Medicine and Health Sciences Commons^™

Screening For Mutants In The Output Pathway Of The Circadian Clock In Chlamydomonas Reinhardtii, Wei Yuan

Masters Theses & Specialist Projects

The circadian clock is a basic component of biological systems and has been found in many organisms. It is composed by three major components: the input pathway, the oscillator and the output pathway. The purpose of this research is to study the output pathway of the circadian clock in Chlamydomonas reinhardtii strain Carnil by screening mutants, which were generated by insertional mutagenesis via glass bead transformation. The plasmid pSP124S containing the ble marker was used to introduce mutations. The CABII-1 gene has been reported to show a circadian rhythm in expression. The reporter gene ARS2 that was transcriptionally fused to …

Genetic Control Of Eye Development, A Case Study Focused On The Murine Mutation Belly Spot And Tail (Bst), Qing Tang

Theses and Dissertations (ETD)

Development of the eye is controlled by a network of genes, often conserved, that regulate the timing and location of cellular differentiation. One approach to understanding this network of genes and their interaction is to focus on mutations, spontaneous or induced, that predictably disrupt the proper function of such networks, and by examining the effect of such disruption on the function of other genes.

The Belly spot and tail (Bst) semi-dominant mutation, mapped to mouse Chromosome 16, leads to developmental defects of the eye, skeleton, and coat pigmentation. In the eye, the mutant phenotype is characterized by the presence of …

A Ligand Binding Domain Mutation In The Mouse Glucocorticoid Receptor Functionally Links Chromatin Remodeling And Transcription Initiation, Lynn A. Sheldon, Catharine L. Smith, Jack E. Bodwell, Allan U. Munck, Gordon L. Hager

Dartmouth Scholarship

We utilized the mouse mammary tumor virus (MMTV) long terminal repeat (LTR) in vivo to understand how the interaction of the glucocorticoid receptor (GR) with a nucleosome-assembled promoter allows access of factors required for the transition from a repressed promoter to a derepressed, transcriptionally competent promoter. A mutation (C644G) in the ligand binding domain (LBD) of the mouse GR has provided information regarding the steps required in the derepression/activation process and in the functional significance of the two major transcriptional activation domains, AF1 and AF2. The mutant GR activates transcription from a transiently transfected promoter that has a disordered nucleosomal …

Monoclonal Antibody 1a7 And Related Polypeptides, Malaya Chatterjee, Kenneth A. Foon, Sunil K. Chatterjee

Microbiology, Immunology and Molecular Genetics Faculty Patents

The present invention relates to monoclonal antibody 1A7. This is an anti-idiotype produced by immunizing with an antibody specific for ganglioside GD2, and identifying a hybridoma secreting antibody with immunogenic potential in a multi-step screening process. Also disclosed are polynucleotide and polypeptide derivatives based on 1A7, including single chain variable region molecules and fusion proteins, and various pharmaceutical compositions. When administered to an individual, the 1A7 antibody overcomes immune tolerance and induces an immune response against GD2, which comprises a combination of anti-GD2 antibody and GD2-specific T cells. The invention further provides methods for treating a disease associated with altered …

Murine Anti-Idiotype Antibody 3h1, Malaya Chatterjee, Heinz Köhler, Sunil K. Chatterjee, Kenneth A. Foon

Microbiology, Immunology and Molecular Genetics Faculty Patents

The present invention provides a monoclonal anti-idiotype antibody 3H1 that escapes immune tolerance and elicits a specific immune response to CEA in mice, rabbits, monkeys, and patients with advanced CEA-associated disease. This invention also provides compositions which can be used in the detection or treatment of CEA-associated tumors mimics a specific epitope on carcinoembryonic antigen and a hybridoma that produces 3H1.

Construct Validity Of The Continuous Recognition Memory Test, K. L. Fuchs, J. H. Hannay, W. M. Huckeba, K. A. Espy

Developmental Cognitive Neuroscience Laboratory: Faculty Publications

A principal factor analysis was performed on variables derived from a neuropsychological battery administered to 100 healthy young adults (aged 17-41 yrs) to investigate the construct validity of the Continuous Recognition Memory test (CRM). It was hypothesized that CRM "hits" and "false alarms" would load on different factors. The factors that emerged in the analysis were labeled Verbal Ability, Divided Attention, Attention to Visual Detail, Visuomotor Integration and Planning, and Learning and Memory. As expected, CRM hits had a significant loading on the Learning and Memory factor. However, CRM false alarms did not have a significant loading on the Divided …

Neuropsychologic Function In Toddlers Exposed To Cocaine In Utero: A Preliminary Study, K. A. Espy, P. M. Kaufmann, M. L. Glisky

Developmental Cognitive Neuroscience Laboratory: Faculty Publications

Patterns of neuropsychological performance on A-not-B, inhibition, motor, cognitive, language, and behavior tasks were examined in 34 toddlers--17 cocaine-exposed (CE) and 17 nonexposed (NE) controls. CE toddlers exhibited greater perseveration, less inhibition, poorer emotional regulation, and less task orientation relative to NE toddlers. Overall cognitive and language skills and motor impairment status were comparable among CE and NE toddlers. Differences in perseveration, emotional regulation, and task orientation between CE and NE toddlers remained significant after statistically controlling for overall cognitive skill. Prenatal cocaine exposure may impart selective vulnerability for deficits in executive function, inhibition, and emotional regulation in toddlers, perhaps …

Executive Functioning In Preschool Children: Performance On A-Not-B And Other Delayed Response Format Tasks, K. A. Espy, P. M. Kaufmann, M. D. Mcdiarmid, M. L. Glisky

Developmental Cognitive Neuroscience Laboratory: Faculty Publications

The A-not-B (AB) task has been hypothesized to measure executive/frontal lobe function; however, the developmental and measurement characteristics of this task have not been investigated. The present study examined performance on AB and comparison tasks adapted from developmental and neuroscience literature in 117 1.9-5.5 yr old preschool children. Age significantly predicted performance on AB, Delayed Alternation, Spatial Reversal, Color Reversal, and Self-Control tasks. A 4-factor analytic model best fit task performance data. AB task indices loaded on 2 factors with measures from the Self-Control and Delayed Alternation tasks, respectively. AB indices did not load with those from the reversal tasks …

The Predictive Use Of Event-Related Potentials In Language Development And The Treatment Of Language Disorders, Dennis L. Molfese, Victoria J. Molfese, K. A. Espy

Developmental Cognitive Neuroscience Laboratory: Faculty Publications

Attempts to relate what is currently known regarding the brain's involvement in language processing during the early years of life. The authors focus on the event related potential (ERP) as a means to study the neuroelectrical correlates of language in the brains of infants and children. After reviewing general information concerning ERPs and language, this presentation relates how neonatal ERP measures of phonetic discrimination predict later language and reading abilities.

Enhanced Apoptosis Mediates Inhibition Of Ebv-Transformed Lymphoblastoid Cell Line Proliferation By Curcumin., D Ranjan, T D Johnston, K S Reddy, G Wu, Subbarao Bondada, C Chen

Microbiology, Immunology, and Molecular Genetics Faculty Publications

BACKGROUND: Epstein-Barr virus (EBV)-associated B-cell lymphomas occur more frequently in immunodeficient states such as organ transplantation and HIV infection. We have previously reported that B cell immortalization with EBV was promoted by cyclosporin A (CyA) and that curcumin (Cur), a natural phenol with known antioxidant and antitumor properties, blocked EBV-induced B cell immortalization. In the following experiments we show that Cur inhibits the proliferation of EBV-transformed lymphoblastoid cell lines (LCL) via enhanced apoptosis.

METHODS: LCL were generated by infecting freshly isolated human B cells with EBV (B95-8) for 12 h and coculturing with predetermined optimal concentrations of CyA (500 ng/ml) …

Immune Cd8+ T Cells Prevent Reactivation Of Toxoplasma Gondii Infection In The Immunocompromised Host, Imtiaz A. Khan, William R. Green, Lloyd H. Kasper, Kathy A. Green, Joseph D. Schwartzman

Dartmouth Scholarship

Toxoplasma gondii remains a serious cause of morbidity and mortality in individuals that are immunosuppressed, patients with AIDS in particular. The cellular immune response, especially by gamma interferon (IFN-γ)-producing CD8⁺ T cells, is an essential component of protective immunity against the parasite. In the present study the role of CD8⁺ T cells during the reactivation of Toxoplasma infection in an immunocompromised murine model was evaluated. Chronically infected mice were challenged with LP-BM5 virus, and the kinetics of CD8⁺ T-cell function was studied. At 10 weeks after viral infection, mice showed obvious signs of systemic illness and began …

Cartilage Regeneration On A Large Articular Surface Facilitated By Stress Shielding, Charles L. Mcdowell, Jennifer S. Wayne, Robert Tuten, Hunter H. Mcguire Jr.

Virginia Journal of Science

An animal model for the study of articular cartilage regeneration in-vivo facilitated by stress-shielding is introduced. The object of the model is to test the hypothesis that some form of cartilaginous tissue will grow upon a large joint surface in vivo with the joint in normal motion. The model utilizes the known capability of immature cells to differentiate. The source of cells is bleeding subchondral bone. In addition, the model provides a mechanically shielded environment in which cell differentiation and maturation can occur. The study showed that a substantial amount of tissue will grow in the animal model only when …

Subependymomas: Clinicopathologic Study Of 14 Tumors, Daniel F. Brown Md, Mba, Elizabeth J. Rushing Md

Department of Pathology & Laboratory Medicine

No abstract provided.

Differential Transcription Of The Tcpph Operon Confers Biotype-Specific Control Of The Vibrio Cholerae Toxr Virulence Regulon, Yvette M. Murley, Patricia A. Carroll, Karen Skorupski, Ronald K. Taylor, Stephen B. Calderwood

Dartmouth Scholarship

Epidemic strains of Vibrio cholerae O1 are divided into two biotypes, classical and El Tor. In both biotypes, regulation of virulence gene expression depends on a cascade in which ToxR activates expression of ToxT, and ToxT activates expression of cholera toxin and other virulence genes. In the classical biotype, maximal expression of this ToxR regulon in vitro occurs at 30 degrees C at pH 6.5 (ToxR-inducing conditions), whereas in the El Tor biotype, production of these virulence genes only occurs under very limited conditions and not in response to temperature and pH; this difference between biotypes is mediated at the …

Immune Responses Induced By Gene Gun Or Intramuscular Injection Of Dna Vaccines That Express Immunogenic Regions Of The Serine Repeat Antigen From Plasmodium Falciparum, Alexia A. Belperron, David Feltquate, Barbara A. Fox, Toshihiro Horii, David J. Bzik

Dartmouth Scholarship

The liver- and blood-stage-expressed serine repeat antigen (SERA) of Plasmodium falciparum is a candidate protein for a human malaria vaccine. We compared the immune responses induced in mice immunized with SERA-expressing plasmid DNA vaccines delivered by intramuscular (i.m.) injection or delivered intradermally by Gene Gun immunization. Mice were immunized with a pcdna3 plasmid encoding the entire 47-kDa domain of SERA (amino acids 17 to 382) or the N-terminal domain (amino acids 17 to 110) of SERA. Minimal antibody responses were detected following DNA vaccination with the N-terminal domain of SERA, suggesting that the N-terminal domain alone is not highly immunogenic …

Intermittent Antiretroviral Therapy (Art) Can Induce Reduction Of Viral Rebounding During Art-Interruption, Joseph L. Yozviak Do, Facp, R Eric Doerfler Np, Cch, William C. Woodward Do

Department of Medicine

No abstract provided.

Vacuole Acidification Is Required For Trans-Snare Pairing, Lma1 Release, And Homotypic Fusion, Christian Ungermann, William Wickner, Zuoyu Xu

Dartmouth Scholarship

Vacuole fusion occurs in three stages: priming, in which Sec18p mediates Sec17p release, LMA1 (low M_r activity 1) relocation, and cis-SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) complex disassembly; docking, mediated by Ypt7p and trans-SNARE association; and fusion of docked vacuoles. Ca²⁺ and calmodulin regulate late stages of the reaction. We now show that the vacuole proton gradient, generated by the vacuolar proton ATPase, is needed for trans-SNARE complex formation during docking and hence for the subsequent LMA1 release. Though neither the vacuolar Pmc1p Ca²⁺-ATPase nor the Vcx1p Ca²⁺/H⁺ exchanger …

Phosphoinositide-Ap-2 Interactions Required For Targeting To Plasma Membrane Clathrin-Coated Pits., I Gaidarov, James H. Keen

Department of Microbiology and Immunology Faculty Papers

The clathrin-associated AP-2 adaptor protein is a major polyphosphoinositide-binding protein in mammalian cells. A high affinity binding site has previously been localized to the NH(2)-terminal region of the AP-2 alpha subunit (Gaidarov et al. 1996. J. Biol. Chem. 271:20922-20929). Here we used deletion and site- directed mutagenesis to determine that alpha residues 21-80 comprise a discrete folding and inositide-binding domain. Further, positively charged residues located within this region are involved in binding, with a lysine triad at positions 55-57 particularly critical. Mutant peptides and protein in which these residues were changed to glutamine retained wild-type structural and functional characteristics by …

Method Of Protecting Against Neuron Loss, Philip W. Landfield

Pharmacology and Nutritional Sciences Faculty Patents

The present invention pertains to a method of protecting against neuron loss in a subject by administering a compound that protects against neuron loss by acting through a vitamin D receptor. Some of these compounds may prevent or retard neuron loss by regulating intraneuronal and/or peripheral calcium and phosphate levels. Other compounds of the invention act through a vitamin D receptor to protect against neuron loss through mechanisms not involving calcium or phosphate regulation. A preferred compound is a biologically active form of vitamin D, a precursor, metabolite, or analog of vitamin D. A preferred form of vitamin D is …

Polynucleotides Related To Monoclonal Antibody 1a7 And Use For The Treatment Of Melanoma And Small Cell Carcinoma, Malaya Chatterjee, Kenneth A. Foon, Sunil K. Chatterjee

Microbiology, Immunology and Molecular Genetics Faculty Patents

The present invention relates to monoclonal antibody 1A7. This is an anti-idiotype produced by immunizing with an antibody specific for ganglioside GD2, and identifying a hybridoma secreting antibody with immunogenic potential in a multi-step screening process. Also disclosed are polynucleotide and polypeptide derivatives based on 1A7, including single chain variable region molecules and fusion proteins, and various pharmaceutical compositions. When administered to an individual, the 1A7 antibody overcomes immune tolerance and induces an immune response against GD2, which comprises a combination of anti-GD2 antibody and GD2-specific T cells. The invention further provides methods for treating a disease associated with altered …

The Effects Of Phosphorus Enrichment On The Dominant Phytoplankton Communities Of Chaney Lake, Kathy Tehrani

Masters Theses & Specialist Projects

Chaney Lake is a temporary karst wetland located in southern Warren County, Kentucky. Because of an impermeable chert layer between the surface and the porous limestone, Chaney Lake fills with water over the winter and spring and then gradually drains over the summer. Three experiments were conducted over the course of the 1998 flooding season to assess the effects of phosphorus addition on the phytoplankton community in Chaney Lake. Ten plastic-sided mesocosms were constructed and placed in the marsh area of the wetland at three different periods: early spring, early summer, and late summer. Phosphorus in the form of K2HPO4 …

The Localization Of Gaba-Like Immunoreactivity In The Optic Lobe Neuropils And Optic Tracts Of The Cockroach Leucophaea Maderae, David Guffey

Masters Theses & Specialist Projects

Gamma aminobutyric acid (GABA) is a prominent neurotransmitter found in both vertebrates and invertebrates. The action of GABA has been accepted to be inhibitory in nature, but recent evidence suggests it is both inhibitory and excitatory, depending upon time of day. The amount of GABA measured in the brain of the cockroach Leucophaea maderae was discovered to fluctuate in a circadian pattern similar to the pattern of electrical output measured from the optic nerve in L. maderae. Considering the circadian oscillator for L. maderae has been localized to the optic lobes of the brain, the discovery of GABA-like immunoreactivity in …

Genetic And Structural Analyses Of Cytoplasmic Filaments Of Wild-Type Treponema Phagedenis And A Flagellar Filament-Deficient Mutant, Jacques Izard, William A. Samsonoff, Mary Beth Kinoshita, Ronald J. Limberger

Department of Food Science and Technology: Faculty Publications

Unique cytoplasmic filaments are found in the treponeme genus of spirochete bacteria. Their function is unknown, but their location underneath the periplasmic flagellar filaments (PFF) suggests a role in motility and/or cell structure. To better understand these unique structures, the gene coding for the cytoplasmic filaments, cfpA, was identified in various treponemal species. Treponema phagedenis cfpA was 2,037 nucleotides long, and the encoded polypeptide showed 78 to 100% amino acid sequence identity with the partial sequence of CfpA from T. denticola, T. vincentii, and T. pallidum subsp. pertenue. Wild-type T. phagedenis and a …

Global Cns Gene Transfer For A Childhood Neurogenetic Enzyme Deficiency: Canavan Disease., Paola Leone, Christopher G Janson, Scott J Mcphee, Matthew J During

Department of Neurosurgery Faculty Papers

The neurogenetic prototypic disease on which we chose to test our gene therapy strategy is Canavan disease (CD). CD is an autosomal recessive leukodystrophy associated with spongiform degeneration of the brain. At present the disease is uniformly fatal in affected probands. CD is characterized by mutations in the aspartoacylase (ASPA) gene, resulting in loss of enzyme activity. In this review, recent evidence is summarized on the etiology and possible treatments for CD. In particular, we discuss two gene delivery systems representing recent advances in both viral and liposome technology: a novel cationic liposome-polymer-DNA (LPD) complex, DCChol/DOPE-protamine, as well as recombinant …

Effects Of Community Structure On The Seagrass Thalassia Testudinum, Craig D. Rose, Clinton J. Dawes

Integrative Biology Faculty and Staff Publications

The influence of community structure on the seagrass Thalassia testudinum was studied in 3 distinct communities: low and high density monocultures, and patches intermixed with Halodule wrightii, in Cockroach Bay, Tampa, Florida. T. testudinum shoot-specific leaf mass and growth were significantly higher in low density monocultures, and both variables were negatively correlated with short-shoot density. Intraspecific, competition-density effects in high-density seagrass beds may be responsible for the relatively lower shoot-specific leaf mass and growth rates, possibly due to the reduction of available light from dense leaf canopies. These observations are supported by significantly higher T. testudinum leaf C:N and …

Protein Adducts Of Iso[4]Levuglandin E2, A Product Of The Isoprostane Pathway, In Oxidized Low Density Lipoprotein, Robert G. Salomon, Wei Sha, Cynthia Brame, Kamaljit Kaur, Ganesamoorthy Subbanagounder, June O'Neil, Henry F. Hoff, L. Jackson Roberts Ii

Pharmacy Faculty Articles and Research

Levuglandin (LG) E₂, a cytotoxic seco prostanoic acid co-generated with prostaglandins by nonenzymatic rearrangements of the cyclooxygenase-derived endoperoxide, prostaglandin H₂, avidly binds to proteins. That LGE₂-protein adducts can also be generated nonenzymatically is demonstrated by their production during free radical-induced oxidation of low density lipoprotein (LDL). Like oxidized LDL, LGE₂-LDL, but not native LDL, undergoes receptor-mediated uptake and impaired processing by macrophage cells. Since radical-induced lipid oxidation produces isomers of prostaglandins, isoprostanes (isoPs), via endoperoxide intermediates, we postulated previously that a similar family of LG isomers, isoLGs, is cogenerated with isoPs. Now …

K-Cl Cotransport: Membrane Immunofluorescence Localization By Confocal Microscopy Of The Kcc1 Protein, Before And After Truncation Of The Cooh-Terminal Domain, In Human Embryonic Kidney (Hek293) Cells, Peter K. Lauf, Kenneth B.E. Gagnon, Jin Zhang, Eric Delpire, Robert E.W. Fyffe, Norma C. Adragna

Pharmacology and Toxicology Faculty Publications

No abstract provided.

Clinical Trials In Alzheimer Disease: Debate On The Use Of Placebo Controls, Peter J. Whitehouse

Faculty Scholarship

During the past 10 years, there has been a rapidly growing number of pharmaceutical industry-sponsored drug trials for treatment of Alzheimer disease (AD) and other neurodegenerative diseases. As public awareness and concerns about AD have grown, so has interest in developing drug therapies for retarding symptom progression, delaying onset, and ultimately curing the disease. Ethical debate on the use of placebo control trials in AD research has come of age in the United States with the availability of treatments approved by the Food and Drug Administration. The experts and the public agree that more effective therapies are necessary, and new …

Dichotomy Between Naïve And Memory Cd4+ T Cell Responses To Fas Engagement, J. Desbarats, T. Wade, W. F. Wade, M. K. Newell

Dartmouth Scholarship

Engagement of Fas (APO-1, CD95), a member of the tumor necrosis factor receptor superfamily, can induce apoptotic cell death. However, Fas engagement also can costimulate lymphocyte proliferation. The physiologic regulation of these two outcomes is poorly understood. Here, we have used two systems, the first in vitro and the second in vivo, to demonstrate that naïve and memory CD4+ T cells display dichotomous responses to Fas ligation. Naïve CD4+ T cells (CD44lo, CD45RB+, CD62L+) die as a consequence of Fas ligation in the presence of anti-CD3 antibody, whereas memory T cells (CD44hi, CD45RB−, CD62L−), freshly isolated from the same …

Optimal Replication Activity Of Vesicular Stomatitis Virus Rna Polymerase Requires Phosphorylation Of A Residue(S) At Carboxy-Terminal Domain Ii Of Its Accessory Subunit, Phosphoprotein P, Leroy N. Hwang, Nathan Englund, Tapas Das, Amiya K. Florida, Asit K. Pattnaik

School of Veterinary and Biomedical Sciences: Faculty Publications

The phosphoprotein, P, of vesicular stomatitis virus (VSV) is a key subunit of the viral RNA-dependent RNA polymerase complex. The protein is phosphorylated at multiple sites in two different domains. We recently showed that specific serine and threonine residues within the amino-terminal acidic domain I of P protein must be phosphorylated for in vivo transcription activity, but not for replication activity, of the polymerase complex. To examine the role of phosphorylation of the carboxy-terminal domain II residues of the P protein in transcription and replication, we have used a panel of mutant P proteins in which the phosphate acceptor sites …