Microbiology Commons^™

Nitric Oxide Production: A Mechanism For Inhibition Of Chlamydia Trachomatis Replication, Bojun Chen

Electronic Theses and Dissertations

Chlamydia trachomatis (CT) replicates in macrophages, but is inhibited by IFN-$\gamma$ or LPS. IFN-$\gamma$ and/or LPS induced nitrite production in mouse peritoneal macrophages, macrophage cell lines (RAW264.7 and J774A.1) and McCoy cells. Kinetic studies indicated that peak production occurred 48 hours post-treatment. CT infection itself was insufficient to induce nitrite production, but resulted in enhancement of nitrite production in IFN-$\gamma$-treated cells. Treatment with IFN-$\gamma$ or LPS resulted in significant inhibition of CT replication in these cells. Strong correlation between nitrite production and inhibition of CT replication was observed in RAW264.7 and J774A.1 cells (correlation coefficients: $-$0.93 and $-$0.94, p $<$ 0.001). N$\sp{\rm g}$- monomethyl-L-arginine (L-NMMA) specifically inhibited nitrite production and partially reversed inhibition of CT replication in macrophage cell lines. NOS mRNA was measured in RAW264.7 cells by Northern blot and Dot blot hybridization. Strong correlation between NOS mRNA expression and inhibition of CT replication (correlation coefficient: $-$0.97, p $<$ 0.05) was observed. Anti-TNF-$\alpha$ antibody completely neutralized the biological activity of TNF-$\alpha$ secreted by LPS-treated RAW264.7 cells, yet the antibody neither reduced nitrite production nor restored CT replication. Combination of the antibody and L-NMMA significantly enhanced restoration of CT replication. In peritoneal macrophages, inhibition of CT replication induced by IFN-$\gamma$ was partially restored by L-NMMA or anti-TNF-$\alpha$ antibody. In McCoy cells, inhibition of CT replication induced by IFN-$\gamma$ and LPS was not significantly restored by L-NMMA. Great restoration of CT replication by 1 mM L-NMMA was observed in LPS-treated J774A.1 cells (31%), but not in IFN-$\gamma$-treated cells (5%). Our data indicate that (1) NO production is one of the mechanisms for inhibition of CT replication in IFN-$\gamma$-activated peritoneal macrophages and RAW264.7 cells; (2) NO plays a significant role in CT inhibition in LPS-treated macrophage cell lines, but not peritoneal macrophages; (3) TNF-$\alpha$ may be associated with inhibition, but the mechanism(s) may not involve NO production; (4) NO production may not be the mechanism for CT inhibition in McCoy cells treated with IFN-$\gamma$ and LPS.

A Characterization Of Extractable, Hydroxylated Fatty Acid Bearing Components In Legionella Pneumophila, Jonathan R. Lane

Electronic Theses and Dissertations

Extraction of the lipids of Legionella pneumophila yields phases unlike those produced from other Gram-negative bacteria. A viscous interface forms between the aqueous (wash) and organic phases. More than half of the hydroxylated fatty acids were found distributed between the aqueous phase and the interfacial material, fractions in which such constituents have not been reported in other Gram-negative species. It was further observed that after the material from the aqueous/interfacial phase was dissolved in methanol or chloroform/methanol (2:1 (V/V)), the addition of acetone would create a white, flocculent precipitate. Analyses showed that the supernatant contained fatty acids that were nonhydroxylated …

Infection Of Macrophages With Lymphotropic Human Immunodeficiency Virus Type 1 Can Be Arrested After Viral Dna Synthesis, Zheng-Bo Huang, Mary Jane Potash, Malgorzata Simm, Wei Chao, Howard Gendelman, Edward Edin, David J. Volsky

Nebraska Center for Virology: Faculty Publications

Lymphotropic strains of human immunodeficiency virus type 1 (HIV-1), including HTLV-IIIB, replicate poorly in macrophages. We have shown previously that lymphotropic HIV-1 fuses equally well with T lymphocytes and macrophages (M. J. Potash, M. Zeira, Z.-B. Huang, T. Pearce, E. Eden, H. Gendelman, and D. J. Volsky, Virology 188:864-868, 1992), suggesting that events in the virus life cycle following virus-cell fusion limit virus replication. We report here that HIV-1 DNA is synthesized effliciently in either ADA or HTLV-IIIB infected alveolar macrophages or monocyte-derived macrophages within 24 h of virus infection, as observed by polymerase chain reaction for amplification of viral …

Efficient Transcriptional Activation Of Many Simple Modular Promoters By Simian Virus 40 Large T Antigen., Philip W. Rice, Charles N. Cole

Dartmouth Scholarship

Simian virus 40 (SV40) large T antigen is a multifunctional protein which plays central roles during both lytic and transforming infections by SV40. It is a potent transcriptional activator and increases expression from the SV40 late promoter and from several cellular promoters. To understand better the transcriptional activation activity of large T antigen, we examined its ability to transactivate a set of simple modular promoters containing one of four upstream activation sequences coupled with one of three different TATA box sequences originally constructed and studied by Taylor and Kingston (Mol. Cell. Biol. 10:165-175, 1990). Large T antigen activated transcription from …

A Comparison Of No And N20 Production By The Autophic Nitrifier Nitrosomonas Europaea And The Heterotrophic Nitrifier Alcaligenes Faecalis, Iris C. Anderson, Mark Poth, Julie Homstead, David J. Burdige

OES Faculty Publications

Soil microorganisms are important sources of the nitrogen trace gases NO and N₂O for the atmosphere. Present evidence suggests that autotrophic nitrifiers such as Nitrosomonas europaea are the primary producers of NO and N₂O in aerobic soils, whereas denitrifiers such as Pseudomonas spp. or Alcaligenes spp. are responsible for most of the NO and N₂O emissions from anaerobic soils. It has been shown that Alcaligenes faecalis, a bacterium common in both soil and water, is capable of concomitant heterotrophic nitrification and denitrification. This study was undertaken to determine whether heterotrophic nitrification might be …

Identification Of A Protein In Several Borrelia Species Which Is Related To Ospc Of The Lyme Disease Spirochetes., Richard T. Marconi, D. Scott Samuels, Tom G. Schwan, Claude F. Garon

Biological Sciences Faculty Publications

Using oligonucleotide probes which have previously been shown to be specific for the ospC gene found in the Lyme disease spirochete species Borrelia burgdorferi, B. garinii, and group VS461, we detected an ospC homolog in other Borrelia species including B. coriaceae, B. hermsii, B. anserina, B. turicatae, and B. parkeri. In contrast to the Lyme disease spirochetes, which carry the ospC gene on a 26-kb circular plasmid, we mapped the gene in other Borrelia species to linear plasmids which varied in size among the isolates tested. Some isolates carry multiple copies of the gene residing on linear plasmids of different …

Antibacterial Properties Of An Egg, Altaf Ahmed

Department of Pathology and Laboratory Medicine

No abstract provided.

Nonenzymatic Glycosylation Of Lepidopteran-Active Bacillus Thuringiensis Protein Crystals, Meenakshi Bhattacharyi, Bradley A. Plantz, Jane D. Swanson-Kobler, Kenneth W. Nickerson

Papers in Microbiology

We used high-pH anion-exchange chromatography with pulsed amperometric detection to quantify the monosaccharides covalently attached to Bacillus thuringiensis HD-1 (Dipel) crystals. The crystals contained 0.54% sugars, including, in decreasing order of prevalence, glucose, fucose, arabinose/rhamnose, galactose, galactosamine, glucosamine, xylose, and mannose. Three lines of evidence indicated that these sugars arose from nonenzymatic glycosylation: (i) the sugars could not be removed by N- or O-glycanases; (ii) the sugars attached were influenced both by the medium in which the bacteria had been grown and by the time at which the crystals were harvested; and (iii) the chemical identity and stoichiometry of the …

Isolation And Characterization Of New Wild-Type Isolates Of Bovine Lentivirus, David Suarez, Martin Van Der Maaten, Charles Wood, Ceclia Whetstone

Nebraska Center for Virology: Faculty Publications

Two new isolates of bovine lentivirus, also known as bovine immunodeficiency-like virus (BIV), were obtained from a seropositive cattle herd in Florida. This is the first report of new isolates of BIV since the original BIV strain, R29, was isolated in 1%9. The two new BIV isolates were derived from blood butFy coat cells cocultivated in vitro with fetal bovine lung cell cultures. The new isolates differed in vitro from the original R29 isolate in replication and syncytium formation in fetal bovine lung cells. Both new isolates were confirmed as BIV by immunofluorescence assay, Western blotting (immunoblotting), and polymerase chain …

Molecular Cloning Of Infectious Ecotropic Murine Leukemia Virus Ak7 From An Emv-14-Positive Akxl-5 Mouse And The Resistance Of Ak7 To Recognition By Cytotoxic T Lymphocytes., Hillary D. White, William R. Green, Nuria R. Giné

Dartmouth Scholarship

The AKXL-5 recombinant inbred mouse strain is positive for the endogenous ecotropic murine leukemia virus emv-14, the only emv present in its germ line. emv-14 is of particular interest because spleen cells expressing emv-14 virus escape recognition by anti-AKR/Gross virus-specific cytotoxic T lymphocytes. We report here the isolation and characterization of a replication-competent emv clone, pAK7, derived from an AKXL-5 mouse. This clone is novel in that it encodes a variant ecotropic murine leukemia virus that, when expressed in SC.Kb target cells, fails to be recognized efficiently by anti-AKR/Gross virus cytotoxic T lymphocytes. The pAK7 clone can therefore be used …

Does Red Blood Cell Distribution Width (Rdw) Improve Evaluation Of Microcytic Anaemias?, Muhammad Sajjad Baqar, Mohammad Khurshid, A Molla

Department of Pathology and Laboratory Medicine

Abstract

The red cell distribution width (RDW) is an index of the variation in red cells size (anisocytosis). A study was conducted to examine the validity of using RDW in improving classification of microcytic anaemias. A total of 300 blood samples collected from a patient population aged 3 months to 55 years who were referred for haemoglobin electrophoresis were examined at The Aga Khan University Hospital (AKUH). On complete blood count, initially 200 patients (66.6%) were found to have hypochromic microcytic anaemia. Following haemoglobin electrophoresis 41% (821200) patients were diagnosed to have thalassemia minor and 59% (1181200) had hypochromic microcytic …

Simian Immunodeficiency Virus Mutants Resistant To Serum Neutralization Arise During Persistent Infection Of Rhesus Monkeys, Dawn P. Wooley, Catherine Collignon, Ronald C. Desrosiers

Neuroscience, Cell Biology & Physiology Faculty Publications

We previously described the pattern of sequence variation in gp120 following persistent infection of rhesus monkeys with the pathogenic simian immunodeficiency virus SIVmac239 molecular clone (D.P.W. Burns and R.C. Desrosiers, J. Virol. 65:1843, 1991). Sequence changes were confined largely to five variable regions (V1 to V5), four of which correspond to human immunodeficiency virus type 1 (HIV-1) gp120 variable regions. Remarkably, 182 of 186 nucleotide substitutions that were documented in these variable regions resulted in amino acid changes. This is an extremely nonrandom pattern, which suggests selective pressure driving amino acid changes in discrete variable domains. In the present study, …

Bacterial Starvation Stress And Contamination Of Beef, James S. Dickson, J. F. Frank

James S. Dickson

Salmonella typhimurium, Listeria monocytogenes, ad Escherichia coli 0157;H7 were starved by suspending washed cells in phosphate buffer for 5 days at 10, 25 or 37 degrees C. Starved bacteria were evaluated for their ability to attach to beef tissue and their sensitivity to acetic acid sanitizing. Starvation stress generally decreased the numbers of bacteria which attached to the tissue when compared to control bacterial cultures grown to late logarithmic growth phase. Starvation temperature was negatively correlated to the numbers of attached bacteria. Starvation slightly increased the sensitivity of attached S. typhimurium and L. monocytogenes cells to acetic acid, but did …

Suggested Reference Ranges In Clinical Chemistry For Apparently Healthy Males And Females Of Pakistan., Ayesha Molla, Mohammad Khurshid, William T. Manser, Rukhsana Lalani, Anis Alam, Zubaida Mohammad

Department of Pathology and Laboratory Medicine

Abstract

Seven hundred and eighty six apparently healthy males (418) and females (368) aged 0-69 years were randomly selected for estimation of reference ranges of 24 serum analytes at the clinical chemistry laboratory of The Ago Khon University Hospital (AKUH). Of the total study samples, 56% (439/786) were in the poediatric age group (0-14 years) and 44% (347/786) in the adult (1 5_60 years) group. Beckman Astra Ideal Autoanalyzer was used for all the estimations. Moon and standard deviations (SD) were calculated for each of the age groups. Reference ranges were calculated following standard methods of the International Federation of …

Suppressive Effects Of Transforming Factor-Β And Interleukin-10 On The Cytolytic Activity Of Murine Macrophages And Reversal By Cytokines, Chin-Hung Lin

Loma Linda University Electronic Theses, Dissertations & Projects

In this study, the suppressive effects of transforming growth factor-β (TGF-β) and interleukin-10 (IL-10) on peritoneal macrophage killing of H238 target cells and the potential for reversal of the immunosuppressive effect by IL-4 and interferon-γ (IFN-γ) were investigated. The responsiveness of naive and peptone-activated macrophages was compared. The cytolytic activity for tumor cells of these effector cells was measured by percent lysis of ³H-thymidine labeled Herpes simplex virus type 2-transformed tumor cells (H238). After 18-24 hours of incubation with TGF-β or IL-10, the cytolytic activity of macro-medium alone. The immunosuppressive effect of TGF-β or IL-10 on non-activated macrophages was …

Thymic Peptide Modulates Glutathione Redox Cycle And Antioxidant Enzymes In Macrophages, Choon Sil Park

Loma Linda University Electronic Theses, Dissertations & Projects

The effect of a 6-kDa thymic peptide (TP) on the oxidative burst of a murine macrophage cell line J774 was determined. TP (12.5-200 μg/ml) was incubated with 5 x 10⁵ J774 cells at 37° C and 5% C0₂ for 18 h and oxidative burst was triggered by zymosan; chemiluminescence was amplified by luminol and measured in an automated luminometer. TP exhibited a concentration-dependent inhibition of oxidative burst. To study the mechanisms involved in TP’s inhibition of oxidative burst, its effect on the glutathione (GSH) redox cycle, GSH biosynthesis, and antioxidant enzymes was investigated. TP was shown to increase …

Identifying Bovine Respiratory Syncytial Virus By Reverse Transcription-Polymerase Chain Reaction And Oligonucleotide Hybridizations, R. D. Oberst, M. P. Hays, K. J. Hennessey, L. C. Stine, J. F. Evermann, Clayton L. Kelling

Nebraska Center for Virology: Faculty Publications

An assay to identify tissue culture cells infected with bovine respiratory syncytial virus (BRSV) that utilizes reverse transcription (RT), the polymerase chain reaction (PCR), and a synthetic oligonucleotide hybridization probe has been developed. The RT-PCR assay uses a BRSV-specific negative-sense oligonucleotide primer to synthesize cDNA from a BRSV fusion protein mRNA template and another BRSV-specific oligonucleotide primer (positive sense) upstream from the negative-sense primer for PCR amplification. In the presence of mRNA templates of BRSV isolates originating from locations throughout the United States, the BRSV RT-PCR assay resulted in amplified products (381 bp) that were specific to BRSV, as demonstrated …

Prevalence Of Thalassemia Minor Trait In Pakistani Population Presented At Akuh For Complete Blood Count Estimation (Cbc)., Alia Shaikh, Mohammad Khurshid

Department of Pathology and Laboratory Medicine

No abstract provided.

Differential Mating Success Of Male Wing Morphs Of The Cricket, Gryllus Rubens, Cami L. Holtmeier, Anthony J. Zera

Anthony Zera Publications

Genetically marked individuals were used to study differential mating success between male wing morphs of the cricket, Gryllus rubens. Previous studies of Gryllus rubens and other wing-dimorphic insects have documented that flightless short-winged or wingless females typically attain reproductive maturity earlier and oviposit more eggs relative to their long-winged counterparts. This study was done to determine if flightless males also exhibit enhanced reproductive characteristics. Segregation analyses documented the genetic basis of allozymes used to infer paternity in subsequent experiments. Control experiments documented the absence of effects on mating success independent of wing morph due to (1) the genetic stock …

Two Factors That Bind To Highly Conserved Sequences In Mammalian Type C Retroviral Enhancers., Nancy R. Manley, Mary M. O'Connell, Wanwen Sun, Nancy A. Speck, Nancy Hopkins

Dartmouth Scholarship

The transcriptional enhancers of the Moloney and Friend murine leukemia viruses (MLV) are important determinants of viral pathogenicity. We used electrophoretic mobility shift and methylation interference assays to study nuclear factors which bind to a region of these enhancers whose sequence is identical between Moloney and Friend viruses and particularly highly conserved among 35 mammalian type C retroviruses whose enhancer sequences have been aligned (E. Golemis, N. A. Speck, and N. Hopkins, J. Virol. 64:534-542, 1990). Previous studies identified sites for the leukemia virus factor b (LVb) and core proteins in this region (N. A. Speck and D. Baltimore, Mol. …

Characterization Of A Protein That Binds Multiple Sequences In Mammalian Type C Retrovirus Enhancers., Wanwen Sun, Mary M. O'Connell, Nancy A. Speck

Dartmouth Scholarship

Mammalian type C retrovirus enhancer factor 1 (MCREF-1) is a nuclear protein that binds several directly repeated sequences (CNGGN6CNGG) in the Moloney and Friend murine leukemia virus (MLV) enhancers (N. R. Manley, M. O'Connell, W. Sun, N. A. Speck, and N. Hopkins, J. Virol. 67:1967-1975, 1993). In this paper, we describe the partial purification of MCREF-1 from calf thymus nuclei and further characterize the binding properties of MCREF-1. MCREF-1 binds four sites in the Moloney MLV enhancer and three sites in the Friend MLV enhancer. Ethylation interference analysis suggests that the MCREF-1 binding site spans two adjacent minor grooves of …

Siderophore Utilization By Bradyrhizobium Japonicum, Ora Plessner, Taryn Klapatch, Mary Lou Guerinot

Dartmouth Scholarship

Bradyrhizobium japonicum USDA 110 and 61A152 can utilize the hydroxamate-type siderophores ferri- chrome and rhodotorulate, in addition to ferric citrate, to overcome iron starvation. These strains can also utilizethepyoverdin-typesiderophorepseudobactinSt3.Theabilitytoutilizeanotherorganism'ssiderophores may confer a selective advantage in the rhizosphere.

Isolation Of Listeria Spp. From Feces Of Feedlot Cattle, James S. Dickson, G. R. Siragusa, E. K. Daniels

James S. Dickson

Healthy feedlot beef cattle were surveyed for the presence of Listeria spp. in fecal grab samples taken over 3 months. Composite samples were made from 224 individual animals each month. Listeria monocytogenes was isolated from one composite sample (4%) from the first sampling and not from the subsequent two. Listeria innocua was found in composite samples from all three samplings at levels of 17, 9, and 35%, respectively. From the individual samples comprising the Listeria spp.—positive composites, L. monoytogenes was isolated from one sample (3%) in the second sampling but not in the first or third samplings. L. innocua was …

Differential Allocation Of Resources Underlies The Dispersal-Reproduction Trade-Off In The Wing-Dimorphic Cricket, Gryllus Rubens, Simon Mole, Anthony J. Zera

Anthony Zera Publications

The cricket, Gryllus rubens (Orthoptera, Gryllidae), exists in natural populations as either a fully-winged (LW), flight-capable morph or as a short-winged (SW) morph that cannot fly. The SW morph is substantially more fecund than the LW morph. In this study we report on the physiological basis of this trade-off between flight capability and fecundity. Results from gravimetric feeding trials indicate that LW and SW morphs are equivalent in their consumption and digestion of food. However, during the adult stage, the LW morph is less efficient in converting assimilated nutrients into biomass. This may be a consequence of the respired loss …

A Continuous Culture Model To Examine Factors That Affect Transduction Among Pseudomonas Aeruginosa, Jean Replicon

Dissertations

No abstract provided.

Investigation Of The Structural Heterogeneity Of Lipooligosaccharides From Pathogenic Haemophilus And Neisseria Species And Of R-Type Lipopolysaccharides From Salmonella Typhimurium By Electrospray Mass Spectrometry, B W. Gibson, William Melaugh, Nancy J. Phillips, M A. Apicella, A A. Campagnari, J M. Griffiss

Chemistry Faculty Publications

Heterogeneity in the lipooligosaccharides (LOS) of pathogenic Haemophilus and Neisseria species is evident from the multiplicity of components observed with electrophoretic analyses. Knowledge of the precise structures that make up these diverse LOS molecules is clearly the key to reaching an understanding of pathogenic processes such as phase variation and molecular mimicry. Except for a few cases, little is known about the specific structural features of LOS that underlie phase variation and molecular mimicry, partly because of the inherent difficulties in the structural elucidation of these complex glycolipids. In the lipopolysaccharides (LPS) from Salmonella typhimurium and Escherichia coli, rough, or …

Cyanonews (Vol. 9, No. 2, July 1993), 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.

A Computer-Controlled Spray Chill Unit For Red Meat Carcasses, Maynard E. Anderson, James S. Dickson, Mark M. Hulse, Scott T. Drummond, John D. Crouse, Robert T. Marshall

James S. Dickson

A computer controlled and operated spray chill unit which can be used in future research studies for both spray chilling and decontamination of red meat carcasses is described. The unit consists of two parts: (1) an electromechanical system, and (2) a data acquisition/control system. Design concepts for the spray chill system are (1) versatility, (2) rapid change of spray parameters, and (3) computer control of the electromechanical system. An added feature is the computer data collection system for recording information related to each carcass (pH of muscle, surface and internal tissue temperatures, and weights of each half carcass during the …

Cyanonews (Vol. 9, No. 1, January 1993), 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.

Mechanisms Of Complement Resistance By Pathogenic Naegleria Fowleri Amoebae, Denise Marie Toney

Theses and Dissertations

The genus Naegleria is composed of a distinct group of free-living amoeboflagellates that include both pathogenic and nonpathogenic species. N. fowleri, the only pathogenic species of Naegleria to be isolated from humans, is the etiological agent of primary amoebic meningoencephalitis, a rare but rapidly fatal disease of the central nervous system in humans and in laboratory animals. The mechanisms of pathogenicity and the determinants of virulence of N. fowleri are unknown. Both pathogenic and nonpathogenic Naegleria activate the alternative complement pathway, however pathogenic N. fowleri are complement-resistant and nonpathogenic N. gruberi are complement-sensitive. The ability to resist complement-mediated lysis …