Medical Sciences Commons^™

Media Components Influence Viral Gene Expression Assays In Human Fetal Astrocyte Cultures, Micheline Mccarthy, Charles Wood, Larisa Fedoseyeva, Scott R. Whittemore

Nebraska Center for Virology: Faculty Publications

In vitro neurovirological studies of viral infectivity or viral gene expression may be confounded by the mulHple neural cell types and/or fibrob last contamination present in early passage cultures prepared from dissociated human central nervous system (eNS) tissue. We have developed highly enriched astrocyte cultures for neurovirological study by culturing in a serum-free defined medium, 816, supplemented with basic fibroblast growth factor (FGF-2). Subculture in this medium selects against fibroblast proliferation and favors sustained proliferation of a highly enriched glial fibrillary acidic protein (GFAP)-positive cell population. These astrocytes support productive replication of cytomegalovirus (CMV) and transient expression of transfected CMVand …

Interleukin 1-Beta In Symptomatic And Asymptomatic Human Periradicular Lesions, Gerald C. Lim

Loma Linda University Electronic Theses, Dissertations & Projects

Interleukin 1-Beta has been shown to be a potent mediator of bone resorption and has been implicated in resorption in periradicular lesions. In this study we compared the local concentrations of IL-1β in symptomatic and asymptomatic human periradicular lesions. Periradicular tissues from patients with symptomatic and asymptomatic lesions were obtained and stored at -70°C. Pulpal tissues from unerupted third molars and chronically inflamed gingival tissues were also obtained and used as negative and positive controls, respectively. Tissue samples were homogenized, and supernatants were assayed for IL-1β by ELISA. Significantly higher levels of IL-1β were found in symptomatic and asymptomatic lesions …

Book Review: The Baculovirus Expression System: A Laboratory Guide (1992) King, L. A. & Possee, R. D., David D. Dunigan

Nebraska Center for Virology: Faculty Publications

The power of molecular biology is unleashed with the ability to clone and sequence genes, and then express these genes in heterologous systems. This sets the stage for the full analysis of proteins that are otherwise difficult to isolate and/or purify, especially when present at very low copy number per cell or when isolated from relatively precious materials. Overexpression of protein is now possible in a number of systems including prokaryotes (e.g., E. coli) and various eukaryotes (yeast, insects, and plants). The issue then becomes, which system (1) most closely reflects the homologous expression with respect to posttranslational modifications, …

Altered Expression Of Adenovirus 12 Dna-Binding Protein But Not Dna Polymerase During Abortive Infection Of Hamster Cells, Lynne A. Lucher, Benjawan Khuntirat, Jiansheng Zhao, Peter C. Angeletti

Nebraska Center for Virology: Faculty Publications

Replication of human adenovirus type 12 DNA is blocked in abortively infected baby hamster kidney cells. The activity and accumulation of adenovirus 12 DNA polymerase is equivalent in infected hamster and human cell extracts. However, the accumulation of adenovirus type 12 DNA-binding protein is approximately 120-fold lower in extracts from infected hamster cells when compared to infected permissive human cells. This difference in accumulation is not because of replication of viral DNA during productive infection, since this difference is observed in the presence of hydroxyurea. The DNA-binding protein from infected hamster cells retains the ability to bind denatured DNA-cellulose. An …

Phosphorylation Of The Rex Protein Of Human T-Cell Leukemia Virus Type 1, Yoshifumi Adachi, Terry D. Copeland, Chiaki Takahashi, Tetsuya Nosaka, Aftab Ahmed, Stephen Oroszlan, Masakazu Hatanaka

Pharmacy Faculty Articles and Research

Rex protein, the posttranscriptional regulator of human T-cell leukemia virus type I (HTLV-I), is required for the control of viral structural protein expression and virus replication. Rex is a phosphoprotein found predominantly in the cell nucleolus, whose function is thought to be regulated by its nucleolar localization and phosphorylation. Therefore, we investigated the in vivo phosphorylation of Rex protein in more detail. Phosphorylation of Rex occurred in all HTLV-I-infected cell lines examined in vivo, primarily at serine residues and to a very small extent at threonine residues. Treatment of cells with 12-O-tetradecanoylphorbol-13-acetate (TPA) led to significant but transient enhancement of …

Maternal Immunomodulation Of Neonatal Alloantigen Response, Leh Chang

Loma Linda University Electronic Theses, Dissertations & Projects

Remarkable success has been achieved in the transplantation of allogeneic cardiac grafts into newborn infants at Loma Linda University Medical Center. The superior graft survival rate documented in these patients has not correlated with the degree of immunosuppression rendered, or the selection of genetically matched donors. However, the clinical success has correlated with the age of the recipient at the time of receiving a transplant. Patients receiving an allograft within the first few weeks of life are unique in that they seem to accept the alloantigens of their cardiac graft while responding aggressively to antigens in their environment. These observations …

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. …

Interaction Of Putative Estrogens And The Estrogen Receptor System In Leydig Cells In The Balb/C Mouse Testis Resulting In The Initiation Of Dna Synthesis, R. Lloyd Juriansz

Loma Linda University Electronic Theses, Dissertations & Projects

Continuous administration of estrogens for 7-9 months, both steroidal and nonsteroidal, to male BALB/c mice, leads to the formation of testicular Leydig cell tumors. Three days following the subcutaneous implantation of a pellet of estrogen in cholesterol, there is a peak in the incorporation of ³H-tymidine into the DNA of the interstitial cells. These effects are hypothesized to be mediated by the estrogen receptor system in the Leydig cell. Common experimental techniques for the measurement of hormone binding, such as dextran coated charcoal treatment, proved to be impossible to employ in this system, therefore a procedure was developed using …

The Effect Of Diet On Neutrophilic Phagocytosis, Linda L. Hoover

Loma Linda University Electronic Theses, Dissertations & Projects

Two series of experiments were conducted in order to determine the effects of diet on neutrophilic phagocytosis. The first series was designed to determine the effects of varying amounts of oral glucose on phagocytosis. Blood samples from 6 to 8 subjects were drawn at fasting, and at various time periods following ingestion of 12.5, 25, 50, 75, and 100 grams of glucose. In the second series of experiments, blood samples were taken from 11 subjects after an overnight fast and 2 hours following ingestion of one of 4 different liquid diets, which included the following: the meal (I), consisting of …

Cardioregulatory Role Of The Abdominal Ganglion Of Aplysia Californica, George Maeda

Loma Linda University Electronic Theses, Dissertations & Projects

An isolated heart-abdominal ganglion preparation of this mollusc was developed and used to study the cardioregulatory function of the ganglion and nerves. The heart was perfused with artificial sea water through an atrial cannula and the gastro-esophageal artery was catheterized for blood pressure monitoring. The presence and cardioregulatory effects of axons in the nerves was investigated by electrical stimulation. Fine-tipped glass microelectrodes inserted into ganglionic neurons were used to determine the relationship between the neuronal and cardiac activities.

The principal results are listed below.

1. Primarily inhibitory responses are obtained from both right and left connective stimulation, with some evidence for …

The Conversion Of Cytidylic Acid To Deoxycytidylic Acid In Rat Kidney, Judith Marie Werderitsch

Master's Essays (1922 - )

Evidence was presented in 1949 by Bendich, Getler, and Brown (2), that the free pyrimidine base, cytosine, was not incorporated into the nucleotides of deoxyribonucleic acid (DNA). Further studies by Hammarsten, Reichard, and Saluste (9) with cytidine and those by Roll, Weinfeld, and Carroll (26) with cytidylic acid, showed that the nucleoside and nucleotide derivatives were well incorporated into DNA. These experiments indicated that the conversion of the riboside (or ribotide) to the deoxyribose form had occurred with the nucleosidic linkage intact. This theory was further substantiated by Rose and Schweigert (29) using cytidylic acid incorporation into the total viscera …

Replicating Single-Cycle Adenovirus Vectors Generate Amplified Influenza Vaccine Responses, Catherine M. Crosby, William E. Matchett, Stephanie S. Anguiano-Zarate, Christopher A. Parks, Eric A. Weaver, Larry R. Pease, Richard J. Webby, Michael A. Barry

Nebraska Center for Virology: Faculty Publications

Head-to-head comparisons of conventional influenza vaccines with adenovirus (Ad) gene-based vaccines demonstrated that these viral vectors can mediate more potent protection against influenza virus infection in animal models. In most cases, Ad vaccines are engineered to be replication-defective (RD-Ad) vectors. In contrast, replication-competent Ad (RC-Ad) vaccines are markedly more potent but risk causing adenovirus diseases in vaccine recipients and health care workers. To harness antigen gene replication but avoid production of infectious virions, we developed “single-cycle” adenovirus (SC-Ad) vectors. Previous work demonstrated that SC-Ads amplify transgene expression 100-fold and produce markedly stronger and more persistent immune responses than RD-Ad vectors …