Life Sciences Commons^™

Chemical Characterization Of Condensate From Steam Classified Municipal Solid Waste, Gregory C. Batson

Honors Capstone Projects and Theses

No abstract provided.

Biosynthesis Of Galactose Β-(1–>4)-Mannose, A Novel Disaccharide, Through Loosened Acceptor Specificity Of Bovine Galactosyltransferase, Darren Schmidt

Honors Capstones

No abstract provided.

Identification Of The Catalytic Residue In Rabbit Muscle Phosphofructokinase By Site-Directed Mutagenesis, Wenjue Hu

Honors Capstones

No abstract provided.

Moraxella (Branhamella) Catarrhalis: A Molecular Epidemiology Study, Lyndell R. Gill

Electronic Theses and Dissertations

Moraxella (Branhamella) catarrhalis is the third-most-frequently isolated microorganism associated with acute exacerbations of chronic bronchitis in patients during their stay at the Mountain Home VA Medical Center (MHVAMC). In order to develop a practical, epidemiologically-meaningful typing method for M. (B.) catarrhalis, we tested two methods based on analysis of chromosomal DNA for typeability, reproducibility, and ability to differentiate between unrelated strains (discriminatory power, D). M. (B.) catarrhalis isolants from MHVAMC from 7/1/87-6/30/88 were grown overnight in broth and embedded in agarose. DNA was isolated by standard methods. The DNA was subjected to: (1) restriction endonuclease digestion (with either Bgl II …

Heterozygosity Mapping Of Partially Congenic Lines: Mapping Of A Semidominant Neurological Mutation, Wheels ( Whl), On Mouse Chromosome 4, Patrick M. Nolan, Patricia J. Sollars, Barbara A. Bohne, Warren J. Ewens, Gary E. Pickard, Maja Bucan

School of Veterinary and Biomedical Sciences: Faculty Publications

We identified a semidominant, chemically induced, mouse use mutation with a complex array of abnormal behaviors including bidirectional circling and hyperactivity, abnormal circadian rhythmicity and abnormal responses to light. In this report, we genetically and phenotypically characterized the circling/ waltzing component of the abnormal behavior. We mapped the locus controlling this trait by heterozygosity mapping of partially congenic lines carrying the mutagenized chromosome outcrossed to different inbred strains for three generations. Analysis of 68 PCR-based markers in 13 affected individuals indicated that the mutant locus, named Wheels (Whl), resides in the subcentromeric portion of mouse chromosome 4. The …

Characterization Of An Axial Ligand Substitution In Sperm Whale Myoglobin, Michael J. Chen

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Of central importance to the study of heme proteins are the effects imposed by axial ligand(s) on the heme structure and, therefore, on the overall activity of the protein. In this study, we confirm and extend the spectroscopic characterization of a mutated sperm whale myoglobin in which the proximal Histidine is replaced with a Tyrosine residue (MbH93Y). The MbH93Y, as well as wild-type sperm whale myoglobin and horse erythrocyte catalase (HEC), was purified and characterized by optical absorption and x-ray absorption (XAS) spectroscopies. Optical absorption spectra of HEC and the metmyoglobin, cyanometmyoglobin, reduced, oxy, and carbon-monoxy forms of both sperm …

Extended X-Ray Absorption Fine Structure And Redox Potential Studies Of Heme-Substituted Horseradish Peroxidase And Myoglobin, Bing He

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Heme-substituted horseradish peroxidases and myoglobins were reconstituted from the apoenzyme using mesoheme and diacetyldeuteroheme. X-ray absorption spectroscopy was used to determine the dimensions of the active sites of these heme-substituted proteins, and were compared with those of the proto-hemeproteins. The change in the active-site structure corresponded with the electron withdrawing and donating effects of the different side chains. The oxidation-reduction potentials of Fe⁴⁺/Fe³⁺ couples of the heme-substituted proteins were measured at pH 7 with K₂IrCl₆. The oxidation-reduction potential sequence for compound I/compound II was diacetyldeutero- > proto- > meso- in horseradish peroxidase. The oxidation-reduction potential …

Identification And Characterization Of Genes Associated With V-Jun Induced Cell Transformation, Martin Toralballa Hadman

Biological Sciences Theses & Dissertations

The v-jun oncogene was initially identified as the causative agent for fibrosarcomas in chickens. Studies show that overexpression of v-Jun proteins transforms chicken embryo fibroblasts (CEF) in vitro, and forms tumors in chickens in vivo. The mechanisms for this are not clearly defined. Conceivably, overexpression of an unregulated transcription factor would cause cell transfonnation by illicit regulation of its target genes. In support of this, we show that in vivo v-Jun complexes exhibit differential binding to in vitro generated AP-1 and 'AP-1 like' target sequences, suggesting that the pattern of target gene expression is altered during cell transformation. …

Pathogenicity Of Murine Cytomegalovirus Mutants, Victoria Jean Cavanaugh

Theses and Dissertations in Biomedical Sciences

The purpose of this study was to identified nonessential murine cytomegalovirus (MCMV) genes involved in pathogenesis in vivo. Our approach to identifyjng these genes consisted of constructing MCMV mutants, and then analyzing these mutants in vitro and in vivo. Recombinant viruses (RV) expressing the β-glucuronidase marker gene were constructed by site-directed insertion and deletion mutagenesis of the MCMV Hind III-J and -I regions of the viral genome. Mutations were targeted to this region of the MCMV genome because the corresponding region of the human CMV genome is nonessential and is involved in down-regulating major histocompatibility complex (MHC) class I expression …

Structural Characterization Of A Novel Inhibitor Of Hiv Reverse Transcriptase (Hiv Rt), Greggory Jon Woitte

Chemistry & Biochemistry Theses & Dissertations

Human immunodeficiency virus (HIV) infections have become a leading cause of death among young people in the United States today. As the number of HIV infections increases, so too does the cost of treatment. Together, these numbers have prompted an increase in the development of pharmaceutical interventions. HIV reverse transcriptase (HIV RT) has become a suitable target for drug therapy because it is the sole enzyme responsible for HIV replication.

Fucoidan, a sulfated polysaccharide isolated from the brown algae Fucus vesiculosus, has been shown to block a variety of cell adhesion related events including metastasis. In addition, fucoidan has also …

A Kinetic Investigation Of The Mechanism Of Muscle Contraction With A Series Of Nucleotides, Wei Jiang

Theses and Dissertations in Biomedical Sciences

Muscle contraction is thought to be accomplished by sliding of myosin filaments along actin filaments. Although actomyosin naturally uses ATP as an energy source, it can also use other nucleoside triphosphates (NTP) as substrates. In this work, the dependence of rate and equilibrium constants of the various steps in the muscle contraction mechanism upon nucleotide structure was investigated to unravel the dependence of the mechanical properties of the muscle upon changes in different biochemical steps of the mechanism.

The experiments measuring the dissociation of actomyosin by NTP (or mant-NTP) shows that both the rate constant of actomyosin-S1 dissociation and second …

The Kinetic Characterization Of Mutant L289p Of Recombinant Hiv-1 Reverse Transcriptase, Tryn Thomas Stimart

Chemistry & Biochemistry Theses & Dissertations

The Human Immunodeficiency Virus, type 1 (HIV-1) is the causative agent of the Acquired immunodeficiency Syndrome (AIDS). Currently, AIDS is the leading cause of death amongst young people in the United States. The large increase of HIV-1 infections over the past decade has spawned considerable research in fighting the spread of this disease. One area of intense HIV-1 research has focused on the enzyme Reverse Transcriptase (RT) which is an essential enzyme involved in the replication and life cycle of HIV-1. HIV-1 RT is composed of p66 and p51 subunits and is only active as a heterodimer in vivo. …

The Crystal Structure Of Recombinant Human Neutrophil-Activating Peptide-2 (M6l) At 1.9-Å Resolution, Michael G. Malkowski, Jean Yang Wu, Jerome B. Lazar, Paul H. Johnson, Brian Fp Edwards

Biochemistry and Molecular Biology Faculty Publications

Neutrophil-activating peptide-2 (NAP-2) is a 70-residue carboxyl-terminal fragment of platelet basic protein, which is found in the a-granules of human platelets. NAP-2, which belongs to the CXC family of chemokines that includes Interleukin-B and platelet factor 4, binds to the interleukin-8 type II receptor and induces a rise in cytosolic calcium, chemotaxis of neutrophils, and exocytosis. Crystals of recombinant NAP-2 in which the single methionine at position 6 was replaced by leucine to facilitate expression belong to space group PI (unit cell parameters a = 40.8, b = 43.8, and c = 44.7 A and a = 98.4°, fl = …

Restoration Of Circadian Behavior By Anterior Hypothalamic Heterografts, Patricia J. Sollars, Daniel P. Kimble, Gary E. Pickard

School of Veterinary and Biomedical Sciences: Faculty Publications

The suprachiasmatic nucleus (SCN) of the anterior hypothalamus (AH) is a circadian oscillator and an important component of the mammalian circadian system. To determine whether the SCN is the dominant circadian pacemaker responsible for generating a species-typical characteristic of circadian rhythms [i.e., period length (T)], neural transplantation was conducted using fetal AH donors of different species and SCN-lesioned (SCNx) hosts. The circadian behavior of each of the three donor species is clearly distinguishable by its species-typical T. The extent of SCN pacemaker autonomy was assessed by noting whether the period of the restored circadian rhythm following heterograft transplantation was characteristic …

Conversion Of Bovine Pancreatic Phospholipase A At A Single Site Into A Competitor Of Neurotoxic Phospholipases A By Site-Directed Mutagenesis, Mu-Chin Tzeng, Chon-Ho Yen, Ming-Jhy Hseu, Cynthia M. Dupureur, Ming-Daw Tsai

Educator Preparation & Leadership Faculty Works

A 45-kDa polypeptide preferentially present in neuronal membranes was previously identified as a subunit of a binding (or receptor) protein for several phospholipase A2 variants with neurotoxicity, including crotoxin, by chemical cross-linking experiments (Yen, C.-H., and Tzeng, M.-C.(1991) Biochemistry 30, 11473-11477). The binding of crotoxin to this receptor protein was completely suppressed by sufficient F22Y, a mutated bovine pancreatic phospholipase A2 generated by site-directed mutagenesis of Phe of the wild-type enzyme to Tyr. The IC of this inhibition was estimated to be 1 μM. In sharp contrast, the wild-type enzyme gave no effect even at 50 μM. This …

Sensitization To Self (Virus) Antigen By In Situ Expression Of Murine Interferon-Gamma., Myung-Shik Lee, Matthias Von Herrath, Hans Reiser, Michael B.A. Oldstone, Nora Sarvetnick

Journal Articles: Regenerative Medicine

Autoimmune disease results from inflammatory destruction of tissues by aberrant self-reactive lymphocytes. We studied the autoimmune potential of T lymphocytes immunologically ignorant of viral antigens acting as self antigens and whether the host defense molecule IFN-gamma could stimulate these cells to cytotoxic competency. For this purpose, we produced double transgenic mice expressing pancreatic IFN-gamma as well as lymphocytic choriomeningitis virus (LCMV) nucleoprotein (NP) or glycoprotein (GP) antigen. 100% of the NP+/IFN-gamma+ mice became diabetic before 2 mo of age, while none of the NP single transgenic littermates and only 10% of IFN-gamma single transgenic littermates did. Strikingly, NP+/IFN-gamma+ mice spontaneously …

Myasthenia Gravis-Like Syndrome Induced By Expression Of Interferon Gamma In The Neuromuscular Junction., Danling Gu, Lise Wogensen, Nigel A. Calcutt, Chunyao Xia, Simin Zhu, John P. Merlie, Howard S. Fox, Jon Lindstrom, Henry C. Powell, Nora Sarvetnick

Journal Articles: Regenerative Medicine

Abnormal humoral responses toward motor end plate constituents in muscle induce myasthenia gravis (MG). To study the etiology of this disease, and whether it could be induced by host defense molecules, we examined the consequences of interferon (IFN) gamma production within the neuromuscular junction of transgenic mice. The transgenic mice exhibited gradually increasing muscular weakness, flaccid paralysis, and functional disruption of the neuromuscular junction that was reversed after administration of an inhibitor of acetylcholinesterase, features which are strikingly similar to human MG. Furthermore, histological examination revealed infiltration of mononuclear cells and autoantibody deposition at motor end plates. Immunoprecipitation analysis indicated …

Complementary Gradients In Expression And Binding Of Elf-1 And Mek4 In Development Of The Topographic Retinotectal Projection Map, Masaru Nakamoto

Biology Faculty Publications

Topographic maps with a defined spatial ordering of neuronal connections are a key feature of brain organization. Such maps are believed to develop in response to complementary position-specific labels in presynaptic and postsynaptic fields. However, the complementary labeling molecules are not known. In the well studied visual map of retinal axons projecting to the tectum, the labels are hypothesized to be in gradients, without needing large numbers of cell-specific molecules. We recently cloned ELF-1 as a ligand for Eph family receptors. Here, RNA hybridization shows matching expression gradients for ELF-1 in the tectum and its receptor Mek4 in the retina. …

Antifungal Activity Of Halophenols And Halonitrophenols / H. Gershon, D. D. Clarke, And M. Gershon Department Of Chemistry, Fordham University, Bronx, Ny 10458, Usa New York Botanical Garden, Bronx, Ny 10458, Usa, Herman Gershon, Donald Dudley Clarke Phd, Muriel Gershon

Chemistry Faculty Publications

Thirty one compounds (phenol; its 12 possible monohalo analogues; 18 nitrophenols (2- and 4-nitrophenols, 4-, 5-, and 6-halo-2-nitrophenols, 3-halo-4-nitrophenols)) were tested for antifungal activity against six fungi (A. niger, A. oryzae, M. verrucaria, T. viride, M. cirinelloides, and T. mentagrophytes) in Sabouraud dextrose broth. The two most fungitoxic compounds of those studied were 5-fluoro- and 5-iodo-2-nitrophenols which inhibited all the fungi at concentrations under 10 μg/ml. 6-Iodo-2-nitrophenol inhibited five fungi at a concentration below 10 μg/ml and M. cirinelloides at 10- 100 μg/ ml

Biotransformation Of Bile Acids, Cholesterol And Steroids. Chapter 13 In: The Ecology And Physiology Of Gastrointestinal Microbes. Vol. 1, New York, Chapman And Hall., Stephen Baron, Phillip B. Hylemon

Biology Faculty Scholarship

No abstract provided.

Substituted 8-Quinolinols: Halo, Nitro, And Sulfonic Acids / Herman Gershon, Donald D. Clarke, And Muriel Gershon Department Of Chemistry, Fordham University, Bronx, New York 10458 U.S.A., New York Botanical Garden, Bronx, New York 10458, U.S.A., Herman Gershon, Donald Dudley Clarke Phd, Muriel Gershon

Chemistry Faculty Publications

Methods have been systematized for preparing substituted 8-quinolinols. The 5 and 7 positions are those available for electrophilic substitution. The entry position of the electrophile can be controlled by controlling the prototropic form of the 8-quinolinol. Under acidic conditions the 5 position is attacked first and under basic conditions the electrophile is directed to the 7 position. Iodination is the reverse. Regiospecificity also can be achieved by blocking the 5 or 7 position with sulfonic acid groups followed by addition of the electrophile and deblocking by acid hydrolysis, providing the new substituent is stable to acid hydrolysis. When compounds containing …

A Small Modified Hammerhead Ribozyme And Its Conformational Characteristics Determined By Mutagenesis And Lattice Calculation, Brooke Lustig, N H. Lin, S M. Smith, R L. Jernigan, K.-T Jeang

Faculty Publications, Chemistry

A prototypic hammerhead ribozyme has three helices that surround an asymmetrical central core loop. We have mutagenized a hammerhead type ribozyme. In agreement with previous studies, progressive removal of stem-loop II from a three stemmed ribozyme showed that this region is not absolutely critical for catalysis. However, complete elimination of stem II and its loop did reduce, but did not eliminate, function. In a stem-loop II-deleted ribozyme, activity was best preserved when a purine, preferably a G, was present at position 10.1. This G contributed to catalysis irregardless of its role as either one part of a canonical pair with …

Molecular Mimicry Between Fc Receptor And S Peplomer Protein Of Mouse Hepatitis Virus, Bovine Corona Virus, And Transmissible Gastroenteritis Virus [Abstract], Emilia L. Oleszak, Jacek Kuzmak, Brenda Hogue, Rebecca D. Parr, Ellen W. Collisson, L. Scott Rodkey, Julian L. Leibowitz

Faculty Publications

No abstract provided.

Kinetic Analysis Of The Non-Phosphorylated, In Vitro Phosphorylated, And Phosphorylation-Site-Mutant (Asp8) Forms Of Intact Recombinant C4 Phosphoenolpyruvate Carboxylase From Sorghum, Stephen M. G. Duff, Carlos S. Andreo, Valérie Pacquit, Loïc Lepiniec, Gautam Sarath, Shirley A. Condon, Jean Vidal, Pierre Gadal, Raymond Chollet

Gautam Sarath Publications

Steady-state kinetic analyses were performed on the non-phosphorylated, in vitro phosphorylated and phosphorylation-site mutant (Ser8→Asp) forms of purified recombinant sorghum C₄ phosphoenolpyruvate (P-pyruvate) carboxylase (EC 4.1.1.3 1) containing an intact N-terminus. Significant differences in certain kinetic parameters were observed between these three enzyme forms when activity was assayed at a suboptimal but near-physiological pH (7.3), but not at optimal pH (8.0). Most notably, at pH 7.3 the apparent K_i/ for the negative allosteric effector l-malate was 0.17 mM, 1.2 mM and 0.45 mM while the apparent K_a for the positive allosteric effector glucose 6-phosphate (Glc6P) at 1mM …

Consistencies Of Individual Dna Base-Amino Acid Interactions In Structures And Sequences, Brooke Lustig, R L. Jernigan

Faculty Publications, Chemistry

Amino acid-amino acid interaction energies have been derived from crystal structure data for a number of years. Here is reported the first derivation of normalized relative interaction from binding data for each of the four bases interacting with a specific amino acid, utilizing data from combinatorial multiplex ONA binding of zinc finger domains [Desjarlais, J. R. and Berg, J. M. (1994) Proc. Natl. Acad. Sci. USA, 91, 11099–11103]. The five strongest Interactions are observed for lysine-guanine, lysine-thymine, arginine- guanine, aspartic acid-cytosine and asparagineadenine. These rankings for interactions with the four bases appear to be related to base-amino acid partial charges. …

Fish Analysis On Spontaneously Arising Micronuclei In The Icf Syndrome, Michael W. Stacey, M. S. Bennett, M. Hulten

Bioelectrics Publications

The ICF syndrome is a rare disorder where patients show undercondensation of the heterochromatic blocks of chromosomes 1, 9, and 16 along with variable immunodeficiency. The undercondensation of the heterochromatic block appears to be restricted to a portion of PHA stimulated T cells. Patients with this syndrome also show an increase in micronuclei formation. We have used dual colour FISH to investigate the chromosomal content of these micronuclei in PHA stimulated peripheral blood cultures, an EBV transformed B cell line, and also micronuclei observed in vivo from peripheral blood smears. Chromosome 1 appears to be present in a higher proportion …

New Optical Cell Design For Laser Flash Photolysis Studies In Supercritical Fluids, Steven R. Goates, Milton L. Lee, Qin Ji, Edward M. Eyring, Rudi Van Eldik, Kedika Bal Reddy

Faculty Publications

A high-pressure optical cell has been designed that achieves an effective separation between the chemical sample and the pressurizing medium and system. This design limits possible sample contamination and catalytic effects under supercritical fluid sample conditions. Laser flash photolysis experiments were carried out on molybdenum hexacarbonyl dissolved in supercritical CO2. The thermal ring closure reaction of the species Mo(CO)5L, where L is 2,2- bipyridine was found to proceed at rates comparable to those measured previously in liquid benzene or toluene. Much larger activation volumes were found for the reaction in supercritical CO2 than in liquid toluene.

The Expression Of Early Insulin Signaling Components In An Animal Model Of Insulin-Resistant Diabetes Mellitus, James A. Bonini

Master's Theses

No abstract provided.

Activation Of Transfer Rna-Guanine Ribosyltransferase By Protein Kinase C, Rana C. Morris, Bonnie J. Brooks, Panayota Eriotou, Deborah F. Kelly, Sandeep Sagar, K. Lenore Hart, Mark S. Elliot

Chemistry & Biochemistry Faculty Publications

Transfer RNA-guanine ribosyltransferase (TGRase) irreversibly incorporates queuine into the first position in the anticodon of four tRNA isoacceptors. Rat brain protein kinase C (PKC) was shown to stimulate rat liver TG Rase activity, TGRase preparations derived from rat liver have been observed to decrease in activity over time in storage at -20 or -70°C, Contamination of the samples by phosphatases was indicated by a p-nitrophenylphosphate conversion test, The addition of micromolar concentrations of the phosphatase inhibitors sodium pyrophosphate and sodium fluoride into TGRase isolation buffers resulted in a greater return of TGRase activity than without these inhibitors, Inactive TGRase preparations …

The Development Of Methodology For The Examination Of Interhelical Distances Of A Bacteriorhodopsin Mutant Using The Heterobifunctional Cross-Linker 4-(N-Maleimido) Benzophenone, Tracie L. Bishop

MUSC Theses and Dissertations

Bacteriorhodopsin (BR) is the sole protein found in the purple membrane of Halobacteria salinarium. The chromophore of BR, all-trans retinal, is bound to the apoprotein of BR via a protonated Schiff base linkage to the E-amino group of Lys216. The protonated all-trans retinal isomerizes to the deprotonated I3-cis retinal upon the absorption of light. This light induced chromophore isomerization triggers a cycle of proton pumping from the cytosol to the extracellular side of the membrane (one H+ per trans to cis isomerization). This proton pumping creates a gradient across the membrane which can be used by the bacterium to synthesize …