Life Sciences Commons^™

Yeast And Human Frataxin Are Processed To Mature Form In Two Sequential Steps By The Mitochondrial Processing Peptidase, Steven S. Branda, Patrizia Cavadini, Jiri Adamec, Frantisek Kalousek, Franco Taroni, Grazia Isaya

Department of Biochemistry: Faculty Publications

Frataxin is a nuclear-encoded mitochondrial protein which is deficient in Friedreich’s ataxia, a hereditary neurodegenerative disease. Yeast mutants lacking the yeast frataxin homologue (Yfh1p) show iron accumulation in mitochondria and increased sensitivity to oxidative stress, suggesting that frataxin plays a critical role in mitochondrial iron homeostasis and free radical toxicity. Both Yfh1p and frataxin are synthesized as larger precursor molecules that, upon import into mitochondria, are subject to two proteolytic cleavages, yielding an intermediate and a mature size form. A recent study found that recombinant rat mitochondrial processing peptidase (MPP) cleaves the mouse frataxin precursor to the intermediate but not …

Fabrication Of Protein Arrays Using Electron Beam Lithography, Duy-Bao Phan Dinh

Theses, Dissertations and Capstones

Very few electron beam based methods for producing high-resolution patterns of surface bound proteins on glass have been developed.

There are only two types of end groups at the surface of glass, they are silanol and siloxane groups. An estersil layer can be formed on the surface of glass by treatment with hot alcohol creating a hydrophobic layer. The reaction is depicted as follows.

-Si-OH + HOR -SiO-R +H2O R = alkyl chain

The proposal that protein arrays could be fabricated by patterned electron bombardment using estersils as a negative resist was tested in this research. The hydrophobic estersil layer …

Cyclopropane Fatty Acid Expression In Plants, Katherine M. Schmid

Scholarship and Professional Work - LAS

Pants [sic] are transformed with a bacterial cyclopropane fatty acid synthase gene to produce lipids containing cyclopropane fatty acids. Using this technology dihydrosterculate is produced in oilseed crops such as rape.

Carmoviruses (Tombusviridae), Feng Qu, Thomas Jack Morris

School of Biological Sciences: Faculty Publications

Carmovirus is one of two officially recognized genera of Tombusviridae. Members of Tombusviridae all have icosahedral virions of about 30 nm in diameter with T = 3 symmetry that consists of 180 coat protein (CP) subunits of about 38- 43 kDa and a single-stranded (ss) RNA genome ranging in size from 4.0 to 4.7 kb. Carmoviruses share recognizable sequence similarity in both the polymerase and structural genes with members of the other genus, Tombusvirus. Sequence comparisons and phylogenetic analysis of these genes support the view that the genera are distinct but closely related. In addition, the genome organization of the …

A Biogeochemical Comparison Of Fossil (Carboniferous) And Modern Crustose Red Algae, Michael A. Kruge, John E. Utgaard, William Ferry

Department of Earth and Environmental Studies Faculty Scholarship and Creative Works

The nature of the contribution of the various types of algae to sedimentary organic matter continues to be a topic of research interest. Crustose red algae have however received less attention than other types. The fossil calcareous red algae (Rhodophyta) analyzed in this study are two relatively unrecrystallized specimens of Parachaetetes (Family Solenoporacea) from the lower part of the Ste. Genevieve Formation (Carboniferous, Visean) in Union County, Illinois, USA. They occurred in the patch reef phase of a small carbonate mudmound-patchreef. The three modern specimens (collected and identified by F. Collier) are the crustose algae Lithothamnion, Clathromorphum and Phymatolithon …

Transcriptional Regulation Of The Bmp2 Gene: Retinoic Acid Induction In F9 Embryonal Carcinoma Cells And Saccharomyces Cerevisiae, Loree C. Heller, Yong Li, Kevin L. Abrams, Melissa B. Rogers

Bioelectrics Publications

Bmp2, a highly conserved member of the transforming growth factor-beta gene family, is crucial for normal development. Retinoic acid, combined with cAMP analogs, sharply induces the Bmp2 mRNA during the differentiation of F9 embryonal carcinoma cells into parietal endoderm. Retinoic acid (RA) also induces the Bmp2 gene in chick limb buds. Since normal Bmp2 expression may require an endogenous retinoid signal and aberrant Bmp2 expression may cause some aspects of RA-induced teratogenesis, we studied the mechanism underlying the induction of Bmp2. Measurements of the Bmp2 mRNA half-life and nuclear run-on assays …

Analysis Of Phytoestrogens By High Performance Liquid Chromatography, Anne A. Grippo, Yan Xie, Benjamin L. Rougeau, William V. Wyatt

Journal of the Arkansas Academy of Science

Phytoestrogens are biochemicals synthesized in plants which mimic steroidal estrogen activity in mammals. Analysis of these compounds in the legumes which produce them and in body fluids is important to the study of their physiological effects. High pressure liquid chromatography (HPLC) has been found to be an efficient and sensitive method of identification and quantitation of isoflavonoids, one class of phytoestrogen. Here we report the separation of three isoflavonoids, biochanin A, genistein and daidzein using an HPLC system with a Cg reverse phase column and a linear gradient mobile phase containing acetonitrile and acetic acid/water (10/90, v/v) over 60 minutes. …

Electron Shock Waves, Frances Terry, Mostafa Hemmati

Journal of the Arkansas Academy of Science

No abstract provided.

Development Of 31p Nuclear Magnetic Resonance Methods For The Study Of Phosphate Metabolisms In E. Coli And B. Subtilis, Fabian M. Dayrit, Emilyn Q. Espiritu, Noreen Gonzalez, Nina Rosario L. Rojas, Jennifer T. Aguilan, Antonio M. Basilio, Edward T. Chainani, Enrico Cruz, Bernadette Matanguihan

Environmental Science Faculty Publications

31P NMR experiments were performed on Escherichia coli and Bacillus subtilis at various temperatures under aerobic and anaerobic conditions. The total soluble intracellular phosphate concentration was estimated to be 2 x 10^-17 mole/cell, while intracellular orthophosphate concentration was around 1 x 10^-17 mole/cell. Addition of glucose resulted in a general decrease in intracellular pH and was accompanied by the formation of sugar monophosphates. The concentrations of soluble intracellular phosphates, both inorganic and organic phosphates, were estimated by integration versus methylene diphosphonic acid (MDPA) standard. Although intracellular and extracellular orthophosphate could be observed, these appear to exchange rapidly on the NMR …

Construction Of An Escherichia Coli Lac Repressor-Based System To Study Human Cytomegalovirus Gene Expression, Laura Fernanda Cageao-Luchetti

Theses and Dissertations in Biomedical Sciences

An Escherichia coli lac repressor-based system was developed to study the roles of human cytomegalovirus (HCMV) genes during viral replication. To this end, a recombinant HCMV expressing the lac repressor was generated (RVlac), and an HCMV-specific promoter was targeted for conditional expression by inserting the lac operator sequence. The promoter of a nonessential gene was chosen in order to be able to assess parameters of repression and derepression of the operator-containing promoter in the endogenous locus, without having virus growth dependent on the specific inducer isopropylthiogalactoside (IPTG). The feasibility of this approach to conditionally express an HCMV promoter …

Early And Late Endosomal Compartments Of Entamoeba Histolytica Are Enriched In Cysteine Proteases, Acid Phosphatase And Several Ras-Related Rab Gtpases, Lesly A. Temesvari, Ed Harris, Samuel L. Stanley Jr., James Cardelli

Department of Biochemistry: Faculty Publications

Pure populations of early and late endosomes of Entamoeba histolytica were isolated by magnetic fractionation and characterized. It was shown that these vesicles were enriched in acid phosphatase and cysteine protease activities. An important virulence factor, a 27-kDa cysteine protease, was also enriched in early and late endosomes of E. histolytica. These data suggest that E. histolytica hydrolases reside in compartments that are part of or communicate with the endosomal pathway. To begin to identify the role of Rab GTPases in E. histolytica, an oligonucleotide approach was employed to screen an E. histolytica cDNA library for genes encoding …

Characterization Of Rat Pulmonary Carboxylesterase, Timothy J. Wallace

Theses and Dissertations

The 1839 bp cDNA for rat pulmonary carboxylesterase was cloned by reverse transcription polymerase chain reaction (RT PCR) from total rat lung RNA using specific primers derived from the 5' and 3' untranslated regions of rat hepatic cholesteryl ester hydrolase (CEH). The unique cDNA was sequenced and found to have 99% homology with hepatic CEH. This homology extends to the predicted amino acid sequences which show only six amino acid residue differences in the coding region: three conserved and three nonconserved changes. However, the catalytic activitites of the two proteins are dramatically different. While CEH hydrolyzes cholesterol oleate, the pulmonary …

Investigating The Allosteric Behavior Of Malate Dehydrogenase From Escherichia Coli, Eman M. Ghanem

Masters Theses

Regulatory mechanisms of malate dehydrogenase from E.coli (eMDH) involving NADH as an allosteric effector were investigated. The reaction was studied in both directions: malate oxidation and oxaloacetate reduction. When malate was the variable substrate, a plot of rate against substrate concentration was sigmoidal in the presence of 0.065 mM NADH, which indicates the presence of an allosteric site for NADH on the enzyme. Binding of NADH at the allosteric site causes conformational changes in the active site and, thereby, changes the catalytic activity of the enzyme. An increase in K_m value, from 1.3 to 3.9 mM malate was observed, …

Detection Of Ehrlichia Chaffeensis In Ticks Of The Georgia Coast And Barrier Islands Using Polymerase Chain Reaction, John Edward Whitlock

College of Graduate Studies: Theses & Dissertations (1964–2006)

Human monocytic ehrlichiosis (HME) is an emerging, zoonosis transmitted to mammals by ixodid ticks. Prevalence of infected ticks and distribution of infection foci indicate relative risk of human exposure to ehrlichiosis and may be influenced by factors such as geographic isolation and human disturbance. To test this, individual and pooled lone star ticks, Amblyomma americanum (L.), collected from 3 populations from the coast and barrier islands of Georgia were screened for Ehrlichia chaffeensis Anderson, Dawson, Jone, and Wilson, the agent of HME. A species-specific, nested polymerase chain reaction (PGR) assay was used to amplify a 572 base pair fragment of …

Hyperinsulinism Caused By Paternal-Specific Inheritance Of A Recessive Mutation In The Sulfonylurea-Receptor Gene, Benjamin Glaser, Fergus Ryan, Marc Donath, Heddy Landau, Charles Stanley, Lester Baker, David Barton, Paul Thornton

Articles

Neonatal hyperinsulinism (HI) is a genetic disorder of pancreatic b-cells characterized by failure to suppress insulin secretion in the presence of hypoglycemia, resulting in brain damage or death if not adequately treated. Germline mutations in four genes have been associated with HI. Some patients have focal regions of b-cell proliferation (focal HI). Seventy HI probands in whom at least one S U R - 1 mutation was identified were studied. Clinical data from patients with two S U R - 1 mutant alleles were compared with those from patients with single paternally inherited mutations. Thirtyseven probands were homozygous or compound …

Temporal And Spatial Variation In Abundance Of Migratory Birds At Kennesaw Mountain, Georgia, Andrew Allen Kinsey

College of Graduate Studies: Theses & Dissertations (1964–2006)

Migratory birds spend a significant part of each year in transition between their breeding and resting grounds. During this transitional period migrants rely on stopover sites to provide them with the necessary habitat to replenish lost energy stores, rest, and wait for favorable weather conditions before continuing migration. The majority of stopover work related to passerines in North America has occurred along coastal migration routes. Meanwhile, relatively little work has been conducted at inland stopover sites, areas which may be equally important to migrating birds. Kennesaw Mountain is an isolated geographically prominent peak located in the foothills of the southern …

The Atomic Force Microscope In The Elucidation Of Leukocyte Abnormality, Derbrenn O'Connor

Theses

The aim of this project was to investigate the novel use of the atomic force microscope (AFM) as a technique in the elucidation of leukocyte abnormality. To this end, comparative AFM imaging and elasticity studies were performed on normal and myeloid leukaemic leukocytes.

Initially, an evaluation of techniques for the isolation of specific leukocyte (e.g. lymphocytes, granulocytes, etc.) populations from whole blood or leukocyte concentrates, was carried out. It was found that a silanised glass surface failed to select and immobilise a leukocyte population. However, centrifugation of whole blood in an Isopaque-ficoll gradient solution yielded a viable mixed leukocyte population …

Affinity Chromatography: A Review Of Clinical Applications, David S. Hage

David Hage Publications

Affinity chromatography is a type of liquid chromatography that makes use of biological-like interactions for the separation and specific analysis of sample components. This review describes the basic principles of affinity chromatography and examines its use in the testing of clinical samples, with an emphasis on HPLCbased methods. Some traditional applications of this approach include the use of boronate, lectin, protein A or protein G, and immunoaffinity supports for the direct quantification of solutes. Newer techniques that use antibody-based columns for on- or off-line sample extraction are examined in detail, as are methods that use affinity chromatography in combination with …

Plasmd Bearing A Cdna Copy Of The Genome Of Bovine Viral Diarrhea Virus, Chimeric Derivatives Thereof, And Method Of Producing An Infectious Bovine Wral Darrheavirus Using Sad Plasmid, Ruben O. Donis, Ventzislav B. Vassilev

School of Veterinary and Biomedical Sciences: Faculty Publications

A plasmid bearing a cDNA copy of the genome of bovine viral diarrhea virus (BVDV), chimeric derivatives of the plasmid and a method of producing an infectious bovine viral diarrhea virus using the plasmid are disclosed. The invention relates to a plasmid DNA molecule that replicates easily in E. coli and contains a sufficient portion of the genome of BVDV, cloned as cDNA, to be a suitable template to produce RNA in vitro which, upon transfection into bovine cells, gives rise to infectious BVDV. The BVDV created by the process of the invention can be engineered for use as a …

In Situ Hybridization For The Detection And Localization Of Swine Chlamydia Trachomatis, C. Chae, D.-S. Cheon, D. Kwon, O. Kim, B. Kim, J. Suh, D. G. Rogers, K. D. E. Everett, A. A. Anderson

School of Veterinary and Biomedical Sciences: Faculty Publications

Gnotobiotic piglets were inoculated intralaryngeally with swine Chlamydia trachomatis strain R33 or orally with swine C. trachmatis strain R27. Archived formalin-fixed, paraffin-embedded tissues from piglets euthanatized 4–7 days postinoculation were examined by in situ hybridization for C. trachomatis nucleic acid using a nonradioactive digoxigenin-labeled DNA probes that targeted specific ribosomal RNA or omp1 mRNA molecules of the swine C. trachomatis strains. Positive hybridization signals were detected in bronchial epithelial cells, bronchiolar epithelial cells, pneumocytes, alveolar and interstitial macrophages, and jejunal and ileal enterocytes. Chlamydia-infected cells had a strong signal that was confined to the intracytoplasmic inclusions. Positive hybridization signals were …

Two-Photon Excitation Of Rhenium Metal-Ligand Complexes, Joseph R. Lakowicz, Felix N. Castellano, Ignacy Gryczynski, Zygmunt Gryczynski, Jonathan D. Dattelbaum

Chemistry Faculty Publications

We describe the emission spectral properties of two rhenium metal-ligand complexes with one and two-photon excitation, Re(bpy)₂(CO)₃Cl and [Re(bpy)(CO)₃CH₃CN]⁺, where bpy is 2,2’-bipyridyl and CH₃CN is acetonitrile. Similar emission spectra and intensity decay times characteristic of the metal-to-ligand charge transfer state were observed for one- and two-photon excitation. The lifetime and quantum yield of the acetonitrile complex are approximately 14-fold higher than that of the chloride complex. Both complexes display high anisotropies near 0.33 in frozen solution with one-photon excitation. Two-photon excitation results in anisotropies about 40% larger, …

Some Aspects Of The Adsorption Of A Lennard-Jones Gas On A Rough Surface, Douglas Henderson, Pawel Bryk, Stefan Sokolowski

Faculty Publications

We study the adsorption, including wetting, of a Lennard-Jones gas on a rough surface consisting of rough layer on a smooth substrate with which the gas interacts via a 9-3 potential. The rough layer is two molecular diameters thick and consists of a disordered quenched matrix of hard spheres. As well as interacting with the other matrix spheres by the hard-sphere potential, the matrix molecules interact with the gas molecules by the hard-sphere potential. Hence, on average, the degree to which the gas molecules can approach the substrate depends only on the density of the matrix layer. The density of …

Archaeal Aminoacyl-Trna Synthesis: Unique Determinants Of A Universal Genetic Code?, Michael Ibba, A. W. Curnow, J. Bono, P. A. Rosa, C. R. Woese, D. Söll

Biology, Chemistry, and Environmental Sciences Faculty Articles and Research

No abstract provided.

Phorbol 12-Myristate 13-Acetate Stimulates Lysophosphatidic Acid Secretion From Ovarian And Cervical Cancer Cells But Not From Breast Or Leukemia Cells, Zhongzhou Shen, Jerome Belinson, Richard E. Morton, Yan Xu

Chemistry Faculty Publications

Lysophosphatidic acid (LPA) is present in ascites from patients with ovarian cancer. It stimulates calcium release and growth of ovarian cancer cells bothin vitroandin vivo.Recently, we found that LPA levels were significantly elevated in plasma from patients with ovarian cancer and other gynecological cancers. In contrast, LPA levels were not elevated in patients with breast cancer and leukemias. In view of this, we investigated whether gynecological cancer cells could produce LPA. LPA was extracted from the supernatant of cells culturedin vitroand purified by thin layer chromatography. After hydrolysis and transmethylation, the fatty acid derivatives were analyzed by gas chromatography. We …

Principal Component Mapping Applied To Raman Microspectroscopy Of Fiber-Reinforced Polymer Composites, C. M. Stellman, K. S. Booksh, A. R. Muroski, M. P. Nelson, Michael L. Myrick

Faculty Publications

We describe the development and implementation of a spectroscopic Raman imaging system using Principal Component data analysis. Raman microspectroscopy followed by multivariate data analysis and principal component mapping has been used to investigate the spatial variations of chemical and physical properties at interfaces in glass-reinforced composites on the micrometer scale. A protocol for monitoring chemical and physical variances via multivariate statistics has been developed and a preliminary study of a "real world" glass-reinforced composite is presented. It is demonstrated that multivariate Raman imaging is more precisely interpretable than traditional univariate methods and that by appropriately choosing distinct spectral regions (such …

Biochemistry Of Hemolysin Toxin Activation By Fatty Acylation: Characterization Of An Internal Protein Acyltransferase, Michael S. Trent

Electronic Theses and Dissertations

Hemolysin toxin produced and secreted by pathogenic Escherichia coli is one of a family of cytolytic, structurally homologous protein toxins known as RTX (repeats in toxin) toxins. RTX toxins are products of a gene cluster, CABD . The A gene product, nontoxic hemolysin (proHlyA) is made toxic by post-translational fatty acylation of two internal lysine residues. HlyC, C gene product, is essential for acylation, and acyl-acyl carrier protein (ACP) is the acyl donor. HlyB and HlyD are involved in secretion of the toxin. HlyC was thought to serve as an internal protein acyltransferase and remained uncharacterized until now. ProHlyA and …

Selenocysteine-Containing Proteins In Mammals, Vadim N. Gladyshev, Dolph L. Hatfield

Vadim Gladyshev Publications

Since the recent discovery of selenocysteine as the 21st amino acid in protein, the field of selenium biology has rapidly expanded. Twelve mammalian selenoproteins have been characterized to date and each contains selenocysteine that is incorporated in response to specific UGA code words. These selenoproteins have different cellular functions, but in those selenoproteins for which the function is known, selenocysteine is located at the active center. The presence of selenocysteine at critical sites in naturally occurring selenoproteins provides an explanation for the important role of selenium in human health and development. This review describes known mammalian selenoproteins and discusses recent …

Dynamics Of Biomolecules: Assignment Of Local Motions By Fluorescence Anisotropy Decay, Carl N. Bialik, Barnabas Wolf, Edward L. Rachofsky, J. B. A. Ross, William Laws

Chemistry and Biochemistry Faculty Publications

Many biological systems have multiple fluorophores that experience multiple depolarizing motions, requiring multiple lifetimes and correlation times to define the fluorescence intensity and anisotropy decays, respectively. To simplify analyses, an assumption often made is that all fluorophores experience all depolarizing motions. However, this assumption usually is invalid, because each lifetime is not necessarily associated with each correlation time. To help establish the correct associations and recover accurate kinetic parameters, a general kinetic scheme that can examine all possible associations is presented. Using synthetic data sets, the ability of the scheme to discriminate among all nine association models possible for two …

Monte Carlo Simulation Of An Ion-Dipole Mixture As A Model Of An Electrical Double Layer, Douglas Henderson, Dezso Boda, Kwong-Yu Chan

Faculty Publications

Canonical Monte Carlo simulations were performed for a nonprimitive model of an electrical double layer. The ions and the solvent molecules are modeled as charged and dipolar hard spheres, respectively, while the electrode as a hard, impenetrable wall carrying uniform surface charge. We found that the ion-dipole model gives a reasonable description of the double layer for partially charged ions with small to moderate dipole moments, or equivalently for an "effective" dielectric constant. Density, polarization and mean electrostatic potential profiles are reported. Strong layering structure, and at higher charges, charge inversion in the second layer were found. With appropriate choices …

Accumulation Of Methotrexate In Human Tissues Following High-Dose Methotrexate Therapy, M P. Iqbal

Department of Biological & Biomedical Sciences

Methotrexate concentration was analyzed in a number of tissues of a patient of osteogenic sarcoma who had been on high-dose methotrexate therapy for nearly 6 months. Gall bladder and kidney contained the highest concentration of the drug, followed by testis, small intestine, skeletal muscle, bone marrow, lung, spleen, heart and liver. Although, compared to kidney the liver contained relatively small amount of the drug, yet nearly 1/5th of the total drug in liver was in bound form. This bound form of methotrexate is most likely associated with multiple forms of dihydrofolate reductase. The total concentration of methotrexate in kidney is …