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Chapter 25. Selenium In Biology And Human Health: Controversies And Perspectives, Vadim Gladyshev

Vadim Gladyshev Publications

Important unresolved questions raised by the contributors of this book and addressing roles of selenium in biology and human health are discussed. Resolving major scientific controversies in the field should further highlight a bright future for selenium in fundamental science, biotechnology and medicine.

Studies On The Formation Of Dna-Cationic Lipid Composite Films And Dna Hybridization In The Composites, Murali Sastry, Vidya Ramakrishnan, Mrunalini Pattarkine, Krishna N. Ganesh

Faculty Works

The formation of composite films of double-stranded DNA and cationic lipid molecules (octadecylamine, ODA) and the hybridization of complementary single-stranded DNA molecules in such composite films are demonstrated. The immobilization of DNA is accomplished by simple immersion of a thermally evaporated ODA film in the DNA solution at close to physiological pH. The entrapment of the DNA molecules in the cationic lipid film is dominated by attractive electrostatic interaction between the negatively charged phosphate backbone of the DNA molecules and the protonated amine molecules in the thermally evaporated film and has been quantified using quartz crystal microgravimetry (QCM). Fluorescence studies …

Mapping The Toxic And Regulatory Domains Of Doc Of The P1 Plasmid Addiction Operon, Darren Ferree

Honors Capstone Projects and Theses

No abstract provided.

Comprehensive Isotachophoresis-Capillary Electrophoresis Coupled To Time-Of-Flight Mass Spectrometry, Christopher Ryan Bowerbank

Theses and Dissertations

Isotachophoresis (ITP) coupled to capillary zone electrophoresis (CE) in a comprehensive manner was used to separate mixture components in both insufficient and sufficient concentrations without heart-cutting or splitting. Examples of comprehensive ITP-CE involving multiple CE injections of preconcentrated ITP zones are demonstrated. In the comprehensive arrangement, all of the sample in the first dimension (ITP) is subjected to analysis in the second dimension (CE), without significant sample loss or decrease in sample detectability resulting from removal of a portion of the sample. This is especially important for analytes at low concentrations which may form a single mixed zone instead of …

Association Between The 15-Kda Selenoprotein And Udp-Glucose:Glycoprotein Glucosyltransferase In The Endoplasmic Reticulum Of Mammalian Cells, Konstantin V. Korotkov, Easwari Kumaraswamy, You Zhou, Dolph L. Hatfield, Vadim N. Gladyshev

Vadim Gladyshev Publications

Mammalian selenocysteine-containing proteins characterized with respect to function are involved in redox processes and exhibit distinct expression patterns and cellular locations. A recently identified 15-kDa selenoprotein (Sep15) has no homology to previously characterized proteins, and its function is not known. Here we report the intracellular localization and identification of a binding partner for this selenoprotein which implicate Sep15 in the regulation of protein folding. The native Sep15 isolated from rat prostate and mouse liver occurred in a complex with a 150-kDa protein. The latter protein was identified as UDP-glucose:glycoprotein glucosyltransferase (UGTR), the endoplasmic reticulum (ER)-resident protein, which was previously shown …

The Characterization Of A Novel Petite Phenotype In Transgenic Arabidopsis Thaliana And The Examination Of The Expression Patterns Of A Gamma Carbonic Anhydrase Homologue In Arabidopsis Thaliana, Sumeet K. Asrani

Honors Theses

No abstract provided.

Evidence For Two Mechanisms Of Amino Acid Osmolyte Release From Hippocampal Slices, R. Franco, M. E. Torres-Marquez, H. Pasantes-Morales

School of Veterinary and Biomedical Sciences: Faculty Publications

A 30% decrease in osmolarity stimulated ³H-taurine, ³H-GABA and glutamate (followed as ³H-D-aspartate) efflux from rat hippocampal slices. ³H-taurine efflux was activated rapidly but inactivated slowly. It was decreased markedly by 100 μM 5-nitro-(3-phenylpropylamino) benzoic acid (NPPB) and 600 μM niflumic acid and inhibited strongly by tyrphostins AG18, AG879 and AG112 (25–100 μM), suggesting a tyrosine kinase-mediated mechanism. Hyposmolarity activated the mitogen-activated protein kinases (MAPK) extracellular-signal-related kinase-1/2 (ERK1/ERK2) and p38, but blockade of this reaction did not affect ³H-taurine efflux. Hyposmosis also activated phosphatidyl-inositide 3-kinase (PI3K) and its prevention by wortmannin (100 nM) essentially abolished …

The Examination Of The Expression Patterns Of A Gamma Carbonic Anhydrase Homologue In Arabidopsis Thaliana, Eric Thomas Dane Nielsen Iii

Honors Theses

No abstract provided.

Detection Of Aneuploidy For Chromosomes 7 And 8 Using Fluorescence In Situ Hybridization In Patients With Aplastic Anemia And Sequencing Of The Mitotic Checkpoint Gene Hbub1, Laura Jane Aridgides

Theses and Dissertations in Biomedical Sciences

Aplastic anemia (AA) is characterized by complete bone marrow failure. Progression to myelodysplastic syndromes (MDS) and acute nonlymphocytic leukemia (ANLL) occurs frequently. At the time of transformation, cytogenetic abnormalities are common. Detection of cytogenetic abnormalities prior to leukemic transformation may indicate future disease progression. Karyotype analysis is the current method of choice to evaluate chromosome aberrations. However, fluorescence in situ hybridization (FISH) is more sensitive in detecting these abnormalities.

hBUB1, a mitotic spindle checkpoint gene, was shown to be mutated in two colorectal cancer cell lines with high levels of aneuploidy (Cahill, et al., 1998). Although theoretically possible, conclusive …

Phase Modulation At 125 Khz In A Michelson Interferometer Using An Inexpensive Piezoelectric Stack Driven At Resonance, Barbara M. Hoeling, Andrew D. Fernandez, Richard C. Haskell, Daniel C. Petersen

All HMC Faculty Publications and Research

Fast phase modulation has been achieved in a Michelson interferometer by attaching a lightweight reference mirror to a piezoelectric stack and driving the stack at a resonance frequency of about 125 kHz. The electrical behavior of the piezo stack and the mechanical properties of the piezo-mirror arrangement are described. A displacement amplitude at resonance of about 350 nm was achieved using a standard function generator. Phase drift in the interferometer and piezo wobble were readily circumvented. This approach to phase modulation is less expensive by a factor of roughly 50 than one based on an electro-optic effect.

Structural Studies Of Scallop Abductin, A Compressible Elastomer That Translates Hydrophobic Interactions To Elasticity, Felipe Jimenez Jr.

Loma Linda University Electronic Theses, Dissertations & Projects

Abductin is a natural elastomer found as the major component of the elastic hinge ligament in marine scallops. Until now, very little was known about the structure and function of this compressible elastomer. Biochemical studies of a synthetic 25 amino acid peptide of abductin produces fibers that assemble into a gel matrix at room temperature. Further investigation of this phenomenon via temperature-dependent turbidimetry revealed that this peptide self-assembles or aggregates at 23 °C. Secondary structural studies of this peptide at different temperatures revealed that it undergoes an inverse temperature transition much like soluble elastin peptides. Replacing phenylalanine with alanine in …

Plant-Based Multicomponent Vaccines Against Enteric Diseases, Jie Yu

Loma Linda University Electronic Theses, Dissertations & Projects

To generate a plant-based multicomponent oral vaccine against cholera, rotavirus and enterotoxigenic E.coli (ETEC), we have constructed transgenic potato plants that synthesize the cholera toxin B and A2 subunits, the rotavirus enterotoxin NSP4 22 amino acid epitope and the ETEC fimbrial antigen CFA/L To use cholera toxin as a carrier molecule for delivery of multiple antigens to the gut-associated lymphoid tissues, the rotavirus NSP4 epitope was linked to the cholera toxin B subunit (CTB) and the ETEC fimbrial antigen CFA/I was linked to the cholera toxin A2 subunit. Cholera toxin-antigen fusion proteins were detected in transformed tuber tissues by immunoblot …

Nuclear Export Of 60s Ribosomal Subunits Depends On Xpo1p And Requires A Nuclear Export Sequence-Containing Factor, Nmd3p, That Associates With The Large Subunit Protein Rpl10p, Olivier Gadal, Daniela Strau, Jacques Kessl, Bernard Trumpower

Dartmouth Scholarship

Nuclear export of ribosomes requires a subset of nucleoporins and the Ran system, but specific transport factors have not been identified. Using a large subunit reporter (Rpl25p-eGFP), we have isolated several temperature-sensitive ribosomal export (rix) mutants. One of these corresponds to the ribosomal protein Rpl10p, which interacts directly with Nmd3p, a conserved and essential protein associated with 60S subunits. We find that thermosensitive nmd3 mutants are impaired in large subunit export. Strikingly, Nmd3p shuttles between the nucleus and cytoplasm and is exported by the nuclear export receptor Xpo1p. Moreover, we show that export of 60S subunits is Xpo1p dependent. We …

Heterogeneity Within Animal Thioredoxin Reductases, Qi-An Sun, Francesca Zappacosta, Valentina M. Factor, Peter J. Wirth, Dolph L. Hatfield, Vadim N. Gladyshev

Vadim Gladyshev Publications

Animal thioredoxin reductases (TRs) are selenocysteine- containing flavoenzymes that utilize NADPH for reduction of thioredoxins and other protein and nonprotein substrates. Three types of mammalian TRs are known, with TR1 being a cytosolic enzyme, and TR3, a mitochondrial enzyme. Previously characterized TR1 and TR3 occurred as homodimers of 55–57-kDa subunits. We report here that TR1 isolated from mouse liver, mouse liver tumor, and a human T-cell line exhibited extensive heterogeneity as detected by electrophoretic, immunoblot, and mass spectrometry analyses. In particular, a 67-kDa band of TR1 was detected. Furthermore, a novel form of mouse TR1 cDNA encoding a 67-kDa selenoprotein …

Induction Of White Cell Proliferation Due To Haematopoietic Growth Factors Is Associated With An Increase In Multiple Forms Of Dihydrofolate Reductase In Non-Neutropenic Cancer Patients, M P. Iqbal, I A. Burney, F Sultana, N Mehboobali

Department of Biological & Biomedical Sciences

Objective: Granulocyte-colony stimulating factor (G-CSF) and granulocyte macrophage-colony stimulating factor (GM-CSF) are frequently used in cancer patients to overcome the granulocytopenic effects of chemotherapy, and also to mobilize the stem cells. The mobilized stem cells are collected from the peripheral blood and used for transplantation following high doses of chemotherapy. However, the molecular mechanism by which these colony stimulating factors (CSFs) bring about proliferation of myeloid precursor cells is not clearly known. Dihydrofolate reductase (DHFR), which has an established role in DNA synthesis, could be a link between administration of CSF and stem cell proliferation. The purpose of this study …

Process For Immobilizing Waste Using Bagasse (Us Patent 6174275), Michael A. Janusa

Faculty Publications

No abstract provided.

A Crystallographic Map Of The Transition From B-Dna To A-Dna, Jeff Vargason, Keith Henderson, P. Shing Ho

Faculty Publications - Department of Biological & Molecular Science

No abstract provided.

The Cia5 Gene Controls Formation Of The Carbon Concentrating Mechanism In Chlamydomonas Reinhardtii, Youbin Xiang, Jun Zhang, Donald P. Weeks

Department of Biochemistry: Faculty Publications

Wild-type Chlamydomonas reinhardtii cells shifted from high concentrations (5%) of CO2 to low, ambient levels (0.03%) rapidly increase transcription of mRNAs from several CO₂-responsive genes. Simultaneously, they develop a functional carbon concentrating mechanism that allows the cells to greatly increase internal levels of CO₂ and HCO₃. The cia5 mutant is defective in all of these phenotypes. A newly isolated gene, designated Cia5, restores transformed cia5 cells to the phenotype of wild-type cells. The 6,481-bp gene produces a 5.1-kb mRNA that is present constitutively in light in high and low CO₂ both in …

Protein Targeting And Translocation, Barry D. Bruce, D.A. Phoenix

Faculty Publications and Other Works -- Biochemistry, Cellular and Molecular Biology

No abstract provided.

X-Ray Diffraction Structures Of Some Phosphatidylethanolamine Lamellar And Inverted Hexagonal Phases, Paul E. Harper, David A. Mannock, Ruthven Lewis, Ronald N. Mcelhaney

University Faculty Publications and Creative Works

X-ray diffraction is used to solve the low-resolution structures of fully hydrated aqueous dispersions of seven different diacyl phosphatidylethanolamines (PEs) whose hydrocarbon chains have the same effective chain length but whose structures vary widely. Both the lower-temperature, liquid-crystalline lamellar (Lα) and the higher-temperature, inverted hexagonal (H11) phase structures are solved, and the resultant internal dimensions (d-spacing, water layer thickness, average lipid length, and headgroup area at the lipid-water interface) of each phase are determined as a function of temperature. The magnitude of the Lα and H11 phase d-spacings on either side of the Lα/H11 phase transition temperature (Tη) depends significantly …

Cloning And Expression Analysis Of Hemoglobin Genes From Maize (Zea Mays Ssp. Mays) And Teosinte (Zea Mays Ssp. Parviglumis), Elena Aréchaga-Ocampo, Juan Saenz-Rivera, Gautam Sarath, Robert V. Klucas, Raúl Arredondo-Peter

Gautam Sarath Publications

With the exception of barley and rice, little is known about the existence of hemoglobins (Hbs) in cereals. This work reports the cloning and analysis of hb genes from maize (Zea mays ssp. mays) and teosinte (Zea mays ssp. parviglumis). Coding sequences of maize and teosinte hb genes (hbm and hbt, respectively) are highly similar to each other and are interrupted by three introns located at identical positions as other plant hb genes. Sequences of predicted Hbm and Hbt proteins are identical. The hydropathic profile of Hbm and Hbt is highly similar to that …

C3 Origins Of The C4 Pathway Regulatory Enzyme, Ppdk-Rp, C. J. Chastain, Gautam Sarath, Raymond Chollet

Gautam Sarath Publications

A current view of C₄ and CAM evolution holds that all of the constituent enzymes of the C₄ pathway preexisted in C₃ plants, albeit functioning in nonphotosynthetic capacities.

Phylogeographic Analysis Of The Threatened And Endangered Superconglutinate-Producing Mussels Of The Genus Lampsilis (Bivalvia: Unionidae), Kevin J. Roe, Charles Lydeard, Paul D. Hartfield

Kevin J. Roe

Several species of freshwater unionid mussels in the genus Lampsilis exhibit a remarkable reproductive strategy. Female mussels of these species enclose their larvae in a minnow-like lure, called a ‘superconglutinate’, to attract piscivorous fishes. When a fish attempts to ingest the superconglutinate the lure ruptures and the larvae are released to parasitize the fish. Of the four species of mussel which exhibit this strategy and are endemic to the Gulf Coast drainages of the southeastern United States, three are protected under the Endangered Species Act, and one is recognized as imperilled. Phylogenetic analysis of nucleotide sequences of the mitochondrial 16S …

Abnormal Expression Of P27kip1 Protein In Levator Ani Muscle Of Aging Women With Pelvic Floor Disorders – A Relationship To The Cellular Differentiation And Degeneration, Antonin Bukovsky, Pleas Copas, Michael R. Caudle, Maria Cekanova Ms, Rndr, Phd, Tamara Dassanayake, Bridgett Asbury, Stuart E. Van Meter, Robert F. Elder, Jeffrey B. Brown, Stephanie B. Cross

Maria Cekanova MS, RNDr, PhD

Background Pelvic floor disorders affect almost 50% of aging women. An important role in the pelvic floor support belongs to the levator ani muscle. The p27/kip1 (p27) protein, multifunctional cyclin-dependent kinase inhibitor, shows changing expression in differentiating skeletal muscle cells during development, and relatively high levels of p27 RNA were detected in the normal human skeletal muscles.

Methods Biopsy samples of levator ani muscle were obtained from 22 symptomatic patients with stress urinary incontinence, pelvic organ prolapse, and overlaps (age range 38–74), and nine asymptomatic women (age 31–49). Cryostat sections were investigated for p27 protein expression and type I (slow …

Protein Targeting And Translocation, Barry D. Bruce, D.A. Phoenix

Barry D. Bruce

No abstract provided.

Genomic Diversity Of Pseudomonas Aeruginosa In Cystic Fibrosis, Nathaniel Edwards Head

Theses, Dissertations and Capstones

Pseudomonas aeruginosa is a common environmental microorganism. However, it has the genetic capacity to cause diseases in patients with compromised host defense systems. Cystic fibrosis (CF) is one of the major hereditary diseases among Caucasian populations. CF patients are born with a defective chloride channel that is responsible for maintaining fluid and electrolyte balance across the lumen of the lung. This imbalance leads to the production of an abnormal dehydrated viscous mucous, prohibiting the patient from normally clearing the respiratory airway. Chronic pulmonary infections with P. aeruginosa are the major causes of high morbidity and mortality in CF. Two of …

Methods Development For The Identification Of Unknown Proteins, Laura Brumfield Neal

Theses, Dissertations and Capstones

The chemistry of transport, storage, energy production, and reproduction in physiological systems is controlled by the actions and interactions of proteins. A species full complement of these amazing machines is often referred to as its proteome. The new science of proteomics utilizes a set of powerful and evolving tools to characterize the nature of the proteome, and further to fully elucidate the functional attributes of the proteins and their associations.

A methodology was developed by combining several proteomic techniques, such as electrophoresis, RP-HPLC, and mass spectrometry, to correctly identity proteins. Three known proteins (myoglobin, albumin, and actin) were employed for …

Characterization And Regulation Of Skn-Directed Autoimmune Skin Lesions, Pamela Jane Staton

Theses, Dissertations and Capstones

In the Skn model of autoimmunity, pathogenic T cells sensitized to Skn epidermal antigens (Skn-immune spleen cells or SIS cells) elicited disease in the form of skin lesions when adoptively transferred into shaved immunoincompetent syngeneic adult recipients (‘lesion-forming animals’) while concomitant injection of immunoregulatory normal spleen cells at twice the concentration (2 x normal spleen cells or 2XNS cells) of SIS cells significantly lessened lesion development (‘lesion-controlling animals’) by Day 7 post cell transfer. In a time-course RT-PCR cytokine analysis of skin from lesion-forming and lesion-controlling animals, IL-7 mRNA was significantly elevated in lesion-controlling skin on Day 7 while no …

Effect Of Forskolin On Actin Polymerization In Cultured Vascular Smooth Muscle, Mary Janay Bailey

Theses, Dissertations and Capstones

Increases in intracellular cyclic AMP levels have been shown to affect vascular smooth muscle morphology. In the present study, effects of elevated cyclic AMP levels on actin polymerization were assessed to determine whether cyclic AMP induces specific changes in cytoskeletal proteins of vascular smooth muscle cells. Forskolin was used to increase intracellular cyclic AMP levels through direct activation of adenylate cyclase. Light and Electron microscopy was employed to appraise changes in the shape of vascular smooth muscle cells. Confocal microscopy was used to reveal cytoskeletal changes after exposure to M forskolin for increasing intervals of time. Differences in the expression …

Nitric Oxide Mediated Effect Of Cyclo-Oxygenase Inhibitors, M Mahmood, H S. Zuberi, M K. Ashfaq

Department of Biological & Biomedical Sciences

Objectives: Non-Steroidal anti-inflammatory drugs (NSAIDS) have long been used as anti-inflammatory agents, yet their mode of action is not entirely clear. The inhibitory effects of NSAIDS on prostaglandin production can only partly explain their anti-inflammatory actions. This study was aimed at defining the role of cycl-oxygenase (COX) inhibitors on nitric oxide (NO) production in murine macrophages in vitro.
Methods: Murine macrophages were obtained from the peritoneum and after exposure, in vitro to lipopolysaccharide (LPS) produced nitrite, measured after 24 hours by Griess reaction. The macrophages were pre-incubated with aspirin or indomethacin before activation with LPS.
Results: Treatment with aspirin resulted …