Microbiology Commons^™

Inhibition Of Murine Lymphokine Activated Killer (Lak) Cell Cytotoxicity By Adherent Cells And Prostaglandin E2, Kimberly G. Roe

Retrospective Theses and Dissertations

The effect of adherent cells, prostaglandin E₂ (PGE₂), and indomethacin on LAK cell activity of C57BL/6J (B6) mice was investigated. Depletion of adherent cells from splenocyte cultures prior to LAK cell generation increased LAK cell cytotoxicity by 60% compared to non-depleted splenocyte cultures. Depletion of adherent cells from splenocyte cultures following five days of incubation and prior to the LAK cell assay resulted in a similar enhancement. Indomethacin, an inhibitor of prostaglandin synthesis, enhanced LAK cell cytotoxicity in splenocyte cultures. PGE₂ inhibited LAK cell cytotoxicity of adherent cell depleted splenocytes 28% and 88% at l0^{-6 …}

T Cell Subset Analysis In The C57bl/6j Mouse, Bonny Bass

Retrospective Theses and Dissertations

Functional studies of murine T cells were performed to verify the role of T cells and adherent cells in the production of IL-2 and to clarify the role of the L3T4 receptor. Whole spleen cells were passed over nylon wool columns to obtain purified populations of cells for this purpose. It was found upon reconstitution of the T cells and adherent cells that a collaboration between the two was necessary for optimal IL-2 production. Their level of IL-2 production increased above the amount produced by the whole spleen cells. Antibody mediated cytotoxicity using L3T4 antisera identified T helper cells exclusively. …

Comparative Extracellular Killing Of Candida And Tumor Cells By Macrophages At Varying Stages Of Activation, David W. A. Beno

Master's Theses

No abstract provided.

Analysis Of Factors Affecting Germ Tube Formation In Candida Albicans, Jordan H. Pollack

Dissertations

No abstract provided.

Selenium-Induced Enhancement Of Hematoporphyrin Derivative Phototoxicity In Murine Bladder Tumor Cells, Jeffrey M. Tosk

Loma Linda University Electronic Theses, Dissertations & Projects

The phototoxicity of hematoporphyrin derivative (Hpd) to murine bladder tumor (MBT-2) cells was studied in vitro. It was observed that selenium in the form of sodium selenite enhanced Hpd-sensitized photodamage in MBT-2 cells under conditions where the selenite alone was non-toxic. Sodium selenite was found to enhance the fluorescence emission of Hpd and augment the Hpd-sensitized photooxidation of tryptophan. The data indicate that sodium selenite is able to disaggregate Hpd, thereby enhancing Hpd-sensitized phototoxicity.

Immunological Relationships Among Proteins Making Up The Bacillus Thuringiensis Subsp. Israelensis Crystalline Toxin, Mary Ann Pfannenstiel, Graham A. Couche, Elise J. Ross, Kenneth W. Nickerson

Papers in Microbiology

The immunological relationships among the proteins of the mosquito larvicidal toxin produced by Bacillus thuringiensis subsp. israelensis have been investigated by using polyclonal antisera specific for the 28-, 70-, and 135-kilodalton proteins. Each of these proteins was immunologically distinct. There was no cross-reaction among the three proteins and the two non-homologous antisera. Treatment of toxin proteins with larval gut enzymes for 20 h identified protease-resistant domains at approximately 65, 38, and 22 kilodaltons. Similar domains were generated by treatment with trypsin and chymotrypsin. Our immunological and kinetic data indicate that the 28-kilodalton protein is degraded successively to protein bands at …

Analysis Of Mosquito Larvicidal Potential Exhibited By Vegetative Cells Of Bacillus Thuringiensis Subsp. Israelensis, Corey J. Walther, Graham A. Couche, Mary Ann Pfannenstiel, Sara E. Egan, Lisa A. Bivin, Kenneth W. Nickerson

Papers in Microbiology

Vegetative Bacillus thuringiensis subsp. israelensis cells (6 x 10⁵/ml) achieved 100% mortality of Aedes aegypti larvae within 24 h. This larvicidal potential was localized within the cells; the cell-free supernatants did not kill mosquito larvae. However, they did contain a heat-labile hemolysin which was immunologically distinct from the general cytolytic (hemolytic) factor released during solubilization of B. thuringiensis subsp. israelensis crystals. The larvicidal potential of the vegetative cells was not due to poly-3-hydroxybutyrate. Instead, it correlated with the ability of vegetative cells to sporulate during the bioassays. No toxicity was observed when bioassays were conducted in the presence …

The Effects Of Certain Oral Bacteria On The In Vitro Growth Of Bacteroides Gingivalis, Steven G. Morrow

Loma Linda University Electronic Theses, Dissertations & Projects

The effect of nine gram positive and grain negative cocci and bacilli, which are indigenous to the oral cavity, on the in vitro growth of Bacteroides gingivalis was examined in this study. A pure culture of B. gingivalis is was obtained from the American Type Culture Collection. The nine oral test organisms were Streptococcus mitis, Streptococcus mutans, Streptococcus faecalis, Staphylococcus epidermidis, Staphylococcus aureus, Corynebacteriurn hofmanii, Lactobacillus acidophilus, Branhamella catarrhalis, and Escherichia coli. The effect on the growth of B. gingivalis of the nine test organisms was evaluated on a solid medium by use of …

The Effect Of Dietary Fat Level On Tumor Development And Immunity In Balb/C Mice Fed Casein Of Wheat Gluten As Protein And Challenged With Herpes Type 2-Transformed Cells, Helen Drakou

Loma Linda University Electronic Theses, Dissertations & Projects

A high percentage of fat in the diet has been implicated through epidemiological studies in several human cancers. Studies in experimental animals have produced evidence that increasing the fat in the diet decreases the animal’s ability to restrict tumor development and growth. In previous work done in this laboratory with BALB/c mice injected with herpes simplex Type 2-transformed mouse cells (H238), the effects of a diet which greatly enhanced tumor growth was compared with one which restricted tumor growth. The latter, a low (5%) fat, low protein diet, in which the protein was casein, was also found to produce a …

The Expression Of Dna Restriction-Modification Genes Of Escherichia Coli K12, Ainu Prakash

Loma Linda University Electronic Theses, Dissertations & Projects

To facilitate the study of the expression of hsd genes in Escherichia coli K, hsdK::MudX (amp crop lac) operon fusion mutants have successfully been constructed and characterized for the first time. The results indicate that Promoters 1 and 2 for hsdK genes appear to be relatively weak, as reflected by about 50 units of beta-gal activity. Both promoters were active in all the conditions tested except during Beta-gal activity was higher in aerobic than in anaerobic growth; at 30°C and 37°C than at 42°C; and in L-broth early log phase of the culture. than in minimal media. Potential future uses …

Sprague-Dawley Rats Fed Milk Or Beef Protein : Differences In Response To 1,2-Dimethylhydrazine Carcinogenesis, Stanley W. L. Ng

Loma Linda University Electronic Theses, Dissertations & Projects

Colon cancer is responsible for a high percentage of cancer deaths in developing countries, and there is convincing epidemiological evidence that meat protein and fat in the diet increase the incidence of this form of cancer. An animal tumor model which has been used by many investigators interested in colon cancer is the 1,2-dimethylhydrazine (DMH)-rat or DMH-mouse tumor model. In previous work done in this laboratory the DMH-BALB/c mouse tumor model was used to compare the production of colon tumors in mice fed a diet using non-fat powdered milk as a source of protein with those in mice fed beef …

Production Of Acquired Immunodeficiency Syndrome-Associated Retrovirus In Human And Nonhuman Cells Transfected With An Infectious Molecular Clone, Akio Adachi, Howard E. Gendelman, Scott Koenig, Thomas Folks, Ronald Willey, Arnold Rabson, Malcom A. Martin

Nebraska Center for Virology: Faculty Publications

We constructed an infectious molecular clone of acquired immunodeficiency syndrome-associated retrovirus. Upon transfection, this clone directed the production of infectious virus particles in a wide variety of cells in addition to human T4 cells. The progeny, infectious virions, were synthesized in mouse, mink, monkey, and several human non-T cell lines, indicating the absence of any intracellular obstacle to viral RNA or protein production or assembly. During the course of these studies, a human colon carcinoma cell line, exquisitely sensitive to DNA transfection, was identified.

Restriction Analysis Of Bacteriophage, Ali Reza Mostaghimi

Loma Linda University Electronic Theses, Dissertations & Projects

A restriction map of DNA from bacteriophage P3-C₁ for the endonucleases BamH I, Bgl II, EcoR I, Hind III, Sac I and Sca I is described in this study. The phage DNA was determined to be a linear double stranded molecule with a unit genomic length of 31 kb. After electrophoresis in 0.5% agarose gels, the undigested DNA appeared as three molecules of 31, 62 and 93 kb in length. The Hind III digestion pattern of the individual 31 kb molecule was identical to the 62 kb and 93 kb Hind III pattern. With all enzymes except Hind III, …

Augmentation Of Natural Killer Cell Activity By Corynebacterium Parvum: Involvement Of Lipoxygenase Pathway, Gregory D. Kuo

Loma Linda University Electronic Theses, Dissertations & Projects

The ⁵¹Chromium release assay was used to measure natural killer (NK) cell activity associated with Corynebacterium parvum (CP) injection against YAC-1 and MBT-2 targets. The peritoneal exudate cells from intraperitoneal treatment of CP at different dosages consistently gave higher cytotoxicity than their respective spleen cell fractions. In contrast, CP given intravenously showed the reversed effect. Our results demonstrate that the route of administration and dosage of CP are two crucial variables for determining the effectiveness of this immunostimulant. In vitro manipulations of nordihydroguaiaretic acid, indomethacin, and prostaglandin E2 on CP-stimulated NK cells suggest that the lipoxygenase pathway is involved …

Comparison Of Membrane Filter, Multiple-Fermentation-Tube, And Presence-Absence Techniques For Detecting Total Coliforms In Small Community Water Systems., Nicholas J. Jacobs, Wanda L. Zeigler, Frank C. Reed, Therese A. Stukel, Eugene W. Rice

Dartmouth Scholarship

Methods for detecting total coliform bacteria in drinking water were compared using 1,483 different drinking water samples from 15 small community water systems in Vermont and New Hampshire. The methods included the membrane filter (MF) technique, a 10-tube fermentation tube (FT) technique, and the presence-absence (P-A) test.

Purification Of Anthrax Toxin Protective Antigen Component And Characterization Of Its Binding Interaction With Bovine Kidney Cells, Daniel Dalton Martin

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Protective antigen component of B. anthracis toxin was produced and purified to the >99% level. Toxin was purified from culture supernatant utilizing concentration and liquid chromatography techniques. Purity was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

The purified protective antigen retained biological and antigenic activity as evidenced respectively by lethality in Fischer 344 rats when injected in combination with lethal factor, and by positive results on the Ouchterlony double diffussion assay.

Radioiodinated protective antigen was used both in the in vivo and the in vitro experiments.

In vivo distribution of labelled protective antigen was determined in Fischer 344 rats. Assay …

Tropism Of Sheep Lentiviruses For Monocytes: Susceptibility To Infection And Virus Gene Expression Increase During Maturation Of Monocytes To Macrophages, Howard E. Gendelman, Opendra Narayan, Suzanne Kennedy-Stoskopf, Peter G. E. Kennedy, Zahra Ghotbi, J. E. Clements, Jeff Stanley, Gholam Pezeshkpour

Nebraska Center for Virology: Faculty Publications

Visna lentiviruses have a natural tropism for cells of the macrophage lineage of sheep and goats, but virus replication in these cells in vivo is restricted so that only small quantities of virus are produced. One restricting factor suggested in previous studies is that virus replication is dependent on the maturity of the cells: the more mature the cell, the less restrictive the replication of the virus. Since monocytes in peripheral blood are precursors of macrophages, we investigated the effect of cell maturation on virus replication under limited control conditions in vitro by inoculating blood leukocytes with virus and retarding …

Absence Of A Structural Basis For Intracellular Recognition And Differential Localization Of Nuclear And Plasma Membrane-Associated Forms Of Simian Virus 40 Large Tumor Antigen., Donald L. Jarvis, Charles N. Cole, Janet S. Butel

Dartmouth Scholarship

The simian virus 40 large tumor antigen (T-ag) is found in both the nuclei (nT-ag) and plasma membranes (mT-ag) of simian virus 40-infected or -transformed cells. It is not known how newly synthesized T-ag molecules are recognized, sorted, and transported to their ultimate subcellular destinations. One possibility is that these events depend upon structural differences between nT-ag and mT-ag. To test this possibility, we compared the structures of nT-ag and mT-ag from simian virus 40-infected cells. No differences between the two forms of T-ag were detected by migration in polyacrylamide gels, by Staphylococcus aureus V8 partial proteolytic mapping of methionine- …

Plant Viruses., R. A.C. Jones

Experimental Summaries - Plant Research

Cucumber mosaic virus in legumes, 85A024, 85A122, 85A123, 85A153, 85A163, 85A217, 85A219. Alfalfa mosaic virus in annual medics.

Cyanonews (Vol. 2, No. 1, March 1986), 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.

Cyanonews (Vol. 2, No. 3, December 1986), 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.

Cyanonews (Vol. 2, No. 2, July 1986), 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.

Htlv-Iii Serology In Hemophilia: Relationship With Immunologic Abnormalities, Joan C. Gill, Jay E. Menitove, Philip R. Anderson, James T. Casper, Sushilkumar G. Devare, Charles Wood, Stuart Adair, James Casey, Christi Scheffel, Robert R. Montgomery

Nebraska Center for Virology: Faculty Publications

We investigated the relationship of the presence of antibodies to HTLV-III and immunologic abnormalities in patients with hemophilia. Serum antibodies to HTLV-III were analyzed by ELISA assay, immunoprecipitation of labeled cell extracts, and immunoprecipitation of purified HTLV-III p24. Thirty-four (61%) of the total group (n = 56) had antibody to HTLV-III; 34 (76%) of 45 patients given commercial factor VIII preparations were seropositive, compared with none of 11 patients treated exclusively with cryoprecipitate obtained from volunteer blood donors. Of patients who were seropositive for HTLV-III antibody, 94% had abnormal T4/T8 ratios, and 33% of those whose serum was antibody …

Rhizoctonia Patch Of Cereals, G C. Macnish

Journal of the Department of Agriculture, Western Australia, Series 4

Rhizoctonia patch is now widespread within Western Australia, having been recorded on all cereals and on a wide variety of other crops and pasture plants. Although the disease-as its name implies - looks spectacular in the field, the overall reduction in crop yield is probably small. However, its increasing frequency is causing considerable concern as the disease appears to be associated with minimum or reduced tillage systems.

The Effect Of A Bacteriophage On The Competition Of Rhizobium Japonicum Strains For Nodulation Of Glycine Max (L.) Merr, Lori Kelley

Masters Theses

The symbiotic relationship between Rhizobium japonicum (Kir.) Buch. and Glycine max (L.) Merr. was the concern of this thesis. This relationship occurs naturally in soybean fields. In the field there are many different strains of Rhizobium. Some of these strains fix atmospheric nitrogen more efficiently than others resulting in a higher crop yield. The problem is the more efficient nitrogen fixers are out-competed by less efficient nitrogen fixers. In order to solve this problem the more competitive strains have to be controlled. There are two main ways to do this. One is to increase the number of the desired …

Analysis Of The Charleston, Illinois Water Supply For The Presence Of Organic Mutagens Utilizing The Ames Salmonella/Microsome Assay, Steven Lee Washburn

Masters Theses

Water samples from the Charleston, Illinois water supply and selected agricultural run-off areas were tested for the presence of organic mutagenic chemicals. The Ames Salmonella/microsome mutagenicity assay utilizing strains TA98 and TA100 was used to analyze concentrated water samples. A special set of samples was exposed to the S9 activation system coupled with the TA98 strain to simulate the mammalian metabolic system.

No mutagens were detected in any of the samples tested. Several positive and negative control systems were included in the testing system. Negative control systems were used to determine the average spontaneous reversion rates for each strain for …

Characterization Of A Staphylococcal Trimethoprim Resistance Gene, Jerald Preston Coughter

Theses and Dissertations

Trimethoprim resistance(Tp r) is encoded by conjugative plasmids in clinically significant staphylococcal isolates. Two genetically and physically similar plasmids from S. aureus, pG01 and pGOS, have Tp r genes that map in different locations on these plasmids. In order to study the relatedness of the Tp r genes and their products to other known Tp r genes, a 1.2 kb fragment of pG01 and a 4.2 kb fragment of pGOS were cloned in E. coli and used as probes for in situ filter hybridization experiments.

A 500 base pair subclone of the original 1.2 kb fragment containing only the staphylococcal …

Degradation Of Human Anaphylatoxin C3a By Rat Peritoneal Mast Cells: A Role For The Secretory Granule Enzyme Chymase And Heparin Proteoglycan, James Edmund Gervasoni Jr

Theses and Dissertations

Purified human C3a was iodinated (¹²⁵I-C3a) and used to study the interaction of labeled peptide with rat peritoneal mast cells (RMC). Cellular binding of ¹²⁵I-C3a occurred within 30 sec, followed by a rapid dissociation from the cell. Both the binding of ¹²⁵I-C3a and the rate of dissociation from the cell were temperature dependent. At 0°C, the binding of ¹²⁵I-C3a was increased and the rate of dissociation reduced, as compared to 37°C. Once ¹²⁵I-C3a was exposed to RMC, it lost the ability to rebind to a second batch of RMC. Analysis of the supernatants by …

Restriction Mapping And Expression Of Recombinant Plasmids Containing The Arsenic Resistance Genes Of The Plasmid R45, Terry M. Coons

Dissertations and Theses

The trivalent (arsenite) and pentavalent (arsenate) forms of arsenic are introduced into the environment through the use of arsenic in herbicides, pesticides, fertilizers, and the smelting of arsenic-bearing ores. Bacteria resistant to arsenic are readily isolated from surface waters, sewage, and clinical infections. Although some bacterial resistance is provided by inducible phosphate transport systems that discriminate against arsenate, marked resistance is carried on bacterial plasmids.

A 6.9 kilobase fragment previously derived from one such plasmid, R45, and containing the genes for inducible resistance to arsenite and arsenate was ligated into the cloning vectors puce and pUC9 in opposite orientations and …

Stability Of The Larvicidal Activity Of Bacillus Thuringiensis Subsp. Israelensis: Amino Acid Modification And Denaturants, Mary Ann Pfannenstiel, Graham A. Couche, Ganapathy Muthukumar, Kenneth Nickerson

Papers in Microbiology

The Bacillus thuringiensis subsp. israelensis mosquito larvicidal toxin is not a sulfhydryl-activated toxin. The protein disulfide bonds were cleaved and blocked without loss of toxicity. In contrast, modification of the lysine side chains eliminated toxicity. Additionally, the toxin was resistant to high concentrations of salt (8 M NaBr), organic solvents (40% methanol), denaturants (4 M urea), and neutral detergents (10% Triton X-100). However, it was inactivated by both positively and negatively charged detergents and by guanidine hydrochloride.