DOCSLIB.ORG

And URSPRUNG(1966)

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
And URSPRUNG(1966) GENETICS OF OCTANOL DEHYDROGENASE IN DROSOPHILA METZIP+ SARAH BEDICHEK PIPKIN Homrd Uniuersity, Washington, D.C.20001 Received October 11, 1967 CTANOL dehydrogenase (ODH) was studied in Drosophila melanogaster first by URSPRUNGand LEONE(1965) and distinguished from alcohol de- hydrogenase by COURTRIGHT,IMBERSKI, and URSPRUNG(1966). The neotropical species Drosophila metzii is polymorphic for complex octanol dehydrogenase patterns which will be shown to depend on two distinct structural genes, ODH,, apparently homologous with the locus studied by COURTRIGHTet al. (1966), and ODH,, responsible for a more slowly migrating isozyme. The ODH loci are un- linked, and variants display unifactorial inheritance. The ODH molecule is con- sidered at least a dimer but probably a tetramer. Isozyme formation can involve combinations of polypeptides produced by either or both of the two structural genes. Genetic evidence will be presented indicating that egg or embryonic and imaginal ODH isozyme patterns depend on the same structural genes. MATERIALS AND METHODS Single flies were assayed in crude homogenates with 1-octanol as substrate, using the agar gel electrophoresis method, with formazan staining according to the method of DR.H. URSPRUNG (1x5). Modifications of the method for the present work included grinding of single flies in a drop of glass distilled water in specially prepared small homogenizers and allowing the electro- phoresis to proceed at 25Ov for forty minutes instead of half an hour. Enzyme assays for the genetic analysis were made on single females aged 4 to 6 days. The smaller males do not provide sufficient enzyme for reliable single fly analysis. Flies for experimental crosses were reared in pair matings on a medium of corn meal-Karo-Brewer’s yeast #2019 (Standard Brands). Isozyme patterns of extracted lines were also studied in flies reared on the usual medium enriched by the addition of Kellogg K breakfast food. ODH variants were isolated from strains of D. metzii from Trinidad, West Indies; Barro Colorado Island, Canal Zone; Turrialba, Costa Rica; Darien, Panama; and Almirante, Panama. Eggs were collected by allowing fertilized females to lay for thirty hours in small Petri plates containing a solution of Fleischmann’s yeast sprinkled with dilute acetic acid. Contents of the Petri plates were washed onto a Buchner funnel lined with a black cloth overlaid with cheesecloth. The latter strained out drowned flies and debris. Eggs were taken from the black cloth after repeated washing. These were ground in small homogenizers with a drop of glass distilled water. RESULTS Upon inbreeding with pair matings, lines with true breeding isozyme patterns This work was supported by the Division of General Medical Sciences, National Institutes of Health, Bethesda, blaryland, RG-12018 to the Johns Hopkins University, Baltimore, Maryland, and biM-14937 to Howard University, Washington, D.C. + This paper is dedicated to the memory of Dr. WIXSONS. STONE,the Genetics Foundation, the University of Texas, Austin, Texas. Genetics 60: 81-92 September 1968. a2 SARAH BEDICHEK PIPKIN +a b C FIGURE1 .-Zymograms of ODH isozymes of single adult females of (a) a pattern 5 extracted line (from left to right, first two slits); (b) a pattern 3 extracted line (from left, third and fourth slits); (c) a pattern 1, or pattern 1, extracted line (from left, fifth through eleventh slits). were extracted: (1) those showing a strong band at position 5 and sometimes weak bands at positions 3 and 4, with sometimes also a strong band at position 6 and/or 7 (called pattern 5 and shown in Figure la); (2) those showing a single formazan band at position 3 (called pattern 3 and shown in Figure lb); (3) those OCTANOL DEHYDROGENASE IN D.nzetzii 83 showing a single fomazan band at position 1 (called pattern 1 and shown in Figure IC).The pattern 5 line could be extracted from the Turrialba, Almirante, and Trinidad strains, but most of the breeding work was done with the line ex- tracted from the Turrialba strain in two generations of inbreeding. Without selection the Barro Colorado Island strain was true breeding for pattern 3, al- though this strain originated from 8 females collected in nature (PIPKIN1968). Pattern 3 lines could be extracted in one generation of inbreeding from the Darien strain and in two generations from the Trinidad strain. An almost true breeding line with isozymes in positions 1, 2, 3, 4, 5, and sometimes 6 and 7 (called pattern 1-5) was isolated in two generations from the Trinidad strain and used in early breeding work. Occasionally an individual with isozymes only at positions 1, 2, and 3 would be seen, and in the course of a year, the pattern 1-5 line showed just one isozyme at position 1 (Figure IC).A photograph from single females of the pattern 1-5 line is given in PIPKIN(1968). Sublines from the original pattern 1-5 line were maintained which subsequently showed only a single isozyme at position 1 but exhibited different breeding behavior in crosses with individuals of the pattern 3 line as will be described later. Most of the breeding work was carried out using ordinary corn meal-Brewer's yeast medium. When cultured on medium enriched by Kellogg K breakfast food, isozyme patterns of the Barro Colorado Island pattern 3 line, the Turrialba pat- tern 5 line, and the Trinidad pattern 1 line, which were used for most of the breeding work, remained unchanged except for stronger staining indicating a higher enzyme level. The pattern 1-5 line had become the pattern 1 line by the time the enriched medium was being used. On the other hand, progeny of a single female of the Trinidad pattern 3 line cultured on the enriched medium showed four isozymes at positions 3,4, 5, and 6 in eight individuals and at posi- tion 3 only in three individuals. This was taken at first as a sign of heterogeneity in the line, but efforts to isolate a pattern 3 line by inbreeding further single females failed 'when the enriched medium was used. An explanation will be offered after the breeding results are presented. Males of the freshly extracted pattern 1-5 line were mated singly with pattern 3 females of the Barro Colorado Island strain. Twelve F, females assayed showed isozymes at positions 1, 2, 3, 4, 5, and sometimes 6 and 7. The F, males were backcrossed individually to females of the Barro Colorado Island pattern 3 line. Fifteen backcross progenies appeared to be segregating for the two pairs of alleles because they consisted of four different isozyme patterns in equal numbers as shown in Table 1 (see also diagram, Figure 2). A photograph of these segregating progeny appeared in a preliminary research note (PIPKIN1968). The four pat- terns occurred in a 1: 1: 1: 1 ratio, and frequencies of the four patterns in separate sibships were homogeneous. The four isozyme patterns were found in similar backcross progenies (cultured on ordinary corn meal medium), in which the pattern 3 PI parent had been extracted from the Trinidad strain. Thus, with either a similar or diverse genetic background, backcross sibships were segregat- ing for four isozyme patterns. There is no possibility that any of the ODH isozymes seen here are caused by 84 SARAH EEDICHEK PIPKIN TABLE 1 Inheritance of ODH isozyme patterns: backcrosses and F, X F, Patterns of Patterns of progeny Chi parental lines 1 3 1.3.5.5 3.4.5 1-5 5 1-3 square P 1-5/3 X 3 .. 48 .. 45 57 43 2.378 0.5>P>0.3 3/5 x 3/5 . 11 . 24 . 8 . 0.999 0.7>P>0.5 1B/3 X 1B/3 9 9 .. .. .. , . 15 0.272 0.9>P>0.8 1A/5 X 1A/5 Trinidad/Turrialba 15 . 54 . 17 . 3.22 0.2>P>O.I Trinidad/Trinidad 16 .. 46 .. 22 . 1.61 O.S>P>O.3 IA/5 x 5 .. .. 21 .. .. 14 . 1.40 0.3>P>0.2 lA/5 x 3 .. .. 21 19 .. 0.10 0.8>P>O.9 the alcohol dehydrogenase locus. PIPKIN,in COURTRIGHT,IMEERSKI, and UR- SPRUNG 1966, reported that Drosophila metzii and other members of the tri- punctata species group as well as members of the related cardini and calloptera species groups show little ADH activity using ethanol, primary hexanol, heptanol, or octanol as substrates. A faint ADH band is not uncommonly seen in D. metzii gels four centimeters cathodal to the fastest ODH band. A graduate student, Mr. WILLIAMDOUGLAS, has confirmed the observation that in D. metzii there is FIGURE2.-Segregating progeny of the backcross pattern 1-5/ pattern 3 F, male x pattern 3 female. Assuming a tetramer subunit structure of ODH, the genotypes of parents of the backcross we ODHF,/ODHS,; ODHF,/ODHS, males x ODHs,/ODHS,; ODHs2/ODHs2,where the fast alleles, ODHF, and ODHF,, either produce subunits earlier in development or produce more subunits than the slow alleles, ODHS, and ODHS,. Four classes of segregating progeny of the backcross include: pattern 1-5 individuals (genotype ODHF,/ODH",; ODHF,/ODHs2); pattern 3,4,5 individuals (genotype ODHF1/ODHsl; ODHS2/ODHs,) ; pattern 3 individuals (genotype ODHs,/ODHs,; ODHs,/ODHs2); pattern 1-3 individuals (genotype ODHs,/ODHs,; ODHFz/ ODHS,). From left to right, the first three slits contain single female backcross progeny of pattern 3,4,5; the fourth, fifth, seventh, eighth, and ninth slits, single females of pattern 1-3; the sixth slit, a single female of pattern 3, and the tenth and eleventh slits, single females of pattern 1-5. OCTANOL DEHYDROGENASE IN D. mtzii 85 weak or undetectable ADH formazan staining since he has used the species as a control in testing substrate specificity of ODH and ADH isozymes in a number of neotropical species, using a variety of primary and secondary alcohols (includ- ing 2-butanol) as a substrate.
Load more

Recommended publications
  • Systems and Chemical Biology Approaches to Study Cell Function and Response to Toxins
    Dissertation submitted to the Combined Faculties for the Natural Sciences and for Mathematics of the Ruperto-Carola University of Heidelberg, Germany for the degree of Doctor of Natural Sciences Presented by MSc. Yingying Jiang born in Shandong, China Oral-examination: Systems and chemical biology approaches to study cell function and response to toxins Referees: Prof. Dr. Rob Russell Prof. Dr. Stefan Wölfl CONTRIBUTIONS The chapter III of this thesis was submitted for publishing under the title “Drug mechanism predominates over toxicity mechanisms in drug induced gene expression” by Yingying Jiang, Tobias C. Fuchs, Kristina Erdeljan, Bojana Lazerevic, Philip Hewitt, Gordana Apic & Robert B. Russell. For chapter III, text phrases, selected tables, figures are based on this submitted manuscript that has been originally written by myself. i ABSTRACT Toxicity is one of the main causes of failure during drug discovery, and of withdrawal once drugs reached the market. Prediction of potential toxicities in the early stage of drug development has thus become of great interest to reduce such costly failures. Since toxicity results from chemical perturbation of biological systems, we combined biological and chemical strategies to help understand and ultimately predict drug toxicities. First, we proposed a systematic strategy to predict and understand the mechanistic interpretation of drug toxicities based on chemical fragments. Fragments frequently found in chemicals with certain toxicities were defined as structural alerts for use in prediction. Some of the predictions were supported with mechanistic interpretation by integrating fragment- chemical, chemical-protein, protein-protein interactions and gene expression data. Next, we systematically deciphered the mechanisms of drug actions and toxicities by analyzing the associations of drugs’ chemical features, biological features and their gene expression profiles from the TG-GATEs database.
    [Show full text]
  • Preclinical Evaluation of Protein Disulfide Isomerase Inhibitors for the Treatment of Glioblastoma by Andrea Shergalis
    Preclinical Evaluation of Protein Disulfide Isomerase Inhibitors for the Treatment of Glioblastoma By Andrea Shergalis A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy (Medicinal Chemistry) in the University of Michigan 2020 Doctoral Committee: Professor Nouri Neamati, Chair Professor George A. Garcia Professor Peter J. H. Scott Professor Shaomeng Wang Andrea G. Shergalis [email protected] ORCID 0000-0002-1155-1583 © Andrea Shergalis 2020 All Rights Reserved ACKNOWLEDGEMENTS So many people have been involved in bringing this project to life and making this dissertation possible. First, I want to thank my advisor, Prof. Nouri Neamati, for his guidance, encouragement, and patience. Prof. Neamati instilled an enthusiasm in me for science and drug discovery, while allowing me the space to independently explore complex biochemical problems, and I am grateful for his kind and patient mentorship. I also thank my committee members, Profs. George Garcia, Peter Scott, and Shaomeng Wang, for their patience, guidance, and support throughout my graduate career. I am thankful to them for taking time to meet with me and have thoughtful conversations about medicinal chemistry and science in general. From the Neamati lab, I would like to thank so many. First and foremost, I have to thank Shuzo Tamara for being an incredible, kind, and patient teacher and mentor. Shuzo is one of the hardest workers I know. In addition to a strong work ethic, he taught me pretty much everything I know and laid the foundation for the article published as Chapter 3 of this dissertation. The work published in this dissertation really began with the initial identification of PDI as a target by Shili Xu, and I am grateful for his advice and guidance (from afar!).
    [Show full text]
  • Degruyter Chem Chem-2021-0032 347..357 ++
    Open Chemistry 2021; 19: 347–357 Research Article Belgin Sever, Mehlika Dilek Altıntop*, Yeliz Demir, Cüneyt Türkeş, Kaan Özbaş, Gülşen Akalın Çiftçi, Şükrü Beydemir*, Ahmet Özdemir A new series of 2,4-thiazolidinediones endowed with potent aldose reductase inhibitory activity https://doi.org/10.1515/chem-2021-0032 received December 2, 2020; accepted February 9, 2021 1 Introduction Abstract: In an effort to identify potent aldose reductase Type 2 diabetes (T2D) is a chronic life-threatening disease (AR) inhibitors, 5-(arylidene)thiazolidine-2,4-diones (1–8), characterized by abnormally high blood glucose levels which were prepared by the solvent-free reaction of 2,4- resulting from impaired response of target tissues to thiazolidinedione with aromatic aldehydes in the presence insulin (insulin resistance) and/or progressively reduced in vitro of urea, were examined for their AR inhibitory function of pancreatic β cells. The global burden of T2D is -( - - - - activities and cytotoxicity. 5 2 Hydroxy 3 methylbenzyli increasing considerably, and therefore there is an urgent ) - - (3) dene thiazolidine 2,4 dione was the most potent AR need to develop safe and potent antidiabetic agents [1–5]. inhibitor in this series, exerting uncompetitive inhibition Polyol pathway is a two-step metabolic pathway K ± with a i value of 0.445 0.013 µM. The IC50 value of in which glucose is reduced to sorbitol, which is then 3 fi - compound for L929 mouse broblast cells was deter converted to fructose. The abnormally activated polyol mined as 8.9 ± 0.66 µM, pointing out its safety as an AR pathway has been reported to participate in the patho- inhibitor.
    [Show full text]
  • Gene Expression in Buccal Keratinocytes with Emphasis on Carbonyl Metabolism
    From the Department of Medical Biochemistry and Biophysics Karolinska Institutet, Stockholm, Sweden GENE EXPRESSION IN BUCCAL KERATINOCYTES WITH EMPHASIS ON CARBONYL METABOLISM Claudia A. Staab Stockholm 2008 All previously published papers were reproduced with permission from the publisher. Published by Karolinska Institutet. Printed by [name of printer] © Claudia A. Staab, 2008 ISBN 978-91-7409-127-4 Printed by 2008 Gårdsvägen 4, 169 70 Solna Une sortie, c'est une entrée que l'on prend dans l'autre sens. Boris Vian ABSTRACT The inner lining of the cheek, the buccal mucosa, is a target for air-borne, dietary and tobacco usage-derived carcinogens, but also interesting from a drug delivery point of view. Cancer arising in the buccal epithelium, buccal squamous cell carcinoma (BSCC), often diagnosed at a late disease stage, is highly aggressive and recurrent, emphasizing the need for novel approaches in diagnosis and therapy. An in vitro model for human buccal carcinogenesis consisting of normal buccal keratinocytes (NBK) and two transformed cell lines of buccal origin was applied to explore mechanisms of buccal carcinogenesis, tumor marker and drug target expression. Two-dimensional gel electrophoresis, DNA microarray analysis, and the application of three bioinformatics programs for data mining allowed for the identification of multiple established and potential novel markers for BSCC, including tumor promoter/suppressor genes. Furthermore, post-confluent culture of NBK in absence and presence of fetal bovine serum was successfully used to induce terminal squamous differentiation (TSD) to various extents and thus enrich for different strata of the epithelium. Here, expression and activity of carbonyl-metabolizing enzymes (CMEs) were assessed in view of their multiple roles in phase I biotransformation.
    [Show full text]
  • Dehydrogenase Classes
    Proc. Nati. Acad. Sci. USA Vol. 91, pp. 4980-4984, May 1994 Biochemistry Fundamental molecular differences between alcohol dehydrogenase classes (Drosophila octano dehydrogenase/class m alcohol dehydrogenase/mo ur patterns/zinc enyme famy) OLLE DANIELSSON*, SILVIA ATRIANt, TERESA LUQUEt, LARS HJELMQVIST*, ROSER GONZALEZ-DUARTEt, AND HANS J6RNVALL*f *Department of Medical Biochemistry and Biophysics, Karolinska Institutet, S-171 77 Stockholm, Sweden; tCenter for Biotechnology, Karolinska Institutet, S-141 86 Huddinge, Sweden; and tDepartment of Genetics, University of Barcelona, E-08071 Barcelona, Spain Communicated by Sune Bergstrom, January 18, 1994 ABSTRACT Two types of alcohol dehydrogenase in sepa- ary patterns, with class III being "constant" and class I rate protein families are the "medium-chain" zinc enzymes "variable" (10), result in a consistent picture of the enzyme (including the classical liver and yeast forms) and the "short- system and place the classes of medium-chain alcohol dehy- chain" enzymes (including the insect form). Although the drogenases as separate enzymes in the cellular metabolism. medium-chain family has been characterized in prokaryotes Similarly, another protein family, short-chain dehydroge- and many eukaryotes (fungi, plants, cephalopods, and verte- nases, has also evolved into a family comprising many brates), insects have seemed to possess only the short-chain different enzyme activities, including an alcohol dehydroge- enzyme. We have now also characterized a medium-chain nase (11). This form operates by means of a completely alcohol dehydrogenase in Drosophila. The enzyme is identical different catalytic mechanism and is related to mammalian to insect octanol dehydrogenase. It Is a typical class m alcohol prostaglandin dehydrogenases/carbonyl reductase (12).
    [Show full text]
  • A Phase II Neoadjuvant Study of Apalutamide, Abiraterone Acetate, Prednisone, Degarelix and Indomethacin in Men with Localized Prostate Cancer Pre-Prostatectomy
    CC Protocol Number: 9628 PI: Michael T. Schweizer, MD Neoadjuvant therapy in high-risk Prostate Cancer Protocol Version: 4.0; March 23, 2018 A Phase II neoadjuvant study of Apalutamide, abiraterone acetate, prednisone, degarelix and indomethacin in men with localized prostate cancer pre-prostatectomy University of Washington / Seattle Cancer Care Alliance Cancer Consortium Protocol Number: 9628 IND Number: 129692 ClinicalTrials.gov: NCT02849990 Protocol Version Number: 4.0 March 23, 2018 Sponsor-Investigator / Principal Investigator: Site: Michael T. Schweizer, MD University of Washington / Seattle University of Washington / Seattle Cancer Care Alliance Cancer Care Alliance Email: [email protected] Medication Support Provided by: Biostatistician: Janssen Scientific Affairs, LLC Roman Gulati Fred Hutchinson Cancer Research Center Email: [email protected] 1 CC Protocol Number: 9628 PI: Michael T. Schweizer, MD Neoadjuvant therapy in high-risk Prostate Cancer Protocol Version: 4.0; March 23, 2018 Title: A Phase II neoadjuvant study of Apalutamide, abiraterone acetate, prednisone, degarelix and indomethacin in men with localized prostate cancer pre-prostatectomy Objectives: To assess the pathologic effects of 3-months (12 weeks) of neoadjuvant apalutamide, abiraterone acetate, degarelix and indomethacin in men with localized prostate cancer pre-prostatectomy. Study Design: Open label, single-site, Phase II study designed to determine the pathologic effects that 3-months (12 weeks) of neoadjuvant therapy has on men with localized prostate cancer. Primary Center: University of Washington/Seattle Cancer Care Alliance Participating Institutions: 1 site in the United States. Medication Support: Janssen Scientific Affairs, LLC Timeline: This study is planned to complete enrollment in one year, with 2-years of additional follow up following accrual of the last subject.
    [Show full text]
  • Bupropion's Bioinequivalence
    Bupropion’s Bioinequivalence: Patient Variability, Absorption, and Metabolism by Jamie Nicole Connarn A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy (Pharmaceutical Sciences) in the University of Michigan 2015 Doctoral Committee: Professor Duxin Sun, Chair Professor Gordon L. Amidon Professor Vicki L. Ellingrod Professor David E. Smith ©Jamie N. Connarn 2015 Dedication To my family for all their love, prayers, and support; especially my Mom. ii Acknowledgements First and foremost, I would like to thank my advisor, Dr. Duxin Sun for all the remarkable opportunities that he gave me during my graduate career as well as the full support and training. Without all his support, knowledge, and dedication to producing successful graduate students, this would not be possible. I would like to thank my committee members; Dr. David Smith, Dr. Gordon Amidon, and Dr. Vicki Ellingrod for all their insightful expertise in guiding my research smoothly. I would also like to thank Dr. Jason Gestwicki, who was a huge help on my Heat Shock Protein studies. In addition, I would like to thank Dr. Rose Feng, Dr. Simon Zhou, and Dr. Yan Li for collaborative projects and all the assistances with my PK analysis. I am grateful for all my lab mates and staff; Hayley Paholak, Joe Burnett, Becky Reed, Xiaoqing Ren, Kanokwan Sansanaphongpricha, Albert Lin, Mari Gasparyan, Alex Yu, Ila Myers, Nathan Truchan, Huixia Luo, Dr. Hongwei Chen, Dr. Ruijuan Luo, Dr. Ting Zhao, Dr. Bo Wen, Dr. Siwei Li, Dr. Xiaoqin Li, Dr. Yasuhiro Tsume, Marisa Gies, and Gail Benninghoff. In addition, I would like to thank those who were a part of our large clinical team; Marisa Kelly, Gloria Harrington, Stephanie Flowers, Kirsten Weiss, Xinyuan (Susie) Zhang, and Andrew Babiskin I would like to express how grateful I am for all of my friends I have met here; Xiaomei Chen, Amy Doty, Morgan Giles, Kelly Hansen, Eric Lachacz, Maya Lipert, Max Mazzara, Allison Maytas, Max Stefan, Charlie Steffens, Arjang Talattof, Karthik Pisupati, Maria Posada, and too many more.
    [Show full text]
  • Development of Novel Oxotriazinoindole Inhibitors Of
    Article Cite This: J. Med. Chem. 2020, 63, 369−381 pubs.acs.org/jmc Development of Novel Oxotriazinoindole Inhibitors of Aldose Reductase: Isosteric Sulfur/Oxygen Replacement in the Thioxotriazinoindole Cemtirestat Markedly Improved Inhibition Selectivity † ‡ ̌ ‡ ̌ † § Matuś̌Hlavać,̌Lucia Kovaćikovǎ ,́Marta Soltesová ́Prnova,́Peter Sramel, Gabriela Addova,́ ‡ ∥ †,⊥ ̌ ,‡ Magdalená Majeková ,́Gilles Hanquet, Andrej Bohać,̌ and Milan Stefek* † Department of Organic Chemistry, Faculty of Natural Sciences, Comenius University in Bratislava, Ilkovicovǎ 6, 842 15 Bratislava, Slovakia ‡ Institute of Experimental Pharmacology and Toxicology, CEM, SAS, Dubravská ́cesta 9, 841 04 Bratislava, Slovakia § Institute of Chemistry, Faculty of Natural Sciences, Comenius University in Bratislava, Ilkovicovǎ 6, 842 15 Bratislava, Slovakia ∥ Universitéde Strasbourg, Universitéde Haute-Alsace, CNRS, UMR 7042-LIMA, ECPM, 25 rue Becquerel, 67087 Strasbourg, France ⊥ Biomagi, Inc., Mamateyova 26, 851 04 Bratislava, Slovakia *S Supporting Information ABSTRACT: Inhibition of aldose reductase (AR), the first enzyme of the polyol pathway, is a promising approach in treatment of diabetic complications. We proceeded with optimization of the thioxotriazinoindole scaffold of the novel AR inhibitor cemtirestat by replacement of sulfur with oxygen. A series of 2-(3-oxo-2H-[1,2,4]triazino[5,6-b]indol-5(3H)-yl)acetic acid derivatives (OTIs), designed by molecular modeling and docking, were synthesized. More electronegative and less bulky oxygen of OTIs compared to the sulfur of the original thioxotriazinoindole congeners was found to form a stronger H- bond with Leu300 of AR and to render larger rotational flexibility of the carboxymethyl pharmacophore. AR inhibitory activities of the novel compounds were characterized by the IC50 values in a submicromolar range.
    [Show full text]
  • Download (8Mb)
    A Thesis Submitted for the Degree of PhD at the University of Warwick Permanent WRAP URL: http://wrap.warwick.ac.uk/111600/ Copyright and reuse: This thesis is made available online and is protected by original copyright. Please scroll down to view the document itself. Please refer to the repository record for this item for information to help you to cite it. Our policy information is available from the repository home page. For more information, please contact the WRAP Team at: [email protected] warwick.ac.uk/lib-publications > 5 0 0 4 ^ /g 4 THE UTILIZATION OF CL.-C,,, 14 16 Le 6EX>S M •£• N-ALKANES BY MICROORGANISMS by M . R . Lebens B .Sc. fsfbf 6 *0 &>fib»riGr. A thesis submitted in fulfilment of the requirements for the degree of Ph .D. Department of Biological Sciences, University of Warwick, Coventry CV4 7AL U.K. January 1983 Summary Microorganisms capable of growth using n-alkanes as sole carbon and energy source were isolated from the environment. Subsequently selected organisms from those isolated were subjected to morphological and biochemical surveys that involved electron microscopy and assays of enzymes concerned with n-alkane degradation and assimilation. The alcohol and aldehyde dehydrogenases associated with the growth of a pseudomonad on n-alkanes were partially purified and shown to be NAD(P) independent. Genetic manipulations were attempted to produce mutants of a coryneform bacterium, by chemical and U .V. mutagenesis, that were blocked at specific points in the n-alkane degradation pathway which were shown to be chromosomally borne. Appropriate selection procedures were devised and mutations identified by product accumulation by whole cells .
    [Show full text]
  • Lens Adaptation to Glutathione Deficiency
    LENS ADAPTATION TO GLUTATHIONE DEFICIENCY: IMPLICATIONS FOR CATARACT by JEREMY A. WHITSON Submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy Department of Pathology CASE WESTERN RESERVE UNIVERSITY May, 2017 1 Table of Contents List of Tables 6 List of Figures 7 Acknowledgements 11 List of Abbreviations 12 Abstract 17 1. Introduction 19 1.1. The Crystalline Lens - Biological Glass 20 1.1.1. Anatomy of the Lens 20 1.1.2. Functions of the Lens 21 1.1.3. Lens Cell Biology 22 1.1.4. Transport and Homeostasis in the Lens 23 1.1.5. Conclusions 25 1.2. Age-Related Cataract - A Disease of Protein Aggregation 26 1.2.1. Pathology of Age-Related Cataract 26 1.2.2. Risk Factors for and Impact of Age-Related Cataract 27 1.2.3. Clinical Treatment of Cataract 28 1.2.4. Posterior Capsular Opacification 29 1.2.5. Conclusions 30 1.3. Glutathione - The Guardian of Lens Redox Homeostasis 30 1.3.1. Structure and Biosynthesis of Glutathione 30 1.3.2. Regulation of Cellular Glutathione Content 32 1.3.3. The Essential Role of Glutathione in the Lens 32 1.3.4. Lenticular Glutathione Declines with Age 34 1.3.5. A Loss of Glutathione Activity Leads to Cataract 34 1.3.6. The LEGSKO Mouse Model 35 1.3.7. A Salvage Pathway for Glutathione Synthesis 36 1.3.8. Conclusions 37 1.4. Glutathione Transport – Known Mechanisms 38 1.4.1. Glutathione Transport in the Lens 38 1.4.2.
    [Show full text]
  • Analysis of Catecholamines and Pterins in Inborn Errors of Monoamine Neurotransmitter Metabolism—From Past to Future
    cells Review Analysis of Catecholamines and Pterins in Inborn Errors of Monoamine Neurotransmitter Metabolism—From Past to Future Sabine Jung-Klawitter * and Oya Kuseyri Hübschmann Department of General Pediatrics, Division of Neuropediatrics and Metabolic Medicine, University Hospital Heidelberg, 69120 Heidelberg, Germany * Correspondence: [email protected]; Tel.: +49-(0)6221-5639586 Received: 30 June 2019; Accepted: 4 August 2019; Published: 9 August 2019 Abstract: Inborn errors of monoamine neurotransmitter biosynthesis and degradation belong to the rare inborn errors of metabolism. They are caused by monogenic variants in the genes encoding the proteins involved in (1) neurotransmitter biosynthesis (like tyrosine hydroxylase (TH) and aromatic amino acid decarboxylase (AADC)), (2) in tetrahydrobiopterin (BH4) cofactor biosynthesis (GTP cyclohydrolase 1 (GTPCH), 6-pyruvoyl-tetrahydropterin synthase (PTPS), sepiapterin reductase (SPR)) and recycling (pterin-4a-carbinolamine dehydratase (PCD), dihydropteridine reductase (DHPR)), or (3) in co-chaperones (DNAJC12). Clinically, they present early during childhood with a lack of monoamine neurotransmitters, especially dopamine and its products norepinephrine and epinephrine. Classical symptoms include autonomous dysregulations, hypotonia, movement disorders, and developmental delay. Therapy is predominantly based on supplementation of missing cofactors or neurotransmitter precursors. However, diagnosis is difficult and is predominantly based on quantitative detection of neurotransmitters, cofactors, and precursors in cerebrospinal fluid (CSF), urine, and blood. This review aims at summarizing the diverse analytical tools routinely used for diagnosis to determine quantitatively the amounts of neurotransmitters and cofactors in the different types of samples used to identify patients suffering from these rare diseases. Keywords: inborn errors of metabolism; catecholamines; pterins; HPLC; fluorescence detection; electrochemical detection; MS/MS 1.
    [Show full text]
  • Novel Substituted N-Benzyl(Oxotriazinoindole
    Novel substituted N-benzyl(oxotriazinoindole) inhibitors of aldose reductase exploiting ALR2 unoccupied interactive pocket Matúš Hlaváč, Lucia Kováčiková, Marta Prnová, Gabriela Addová, Gilles Hanquet, Milan Štefek, Andrej Boháč To cite this version: Matúš Hlaváč, Lucia Kováčiková, Marta Prnová, Gabriela Addová, Gilles Hanquet, et al.. Novel substituted N-benzyl(oxotriazinoindole) inhibitors of aldose reductase exploiting ALR2 unoccupied interactive pocket. Bioorganic and Medicinal Chemistry, Elsevier, 2020. hal-03070904 HAL Id: hal-03070904 https://hal.archives-ouvertes.fr/hal-03070904 Submitted on 16 Dec 2020 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Novel substituted N - benzyl(oxotriazinoindole) inhibitors of aldose reductase exploiting ALR2 unoccupied interactive pocket Matúš Hlaváč 1* , Lucia Kováčiková 2 , Marta Šoltésová Prnová 2 , Gabriela Addová 1 , Gilles Hanquet 3 , Milan Štefek 2 , Andrej Boháč 1, 4 1 Department of Organic Chemistry, Faculty of Natural Sciences, Comenius University in Bratislava, Ilkovičova 6, 842 15 Bratislava,
    [Show full text]