Life Sciences Commons^™

Interactions Of Deet And Novel Repellents With Mosquito Odorant Receptors, Gariel G. Grant, Rachel R. Estrera, Narendra Pathak, C. Dennis Hall, Maia Tsikolia, Kenneth J. Linthicum, Ulrich R. Bernier, Adam C. Hall

Neuroscience: Faculty Publications

The carboxamide N,N-di-ethyl-meta-Toluamide (DEET) is the most effective and widely used insect repellent today. However, drawbacks concerning the efficacy and the safety of the repellent have led to efforts to design new classes of insect repellents. Through quantitative structure-Activity relationships, chemists have discovered two chemical groups of novel repellents: The acylpiperidines and the carboxamides, with the acylpiperidines generally more potent in biological assays. Although the exact mechanism of action of DEET and other repellents has not yet been thoroughly elucidated, previous research shows that the activity of insect odorant receptors are inhibited in the presence of repellents. The present electrophysiological …

Synthesis Of Supramolecular Complex L– (-) – Menthol, Ziyada Djumanova Dr, Lolaxan Ettibaeva, Ugilay Abduraxmonova, Zulfiya Khalmuratova

Karakalpak Scientific Journal

In this article glycyrrhizic acid content with menthol in several various proportions (2:1; 4:1; 9:1), information was given on synthesis and physical and chemical properties of new supermolecular complexes. Received supramolecular complexes concerning 4:1 it was chemically defined and also studied the structure of supramolecular complexes by physical methods on the basis of interaction of organic molecules with electromagnetic radiation, in particular their range IR – of spectroscopy. Our work of the near future will be turned to check its recent supramolecular complexes on the basis of GA: Menthol for growth of a plant and biotic elasticity of tension …

Structural And Functional Determinants Of Rodent And Human Surfactant Protein A: A Synthesis Of Binding And Computational Data, Armen Nalian, Todd M. Umstead, Ching-Hui Yang, Patricia Silveyra, Neal J. Thomas, Joanna Floros, Francis X. Mccormack, Zissis C. Chroneos

Faculty Publications

Surfactant protein A (SP-A) provides surfactant stability, first line host defense, and lung homeostasis by binding surfactant phospholipids, pathogens, alveolar macrophages (AMs), and epithelial cells. Non-primates express one SP-A protein whereas humans express two: SP-A1 and SP-A2 with core intra- and inter-species differences in the collagen-like domain. Here, we used macrophages and solid phase binding assays to discern structural correlates of rat (r) and human (h) SP-A function. Binding assays using recombinant rSP-A expressed in insect cells showed that lack of proline hydroxylation, truncations of amino-terminal oligomerization domains, and site-directed serine (S) or alanine (A) mutagenesis of cysteine 6 (C6S), …

A Magnetic Affinity Approach To Identify Plant Gaba-Binding Proteins, Jie Zou, Jingzhe Guo, Shisheng Li

Turkish Journal of Biology

In plants, GABA plays a critical role in sexual plant reproduction; however, GABA receptors and the associated detailed signaling mechanisms remain to be elucidated. Our experiments show that the proposed technique is reliable and convenient for probing GABA-binding proteins and could be applicable in similar projects by covalently immobilizing the free carboxylic group of GABA on magnetic beads (SiMAG-Carboxyl). New probes produced by covalently immobilizing the free carboxylic group of GABA on magnetic beads (SiMAG-Carboxyl) can obtain useful information on GABA receptors in plants.

Examining The Role Of Hres On The Regulation Of Opioid Receptor Gene Expression In Neuronal Cells Undergoing Hypoxic Mimic Condition, Alberto Herrera

Seton Hall University Dissertations and Theses (ETDs)

Hypoxia is a condition of inadequate oxygen supply, which can induce cell death. Using human neuronal cells treated with a hypoxia memetic compound, desferoxamine (DFO), a hypoxic cell model system was created. Our lab reported previously that treatment with DFO resulted in the decrease of cell viability. However, there were still surviving neurons. The surviving cells did not exhibit significant morphological changes, as compared to the control cells, under confocal microscopy analysis using annexin-V-FLUOS and propidium iodide staining, indicating that they were not at apoptotic or necrotic stages. These surviving neurons, therefore, developed adaptive responses under hypoxic challenge. Several changes …

Utilizing Phluorin-Tagged Receptors To Monitor Subcellular Localization And Trafficking, Ashley M. Fox-Loe, Brandon J. Henderson, Christopher I. Richards

Chemistry Faculty Publications

Understanding membrane protein trafficking, assembly, and expression requires an approach that differentiates between those residing in intracellular organelles and those localized on the plasma membrane. Traditional fluorescence-based measurements lack the capability to distinguish membrane proteins residing in different organelles. Cutting edge methodologies transcend traditional methods by coupling pH-sensitive fluorophores with total internal reflection fluorescence microscopy (TIRFM). TIRF illumination excites the sample up to approximately 150 nm from the glass-sample interface, thus decreasing background, increasing the signal to noise ratio, and enhancing resolution. The excitation volume in TIRFM encompasses the plasma membrane and nearby organelles such as the peripheral ER. Superecliptic …

Regulation Of Ampa-Type Glutamate Receptor Homolog Glr-1 By Erad Ubiquitin Ligases In C. Elegans, Sam Witus

Scholars Week

Endoplasmic reticulum-associated degradation (ERAD) maintains cellular health by removing misfolded proteins from the endoplasmic reticulum (ER). ERAD is ubiquitin-dependent, and ubiquitination of target proteins can be catalyzed by ER-resident E3 ubiquitin ligases. In C. elegans, genes for three putative ERAD E3 ubiquitin ligases have been identified: hrd-1, hrdl-1, and marc-6 (HRD-1, GP78/AMFR, and MARCH-6 in mammalian systems). In C. elegans, these three genes cooperate to maintain the overall health of animals during ER stress. We are testing the roles of hrd-1, hrdl-1, and marc-6 in the neurons of C. elegans. GLR-1 is a glutamate receptor that is expressed in a …

Xenoestrogens From Household Plastics Bind Estrogen Receptors And Affect Cell Proliferation, Susan G. Hurley, Brett R. Lackey, Patricia L. Tate, Sandra L. Gray

Journal of the South Carolina Academy of Science

Xenoestrogens are “foreign” chemicals or compounds that interact with estrogen receptors as either agonists or antagonists to interfere with endocrine system function. Xenoestrogens include phytoestrogens, pesticides, industrial by- products and synthetic estrogens. In this study, consumer plasticware, including baby bottles and food storage containers, were found to release xenoestrogenic compounds that bound to ERα and ERβ and affected cell proliferation.

Effects Of Melatonin On Heartbeat And Possible Identification Of A Melatonin Receptor In Drosophila Melanogaster, Tricia L. Vankirk

Electronic Theses and Dissertations

Chapter 1 of this manuscript is a literature review that serves as an introduction to the entire dissertation. Chapter 2 examines the effects of the melatonin injection on heart rate and rhythmicity in Drosophila melanogaster Canton-S (wild-type) pupae and pupae bearing a variety of heart mutations. Chapter 3 investigates further the possible mechanisms of melatonin’s ability to increase heart rhythmicity without significantly affecting heart rate. A melatonin antagonist, luzindole; a high-affinity melatonin agonist, 2-iodomelatonin and RNAi techniques are used to identify a possible melatonin receptor in Drosophila melanogaster.

An appendix contains a previously published manuscript detailing experiments performed at the …

Importance Of Ubiquitin-Mediated Degradation On Diacetyl Chemosensation In C. Elegans, Ellen Zocher, Nelson Ruth

Scholars Week

Ubiquitin is a small regulatory protein that can be attached to other proteins in a cell, tagging them for destruction. Ubiquitin plays a critical role in regulating the abundance and activity of many proteins. We examined the role of ubiquitin and the cellular pathway it follows in olfactory neurons in the model organism C. elegans. C. elegans senses and moves towards sources of diacetyl, a volatile compound generated by the bacteria it consumes. This behavior is dependent on the diacetyl receptor, ODR-10. We hypothesized that the ubiquitin-mediated degradation system is involved in the regulation of this sensory receptor. Using transgenic …

Interaction Of Human Follicle Stimulating Hormone Receptor With Caveolin Within The Cell Membrane, Jordan Pereira

Honors Theses

The human FSH receptor is a g protein-coupled receptor (GPCR) expressed on the surface of granulosa cells in the ovary and Sertoli cells in the testes. FSHR has a sequence of amino acids consistent with a caveolin interaction motif (φXφXXXXφXXφ) found between amino acids 479-489 in the primary receptor sequence in the putative 4th transmembrane domain. Caveolin is a protein found in cell membrane micro domains such as lipid rafts. These densely packed regions of the membrane are enriched for sphingolipids and cholesterol and are thought to be involved in signal transduction. Caveolin can be found in caveolae, flask-like structures …

Localization Of The Follicle-Stimulating Hormone Receptor In Lipid Raft Domains, Tyler Esposito

Honors Theses

Lipid raft domains have been shown to be important in receptor signaling, especially for G Protein Coupled Receptors (GPCR). Follicle stimulating hormone is a vital reproductive signal that relies on effective signaling of its GPCR. The goal of this study was to test whether or not the follicle stimulating hormone receptor (FSHR) utilizes lipid raft domains when signaling. The main methods used were fluorescent antibodies and various forms of microscopy, including confocal microscopy. A fluorescent antibody for the FSHR (mAb 106.105) allowed us to perform these studies. In the first experiment, cells treated with FSH and stained with the receptor …

Tissue-Specific Thyroid Hormone Regulation Of Gene Transcripts Encoding Iodothyronine Deiodinases And Thyroid Hormone Receptors In Striped Parrotfish (Scarus Iseri), Kaitlin M. Johnson, Sean C. Lema

Biological Sciences

In fish as in other vertebrates, the diverse functions of thyroid hormones are mediated at the peripheral tissue level through iodothyronine deiodinase (dio) enzymes and thyroid hormone receptor (tr) proteins. In this study, we examined thyroid hormone regulation of mRNAs encoding the three deiodinases dio1, dio2 and dio3 – as well as three thyroid hormone receptors trαA, trαB and trβ – in initial phase striped parrotfish (Scarus iseri). Parrotfish were treated with dissolved phase T₃ (20 nM) or methimazole (3 mM) for 3 days. Treatment with exogenous T₃ elevated circulating T₃, while the …

Mutational-Kinetic Analysis Reveals The Roles Of Arginines And Aromatics In The Core Of The Gabaa Receptor Ligand-Binding Pocket, Phu Tran

Dissertations (1934 -)

The γ-aminobutyric acid type A (GABA) receptor is a member of the cys-loop family of ligand-gated ion channels that plays a crucial role in normal brain function by transducing the majority of inhibitory neurotransmission in the central nervous system. The studies documented in this dissertation were aimed at validating and refining the current best model for the interaction of GABA with the GABAA receptor via structure-function perturbation analysis. Mutational-kinetic data was used in conjunction with homology modeling knowledge to draw up architectural and functional roles of the arginines and aromatics in GABA-binding pocket. The results provide interesting new insights.

Two …

Diurnal Rhythms Of Behavior And Brain Mrna Expression For Arginine Vasotocin, Isotocin, And Their Receptors In Wild Amargosa Pupfish (Cyprinodon Nevadensis Amargosae), Sean C. Lema, Lauren J. Wagstaff, Nina M. Gardner

Biological Sciences

Amargosa pupfish (Cyprinodon nevadensis amargosae) occupy remote desert habitats that vary widely in environmental conditions from day to night. In this study, die! patterns of behavior were documented for pupfish in their natural habitat, and examined relative to changes in the abundance of mRNAs encoding prepro-arginine vasotocin (pro-VT), prepro-isotocin (pro-IT), three distinct vasotocin receptors (V1a1, V1a2, and V2), and an isotocin receptor (ITR) in the brain. The behavior of wild pupfish varied diurnally, with frequent aggression from 12:00 to 15:00 h and courtship and spawning most common between 15:00 and 19:00h. Transcript abundance for pro-VT in the brain …

Identification Of Multiple Vasotocin Receptor Cdnas In Teleost Fish: Sequences, Phylogenetic Analysis, Sites Of Expression, And Regulation In The Hypothalamus And Gill In Response To Hyperosmotic Challenge, Sean C. Lema

Biological Sciences

Vasopressin and its homolog vasotocin regulate hydromineral balance, stress responses, and social behaviors in vertebrates. In mammals, the functions of vasopressin are mediated via three classes of membrane-bound receptors: V1a-type, V1b-type and V2-type. To date, however, only a single class of vasotocin receptor has been identified in teleost fish. Here, cDNAs encoding three putative vasotocin receptors – two distinct V1a-type receptor paralogs (V1a1 and V1a2) and a previously undescribed V2- type receptor (V2) – and a single isotocin receptor were isolated and sequenced from the Amargosa pupfish (Cyprinodon nevadensis amargosae). RT-PCR revealed that mRNAs …

Ligand-Induced Conformations Of Extracellular Loop 2 Of At1r, Hamiyet Unal

ETD Archive

Angiotensin II type 1 receptor (AT1R) is a G-protein coupled receptor (GPCR) and an important regulator of blood pressure. It is a target for drug development, because abnormalities in its function are linked to hypertension, cardiac hypertrophy and heart failure. AT1R is composed of seven transmembrane helices connected by three extracellular loops and three intracellular loops. The extracellular loop 2 (ECL2) of AT1R directly interacts with the ligands. This loop is targeted by autoantibodies that activate AT1R in several pathologies such as preeclampsia, malignant hypertension and vascular allograft rejection. Therefore, we proposed that the conformation of ECL2 in AT1R is …

Characterizing The Polycation Receptor Of Paramecium, Eric D. Robinette, Heather G. Kuruvilla

Science and Mathematics Faculty Presentations

Unicellular eukaryotes are complex systems, performing all the tasks needed for survival within the context of a single cell. Protozoans, such as Tetrahymena and Paramecium, use chemosensory systems to detect food and to avoid predation.

Both Tetrahymena and Paramecium have been used as models for studying chemorepellents. Lysozyme, ATP, and GTP have been found to have chemorepellent activity in both ciliates. In Tetrahymena, several PACAP isoforms have been shown to bind to the same receptor as lysozyme, indicating that this receptor may be a more general “polycation receptor” (Keedy et al., 2003). The polycation receptor in Tetrahymena appears to be …

Autocrine Effects Of Catecholamines On Macrophage Release Of Interleukin-6 (Il-6), Shaunta D. Poe

Theses and Dissertations

Effects of norepinephrine (NE) on macrophage cytokine release are complex because the cells have both α2 and β2 adrenergic receptors, which mediate opposing actions. Furthermore, β2-adrenergic agonists are reported to have both stimulatory and inhibitory effects on interleukin-6 (IL-6). This study was designed to clarify the autocrine role of macrophage-derived NE on IL-6 production in activated peritoneal macrophages. Effects of NE on IL-6 production in the RAW264.7 macrophage cell-line also were investigated. Treatment of activated peritoneal macrophages with endotoxin, the α2-adrenergic antagonists yohimbine or RS79948 revealed that the α2-adrenergic receptor mediates a stimulatory autocrine action of catecholamines on IL-6 production. …

Novel Characteristics Of Murine Bone Marrow-Derived Macrophages And Human Macrophage-Like Cells, George Tharwat Georges

Theses and Dissertations

These studies provide evidence for novel properties of macrophages derived from bone marrow stem cells. In study 1, treatment of activated mouse bone marrow-derived macrophages (BMM) with either catecholamine synthesis inhibitors (α-methyl-para-tyrosine and fusaric acid) or the β2 adrenergic receptor antagonist ICI 118,551 demonstrated that BMM produce catecholamines. The catecholamines modulated macrophage cytokine production through autocrine actions on adrenergic receptors. In study II, undifferentiated human bone marrow cells were incubated in 30% mouse L929 fibroblast conditioned medium and generated adherent cells within three days. The cells were clearly identifiable as macrophages based on surface proteins and phagocytic activity but produced …

Two Drosophila Suppressors Of Cytokine Signaling (Socs) Differentially Regulate Jak And Egfr Pathway Activities, Jason S. Rawlings, Gabriela Rennebeck, Susan M.W. Harrison, Rongwen Xi, Douglas A. Harrison

Biology Faculty Publications

BACKGROUND: The Janus kinase (JAK) cascade is an essential and well-conserved pathway required to transduce signals for a variety of ligands in both vertebrates and invertebrates. While activation of the pathway is essential to many processes, mutations from mammals and Drosophila demonstrate that regulation is also critical. The SOCS (Suppressor Of Cytokine Signaling) proteins in mammals are regulators of the JAK pathway that participate in a negative feedback loop, as they are transcriptionally activated by JAK signaling. Examination of one Drosophila SOCS homologue, Socs36E, demonstrated that its expression is responsive to JAK pathway activity and it is capable of downregulating …

Purification Of Anthrax Toxin Protective Antigen Component And Characterization Of Its Binding Interaction With Bovine Kidney Cells, Daniel Dalton Martin

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Protective antigen component of B. anthracis toxin was produced and purified to the >99% level. Toxin was purified from culture supernatant utilizing concentration and liquid chromatography techniques. Purity was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

The purified protective antigen retained biological and antigenic activity as evidenced respectively by lethality in Fischer 344 rats when injected in combination with lethal factor, and by positive results on the Ouchterlony double diffussion assay.

Radioiodinated protective antigen was used both in the in vivo and the in vitro experiments.

In vivo distribution of labelled protective antigen was determined in Fischer 344 rats. Assay …