Microbiology Commons^™

Transactivation Of The Moloney Murine Leukemia Virus And T-Cell Receptor Beta-Chain Enhancers By Cbf And Ets Requires Intact Binding Sites For Both Proteins., Wanwen Sun, Barbara J. Graves, Nancy A. Speck

Dartmouth Scholarship

The Moloney murine leukemia virus (Mo-MLV) enhancer contains binding sites (LVb and LVc) for the ets gene family of proteins and a core site that binds the polyomavirus enhancer-binding protein 2/core-binding factor (cbf) family of proteins. The LVb and core sites in the Mo-MLV enhancer contribute to its constitutive activity in T cells. All three binding sites (LVb, LVc, and core) are required for phorbol ester inducibility of the Mo-MLV enhancer. Adjacent binding sites for the ets and cbf proteins likewise constitute a phorbol ester response element within the human T-cell receptor beta-chain (TCR beta) enhancer and contribute to constitutive …

The Kinetic Characterization Of Mutant L289p Of Recombinant Hiv-1 Reverse Transcriptase, Tryn Thomas Stimart

Chemistry & Biochemistry Theses & Dissertations

The Human Immunodeficiency Virus, type 1 (HIV-1) is the causative agent of the Acquired immunodeficiency Syndrome (AIDS). Currently, AIDS is the leading cause of death amongst young people in the United States. The large increase of HIV-1 infections over the past decade has spawned considerable research in fighting the spread of this disease. One area of intense HIV-1 research has focused on the enzyme Reverse Transcriptase (RT) which is an essential enzyme involved in the replication and life cycle of HIV-1. HIV-1 RT is composed of p66 and p51 subunits and is only active as a heterodimer in vivo. …

Structural Characterization Of A Novel Inhibitor Of Hiv Reverse Transcriptase (Hiv Rt), Greggory Jon Woitte

Chemistry & Biochemistry Theses & Dissertations

Human immunodeficiency virus (HIV) infections have become a leading cause of death among young people in the United States today. As the number of HIV infections increases, so too does the cost of treatment. Together, these numbers have prompted an increase in the development of pharmaceutical interventions. HIV reverse transcriptase (HIV RT) has become a suitable target for drug therapy because it is the sole enzyme responsible for HIV replication.

Fucoidan, a sulfated polysaccharide isolated from the brown algae Fucus vesiculosus, has been shown to block a variety of cell adhesion related events including metastasis. In addition, fucoidan has also …

Characterization Of An Antiviral Agent Based On Nonionic Surfactants And It's Effects On Human Dermal Fibroblasts, Ji Young Li

Biological Sciences Theses & Dissertations

The AIDS ·pandemic has directed various research endeavors towards finding an appropriate method for eliminating all potentially infectious material within bone allografts before implantation into a recipient. To that effect, Panavirocide was conceived by Medicine and Applied Science·s, Inc. This compound, in which three nonionic surfactants serve as the active agents, has been shown to inactivate HIV-1 particles within blood and blood products. Because Panavirocide has never been used with allografts, the purpose of the present research was to characterize the nonionic surfactants for the putative alteration of the formulation by determining the critical micelle concentration values and to determine …

The Amino-Terminal Functions Of The Simian Virus 40 Large T Antigen Are Required To Overcome Wild-Type P53-Mediated Growth Arrest Of Cells., Robin S. Quartin, Charles N. Cole, James M. Pipas, Arnold J. Levine

Dartmouth Scholarship

High levels of the p53 tumor suppressor protein can block progression through the cell cycle. A model system for the study of the mechanism of action of wild-type p53 is a cell line (T64-7B) derived from rat embryo fibroblasts transformed by activated ras and a temperature-sensitive murine p53 gene. At 37 to 39 degrees C, the murine p53 protein is in a mutant conformation and the cells actively divide, whereas at 32 degrees C, the protein has a wild-type conformation and the cells arrest in the G1 phase of the cell cycle. Wild-type simian virus 40 large T antigen and …

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 …

Kinetic Characterization Of A Recombinant C-Terminal Mutant Of Reverse Transcriptase From The Human Immunodeficiency Virus, Thomas S. Heard

Chemistry & Biochemistry Theses & Dissertations

The human immunodeficiency virus (HIV) reverse transcriptase (RT) (EC 2.7.7.49) is the central replication enzyme for HIV. In general, the kinetic mechanism for this and all other polymerases involves the ordered binding of two substrates: a primer-template (PT) followed by a deoxyribonucleoside triphosphate (dNTP). Previous investigations prompted this research when it was discovered that the substrate dNTP, in absence of PT, could protect a recombinant c-terminal mutant HIV-1 RT from inhibition by pyridoxal-5'-monophosphate (PLP), an active-site dNTP inhibitor. In contrast, the non-mutant recombinant HIV-1 RT required both substrates for protection from PLP inhibition. This investigation sought to determine if this …

Cytolytic T Lymphocytes Specific For Tumors And Infected Cells From Mice With A Retrovirus-Induced Immunodeficiency Syndrome., Jennifer G. Erbe, Kathy A. Green, Karen M. Crassi, Herbert C. Morse, W R. Green

Dartmouth Scholarship

LP-BM5 retrovirus complex-infected C57BL/6 mice develop immunodeficiency, somewhat analogous to AIDS, termed murine AIDS (MAIDS). After secondary stimulation with syngeneic B-cell lymphomas from LP-BM5-infected mice, C57BL/6 mice produced vigorous CD8+ cytotoxic T lymphocytes specific for MAIDS-associated tumors. An anti-LP-BM5 specificity was suggested because spleen and lymph node cells from LP-BM5-infected mice served as target cells in competition assays, and cells from LP-BM5, but not ecotropic, virus-infected mice functioned as secondary in vitro stimulators to generate cytotoxic T lymphocytes to MAIDS tumors.

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. …

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 …

Simian Virus 40 Host Range/Helper Function Mutations Cause Multiple Defects In Viral Late Gene Expression., Terryl Stacy, Michele Chamberlain, Charles N. Cole

Dartmouth Scholarship

Simian virus 40 (SV40) deletion mutants dlA2459 and dlA2475 express T antigens that lack the normal carboxy terminus. These mutants are called host range/helper function (hr/hf) mutants because they form plaques at 37 degrees C on BSC-1 and Vero monkey kidney cell lines but not on CV-1p monkey kidney cells. Wild-type SV40 can provide a helper function to permit growth of human adenoviruses in monkey kidney cells; the hr/hf mutants cannot. Progeny yields of hr/hf mutants are also cold sensitive in all cell lines tested. Patterns of viral macromolecular synthesis in three cell lines (Vero, BSC-1, and CV-1) at three …

In Vitro Evaluation Of Cancer Patient Immune Responses Following Infusion Of Radiolabeled Murine Monoclonal Antibody, Kelly A. King

Loma Linda University Electronic Theses, Dissertations & Projects

Several monoclonal antibody (MAb) types derived from mice have been developed specifically for the use of targeting human tumors. The specificity of these murine MAb’s for their respective tumor antigens is very high making them potentially good immunotherapeutic agents for cancer treatment and diagnosis. A limiting factor in using these murine MAb’s in patients is the development of human anti-mouse antibody (HAMA). The presence of HAMA may reduce the effectiveness of MAb for tumor targeting. If patient sensitization could be quickly detected following the initial infusion of MAb, then future infusion could be altered or terminated to prevent further sensitization. …

Enhancement Of Intratumor Antibody Distribution With A Bifunctional Antibody Delivery System And With Hyperthermia, Karen Lynn King

Loma Linda University Electronic Theses, Dissertations & Projects

The intratumor distribution of a new monoclonal antibody-hapten-based delivery system was examined. From two marine antibodies, Hybritech, Inc. has developed a bifunctional antibody (BPA) with specificities against CEA (ZCE-025 or GEM 231) and against one of two derivatives of [¹¹¹In]-benzyl-EDTA (CHA 255), EOTUBE or BLEDTA IV, to form ZCE/CHA or CEM/CHA. BFA accumulates in CEA-expressing tissue and clears from non-antigenic tissues prior to administration of an [¹¹¹In]-hapten.

Using five groups of nude mice bearing human colon tumor xenografts (T380), we evaluated different carriers of [¹¹¹In]-EOTUBE to prelocalized ZCE/CHA. We administered all antibodies and haptens …

Effects Of Corticosteroids And Of Cyclosporin A On Trypanosoma Musculi In Mice And In Culture, Philip Stephen Schuyler

Loma Linda University Electronic Theses, Dissertations & Projects

The in vivo and in vitro effects of cyclosporin A and corticosteroids on Trypanosoma musculi were examined. Mice that received continuous administration of 6 mg/liter/day of dexamethasone in drinking water and 200 mg/kg hydrocortisone acetate as a single dose 24 hrs prior to injection with T. musculi had significantly higher and longer parasitemias, than controls. Body and spleen weights and B cell responsiveness were lower in test animals and there were some deaths. Mice injected with T. musculi that received 100 mg/kg/day cyclosporin A had longer, but not higher, parasitemias than controls. Corticosteroids appear to partially inactive antibody production (i.e. …

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- …

A Rapid Concentration And Identification Technique For Human Malaria Infections Using A Mouse Model, Donald D. Barton

Loma Linda University Electronic Theses, Dissertations & Projects

Red blood cells infected with malaria parasites experience a change in density making them lighter (less dense). This paper reports a new, rapid diagnostic technique using this change in density in the infected red blood cell. It is not only easy to use, but it is also compatible with the technological conditions found where the malaria problem is most acute - in many developing countries of the world. A small sample of blood drawn from a simple finger stick can be concentrated and examined microscopically for malaria parasites within 10 minutes. The thick film slide preparation, presently used, takes a …

Cryptosporidiosis In The Immunosuppressed Mouse And In Cell Culture, Mercy Prabhu Das

Loma Linda University Electronic Theses, Dissertations & Projects

Cryptosporidiosis is a parasitic infection primarily of animals that has been recently recognized as an important disease in immunocompromised humans. Most cases to date have occurred in patients with acquired immunodeficiency syndrome (AIDS). Persons with abnormal humoral or cellular immune function, and even normal subjects, have also been affected.

To determine whether or not mice, when experimentally immunosuppressed, would develop cryptosporidiosis, a test was designed using the AKR/J strain. This strain was selected because, it was previously reported that it was the strain shown to respond most readily to the production of disease caused by the opportunistic protozoan, Pneumocystis carinii …

Enzyme Linked Immunosorbent Assay For Anatid Herpesvirus, Margaret Anne Lewis

Biological Sciences Theses & Dissertations

Duck plague virus causes a hemorrhagic disease affecting the family Anatidae, resulting in 60-80% mortality in captive flocks. Plaque and neutralization assays for Anatid herpesvirus (AHV) require 5-10 days for completion and are restricted by the seasonal availability of fertile duck eggs. The enzyme linked innnunosorbent assay (ELISA) is rapid and specific and avoids the use of living cells. The Holland strain of AHV was grown on duck embryo fibroblasts and purified by banding on CsCl gradients. The resulting virus was used to raise antisera in prebled New Zealand white male rabbits. Antibody production was confirmed in double diffusion plates …

Cell Mediated Immunity By Cytotoxicity Assay And The Effect Of Corynebacterium Parvum And Radiation On Mice Bearing Herpes-Induced Tumors, Robert B. Stagg

Loma Linda University Electronic Theses, Dissertations & Projects

Cell mediated immune (CMI) cytotoxic reactivity of Balb/c mice against H238 cells, a Herpes simplex virus (HSV) Type 2 - induced sarcoma, was measured by the ¹²⁵IUdR release assay. The Balb/c mouse response to the growing HSV-induced tumor, treated with radiation and , was measured by survival rate, tumor growth and immune cytotoxicity of spleen and peritoneal exudate cells (PEC) as determined by ¹²⁵IUdR release assay. Subcutaneous (s.c.) inoculation of 1 x 10⁶ H238 cells (high dose) produced progressive tumor growth while s.c. inoculation of 1 x 10⁴ H238 cells (low dose) produced no tumors. A …

Effects Of Trypanosoma Musculi Kendall, 1906 Immune Serum On The Reproduction Of Trichinella Spiralis (Owen, 1835) Railliet, 1896 In Mice, Samuel L. Chafin

Loma Linda University Electronic Theses, Dissertations & Projects

The possible effects of ablastin, a reproduction-inhibiting antibody which is produced during the infection of the blood parasite, Trypanosoma musculi in the mouse, were investigated by fractionating the parasite into a supernatant fraction (designated as Group A test mice) and a pellet fraction (Group B test mice). These groups A and B mice were injected with the respective fractions, plus Freund’s Complete Adjuvant (FCA). Control mice were injected with FCA only. Sera from these three groups of mice were removed after the Group A and Group B mice were shown to be negative to T. musculi challenge, and Group A …

Effect Of Time And Diet On The Immune Status Of Mice Challenged With Herpes-Transformed Cells, Daila S. Gridley

Loma Linda University Electronic Theses, Dissertations & Projects

Human Herpes simplex virus type 2 (HSV 2)-transformed Balb/c mouse cells (H238 cells) are malignant in Balb/c mice. The immune stratus of mice bearing H238 cell-induced tumors was studied and changes in immune response with time and with altered diet of the mice were measured.

In the experiments dealing with the status of immunity with time after tumor cell injection, mice were fed Purina Laboratory Chow (P.L.C.), injected with H238 cells, and sacrificed three to eight weeks post-injection. In the diet experiments, groups of mice were fed either P.L.C. or one of six equicaloric diets, consisting of low or high …

The Fecal Shedding Of Listeria Monocytogenes By Conventional And "Germ Free" Mice, Margaret Joyce Ramsay

Loma Linda University Electronic Theses, Dissertations & Projects

A series of experiments involving Listeria monocytogenes was undertaken to study first the fecal shedding of Listeria by conventional mice challenged intraperitoneally with Listeria and second the fecal shedding and ability to survive of germ free mice with similar challenge. In addition, mites infesting Listeria infected mice were cultured for their possible contamination by Listeria.

Conventional mice were challenged intraperitoneally with a single near LD₅₀ dose of Listeria. Fecal Listeria were detected by collecting specimens from each mouse daily for the first 4 days and every second day thereafter. The feces was held at 7°C for 14 …

Absence Of Pasteurella Pestis In The Wildlife Of Arkansas, R. Reece Corey, Leo J. Paulissen, Delbert Swartz

Journal of the Arkansas Academy of Science

No abstract provided.

The Effect Of Infection With Polyoma Virus On The Protein, Rna, And Dna Of Mouse Embryo Cell Cultures, Lawrence E. Mcconnehey

Loma Linda University Electronic Theses, Dissertations & Projects

In four separate experiments mouse embryo cell monolayers infected with polyoma virus and uninfected control mouse embryo cells were maintained in Eagle’s Minimum Essential Medium for periods of time ranging from 0 to 168 hours. The cell layers were removed from the glass by trypsinization. Samples of the cells were counted in a Coulter Counter and the remainder of the cells frozen after centrifugation, in a button. Later, the buttons were thawed and protein, RNA, and DNA analyses were performed on each sample. The micrograms per million cells of protein. RNA, and DNA were determined for each sample and plotted …

Natural History Of Thelazia Californiensis Price, 1930 (Nematoda: Thelaziidae), A Mammalian Eye Worm, Warren E. Parmelee

Loma Linda University Electronic Theses, Dissertations & Projects

Various factors—embryology, host associations, and seasonal changes In eye worn populations—tending to promote the propagation of T. califomlensls were studied.

The early embryology closely parallels that of other nematode species.

The mature embryos remain alive in saline at 35°C as long as 5 days. Dead embryos can be distinguished from living ones by virtue of the rapid staining by methylene blue by the former. Parturient female thelazias release embryos into saline, and embryos are found regularly in tear secretions of infected laboratory rabbits.

Ovic embryos possess an enveloping egg membrane containing chitin. On this account, this eye worm species is …

Natural And Experimental Infections Of Thelazia Californiensis Price (1930, Nematoda: Thelaziidae) With Descriptions Of The Adult And Larval Parasites, Howard S. Burnett

Loma Linda University Electronic Theses, Dissertations & Projects

The purpose of this work was to determine the extent and effects of infestation of the eye worm of mammals, Thelazia californiensis, and to elucidate through experimentation or observation the mode of transmission.

A survey of literature revealed that thelaziasie (infestation by Thelazia species) may cause serious pathological changes in the eye and its adnexa, sometimes resulting in blindness. It was found that reports of thelaziasis in the United States were (with two exceptions) of cases in California. It was further found that T. californiensis has been reported in a wider variety of host than has any of other …