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Arginine 447 Plays A Pivotal Role In Substrate Interactions In A Neuronal Glutamate Transporter, Annie Bendahan, Ayelet Armon, Navid Madani, Michael Kavanaugh, Baruch I. Kanner

Biomedical and Pharmaceutical Sciences Faculty Publications

Glutamate transporters from the central nervous system play a crucial role in the clearance of the transmitter from the synaptic cleft. Glutamate is cotransported with sodium ions, and the electrogenic translocation cycle is completed by countertransport of potassium. Mutants that cannot interact with potassium are only capable of catalyzing electroneutral exchange. Here we identify a residue involved in controlling substrate recognition in the neuronal transporter EAAC-1 that transports acidic amino acids as well as cysteine. When arginine 447, a residue conserved in all glutamate transporters, is replaced by cysteine, transport of glutamate or aspartate is abolished, but sodium-dependent cysteine transport …

A Simple Method For Generating Full Length Cdna From Low Abundance Partial Genomic Clones, Yongxin Wang, Joseph M. Fugaro, Fauzia Siddiq, Chandra Mouli V. Goparaju, Fulvio Lonardo, Anil Wali, John F. Lechner, Harvey I. Pass

Wayne State University Associated BioMed Central Scholarship

Abstract

Background

PCR amplification of target molecules involves sequence specific primers that flank the region to be amplified. While this technique is generally routine, its applicability may not be sufficient to generate a desired target molecule from two separate regions involving intron /exon boundaries. For these situations, the generation of full-length complementary DNAs from two partial genomic clones becomes necessary for the family of low abundance genes.

Results

The first approach we used for the isolation of full-length cDNA from two known genomic clones of Hox genes was based on fusion PCR. Here we describe a simple and efficient method …

Congenital Chylothorax, Ni̇lgün Köksal, Sultan Demi̇r, Mehmet Okan, Cengi̇z Gebi̇teki̇n, Işik Şenkaya

Turkish Journal of Medical Sciences

No abstract provided.

Biomimetic Preparation Of Ha Powders At 37°C In Urea-And Enzyme Urease-Containing Synthetic Body Fluids, Defne Bayraktar, A. Cüneyt Taş

Turkish Journal of Medical Sciences

An important inorganic phase of synthetic bone applications, calcium hydroxyapatite (HA, Ca_10 (PO_ 4 ) 6_(OH_2 ), was prepared as a single-phase and sub-micron bioceramic powder. Carbonated HA powders were synthesized from calcium nitrate tetrahydrate and di-ammonium hydrogen phosphate salts dissolved in synthetic body fluid (SBF) solutions, containing urea (H_2 NCONH_2 ) and enzyme urease, under the biomimetic conditions of 37°C and pH 7.4, with a novel chemical precipitation technique. These powders were also found to contain trace amounts of Na and Mg ions in them, intentionally incorporated by using SBF solutions, instead of pure water, during their synthesis. The …

Hvj-Liposome-Mediated Gene Transfer To Healing Rat Meniscal Tissue, İlhan Özkan, Norimasa Nakamura, Konsei Shino, Takashi Natsu-Ume, Tetsuya Tomita, Yasufumi Kaneda, Takahiro Ochi

Turkish Journal of Medical Sciences

The success rate of a repaired torn meniscus is not more than 70%. Gene transfer to healing soft tissues appears to be a feasible method for manipulating the healing process. In an attempt to evaluate the feasibility of gene transfer to meniscal tissue we tried to transfer a reporter gene ( Echerichia coli, b-galactosidase gene) by using HVJ-liposome mediated gene transfer. After performing complete radial tear on the medial meniscus of fifty 14-week-old male Wistar rats, HVJ-liposomes with DNA were injected into the femoral artery of 15 rats and into the knee joint of the other 15 rats. HVJ- liposomes …

Role Of End-Tidal Co_2 Monitoring In Patients Intubated And Resuscitated In The Emergency Department, Özgür Karcioğlu, Yasemi̇n Karcioğlu

Turkish Journal of Medical Sciences

Objective: We examined the reliability of end-tidal CO_2 (ETCO_2 ) level in predicting mortality after endotracheal intubation in emergency situations. Methods: In this prospective study, the reliability of ETCO_2 monitoring in the emergency setting as a useful predictor of outcome was investigated in 36 adult patients with pending cardiopulmonary collapse. The cardiopulmonary resuscitation (CPR) procedure was performed as usual and the cardiac rhythm, arterial O_2 saturation, non- invasive blood pressure and ETCO_2 levels were continuously monitored in all the patients. Results: Patients with an ETCO_2 concentration below 0.5% had significantly lower rates of survival. The sensitivity and specificity values of …

Assessment Of Prothrombotic Tendency In Humans Using Functional And Genomic Determinants, Pádraig O'Sullivan

Theses

Thrombophilia describes the familial or acquired disorders of the haemostatic mechanism that are likely to predispose to thrombosis. Venous thrombosis and its associated complications account for a significant number of hospital admissions and deaths annually having a significant health-economic impact on the heath-care industry globally. Mutations in genes that code for proteins involved directly (or indirectly) in blood coagulation have been associated with prothrombotic states. The majority of inherited genetic defects were attributed to polymorphisms in Antithrombin III, Protein C, and Protein S genes until 1994. Together these accounted for only 5-10% of individuals with thrombosis. Subsequently the Factor V …

Isovolumic Regulation In Nervous Tissue: A Novel Mechanism Of Cell Volume Regulation, O. Quesada, R. Franco, K. Hernandez-Fonseca, K. Tuz

School of Veterinary and Biomedical Sciences: Faculty Publications

Cell volume regulation is a property present in most animal cell lineages that allows them to recover their original volume after events of swelling or shrinkage. Such events can be caused by changes in external osmolarity or to osmotic gradients generated during normal cell functioning.4,6 The mechanism of cell volume regulation involves transmembrane fluxes of osmotically active solutes in the necessary direction to counteract the net gain or loss of intracellular water.9 The process through which cells recover their normal volume after swelling is named Regulatory Volume Decrease (RVD). This consists of the efflux of inorganic osmolytes, such as K …

A Cultivated Taste For Yeast., C Brenner

Kimmel Cancer Center Faculty Papers

The availability of complete genomic sequences of Saccharomyces cerevisiae has catalyzed a cultural change in the practice of yeast biology, providing opportunities to develop high throughput techniques to define protein function, to define drug targets, and to discover and characterize drugs.

Faculty Publications 1999, Lsu Health Sciences Center New Orleans

Historical Faculty Publications

This Faculty bibliography contains 871 publications.

Telomerase Activity In Human Preimplantation Embryos, Diane Lynne Wright

Theses and Dissertations in Biomedical Sciences

Telomerase, a ribonucleoprotein, has been described as an essential component of highly proliferative cells, which stabilizes the telomeres and avoids cellular senescence. Telomerase has been identified in various embryonic cell stages, hematopoietic cells, and in >85% of tumor tissue biopsies analyzed. The ability to measure the potential to proliferate successfully could provide an objective measure of an embryo's quality. The objectives of this study were to modify the telomeric repeat amplification protocol (TRAP) assay system for increased sensitivity to allow detection of telomerase activity in the single cell of an oocyte and embryo, obtain telomerase activity levels for the oocyte …

Genetic Control Of Eye Development, A Case Study Focused On The Murine Mutation Belly Spot And Tail (Bst), Qing Tang

Theses and Dissertations (ETD)

Development of the eye is controlled by a network of genes, often conserved, that regulate the timing and location of cellular differentiation. One approach to understanding this network of genes and their interaction is to focus on mutations, spontaneous or induced, that predictably disrupt the proper function of such networks, and by examining the effect of such disruption on the function of other genes.

The Belly spot and tail (Bst) semi-dominant mutation, mapped to mouse Chromosome 16, leads to developmental defects of the eye, skeleton, and coat pigmentation. In the eye, the mutant phenotype is characterized by the presence of …

Murine Anti-Idiotype Antibody 3h1, Malaya Chatterjee, Heinz Köhler, Sunil K. Chatterjee, Kenneth A. Foon

Microbiology, Immunology and Molecular Genetics Faculty Patents

The present invention provides a monoclonal anti-idiotype antibody 3H1 that escapes immune tolerance and elicits a specific immune response to CEA in mice, rabbits, monkeys, and patients with advanced CEA-associated disease. This invention also provides compositions which can be used in the detection or treatment of CEA-associated tumors mimics a specific epitope on carcinoembryonic antigen and a hybridoma that produces 3H1.

Construct Validity Of The Continuous Recognition Memory Test, K. L. Fuchs, J. H. Hannay, W. M. Huckeba, K. A. Espy

Developmental Cognitive Neuroscience Laboratory: Faculty Publications

A principal factor analysis was performed on variables derived from a neuropsychological battery administered to 100 healthy young adults (aged 17-41 yrs) to investigate the construct validity of the Continuous Recognition Memory test (CRM). It was hypothesized that CRM "hits" and "false alarms" would load on different factors. The factors that emerged in the analysis were labeled Verbal Ability, Divided Attention, Attention to Visual Detail, Visuomotor Integration and Planning, and Learning and Memory. As expected, CRM hits had a significant loading on the Learning and Memory factor. However, CRM false alarms did not have a significant loading on the Divided …

The Predictive Use Of Event-Related Potentials In Language Development And The Treatment Of Language Disorders, Dennis L. Molfese, Victoria J. Molfese, K. A. Espy

Developmental Cognitive Neuroscience Laboratory: Faculty Publications

Attempts to relate what is currently known regarding the brain's involvement in language processing during the early years of life. The authors focus on the event related potential (ERP) as a means to study the neuroelectrical correlates of language in the brains of infants and children. After reviewing general information concerning ERPs and language, this presentation relates how neonatal ERP measures of phonetic discrimination predict later language and reading abilities.

Phosphoinositide-Ap-2 Interactions Required For Targeting To Plasma Membrane Clathrin-Coated Pits., I Gaidarov, James H. Keen

Department of Microbiology and Immunology Faculty Papers

The clathrin-associated AP-2 adaptor protein is a major polyphosphoinositide-binding protein in mammalian cells. A high affinity binding site has previously been localized to the NH(2)-terminal region of the AP-2 alpha subunit (Gaidarov et al. 1996. J. Biol. Chem. 271:20922-20929). Here we used deletion and site- directed mutagenesis to determine that alpha residues 21-80 comprise a discrete folding and inositide-binding domain. Further, positively charged residues located within this region are involved in binding, with a lysine triad at positions 55-57 particularly critical. Mutant peptides and protein in which these residues were changed to glutamine retained wild-type structural and functional characteristics by …

Method Of Protecting Against Neuron Loss, Philip W. Landfield

Pharmacology and Nutritional Sciences Faculty Patents

The present invention pertains to a method of protecting against neuron loss in a subject by administering a compound that protects against neuron loss by acting through a vitamin D receptor. Some of these compounds may prevent or retard neuron loss by regulating intraneuronal and/or peripheral calcium and phosphate levels. Other compounds of the invention act through a vitamin D receptor to protect against neuron loss through mechanisms not involving calcium or phosphate regulation. A preferred compound is a biologically active form of vitamin D, a precursor, metabolite, or analog of vitamin D. A preferred form of vitamin D is …

The Effects Of Phosphorus Enrichment On The Dominant Phytoplankton Communities Of Chaney Lake, Kathy Tehrani

Masters Theses & Specialist Projects

Chaney Lake is a temporary karst wetland located in southern Warren County, Kentucky. Because of an impermeable chert layer between the surface and the porous limestone, Chaney Lake fills with water over the winter and spring and then gradually drains over the summer. Three experiments were conducted over the course of the 1998 flooding season to assess the effects of phosphorus addition on the phytoplankton community in Chaney Lake. Ten plastic-sided mesocosms were constructed and placed in the marsh area of the wetland at three different periods: early spring, early summer, and late summer. Phosphorus in the form of K2HPO4 …

Global Cns Gene Transfer For A Childhood Neurogenetic Enzyme Deficiency: Canavan Disease., Paola Leone, Christopher G Janson, Scott J Mcphee, Matthew J During

Department of Neurosurgery Faculty Papers

The neurogenetic prototypic disease on which we chose to test our gene therapy strategy is Canavan disease (CD). CD is an autosomal recessive leukodystrophy associated with spongiform degeneration of the brain. At present the disease is uniformly fatal in affected probands. CD is characterized by mutations in the aspartoacylase (ASPA) gene, resulting in loss of enzyme activity. In this review, recent evidence is summarized on the etiology and possible treatments for CD. In particular, we discuss two gene delivery systems representing recent advances in both viral and liposome technology: a novel cationic liposome-polymer-DNA (LPD) complex, DCChol/DOPE-protamine, as well as recombinant …

Protein Adducts Of Iso[4]Levuglandin E2, A Product Of The Isoprostane Pathway, In Oxidized Low Density Lipoprotein, Robert G. Salomon, Wei Sha, Cynthia Brame, Kamaljit Kaur, Ganesamoorthy Subbanagounder, June O'Neil, Henry F. Hoff, L. Jackson Roberts Ii

Pharmacy Faculty Articles and Research

Levuglandin (LG) E₂, a cytotoxic seco prostanoic acid co-generated with prostaglandins by nonenzymatic rearrangements of the cyclooxygenase-derived endoperoxide, prostaglandin H₂, avidly binds to proteins. That LGE₂-protein adducts can also be generated nonenzymatically is demonstrated by their production during free radical-induced oxidation of low density lipoprotein (LDL). Like oxidized LDL, LGE₂-LDL, but not native LDL, undergoes receptor-mediated uptake and impaired processing by macrophage cells. Since radical-induced lipid oxidation produces isomers of prostaglandins, isoprostanes (isoPs), via endoperoxide intermediates, we postulated previously that a similar family of LG isomers, isoLGs, is cogenerated with isoPs. Now …

Core-Binding Factor Influences The Disease Specificity Of Moloney Murine Leukemia Virus, Amy F. Lewis, Terryl Stacy, William R. Green, Lekidelu Taddesse-Heath, Janet W. Hartley, Nancy A. Speck

Dartmouth Scholarship

The core site in the Moloney murine leukemia virus (Moloney MLV) enhancer was previously shown to be an important determinant of the T-cell disease specificity of the virus. Mutation of the core site resulted in a significant shift in disease specificity of the Moloney virus from T-cell leukemia to erythroleukemia. We and others have since determined that a protein that binds the core site, one of the core-binding factors (CBF) is highly expressed in thymus and is essential for hematopoiesis. Here we test the hypothesis that CBF plays a critical role in mediating pathogenesis of Moloney MLV in vivo. We …

Interaction Between Fgf And Bmp Signaling Pathways Regulates Development Of Metanephric Mesenchyme., Andrew T. Dudley, R. E. Godin, E. J. Robertson

Journal Articles: Genetics, Cell Biology & Anatomy

Nephrogenesis in the mouse kidney begins at embryonic day 11 and ends approximately 10 days postpartum. During this period, new nephrons are continually being generated from a stem-cell population-the nephrogenic mesenchyme-in response to signals emanating from the tips of the branching ureter. Relatively little is known about the mechanism by which the nephrogenic mesenchyme cell population is maintained at the tips of the ureter in the presence of signals promoting tubulogenesis. Previous studies have shown that a loss of Bmp7 function leads to kidney defects that are a likely result of progressive loss of nephrogenic mesenchyme by apoptosis. The studies …

Genetic And Transcriptional Analyses Of The Vibrio Cholerae Mannose-Sensitive Hemagglutinin Type 4 Pilus Gene Locus, Jane W. Marsh, Ronald K. Taylor

Dartmouth Scholarship

The mannose-sensitive hemagglutinin (MSHA) of the Vibrio cholerae O1 El Tor biotype is a member of the family of type 4 pili. Type 4 pili are found on the surface of a variety of gram-negative bacteria and have demonstrated importance as host colonization factors, bacteriophage receptors, and mediators of DNA transfer. The gene locus required for the assembly and secretion of the MSHA pilus has been localized to a 16.7-kb region of the V. cholerae chromosome. Sixteen genes required for hemagglutination, including five that encode prepilin or prepilin-like proteins, have been identified. Examination of MSHA-specific cDNAs has localized two promoters …

Transgenic Mice Which Overexpress Neurotrophin-3 (Nt-3) And Method Of Use, Kathryn M. Albers, Brian M. Davis

Neuroscience Faculty Patents

Transgenic mice express increased levels of neurotrophin-3 (NT-3) in epithelium when their ancestors are microinjected with the NT-3 gene. The NT-3 growth factor expressing transgenic mice are useful in the study of neurodegenerative disorders of the brain such as Parkinson's syndrome and Alzheimer's disease, of the spinal cord motor neurons such as amyotrophic lateral sclerosis, and for testing drug candidates for the treatment of these diseases.

Genetic Monitoring In The Workplace: A Tool Not A Solution, Lillian Trettin, Catherine Musham, Richard Jablonski

RISK: Health, Safety & Environment (1990-2002)

The authors differentiate between genetic monitoring and screening, and discuss the potential risks and benefits of predictive testing technologies.

A Critical Site In The Core Of The Ccr5 Chemokine Receptor Required For Binding And Infectivity Of Human Immunodeficiency Virus Type 1, Salvatore J. Siciliano, Shawn E. Kuhmann, Youmin Weng, Navid Madani, Martin S. Springer, Janet E. Lineberger, Renee Danzeisen, Michael D. Miller, Michael Kavanaugh, Julie A. Demartino, David Kabat

Biomedical and Pharmaceutical Sciences Faculty Publications

Like the CCR5 chemokine receptors of humans and rhesus macaques, the very homologous (∼98–99% identical) CCR5 of African green monkeys (AGMs) avidly binds β-chemokines and functions as a coreceptor for simian immunodeficiency viruses. However, AGM CCR5 is a weak coreceptor for tested macrophage-tropic (R5) isolates of human immunodeficiency virus type 1 (HIV-1). Correspondingly, gp120 envelope glycoproteins derived from R5 isolates of HIV-1 bind poorly to AGM CCR5. We focused on a unique extracellular amino acid substitution at the juncture of transmembrane helix 4 (TM4) and extracellular loop 2 (ECL2) (Arg for Gly at amino acid 163 (G163R)) as the likely …

Research Fundamentals: V. The Use Of Laboratory Animal Models In Research, Brian J. O'Neil, Jeffrey A. Kline, Keith Burkhart, John Younger

Biomedicine and Animal Models in Research Collection

Animal research has provided important information about many aspects of the pathophysiology of human disease. Well-performed animal studies can determine the potential benefit of many proposed therapeutic interventions, and experimental results from animal studies have served as the basis for many landmark clinical trials. Many animal research models are described in the research literature, and choosing the appropriate model to answer a research question can be a daunting task. Even more challenging is developing a new model when none of the existing systems are relevant to the proposed question. This article was prepared by members of the SAEM Research Committee …

Screening For Eating Disorders Utilizing The Minnesota Multiphasic Personality Inventory, Roseann M. Woodka

Dissertations

Problem

Eating disorders, in the form of anorexia nervosa and bulimia nervosa, have been recognized as significant mental-health issues for the last three decades, and the incidence is rising as we approach the new millennium. Currently, many women who come into a mental-health setting due to depression, anxiety, low self-esteem, relationship issues, sexual issues, etc., are also struggling with eating-disordered behaviors, thoughts, and feelings. These behaviors, thoughts, and feelings may remain well hidden from the counselor throughout the course of therapy or until they become severe and more difficult to treat. Because eating disorders are very complex involving psychological, physical, …

Conjugated Bilirubin Measurement Re-Examined: A New 'Approach'., Aidan Kelleher

Theses

Conjugated bilirubin is an important but difficult metabolite to measure accurately, elevated levels of conjugated bilirubin in the serum are indicative of obstructive jaundice and therefore can be used to discriminate this disorder from other forms of unconjugated hyper bilirubinaemia.

Serum D(-)-Lactate And Nitric Oxide (No) Levels Inacute Intestinal Ischemia, Mahmut Basoğlu, Ahmet Balik, Ahmet Kiziltunç, Fatih Akçay, Selçuk Sabri Atamanalp

Turkish Journal of Medical Sciences

To determine serum D(-)-lactate and NO levels in patients with acute mesenteric ischemia. 22 patients (12 with acute mesenteric ischemia, 10 with bowel obstruction) and 12 control patients who were operated due to goitre. Serum D(-)-lactate and NO levels in acute mesenteric ischemia cases compared to small bowel obstruction cases and control patients. Increased serum D(-)-lactate and NO levels in serum levels of ischemic bowel were found to be significantly increased compared to both the cases with small bowel obstruction and control subjects (for D(-)-lactate: p