistently amplify a 205-bp target sequence from BVDV cDNA. BVDV RNAs from cell culture-propagated BVDV reference strains, diverse unrelated cytopathic and noncytopathic field isolates, and clinical serum samples were transcribed to cDNA by using avian myeloblastosis virus reverse transcriptase and further specifically amplified by using the polymerase chain reaction assay. The amplification assay …

Contamination Of Beef Tissue Surfaces By Cattle Manure Inoculated With Salmonella Typhimurium And Listeria Monocytogenes, James S. Dickson

James S. Dickson

Contamination of beef lean and fat tissue surfaces by Salmonella typhimurium and Listeria monocytogenes was evaluated using phosphate buffer or sterilized manure as an inoculation menstruum. Immersion in inoculated phosphate buffer resulted in an increase in numbers of attached cells during the 120 min inoculation for both bacterial species. Tissue immersed in inoculated manure generally showed an increase in cell numbers up to 10 min of immersion with only slight increases in cell numbers from lOto 120 min. Fewer cells attached to either tissue type from the manure inoculum (P<0.05), although actual numerical differences were small.

Quantitation Of Type I Collagen Mrna In Intact Human Fibroblasts, Edward Touma

Master's Theses

No abstract provided.

Interleukin-2 Activated Anti-Fungal Activity Of Lymphocytes, David W.A. Beno

Dissertations

No abstract provided.

Reactivity Of Anti-Group A Type 12 Streptococcal Cell Membrane Monoclonal Antibody To Human Glomerular Basement Membrane, Sally C. Kent

Dissertations

No abstract provided.

Isolation And Immunochemical Characterization Of Streptococcal Cell Membrane Antigens Immunologically Related To Basement Membrane, Mark E. Zelman

Dissertations

No abstract provided.

Detergent (Sodium Dodecyl Sulfate) Shock Proteins In Escherichia Coli, Michael Adamowicz, Philip M. Kelley, Kenneth W. Nickerson

Papers in Microbiology

The protein composition of Escherichia coli W3110 grown in the presence and absence of 5% sodium dodecyl sulfate (SDS) was examined by two-dimensional gel electrophoresis. In SDS-grown cells, at least 4 proteins were turned on, 13 were turned off, 15 were elevated, and 15 were depressed. The 19 unique and elevated SDS-induced spots constituted 7.91% of the total ³⁵S-labeled protein. There was no apparent overlap between these 19 detergent (SDS) stress proteins and those of other known bacterial stress responses. The detergent stress stimulon is a distinct and independent stimulon. Its physiological relevance probably derives from the presence of …

Cyanonews (Vol. 7, No. 1, April 1991), Jeff Elhai

CyanoNews

CyanoNews was a newsletter that served the cyanobacteriological community from 1985 to 2003, with content provided by readers (sort of a blog before there were blogs). The newsletter reported new findings from the lab, summaries of recent meetings (often provided by graduate students and post-docs entering the field), positions sought or available, life transitions, a compendium of recent cyanobacteria-related articles, and other items of interest to those who study cyanobacteria.

Attachment Of Salmonella Typhimurium And Listeria Monocytogenes To Glass As Affected By Surface Film Thickness, Cell Density, And Bacterial Motility, James S. Dickson, E. K. Daniels

James S. Dickson

With equal cell densities, surface film thickness did not influence the numbers &Salmonella typhimurium and Listeria monocytogenes cells which attached to glass. Motile L. monocytogenes cells had a greater cell surface charge and generally attached in higher numbers than non-motile cells.

Control Of Salmonella Typhimurium, Listeria Monocytogenes, And Escherichia Coli 0157:H7 On Beef In A Model Spray Chilling System, James S. Dickson

James S. Dickson

Simulated water spray chilling cycles were modified to include acetic acid as a sanitizer. The modified cycles were tested for effectiveness in reducing levels of S. typhimutium, L. monocyfogenes, and E. coli 0157:H7 on beef lean and fat tissue. Reductions of up to 3 log cycles were obtained for all three bacterial species on fat tissue. Reduction was less on lean tissue with the same treatments, although the modified cycles reduced populations when compared to the control cycle.

Storage And Bacterial Contamination Effects On Myofibrillar Proteins And Shear Force Of Beef, James S. Dickson, John D. Crouse, M. Koohmaraie

James S. Dickson

Thirty-two steaks from the longissimus muscle, fifth rib to third lumbar vertebra, were obtained from youthful carcass beef. Half were sterilized by ultraviolet light and all vacuum packaged and stored for 1, 14, 28 or 57 days at 2°C. After storage, steaks were examined for microbial populations, myofibril fragmentation index (MFI), cooking characteristics and shear force (SF). Aerobic and anaerobic counts decreased during storage. Psychrotrophic counts were low throughout. Sterilization had no effect on SF or MFI. Cooking loss tended (P < 0.09) to increase with time of storage. SF values decreased and MFI values increased through day 14, but remained relatively constant after that. Results of SDS-PAGE, SF and MFI indicate major changes in proteolysis of myofibrils and tenderness were completed by day 14.

Attachment Of Salmonella Typhimurium And Listeria Monocytogenes To Beef Tissue: Effects Of Inoculum Level, Growth Temperature And Bacterial Culture Age, James S. Dickson

James S. Dickson

The effects of inoculum level, growth temperature and culture age on the attachment of Salmonella typhimurium and Listeria monocytogenes to beef tissue surfaces were evaluated. An increase in inoculum level resulted in an increase in the number of attached cells for both bacteria which was proportional to the increase in inoculum. Bacteria grown at 23°C attached in higher numbers flVO5) to fat tissue than bacteria grown at 37°C or at 37°C followed by 24 h at 5°C. Growth temperature did not affect attachment to lean tissue for either bacterium. Overnight cultures of both bacteria attached in greater numbers 0VO.05) to …

In Vivo Activation Of Human Immunodeficiency Virus Type 1 Long Terminal Repeat By Uv Type A (Uv-A) Light Plus Psoralen And Uv-B Light In The Skin Of Transgenic Mice, John D. Morrey, S M. Bourn, T D. Bunch, M K. Jackson, R W. Sidwell, L R. Barrows, R A. Daynes, C A. Rosen

John D. Morrey

UV irradiation has been shown to activate the human immunodeficiency virus type 1 (HIV-1) long terminal repeat (LTR) in cell culture; however, only limited studies have been described in vivo. UV light has been categorized as UV-A (400 to 315 nm), -B (315 to 280 nm), or -C (<280 nm); the longer wavelengths are less harmful but more penetrative. Highly penetrative UV-A radiation constitutes the vast majority of UV sunlight reaching the earth's surface but is normally harmless. UV-B irradiation is more harmful but less prevalent than UV-A. In this report, the HIV-1 LTR-luciferase gene in the skin of transgenic mice was markedly activated when exposed to UV-B irradiation. The LTR in the skin of transgenic mice pretreated topically with a photosensitizing agent (psoralen) was also activated to similar levels when exposed to UV-A light. A 2-h exposure to sunlight activated the LTR in skin treated with psoralen, whereas the LTR in skin not treated with psoralen was activated after 7 h of sunlight exposure. The HIV-1 LTR-β-galactosidase reporter genes have been used to demonstrate the in vivo UV-induced activation of the LTR and might be used to evaluate other environmental factors or pharmacologic substances that might potential activate the HIV-1 LTR in vivo

Cyanonews (Vol. 7, No. 2, September 1991), Jeff Elhai

CyanoNews

CyanoNews was a newsletter that served the cyanobacteriological community from 1985 to 2003, with content provided by readers (sort of a blog before there were blogs). The newsletter reported new findings from the lab, summaries of recent meetings (often provided by graduate students and post-docs entering the field), positions sought or available, life transitions, a compendium of recent cyanobacteria-related articles, and other items of interest to those who study cyanobacteria.