Molecular Genetics Commons^™

Vaccinia Virus-Mediated Glioma Gene Therapy Using P53, Il-2 And Il-12, Bing Chen

Loma Linda University Electronic Theses, Dissertations & Projects

Malignant brain tumors are generally lethal soon after their diagnosis, and the five-year survival rate for the patient is low in spite of neurosurgery, radiation therapy, and/or chemotherapy. Gene therapy has the potential to improve patient survival and quality of life by delivering specific combinations of therapeutic genes directly to the tumor for localized treatment.

We have based our cancer gene therapy for glioma on a highly attenuated variant of the Lister strain of vaccinia virus (VV) which has been proven to be a safe and effective vaccine vector. Other advantages include a wide host range, a strong promoter system, …

Hybridization Of Dna By Sequential Immobilization Of Oligonucleotides At The Air-Water Interface, Murali Sastry, Vidya Ramakrishnan, Mrunalini Pattarkine, Anand Gole, K. N. Ganesh

Faculty Works

The hybridization of DNA by sequential electrostatic and hydrogen-bonding immobilization of single-stranded complementary oligonucleotides at the air-water interface with cationic Langmuir monolayers is demonstrated. The complexation of the single-stranded DNA molecules with octadecylamine (ODA) Langmuir monolayers was followed in time by monitoring the pressure-area isotherms. A large (and slow) expansion of the ODA monolayer was observed during each stage of complexation in the following sequence: primary single-stranded DNA followed by complementary single-stranded DNA followed by the intercalator, ethidium bromide. Langmuir-Blodgett (LB) films of the ODA-DNA complex were formed on different substrates and characterized using quartz-crystal microgravimetry (QCM), Fourier transform infrared …

Accessory Cell Defect In Unresponsiveness Of Neonates And Aged To Polysaccharide Vaccines., Subbarao Bondada, H Wu, D A Robertson, R L Chelvarajan

Microbiology, Immunology, and Molecular Genetics Faculty Publications

T independent antigens elicit antibody responses in the absence of carrier specific T helper cells but require signals from accessory cells (macrophages and dendritic cells) or specific cytokines. They are further subdivided into TI-1 and TI-2 categories based on the ability of TI-1 but not TI-2 antigens to elicit immune responses from neonates. Most bacterial polysaccharides including the pneumococcal polysaccharide vaccines belong to the TI-2 class. It is hypothesized that defects in accessory cell function play a critical role in the failure of neonates to respond to such TI-2 antigens. Immune responses to these TI-2 stimuli are also reduced in …

Connexin-32 Contributes To Radiation Resistance In Thyroid Follicles, Da-Thao Dinh Tran

Loma Linda University Electronic Theses, Dissertations & Projects

The objective of this study was to examine the role of gap junctions in the radiation resistance of thyroid follicles. The question of whether gap junctional intercellular communication plays an important role in the “contact effect”, wherein cells in direct apposition are more resistant to the damaging effects of radiation, has been debated for years. We used the Fischer rat thyroid cell line (FRTL-5) to carry out our experiments. This thesis is divided into two parts: 1) characterization of gap junctional properties in FRTL-5 cells and, 2) exposure of these cells to radiation.

We found that, unlike primary thyroid cells, …

Interactions Of Igf-Ii And Cathepsin D In Mcf-7 Breast Cancer Cells, Jesika S. Faridi

Loma Linda University Electronic Theses, Dissertations & Projects

A primary role of the IGF-II/M6P receptor is to target lysosomal enzymes from the golgi to the lysosomes. This receptor has distinct binding sites for IGF-II and M6P, however, reciprocal interactions between these ligands have been observed (Kiess et al. 1989, 1990). Since IGF-II modulates the routing of cathepsin D in MCF-7 cells by blocking the intracellular binding of cathepsin D to the IGF-II/M6P receptor (De León et al. 1996), we hypothesized that expressing a mutant form of IGF-II that does not bind the IGF-II/M6P receptor will not interfere with lysosomal enzyme trafficking.

In our present study, we report the …

The Occurrence Of The Freshwater Clams, Musculium Partumeium (Say) And Pisidium Casertanum (Poli) (Bivalvia: Sphaeriidae), In The Hawaiian Islands, Albert J. Burky, Carl M. Way, Skippy Hau, M. Eric Benbow

Biology Faculty Publications

Sphaeriid clams were collected from an ancient, continuously cultivated taro pond complex at Ke‘anae Peninsula, Maui. Both species are known for adaptations for temporary pond habitats and could have been initially introduced by the earliest Hawaiian settlers transported with moist taro root stock from other areas of Oceania.

Expression And Requirement Of Epithelial Fatty Acid- Binding Protein In Neuronal Axon Growth, Gregory William Allen

Loma Linda University Electronic Theses, Dissertations & Projects

Epithelial fatty acid-binding protein (E-FABP) is expressed in the dorsal root ganglia following sciatic nerve injury and in migrating and differentiating neurons during neuronal development. It was hypothesized that E-FABP expression is required for the robust outgrowth of axons from developing and regenerating neurons. To test this hypothesis, E-FABP expression in both PC12 cells and primary retinal neurons was examined. In PC12 cells, NGF induces E-FABP mRNA and protein during the period of neurite outgrowth, and E-FABP localizes to the perinuclear cytoplasm, nucleus, and growth cone. Furthermore, E-FABP-deficient cell lines exposed to NGF were less differentiated and had shorter neurites …

Identification And Characterization Of Aer, An Energy Sensor In Escherichia Coli, Anuradha Rebbapragada

Loma Linda University Electronic Theses, Dissertations & Projects

This study investigated the mechanism of signal transduction by Aer, the aerotaxis transducer in E.coli. A computerized search of GenBank revealed an open reading frame at 69.1 min on the E.co//chromosome that had a C-terminal fragment with 96.7% identity to the highly conserved domain of Tsr. The open reading frame was renamed aer for aerotaxis and energy responses. When the aer gene was inactivated, approximately 50% of aerotaxis responses were eliminated. When both aer and fsr were inactivated, aerotaxis, redox taxis and glycerol taxis were completely abolished. Expressing Aer in aer fsr cells restored aerotaxis responses and the duration …

Function And Properties Of Groe Chaperonins In Bacterial And Mammalian Cells, Gregory M. Nelson

Loma Linda University Electronic Theses, Dissertations & Projects

Molecular chaperones play an integral role in the folding of most polypeptides in vivo, and protect proteins against aggregation when a cell is under stress. The GroESL proteins of Escherichia coli are the best characterized of the ringed chaperones, or chaperonins. Chaperonins of the eukaryotic cytoplasm interact with a limited number of polypeptides, whereas GroEL is promiscuous as it binds and mediates the folding of many polypeptides. This feature makes GroEL an attractive protein for investigating various aspects of protein folding in eukaryotic cells because its substrate interaction is diverse. In this work we have expressed the groES and …

Novel Function Of Phosphoinositide 3-Kinase In T Cell Ca2+ Signaling. A Phosphatidylinositol 3,4,5-Trisphosphate-Mediated Ca2+ Entry Mechanism., A L Hsu, T T Ching, G Sen, D S Wang, Subbarao Bondada, K S Authi, C S Chen

Microbiology, Immunology, and Molecular Genetics Faculty Publications

This study presents evidence that phosphoinositide (PI) 3-kinase is involved in T cell Ca(2+) signaling via a phosphatidylinositol 3,4, 5-trisphosphate PI(3,4,5)P(3)-sensitive Ca(2+) entry pathway. First, exogenous PI(3,4,5)P(3) at concentrations close to its physiological levels induces Ca(2+) influx in T cells, whereas PI(3,4)P(2), PI(4, 5)P(2), and PI(3)P have no effect on [Ca(2+)](i). This Ca(2+) entry mechanism is cell type-specific as B cells and a number of cell lines examined do not respond to PI(3,4,5)P(3) stimulation. Second, inhibition of PI 3-kinase by wortmannin and by overexpression of the dominant negative inhibitor Deltap85 suppresses anti-CD3-induced Ca(2+) response, which could be reversed by subsequent …

New World Nine-Primaried Oscine Relationships: Constructing A Molecular Framework, John Klicka, Kevin P. Johnson, Scott M. Lanyon

Ornithology Program (HRC)

Historically, a paucity of comparative morphological characters has led to much debate regarding relationships within and among the major lineages of New World nine-primaried oscines. More recently, DNA-DNA hybridization studies have provided novel and testable hypotheses of relationships, although no consensus has been reached. For 40 songbird taxa, we obtained 1,929 base pairs (bp) of DNA sequence from the mitochondrial cytochrome-b (894 bp) and NADH dehydrogenase subunit 2 (1,035 bp) genes. Phylogenetic analyses confirm the monophyly of this assemblage as traditionally defined. The lineages delineated historically on morphological grounds are retained; finches (Fringillinae) are sister to a well-supported clade …

The Skn7 Response Regulator Of Saccharomyces Cerevisiae Interacts With Hsf1 In Vivo And Is Required For The Induction Of Heat Shock Genes By Oxidative Stress, Desmond C. Raitt, Anthony L. Johnson, Alexander M. Erkine, Kozo Makino, Brian Morgan, David S. Gross, Leland H. Johnston

Scholarship and Professional Work – COPHS

The Skn7 response regulator has previously been shown to play a role in the induction of stress-responsive genes in yeast, e.g., in the induction of the thioredoxin gene in response to hydrogen peroxide. The yeast Heat Shock Factor, Hsf1, is central to the induction of another set of stress-inducible genes, namely the heat shock genes. These two regulatory trans-activators, Hsf1 and Skn7, share certain structural homologies, particularly in their DNA-binding domains and the presence of adjacent regions of coiled-coil structure, which are known to mediate protein–protein interactions. Here, we provide evidence that Hsf1 and Skn7 interact in vitro and …

Cell Cycle-Dependent Binding Of Yeast Heat Shock Factor To Nucleosomes, Christina Bourgeois Venturi, Alexander M. Erkine, David S. Gross

Scholarship and Professional Work – COPHS

In the nucleus, transcription factors must contend with the presence of chromatin in order to gain access to their cognate regulatory sequences. As most nuclear DNA is assembled into nucleosomes, activators must either invade a stable, preassembled nucleosome or preempt the formation of nucleosomes on newly replicated DNA, which is transiently free of histones. We have investigated the mechanism by which heat shock factor (HSF) binds to target nucleosomal heat shock elements (HSEs), using as our model a dinucleosomal heat shock promoter (hsp82-ΔHSE1). We find that activated HSF cannot bind a stable, sequence-positioned nucleosome in G₁-arrested …

Expression And Function Of Hoxc5 And Hoxc6 In The Murine Mammary Gland, Silvia Alejandra Garcia-Gasca

MUSC Theses and Dissertations

Homeotic genes encode transcription factors, believed to regulate cell proliferation, differentiation, as well as cell migration to create positional identities during embryogenesis. In the mouse, the Hox complex consists of at least 39 genes, organized into four clusters, in four different chromosomes. Hoxc5 and Hoxc6 are adjacent genes located in chromosome 15, and are expressed in the developing somites of the thoracic region and central nervous system with overlapping patterns. Both Hoxc5 and Hoxc6 are also expressed in the mammary gland. This study demonstrates that a targeted disruption of the Hoxc6 gene results in a failure of ductal development in …

Haemochromatosis Mutation Analysis In A Normal Irish Population, Fergus Ryan, Joseph Vaughan

Articles

187 Irish blood donors were examined for the presence of C282Y and H63D missense mutations using a multiplex PCR and restriction digest protocol. The allele frequency for C282Y was found to be 12.3% which is higher than reported in other studies and concurs that this mutation is of Celtic origin.

Construction Of Escherichia Coli Arginine Repressor Fusion Protein And Analysis Of Its Function In Xer Site-Specific Recombination, Kholis Abdulrachim

Student Works (2000-2009)

In addition to its role in L-arginine biosynthesis in Escherichia coli, arginine repressor (ArgR), the product of the argR gene, also plays an essential role as an obligate accessory protein in Xer site-specific recombination system. A structure-function relationship study of ArgR was performed to understand more about its role in Xcr site-specific recombination. Fusion proteins between ArgRWT (wild-type ArgR) and a biotinylated peptide as well as between ArgRNV (a mutant ArgR) and a biotinylated peptide were constructed. The biotinylated peptide was fused in frame to the amino-tenninus of ArgR WT and ArgRNV, respectively. Xer recombination assays showed that the ArgRWT-biotinylated …

In Vitro Mutagenesis For Structure/Function Studies Of The Bacteriophage T4 Rega Protein And The Murine Ahr Protein, Estel Rick Barbour

MUSC Theses and Dissertations

The bacteriophage T4 regA protein translationally represses over 30 T4 early genes, including the regA gene, by binding to the translation initiation region of the T4 mRNAs (Karam et al., 1981). Although the recognition sequence has been at least partially characterized in a number regA sensitive mRNAs, there is no consensus sequence or structural feature that is recognized by regA in all transcripts (Winter et al., 1987; Webster et al., 1989; Unnithan et al., 1990). Recent studies have identified an RNA binding site on regA protein, but there remains a large gap in our understanding of how regA protein functions …

Application Of Ai In Medicine, Tarmizi Amirul Hisyam

Student Works (2000-2009)

From the beginning of human civilization, mankind have been trying to fins solution on how to make life easier. They create, modify and adapt, as they would please their need. From the smallest things in our life to the biggest part of all technologies, has improved our life tremendously. Computer technology nowadays has an important role in education, communication, medicine, and many more. In the field of medicine, we have seen many support systems had been developed and use to help human in their daily lives. Application of AI in medicine was proposed in this reason in mind, as a …