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Genetic Determinism Or Genetic Discrimination, George P. Smith Ii

Scholarly Articles

The first part of this essay evaluates the contemporary focus of the genetic revolution seen as such through the Human Genome Initiative - a project which brings not only great hope for new advancements of genetic knowledge designed to control disease and minimize human suffering, but raises real fears of unabated invasions of personal privacy that in turn would lead to discrimination for those found to be genetically handicapped. The extent to which this central fear is justified or rational is justified within the present context of the Genome Initiative, together with the past practices of genetic screening, will then …

Enhanced Immunogenicity Of Protein-Dextran Conjugates: I. Rapid Stimulation Of Enhanced Antibody Responses To Poorly Immunogenic Molecules, Andrew Lees, Fred Finkelman, John K. Inman, Kim Witherspoon, Penny Johnson, Jasiri Kennedy, James J. Mond

Microbiology, Immunology, and Molecular Genetics Faculty Publications

In view of our observation that anti-immunoglobulin antibody conjugated to high-molecular-weight dextran stimulates high levels of B-cell activation (Brunswick et al. J. Immunol . 1989, 143, 1239), we coupled T cell-dependent antigens to dextran. When mice were immunized, in the absence of adjuvant, with a BSA-dextran conjugate (BSA-dex), a persistent, high-titre anti-BSA IgG1 response was induced. Titres were dose-dependent and seen with as little as 10 I~g of conjugated protein. Anti-BSA titres were detected as early as day 7, usually peaked at about day 14 and persisted for at least 4 weeks. Anti-hapten antibodies were also elicited in mice that …

Localisation And Detection Of A Polymorphism In The Human Skeletal Beta-Tropomyosin Gene (Tpm2), Clive C.J. Hunt

Theses : Honours

Tropomyosin is one of the components of the thin filaments of muscle, binding to actin, and, together with troponin, regulating contraction in a calcium-dependent manner (Cho et al.,1990). There are at least four distinct tropomyosin genes in vertebrates and each may encode at least six different isoforms of tropomyosin by alternate splicing (Novy et al, 1993; MacLeod et al., 1988). The alpha-tropomyosin gene TPM1 has recently been localised to 15q22 (Eyre et al, 1994) and has been shown to be mutated in some cases of familial hypertrophic cardiomyopathy (Thierfelder et al., 1994). The alpha-tropomyosin gene TPM3 has been recently localised …

Cooperative Binding Of Ets-1 And Core Binding Factor To Dna., David Wotton, Jacques Ghysdael, Shuwen Wang, Nancy A. Speck, Michael J. Owen

Dartmouth Scholarship

Two phorbol ester-inducible elements (beta E2 and beta E3) within the human T-cell receptor beta gene enhancer each contain consensus binding sites for the Ets and core binding factor (CBF) transcription factor families. Recombinant Ets-1 and purified CBF bound individually to beta E2 and beta E3, in which the Ets and core sites are directly adjacent. In this report, we show that CBF and Ets-1 bind together to beta E2 and beta E3 and that Ets-1-CBF-DNA complexes are favored over the binding of either protein alone to beta E2. Formation of Ets-1-CBF-DNA complexes increased the affinity of Ets-1-DNA interactions and …

Morphometric Analysis Of Enteric Lesions In C3h/Hen Mice Inoculated With Serpulina Hyodysenteriae Serotypes 2 And 4 With Or Without Oral Streptomycin Pretreatment, Jagannatha V. Mysore, Gerald Duhamel

School of Veterinary and Biomedical Sciences: Faculty Publications

The segmental distribution and sequential progression and the role of the indigenous bacterial flora in the development of enteric lesions associated with Serpulina hyodysenteriae infection in laboratory mice have not been defined. We examined the distribution and sequential morphometric changes in the large intestine of mice orally inoculated with S. hyodysenteriae serotypes 2 and 4. To determine the role of colonization resistance conferred by the indigenous bacterial flora, 40 female C3H/HeN mice were administered water alone or water containing 5 mg/mL streptomycin sulfate ad libitum for seven days prior to orogastric inoculation either with S. hyodysenteriae or sterile trypticase soy …

Faculty Publications 1993, Lsu Health Sciences Center New Orleans

Historical Faculty Publications

This Faculty bibliography contains 753 publications.

Antiparasitic And Antiproliferative Effects Of Indoleamine 2,3-Dioxygenase Enzyme Expression In Human Fibroblasts., Sohan L. Gupta, Joseph M. Carlin, Padma Pyati, Wei Dai, Elmer R. Pfefferkorn, Martin J. Murphy Jr

Dartmouth Scholarship

Studies were carried out to evaluate the proposed role of indoleamine 2,3-dioxygenase (INDO) induction in the antimicrobial and antiproliferative effects of gamma interferon (IFN-gamma) in human fibroblasts. The INDO cDNA coding region was cloned in the pMEP4 expression vector, containing the metallothionein (MTII) promoter in the sense (+ve) or the antisense (-ve) orientation. Human fibroblasts (GM637) stably transfected with the sense construct expressed INDO activity after treatment with CdCl2 or ZnSO4, but cells transfected with the antisense construct did not. The growth of Chlamydia psittaci was strongly inhibited in INDO +ve cells but not in INDO -ve cells after treatment …

Use Of Yeast Artificial Chromosomes (Yacs) For Studying Control Of Gene Expression: Correct Regulation Of The Genes Of A Human Beta-Globin Locus Yac Following Transfer To Mouse Erythroleukemia Cell Lines., Kenneth Peterson, Galynn Zitnik, Clare Huxley, Christopher Lowrey

Dartmouth Scholarship

We demonstrate that transfer of a yeast artificial chromosome (YAC) containing 230 kb of the human beta-globin locus into mouse erythroleukemia cells by fusion results in correct developmental regulation of the human beta-like globin genes. Additionally, we show that early after hybrid formation, human embryonic epsilon- and fetal gamma-globin genes are coexpressed with the adult beta gene but that after 10-20 weeks in culture, globin gene expression switches to predominantly adult. Thus, in contrast to shorter gene constructs, the globin genes of the beta-globin locus YAC are regulated like the chromosomal globin genes. These results indicate that transfer of YACs …

Evidence For A Second Type Of Fibril Branch Point In Fibrin Polymer Networks, The Trimolecular Junction, Michael W. Mosesson, James P. Diorio, Kevin R. Siebenlist, Joseph S. Wall, James F. Hainfeld

Biomedical Sciences Faculty Research and Publications

Fibrin molecules polymerize to double-stranded fibrils by intermolecular end-to-middle domain pairing of complementary polymerization sites, accompanied by fibril branching to form a clot network. Mass/length measurements on scanning transmission electron microscopic images of fibrils comprising branch points showed two types of junctions. Tetramolecular junctions occur when two fibrils converge, creating a third branch with twice the mass/length of its constituents. Newly recognized trimolecular junctions have three fibril branches of equal mass/length, and occur when an extraneous fibrin molecule initiates branching in a propagating fibril by bridging across two unpaired complementary polymerization sites. When trimolecular junctions predominate, clots exhibit nearly perfect …

Sds Non-Acrylamide Polymeric Gel-Filled Capillary Electrophoresis For Molecular Size Separation Of Protein, Devon Andres

Honors Theses

Sodium dodecyl sulfide (SDS) non-acrylamide gel-filled capillary columns are a new technology being used for analysis and separation of biotechnology-derived proteins. This research was to compare this new technology to the current methods of SDS polyacrylamide gel electrophoresis (SDS-PAGE) and high-performance size-exclusion chromatography (HPSEC). The molecular mass of four different recombinant proteins were determined by two commercialized SDS non-acrylamide gel-filled capillary columns, SDS-PAGE, and HPSEC. The data obtained showed that the SDS non-acrylamide gel-filled capillary columns were compatible with the SDS-PAGE technique for molecular mass determination. HPSEC was shown to be unreliable for molecular weight determination. SDS non-acrylamide gel-filled capillary …

Molecular Cloning Of Infectious Ecotropic Murine Leukemia Virus Ak7 From An Emv-14-Positive Akxl-5 Mouse And The Resistance Of Ak7 To Recognition By Cytotoxic T Lymphocytes., Hillary D. White, William R. Green, Nuria R. Giné

Dartmouth Scholarship

The AKXL-5 recombinant inbred mouse strain is positive for the endogenous ecotropic murine leukemia virus emv-14, the only emv present in its germ line. emv-14 is of particular interest because spleen cells expressing emv-14 virus escape recognition by anti-AKR/Gross virus-specific cytotoxic T lymphocytes. We report here the isolation and characterization of a replication-competent emv clone, pAK7, derived from an AKXL-5 mouse. This clone is novel in that it encodes a variant ecotropic murine leukemia virus that, when expressed in SC.Kb target cells, fails to be recognized efficiently by anti-AKR/Gross virus cytotoxic T lymphocytes. The pAK7 clone can therefore be used …

Simian Immunodeficiency Virus Mutants Resistant To Serum Neutralization Arise During Persistent Infection Of Rhesus Monkeys, Dawn P. Wooley, Catherine Collignon, Ronald C. Desrosiers

Neuroscience, Cell Biology & Physiology Faculty Publications

We previously described the pattern of sequence variation in gp120 following persistent infection of rhesus monkeys with the pathogenic simian immunodeficiency virus SIVmac239 molecular clone (D.P.W. Burns and R.C. Desrosiers, J. Virol. 65:1843, 1991). Sequence changes were confined largely to five variable regions (V1 to V5), four of which correspond to human immunodeficiency virus type 1 (HIV-1) gp120 variable regions. Remarkably, 182 of 186 nucleotide substitutions that were documented in these variable regions resulted in amino acid changes. This is an extremely nonrandom pattern, which suggests selective pressure driving amino acid changes in discrete variable domains. In the present study, …

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 …

Thymic Peptide Modulates Glutathione Redox Cycle And Antioxidant Enzymes In Macrophages, Choon Sil Park

Loma Linda University Electronic Theses, Dissertations & Projects

The effect of a 6-kDa thymic peptide (TP) on the oxidative burst of a murine macrophage cell line J774 was determined. TP (12.5-200 μg/ml) was incubated with 5 x 10⁵ J774 cells at 37° C and 5% C0₂ for 18 h and oxidative burst was triggered by zymosan; chemiluminescence was amplified by luminol and measured in an automated luminometer. TP exhibited a concentration-dependent inhibition of oxidative burst. To study the mechanisms involved in TP’s inhibition of oxidative burst, its effect on the glutathione (GSH) redox cycle, GSH biosynthesis, and antioxidant enzymes was investigated. TP was shown to increase …

Cloning And Characterization Of Subunits Of The T-Cell Receptor And Murine Leukemia Virus Enhancer Core-Binding Factor., Shuwen Wang, Qing Wang, Barbara E. Crute, Irena N. Melnikova, Susanna R. Keller, Nancy A. Speck

Dartmouth Scholarship

Moloney murine leukemia virus causes thymic leukemias when injected into newborn mice. A major determinant of the thymic disease specificity of Moloney virus genetically maps to the conserved viral core motif in the Moloney virus enhancer. Point mutations introduced into the core site significantly shifted the disease specificity of the Moloney virus from thymic leukemia to erythroid leukemia (N.A. Speck, B. Renjifo, E. Golemis, T.N. Fredrickson, J.W. Hartley, and N. Hopkins, Genes Dev. 4:233-242, 1990). We previously reported the purification of core-binding factors (CBF) from calf thymus nuclei (S. Wang and N.A. Speck, Mol. Cell. Biol. 12:89-102, 1992). CBF binds …

Lithium Phthalocyanine: A Probe For Electron Paramagnetic Resonance Oximetry In Viable Biological Systems., K. J. Liu, P. Gast, M. Moussavi, S. W. Norby, N Vahidi, T Walczak

Dartmouth Scholarship

Lithium phthalocyanine (LiPc) is a prototype of another generation of synthetic, metallic-organic, paramagnetic crystallites that appear very useful for in vitro and in vivo electron paramagnetic resonance oximetry. The peak-to-peak line width of the electron paramagnetic resonance spectrum of LiPc is a linear function of the partial pressure of oxygen (pO2); this linear relation is independent of the medium surrounding the LiPc. It has an extremely exchange-narrowed spectrum (peak-to-peak line width = 14 mG in the absence of O2). Physicochemically LiPc is very stable; its response to pO2 does not change with conditions and environments (e.g., pH, temperature, redox conditions) …

Cryptococcus Neoformans Serotype Groups Found In Clinical And Environmental Isolates, John Clauson

Masters Theses & Specialist Projects

Cryptococcus neoformans is an encapsulated yeast responsible for severe meningoencephalitis. The importance of epidemiological studies on cryptococcosis has increased since the beginning of the AIDS epidemic. C. neoformans exists in two varieties containing four serotypes, C. neoformans var. neoformans (serotypes A and D) and C. neoformans var. gattii (serotypes B and C). Locally C. neoformans var. neoformans has been associated with pigeon feces during those months having an average temperature of 64.2°F j(17.8°C) and above. Clinical and environmental isolates of C. neoformans obtained from regional hospitals and environmental samplings, respectively, have been grouped into their variety status utilizing canavanine-glycine-bromthymol blue …

Common Elements In Interleukin 4 And Insulin Signaling Pathways In Factor-Dependent Hematopoietic Cells., Ling-Mei Wang, Achsah D. A D Keegan, Weiqun Li, Gustav E. Lienhard

Dartmouth Scholarship

Interleukin 4 (IL-4), insulin, and insulin-like growth factor I (IGF-I) efficiently induced DNA synthesis in the IL-3-dependent murine myeloid cell lines FDC-P1 and FDC-P2. Although these factors could not individually sustain long-term growth of these lines, a combination of IL-4 with either insulin or IGF-I did support continuous growth. The principal tyrosine-phosphorylated substrate observed in FDC cells stimulated with IL-4, previously designated 4PS, was of the same size (170 kDa) as the major substrate phosphorylated in response to insulin or IGF-I. These substrates had phosphopeptides of the same size when analyzed by digestion with Staphylococcus aureus V8 protease, and each …

Competitive Enzyme Immunoassay By Polyclonal Antibodies Against Organophosphorus Pesticides, Kuo-Lan Wen

Theses and Dissertations in Biomedical Sciences

Organophosphorus compounds (OPs) comprise one of the major classes of pesticides in use today because they are highly effective and less toxic. Because of their extensive use, there is a need for monitoring OPs in water and vegetables. Gas chromatography (GC) is the usual method for OPs screening, but it is time-consuming and has the high cost of instrumentation. The goal of this project was to find an easy method which was sensitive and had less instrumentation required to test OPs.

Microwells and microtubes were utilized as the solid-phase immobilization for the development of an enzyme immunoassay for OPs. A …

Sequence Specificity Of The Core-Binding Factor, Irena N. Melnikova, Barbara E. Crute, Shuwen Wang, Nancy A. Speck

Dartmouth Scholarship

The core-binding factor (CBF) binds the conserved core motif in mammalian type C retrovirus enhancers. We analyzed the phosphate contacts made by CBF on the Moloney murine leukemia virus enhancer by ethylation interference assay. The phosphate contacts span 9 bp centered around the consensus core site. To examine the sequence preferences for CBF binding, we employed the technique of selected and amplified binding sequence footprinting (T. K. Blackwell and H. Weintraub, Science 250:1104-1110, 1990). The consensus binding site for CBF defined by selected and amplified binding sequence footprinting is PyGPyG GTPy.

Characterization Of A Protein That Binds Multiple Sequences In Mammalian Type C Retrovirus Enhancers., Wanwen Sun, Mary M. O'Connell, Nancy A. Speck

Dartmouth Scholarship

Mammalian type C retrovirus enhancer factor 1 (MCREF-1) is a nuclear protein that binds several directly repeated sequences (CNGGN6CNGG) in the Moloney and Friend murine leukemia virus (MLV) enhancers (N. R. Manley, M. O'Connell, W. Sun, N. A. Speck, and N. Hopkins, J. Virol. 67:1967-1975, 1993). In this paper, we describe the partial purification of MCREF-1 from calf thymus nuclei and further characterize the binding properties of MCREF-1. MCREF-1 binds four sites in the Moloney MLV enhancer and three sites in the Friend MLV enhancer. Ethylation interference analysis suggests that the MCREF-1 binding site spans two adjacent minor grooves of …

Isolation Of Interleukin 2-Induced Immediate-Early Genes., Carol Beadling, Kirk W. Johnson, Kendall A. Smith

Dartmouth Scholarship

Clonal expansion of antigen-reactive T lymphocytes is driven by the lymphokine interleukin 2 (IL-2). To further elucidate the mechanisms of IL-2 action, we have utilized a differential hybridization procedure to clone IL-2-induced immediate-early genes from an IL-2-stimulated human T-cell cDNA library. To increase the frequency of IL-2-induced transcripts represented in the library, the protein synthesis inhibitor cycloheximide was included during the 2-hr IL-2 stimulation to superinduce gene expression, and the uridine analogue 4-thiouridine was utilized to enable selective purification of newly synthesized transcripts. From the enriched library, we have isolated eight IL-2-induced genes, six of which represent previously unrecognized human …

Can An Anesthesia Machine Flush Valve Provide For Effective Jet Ventilation?, S D Gaughan, J L Benumof, G T Ozaki

Faculty, Staff and Student Publications

Transtracheal jet ventilation (TTJV) using a percutaneously inserted intravenous (IV) catheter for the patient who cannot be ventilated or tracheally intubated or, using a jet stylet for changing endotracheal tubes (ETT) in patients for whom subsequent ventilation and/or tracheal reintubation may be difficult, are extremely valuable therapeutic options. The jet ventilation system must have a sufficiently high pressure-oxygen source to drive oxygen through noncompliant tubing and through relatively small IV catheters and/or jet stylets in order to achieve adequate ventilation and oxygenation. There is no evidence that using the common gas outlet of an anesthesia machine by activating the flush …

Update - March 1993, Loma Linda University Center For Christian Bioethics

Update

In this issue:

-- Loma Linda offers Masters of Arts in Biomedical and Clinical Ethics and assists Claremont with Theological degree

[ Depo-Provera: Contraceptive Dream or Nightmare? ]
-- A Gynecologist's Perspective
-- Some Catholic Moral Reflections

-- Doctor of Philosophy at Claremont

Cloning And Expression Of A Human Neutral Amino Acid Transporter With Structural Similarity To The Glutamate Transporter Gene Family, Jeffrey L. Arriza, Michael Kavanaugh, Wendy A. Fairman, Yan-Na Wu, Geoffrey H. Murdoch, R. Alan North, Susan G. Amara

Biomedical and Pharmaceutical Sciences Faculty Publications

A cDNA was isolated from human brain that encodes an amino acid sequence 34-39% identical to previously published glutamate transporter sequences. Injection of RNA transcribed from this cDNA into Xenopus oocytes resulted in expression of a transport activity with the properties of the neutral amino acid uptake system ASC. Superfusion of alanine, serine, and cysteine evoked sodium-dependent inward currents in voltage-clamped oocytes expressing the transporter. These currents were dose-dependent, stereospecific, and saturable, with Km values ranging from 29 to 88 microM. Northern blot analyses revealed ubiquitous expression of this gene, termed ASCT1, consistent with the general metabolic role ascribed to …

In Vivo Spectroscopic And Imaging Studies Of Photosensitizer In Photodynamic Therapy, Y. Liu, Roger M. Hawk, R. K. Pandey, A. H. Fowler, S. Ramaprasad

Journal of the Arkansas Academy of Science

Photodynamic Therapy (PDT) has emerged as a useful cancer treatment modality which utilizes a tumor localizing dye and activating light to selectively destroy neoplastic tissue. In an effort to understand the newly synthesized photosensitizers, we are studying them in a mouse tumor model grown on the dorsal side of the foot by in vivo magnetic resonance techniques. We have synthesized several photosensitizers which are specifically labeled with fluorine. Several coils appropriate for the tumor study by 19F NMR were designed and constructed for this project. The solenoid coil tunable to both 1Hand 19F nuclei was used to monitor the 19F …

Bibliography, George M. Austin, 1993, George M. Austin

The Dr. George M. Austin Neurology Research Collection

Bibliography, 202 entries, 15 pg., last entry dated 1993

Detection Of Point Mutations In The Dystrophin Gene, John Pedretti

Theses : Honours

The dystrophin gene has been localised to Xp 21.1. Mutations of this gene can lead to the clinical manifestations of Duchenne and Becker muscular dystrophies (DMD/BMD). In the majority of DMD and BMD patients the disease-causing mutation is a deletion detectable by southern analysis or multiplex PCR, however in 30% of patients no deletion is observed using these conventional tests. Using PCR amplification of cDNA it was possible to detect a deletion in the product of the dystrophin gene of one such individual affected with BMD. It was then necessary to characterise the mutation in order to determine whether this …

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, …

Detection Of Serum Antibody Responses In Cattle With Natural Or Experimental Neospora Infections, Patricia Conrad, Karen Sverlow, Mark Anderson, Joan Rowe, Robert Bondurant, Gwen Tuter, Richard Breitmeyer, California Department Of Food And Agriculture, Animal Health Branch, Mark Thurmond, Alex Ardans, J. P. Dubey, Gerarld Duhamel, Bradd Barr

School of Veterinary and Biomedical Sciences: Faculty Publications

Parasite-specific antibody responses were detected using an indirect fluorescent antibody (IFA) test in cattle that were naturally or experimentally infected with Neospora parasites. The test was developed using Neospora tachyzoites isolated from an aborted bovine fetus and grown in bovine cell cultures (isolate BPA1). In all cases, infections were confirmed by the identification of Neospora tachyzoites and/or bradyzoite cysts in fetal or calf tissues using an immunoperoxidase test procedure. Fifty-five naturally infected cows that aborted Neospora-infected fetuses had titers of 320-5,120 at the time of abortion. The titer of 6 cows that were serologically monitored over a prolonged period decreased …