Biochemistry, Biophysics, and Structural Biology Commons^™

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 …

Elimination Of Dissolved Sulfide Interference In The Flow-Injection Determination Of Sigma-Co2 By Addition Of Molybdate, Rigel L. Lustwerk, David J. Burdige

OES Faculty Publications

A previously described flow injection method for the analysis of ΣCO₂ included the addition of ZnCl₂ to some samples before analysis in order to precipitate dissolved sulfide (which interferes with the method) as ZnS. However, the use of Zn²⁺ in samples with high concentrations of dissolved sulfide causes the coprecipitation of ZnCO₃, and in our experience with this technique, ZnCO₃ also precipitates even in the absence of dissolved sulfide. The addition of molybdate effectively complexes dissolved sulfide without interfering with the determination of ΣCO2 by this technique.

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 …

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 …

In Vivo Regulation Of Wheat-Leaf Phosphoenolpyruvate Carboxylase By Reversible Phosphorylation, Stephen M. G. Duff, Raymond Chollet

Department of Biochemistry: Faculty Publications

Regulation of C₃ phosphoenolpyruvate carboxylase (PEPC) and its protein-serine/threonine kinase (PEPC-PK) was studied in wheat (Trificum aesfivum) leaves that were excised from low-N-grown seedlings and subsequently illuminated and/or supplied with 40 mM KNO₃. The apparent phosphorylation status of PEPC was assessed by its sensitivity to i-malate inhibition at suboptimal assay conditions, and the activity state of PEPC-PK was determined by the in vitro ³²P labeling of purified maize dephospho-PEPC by [y³²PIATP/ Mg. lllumination (±NO₃^-) for 1 h led to about a 4.5-fold increase in the 50% inhibition constant for i-malate, which …

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.

Molecular Cloning And Rare Cleavage Mapping Of Human 2p, 6q, 8q, 12q, And 18q Telomeres, Roberto A. Macina, Ken Morii, Xue-Lan Hu, Dimitri G. Negorev, Chrysanthe Spais, Lisa A. Ruthig, Harold C. Riethman

School of Medical Diagnostics & Translational Sciences Publications

Large terminal fragments of human chromosomes 2p, 6q, 8q, 12q, and 18q were cloned using yeast artificial chromosomes (YACs). RecA-assisted restriction endonuclease (RARE) cleavage analysis of genomic DNA samples from 11 unrelated individuals using YAC-derived probes confirmed the telomeric localizations of the half-YACs studied. The cloned Fragments provide telomeric closure of maps for the respective chromosome arms and will supply the reagents needed for analyzing and sequencing these distal subtelomeric regions.

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. …

Cloning Of Multiple Novel Human Trinucleotide Repeat Containing Cdna's: A Novel Application Of Rapid Amplification Of Cdna Ends (Race), James P. Carney

Dissertations

No abstract provided.

The Effects Of Ethanol On The Developing Serotonergic Neurons, Jung-Ae Kim

Dissertations

No abstract provided.

Baseline Water Quality Data Inventory And Analysis: Lake Mead National Recreation Area, Volume Ii Of Ii, National Park Service

Publications (WR)

This document presents the results of surface-water-quality data retrievals for Lake Mead National Recreation Area (LAME) from five of the United States Environmental Protection Agency's (EPA) national databases: (1) Storage and Retrieval (STORET) database management system; (2) River Reach File (RF3); (3) Industrial Facilities Discharge (IFD); (4) Drinking Water Supplies (DRINKS); and (5) Flow Gages (GAGES). This document is one product resulting from a cooperative contractual endeavor between the National Park Service's Servicewide Inventory and Monitoring Program, the National Park Service's Water Resources Division (WRD), and Horizon Systems Corporation to retrieve, format, and analyze water quality data for all units …

Factors Influencing The Oxidation Of Lipoproteins And Plasma Lipids, Yanshan Ma

Electronic Theses and Dissertations

The hypothesis that antioxidant vitamins (ascorbate and tocopherols) along with urate protect blood plasma lipids from oxidation was tested. Dietary fat is also an important factor influencing plasma lipid peroxidation. The purpose of this study was to investigate the role of plasma antioxidants and dietary fat on low density lipoprotein (LDL) and plasma lipid oxidation. In the first part of this study, we compared the ability of urate and ascorbate to protect human LDL from in vitro oxidation. LDL oxidation was initiated by 15 mM of a water soluble azo-initiator in the presence or absence of ascorbate or urate. The …

Baseline Water Quality Data Inventory And Analysis: Lake Mead National Recreation Area, Volume I Of Ii, National Park Service

Publications (WR)

This document presents the results of surface-water-quality data retrievals for Lake Mead National Recreation Area (LAME) from five of the United States Environmental Protection Agency's (EPA) national databases: (1) Storage and Retrieval (STORET) database management system; (2) River Reach File (RF3); (3) Industrial Facilities Discharge (IFD); (4) Drinking Water Supplies (DRINKS); and (5) Flow Gages (GAGES). This document is one product resulting from a cooperative contractual endeavor between the National Park Service's Servicewide Inventory and Monitoring Program, the National Park Service's Water Resources Division (WRD), and Horizon Systems Corporation to retrieve, format, and analyze water quality data for all units …

A Study Of Euplotes Crassus Telomere Proteins And Related Genes, John Scott Perez

Department of Chemistry: Dissertations, Theses, and Student Research

A novel PCR technique used to amplify Euplotes crassus macronuclear chromosomes was developed to provide scientific proof that an Euplotes crassus 1.0 kb gene is not genetically related to an Oxytricha nova 1.8 kb β-telomere protein gene. Theories and experimental procedures associated with the development of this PCR technique were the product of research that will have a profound impact on future studies of telomere protein genes and telomeric DNA in a multitude of eukaryotic cells. The hypothesis that a β-telomere protein does not exist in Euplotes crassus was supported by the result of this study.

Expression of the recombinant …

Semiclassical Quantization Of A Nonintegrable System: Pushing The Fourier Method Into The Chaotic Regime, Karl Sohlberg, Randall B. Shirts

Faculty Publications

Semiclassical Einstein–Brillouin–Keller (EBK) quantization of the nonintegrable Hénon–Heiles Hamiltonian succeeds using the Fourier transform method of Martens and Ezra. Two innovations are required for this success: (1) the use of tunneling corrected quantizing actions obtained from an approximate, one-dimensional Hamiltonian and (2) exploitation of intermediate-time approximate quasiperiodicity or "vague tori'' wherein the Fourier transform of chaotic motion over 10–100 vibrational periods allows the determination of frequencies and amplitudes which approximate motion during the time interval. Approximate tori, actions, and EBK energy levels are then straightforward. We use an interpolation method to smooth over small resonance zones that are not expected …

In Situ Regulation Of Cytosolic Phospolipase A₂, Beverly A. Rzigalinski

Theses and Dissertations in Biomedical Sciences

The 85 kDa cytosolic phospholipase A₂ (cPLA₂) is an agonist-responsive effector for intracellular signal transduction through the arachidonate cascade. In vitro studies have demonstrated that this enzyme is regulated by sub-micromolar calcium and is specific for arachidonate as the sn-2 fatty acyl group of phospholipid substrates. However, very little data is available regarding in situ mechanisms which govern the activity of cPLA₂. The primarily objective of these studies was to develop an in situ system for the study of cPLA₂, and investigate mobilization of arachidonate during signal transduction events.

Dimethylsulfoxide differentiation of the …

Sensitivity Of Second Harmonic Generation To Space Charge Effects At Si(111)/Electrolyte And Si(111)/Sio2/Electrolyte Interfaces, P. R. Fischer, J. L. Daschbach, D. E. Gragson, G. L. Richmond

Chemistry and Biochemistry

The potential dependence in the surface second harmonic response from hydrogen terminated n‐Si(111) and oxidized n‐Si(111) surfaces has been examined in aqueous NH₄F and H₂SO₄ solutions. The relative phase of the nonlinear response as measured by rotational anisotropy experiments is found to be highly sensitive to the presence of the oxide and the field applied across the Si(111)/oxide/electrolyte interface. These observations are attributed to field effects within the space–charge region of the semiconductor which vary with the presence and thickness of the insulating oxide layer on the Si(111) surface.

Use Of An Oriented Transmembrane Protein To Probe The Assembly Of A Supported Phospholipid Bilayer, Paul Brian Contino, Carol A. Hasselbacher, J. B. Alexander Ross, Yale Nemerson

Chemistry and Biochemistry Faculty Publications

Planar-supported phopholipid bilayers formed by te adsorpton of vesicles are increasingly used in the investigation of lipK-ependent reactis. We have studied the way in which these bilayers are forned with phopholipid vesicles coaining the btranembrane protein Tssue Factor (TF). TF complexed with te senne protease, factor Vlla, is the primary initiator of bklod coagulation by way of activation of the zymogen factor X. TF has been shown to orient randomly on the inner and outer leaflets of vesicles. We used proteolytic digestion to produce vesicles in which the exracellular domain of TF is located on the inner leaflet These vesicles …

Crystallization Of Hemoglobins Ii And Iii Of The Symbiont-Harboring Clam Lucina Pectinata, M. A. Doyle, Jacqueline Vitali, J. B. Wittenberg, S. N. Vinogradov, D. A. Walz, B. F.P. Edwards, P. D. Martin

Physics Faculty Publications

Diffraction data to 2.7 A resolution were measured on crystals of the homotetramers of components II and III of the cytoplacmic hemoglobin of the symbiont-harboring clam Lucina pectinata. Even though the crystallization conditions are different and the sequence homology of the two hemoglobins is only 63%, the crystals are isomorphous to each other and to the heterotetramer Hb II/III, implying that the residues primarily involved in the intermolecular interactions and responsible for crystal cohesion may be invariant.

The Structure And Dynamics Of The Cs2 Molecular Ion, Karl Sohlberg, Chan Yibai

Faculty Publications

Several common elementary methods of computing molecular properties, prerequisite to molecular dynamics studies, are tested for their validity for CS+2. Judged sufficiently accurate, these methods are then used to aid in investigating the collisionally activated dissociation of CS+2 upon impact with xenon. Rice–Ramsperger–Kassel–Marcus (RRKM) unimolecular decay rate calculations are presented and compared to experimental studies employing collisional activation. RRKM theory is shown to reproduce the experimental results for collision energies near threshold. When corrected for vibrational anharmonicity, the RRKM calculation shows agreement with the experimental results over a slightly wider range of energies. A discussion is given on the applicability …

Dissection Of An Antibody-Catalyzed Reaction, Jon D. Stewart, Joseph F. Krebs, Gary Siuzdak, Anthony J. Berdis, David B. Smithrud, Stephen J. Benkovic

Chemistry Faculty Publications

Antibody 43C9 accelerates the hydrolysis of a p-nitroanilide by a factor of 2.5 x 10(5) over the background rate in addition to catalyzing the hydrolysis of a series of aromatic esters. Since this represents one of the largest rate accelerations achieved with an antibody, we have undertaken a series of studies aimed at uncovering the catalytic mechanism of 43C9. The immunogen, a phosphonamidate, was designed to mimic the geometric and electronic characteristics of the tetrahedral intermediate that forms upon nucleophilic attack by hydroxide on the amide substrate. Further studies, however, revealed that the catalytic mechanism is more complex and involves …

Circadian Clock Locus Frequency: Protein Encoded By A Single Open Reading Frame Defines Period Length And Temperature Compensation., Benjamin D. Aronson, Keith A. Johnson, Jay C. Dunlap

Dartmouth Scholarship

The frequency (frq) locus encodes a key component, a state variable, in a cellular oscillator generating circadian rhythmicity. Two transcripts have been mapped to this region, and data presented here are consistent with the existence of a third transcript. Analysis of cDNA clones and clock mutants from this region focuses attention on one transcript encoding a protein. FRQ, which is a central clock component: (i) mutations in all of the semidominant frq alleles are the result of single amino acid substitutions and map to the open reading frame (ORF) encoding FRQ; (ii) deletion of this ORF, or a frameshift mutation …

Determination Of Pancreatic And Salivary Amylase By Enzyme Immunoassay And Their Prevalence In Hyperamylasemic Patients, Sabdra Borgens Ward

Theses and Dissertations in Biomedical Sciences

Currently, amylase determinations are nonspecific for the organ source and are based entirely on the enzymatic properties of amylase to produce a measurable product or byproduct. The determination of pancreatic amylase is important in the diagnosis of acute pancreatitis. Most commercially available tests for amylase employ the measurement of the change in NADH absorbance at 280 nm or of the p-nitrophenol released from a maltotetrose substrate. These are nonspecific measurements of pancreatic amylase and often necessitate other tests to be run such as a serum lipase.

The two predominant isoenzymes of amylase are pancreatic (p-amylase) and salivary (s-amylase); the most …

Structure-Substrate Binding Relationships Of Hiv-1 Reverse Transcriptase, Steve Chien-Wen Huang

Theses and Dissertations in Biomedical Sciences

Human Immunodeficiency Virus, type 1 (HIV-1), is the causative agent of the Acquired Immunodeficiency Syndrome (AIDS). HIV-1 reverse transcriptase (RT), a heterodimer p66/p51, has been the major target for treatment of AIDS. The significance of the p51 subunit and the RNase H domain of p66 in terms of their influence on the RNA-dependent DNA synthesis was investigated. Clones of the wildtype HIV-1 RT subunits, p66 and p51, and a recombinant C-terminal deletion mutant, p64, [Barr, P. J. (1987) Bio/Technoloav 5, 486-489] were employed to study the structure-substrate binding relationships of HIV-1 RT. The activity assays of RNA-dependent DNA synthesis on …

The Invariant Arginine In Motif 2 Of Escherichia Coli Alanyl-Trna Synthetase : Is Important For Catalysis But Not For Substrate Binding, Ying Lu

Loma Linda University Electronic Theses, Dissertations & Projects

Structural motifs 2 and 3, located in the active site of class II aminoacyl-tRNA synthetases, each contain an invariant arginine thought to participate in interactions with ATP. For Escherichia coli alanyl-tRNA synthetase (AlaRS), sequence comparisons indicate that Arg69 should be aligned with the invariant arginine in motif 2 of other class II synthetases. Site-directed random mutagenesis has been employed to generate a set of proteins containing amino acid substitutions in a portion of motif 2 of AlaRS. In this set, only mutations at position 69 caused the enzyme to lose ability to complement growth of an alaS deletion strain, …

Mechanism Of Mitogenic Action Of Aluminum Ion On Bone Cells : Potential Involvement Of The Insulin-Like Growth Factor Regulatory System, Aera Yoo

Loma Linda University Electronic Theses, Dissertations & Projects

Micromolar concentrations of aluminum ion (oxidation state of 3 +) consistently stimulated [3H]thymidine incorporation into cell DNA and increased cellular alkaline phosphatase activity (an osteoblastic differentiation marker) in osteoblast-like cells of chicken and human origin. Although biphasic, dose-dependent stimulations were highly reproducible, the maximal stimulatory dose varied among different experiments. Mitogenic doses of aluminum ion also stimulated collagen synthesis in cultured human osteosarcoma TE85 cells. In addition, the same mitogenic concentrations of aluminum ion enhanced the 1,25 dihydroxy vitamin D3-dependent stimulation of osteocalcin secretion. Together, these findings indicate that aluminum at micromolar concentrations, stimulates the proliferation, differentiation, and activity of …

Signal Transduction In Bacterial Chemotaxis, Edward Heath Rowsell

Loma Linda University Electronic Theses, Dissertations & Projects

This study investigated the site of ATP utilization in the signal transduction pathway of bacterial chemotaxis and localized the point of convergence of a methylation-independent system of chemotaxis with the methylation-dependent system. The identity of the signal originating from the phosphotransferase system was investigated by substituting the fructose-inducible HPr-like protein FPr for HPr in transport and chemotaxis. In addition, a novel chemoattractant, glycerol, was identified for Salmonella typhimurium. Histidine-auxotrophic S. typhimurium strains ST23 (hisF) and ST171 (hisF cheB) were depleted for ATP. The times required for the bacteria to adapt to a step increase in serine, …

The Effects Of Unpaired Bases In An Oligo Deoxy (A) * Deoxy (T) Tract On Dna Bending, Vasavi Malineni

Honors Capstones

No abstract provided.

The Role Of Tocopherol, Cholesterol, And Oleic Acid In Lipid Peroxidation, Jherie Jean Demouy Blazier

Honors Capstones

No abstract provided.

Investigation Of Selenium Status In Hypo-, Hyper- And Euthyroid Children, Ping Sun

Chemistry & Biochemistry Theses & Dissertations

Recently, it has been reported that human type I iodothyronine deiodinase, an enzyme important in the conversion of T₄ to T₃ , is a selenoenzyme. Several studies have reported alterations in plasma selenium level in hypo- and hyperthyroid patients.

Using polarized Zeeman-effect atomic absorption spectroscopy, we measured selenium, zinc, copper and manganese concentrations in the plasma and/or red blood cells in children with or without thyroid disease being seen at the outpatient clinic at Children's Hospital of the King's Daughters. Children with thyroid disease were subdivided into untreated, treated and nonresponsive groups. Data were analyzed using Student's t-test …