Life Sciences Commons^™

An Enzymatic Method For The Detection Of Human Serum Albumin, M U. Javed, S N. Waqar

Department of Biological & Biomedical Sciences

Albumin is the most abundant protein in human serum. A dye-binding method is commonly used in clinical laboratories for its estimation using different types of dyes. However, all these dye methods were interfered by a variety of compounds. Here we present a method for the detection of albumin in human serum and other biological fluids. The principle is based on the fact that lactate dehydrogenase isoenzyme-5 (LDH-5) binds specifically to Dextran-Blue (DB). Albumin inhibits the binding of LDH-5 with DB. Absence of LDH activity in DB fraction after gel filtration indicates the presence of albumin in sample and vice versa.

Complex Effects Arising In Smoke Plume Simulations Due To Inclusion Of Direct Emissions Of Oxygenated Organic Species From Biomass Combustion, Sherri A. Mason, Richard J. Field, Robert J. Yokelson, Michael A. Kochivar, Mark R. Tinsley, Darold E. Ward, Wei Min Hao

Chemistry and Biochemistry Faculty Publications

Oxygenated volatile organic species (oxygenates), including HCOOH, H₂CO, CH₃OH, HOCH₂CHO (hydroxyacetaldehyde), CH₃COOH, and C₆H₅OH, have recently been identified by Fourier transform infrared measurements as a significant component of the direct emissions from biomass combustion. These oxygenates have not generally been included in the hydrocarbon-based initial emission profiles used in previous photochemical simulations of biomass combustion smoke plumes. We explore the effects of oxygenates on this photochemistry by using an established initial emission hydrocarbon profile and comparing simulation results obtained both with and without addition of the above six …

Shape Imprinting Due To Variable Disulfide Bonds In Polyacrylamide Gels, Andrew B. Greytak, Alexander Y. Grosberg, Toyoichi Tanaka

Faculty Publications

Through the use of variable disulfide crosslinkers, we have created polyacrylamide gels whose shape can be altered after polymerization. N,N'-bisacryloylcystamine is incorporated as a crosslinker, along with a smaller amount of a permanent crosslinker. After polymerization, the disulfide bonds are cleaved into thiols through reduction. By reoxidizing the thiols with the gel held in a new macroscopic shape, a new set of disulfide bonds is formed, and the gel is forced to adopt the new shape. Retension of the new shape improves with greater distortion from the original shape, as well as with increased concentration of variable …

Nonsymbiotic Hemoglobins In Rice Are Synthesized During Germination And In Differentiating Cell Types, E. J. H. Ross, L. Shearman, M. Mathiesen, Y.J. Zhou, R. Arredondo-Peter, Gautam Sarath, R. V. Klucas

Gautam Sarath Publications

Nonsymbiotic hemoglobins (ns-Hbs) previously have been found in monocots and dicots; however, very little is known about the tissue and cell type localization as well as the physiological function(s) of these oxygen-binding proteins. We report the immunodetection and immunolocalization of ns-Hbs in rice (Oryza sativa L.) by Western blotting and in situ confocal laser scanning techniques. Ns-Hbs were detected in soluble extracts of different tissues from the developing rice seedling by immunoblotting. Levels of ns-Hbs increased in the germinating seed for the first six days following imbibition and remained relatively constant thereafter. In contrast, ns-Hb levels decreased during leaf …

Selective Inhibition Of Selenocysteine Trna Maturation And Selenoprotein Synthesis In Transgenic Mice Expressing Isopentenyladenosine-Deficient Selenocysteine Trna, Mohamed E. Moustafa, Bradley A. Carlson, Muhammed A. El-Saadani, Gregory V. Kryukov, Qi-An Sun, John W. Harney, Kristina E. Hill, Gerald F. Combs, Lionel Feigenbaum, David B. Mansur, Raymond F. Burk, Marla J. Berry, Alan M. Diamond, Byeong Jae Lee, Vadim Gladyshev, Dolph A. Hatfield

Vadim Gladyshev Publications

Selenocysteine (Sec) tRNA (tRNA ^[Ser]Sec) serves as both the site of Sec biosynthesis and the adapter molecule for donation of this amino acid to protein. The consequences on selenoprotein biosynthesis of overexpressing either the wild type or a mutant tRNA ^[Ser]Sec lacking the modified base, isopentenyladenosine, in its anticodon loop were examined by introducing multiple copies of the corresponding tRNA ^[Ser]Sec genes into the mouse genome. Overexpression of wild-type tRNA ^[Ser]Sec did not affect selenoprotein synthesis. In contrast, the levels of numerous selenoproteins decreased in mice expressing isopentenyladenosine-deficient (i⁶A^-) tRNA ^[Ser]Sec in a protein- and …

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.

Chapter 13. The 15 Kda Selenoprotein (Sepl5): Functional Studies And A Role In Cancer Etiology, Vadim Gladyshev, Alan M. Diamond, Dolph L. Hatfield

Vadim Gladyshev Publications

The 15 kDa selenoprotein (Sepl5) is one of several recently identified selenoproteins. It contains a single selenocysteine residue in the middle of a 162-amino acid open reading frame and has no detectable homology to known proteins. The human Sepl5 gene spans 5 1 kb, has 5 exons and is located on chromosome 1 at position p3 1. The gene contains two single nucleotide polymorphisms in the 3'-untranslated region (3'-UTR) including one in the SECIS element, that are distributed differently between Caucasians and African Americans. Sep15 localizes to the endoplasmic reticulum where it is tightly bound to UDP-glucose:glycoprotein glucosyltransferase, a protein …

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 Capstones

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 Capstones

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.

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 …

The Algebraic Application Of Lewis Structures In Conjunction With Combinatorial Analysis In Global Molecular Identification From Graphs, Jonathan Geach

Capstone Research Projects

Closed shell bonding can be described by a theory that relates the manner that atoms fill their outer shells to algebraic formulas. Equations can be derived that are capable of deciding whether a given electron configuration gives a closed shell electron structure or not. A combinatorial analysis can be used to try every possible electron configuration for a given structure. The equations are then used to determine whether each structure gives a closed shell electron configuration. Some of these molecules may be considered impossible because they are bonded to noble gases. However, a list of molecules that contain a noble …

Antifungal Activity Of Substituted 8-Quinolinol-5- And 7-Sulfonic Acids: A Mechanism Of Action Is Suggested Based On Intramolecular Synergism / Hermon Gershon, Muriel Gershon, & Donald D. Clarke Harding Laboratory, The New York Botanical Garden, Bronx, New York 10458, Usa; Department Of Chemistry, Fordham University, Bronx, New York 10458, Usa, Herman Gershon, Muriel Gershon, Donald Dudley Clarke Phd

Chemistry Faculty Publications

8-Quinolinol-5-sulfonic acid was nearly devoid of antimicrobial activity, due to what was believed to be an unfavorable partition coefficient. Since twenty six 8-quinolinol-5- and 7 -sulfonic acids were available from our previous work, they were tested against six fungi. The 7 -chloro and 7 -bromo-5-sulfonic acids and the 5-chloro and 5-bromo-7 -sulfonic acids showed fungal inhibition within one order of magnitude of that of 8-quinolinol. It is suggested that a nonchelating mechanism is in part responsible for this fungitoxicity. Five additional 5-sulfonic acids with chlorine in positions 3-, 6-, 3,6-, 3,7-, and 6,7- that were suitable for studies in synergism …

Preparation And Fungitoxicity Of Some Trichloro-, Tribromo-, Tetrachloro-, And Tetrabromo-8-Quinolinols / Herman Gershon, Donald D. Clarke, And Muriel Gershon Department Of Chemistry, Fordham University, Bronx, New York 10458-9993 Usa, New York Botanical Garden, Bronx, New York 10458-9993 Usa, Herman Gershon, Donald Dudley Clarke Phd, Muriel Gershon

Chemistry Faculty Publications

3,5,6-, 3,5,7-, 4,5,7-, and 5,6,7-trichloro- and -tribromo-8-quinolinols as well as 3,5,6,7- tetrachloro- and -tetrabromo-8-quinolinols were prepared and tested against six fungi (Aspergillus niger, Aspergillus oryzae, Myrothecium verrucaria, Trichoderma viride, Mucor cirinelloides, and Trichophyton mentagrophytes) in Sabouraud dextrose broth. The compounds strongly inhibit five fungi but not M. cirinelloides. They are less active than the related dichloro-8-quinolinols which is attributed to steric hindrance

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 …

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 …