DOCSLIB.ORG

Antibacterial Agents: 1,4-Disubstituted 1,2,3-Triazole Analogs of the Oxazolidinone a Dissertation Presented to the Faculty Of

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Antibacterial Agents: 1,4-Disubstituted 1,2,3-Triazole Analogs of the Oxazolidinone a Dissertation Presented to the Faculty Of Antibacterial Agents: 1,4-Disubstituted 1,2,3-Triazole Analogs of the Oxazolidinone A dissertation presented to the faculty of the College of Arts and Sciences of Ohio University In partial fulfillment of the requirements for the degree Doctor of Philosophy George Acquaah-Harrison June 2010 © 2010 George Acquaah-Harrison. All Rights Reserved. 2 This dissertation titled Antibacterial Agents: 1,4-Disubstituted 1,2,3-Triazole Analogs of the Oxazolidinone by GEORGE ACQUAAH-HARRISON has been approved for the Department of Chemistry and Biochemistry and the College of Arts and Sciences by Stephen C. Bergmeier Professor of Chemistry and Biochemistry Benjamin M. Ogles Dean, College of Arts and Sciences 3 ABSTRACT ACQUAAH-HARRISON, GEORGE, Ph.D., June 2010, Chemistry and Biochemistry Antibacterial Agents: 1,4-Disubstituted 1,2,3-Triazole Analogs of the Oxazolidinone ) (235 pp.) Director of Dissertation: Stephen C. Bergmeier The rational design, development and synthesis of structurally diverse small molecule that target RNA is immensely important in antibacterial therapy. Utilizing rational design approach to drug discovery, two lead 4,5-disubstituted 2-oxazolidinone compounds that bind to the highly conserved region of bacterial RNA with high affinity and specificity had been previously identified. This biological target called the T box antiterminator system regulates the expression of many genes including aminoacyl synthethase genes and is found predominantly in Gram-positive pathogens. But, owing to the moderate solubilities of these leads, the focus was directed to improving the solubility challenges without compromising biological activity. To address the solubility challenges with the intent of enhancing or maintaining biological activity, a library of one hundred eight 1,4-disubstituted 1,2,3-triazole compounds that encompasses the diversity elements of the oxazolidinones were developed. This library, which entailed the bioisosteric replacement of the oxazolidinone scaffold was afforded in high yield and purity by employing the regioselective copper(I)-catalyzed azide/alkyne cycloaddition reaction. Three lead compounds, GHB-7, GHB-9 and GHB-16 with enhanced biological activity were identified that rendered them important candidates for structural activity 4 relationship studies (SAR). Besides the SAR studies, few 1,5-regioisomers were prepared to investigate the effect of regioselectivity on biological activity. By embarking on empirical observations, thirty-two structurally relevant analogs were prepared for the SAR and other structural elaboration studies. While biological evaluation is currently ongoing, the preliminary data of the analogs, GHB-144, GHB- 151, GHB-153, GHB-156 and GHB-157 coupled with the enhanced biological activity of GHB-7 relative to the lead oxazolidinone compounds re-inforce the plausibility of finding new 1,4-substituted 1,2,3-triazole compounds with improved antibacterial activity. Approved: _____________________________________________________________ Stephen C. Bergmeier Professor of Chemistry and Biochemistry 5 ACKNOWLEDGMENTS I would like to express my sincere gratitude to my academic advisor Dr. Stephen C. Bergmeier for his mentorship, guidance, insightful discussions and encouragements during my study. I would also like to thank the members of my dissertation committee Dr. Mark C. McMills, Dr. Glen Jackson and Dr. Xiaozhuo Chen for accepting my invitation to serve on my committee. Special thanks go to the faculty of the Department of Chemistry and Biochemistry, especially Dr Jared Butcher, Dr. Klaus Himmeldirk and Dr. Kumar Pichumani as well as the administrative staff of the department of Chemistry and biochemistry for their suggestions and support during my study. I am thankful to the members of Dr. Bergmeier’s research group both past (Dr. Pulipaka Aravinda, Dr. Junfeng Huang, Dr. Ahbigit Nayek, Nova Emerald and Dr. Iwona Maciagiewicz) and present (Dr. Crina M. Orac, Weihe Zhang, Fang Fang, Susann Krake, Gregg Wells and Dennis Roberts) for their suggestions and constructive criticisms as well as promoting a conducive laboratory atmosphere. I am also thankful to Shu Zhou in Dr. Hines laboratory for testing all triazole analogs. I am greatly indebted to my family: Harris Osafo-Acquaah, Mary F. Kusi- Appouh, Franklin Effah, Victoria Dankwa & family, Gina Peters & family, Kwabena Appiah Peprah, Mary Appiah Peprah and the rest of Peprah family as well as Dr. Isaac Blankson and Joseph Blankson for their incessant support, advice and prayers during my doctoral studies. 6 Finally to my wife, Vivian Acquaah-Harrison and three wonderful daughters, Ama Afrakomah Acquaah-Harrison, Nana-Afia Acquaah-Harrison and Nana-Akua Acquaah-Harrison, thank you for recognizing the importance of education. Your love, Charisma, encouragement, patience and motivation is what energized me through this study. 7 Dedicated to my wife Vivian A. Acquaah-Harrison and Three wonderful daughters Ama Afrakomah Acquaah-Harrison, Nana-Afia Antwiwaa Acquaah-Harrison and Nana-Akua Osafoaa Acquaah-Harrisonson 8 TABLE OF CONTENTS Page Abstract................................................................................................................................3 Acknowledgments................................................................................................................5 Dedication ...........................................................................................................................7 List of Tables .......................................................................................................................9 List of Figures....................................................................................................................10 List of Schemes..................................................................................................................12 Chapter 1.Introduction......................................................................................................15 1.1 Statement of purpose ........................................................................................... 15 1.2 Introduction.......................................................................................................... 15 Chapter 2.Background......................................................................................................23 2.1 History of antibiotics ........................................................................................... 23 2.1.1 Natural antibiotics......................................................................................... 24 2.1.2 Synthetic antibiotics...................................................................................... 32 2.1.3 Antibiotics: problems and challenges ........................................................... 35 2.2 Antibacterial drug discovery................................................................................ 37 2.3 RNA domain as a novel target for antimicrobial therapy.................................... 38 2.3.1 T box antiterminator RNA System ............................................................... 40 2.4 Identification of lead compound.......................................................................... 42 2.4.1 Lead compound modification/optimization.................................................. 44 2.5 Oxazolidinone compounds .................................................................................. 46 2.5.1 Oxazolidinone library generation ................................................................. 47 2.6 Significance of research....................................................................................... 51 Chapter 3.Synthesis and Preliminary Results of 1,4-Disubstituted 1,2,3-Triazole Library52 3.1 Introduction.......................................................................................................... 52 3.2 Retrosynthetic analysis of the triazole library ..................................................... 56 3.3 Synthesis of azide components............................................................................ 57 3.4 Synthesis of alkyne components.......................................................................... 65 3.5 Determination of reaction condition for 1,2,3-triazole library synthesis............. 68 9 3.5.1 Model studies ................................................................................................ 71 3.5.2 Determination of workup conditions ............................................................ 72 3.5.3 Generation of 1,4-disubstituted 1,2,3-triazole library................................... 74 3.6 Biological evaluation of 1,4-disubstituted 1,2,3-triazole compounds ................. 81 3.6.1 Fluorescence resonance energy transfer assay.............................................. 81 3.6.2 Antibacterial Assay....................................................................................... 87 3.7 Synthesis of 1,5-disubstituted 1,2,3-triazoles analogs......................................... 89 3.7.1 Biological evaluation of 1,5-analogs ............................................................ 94 3.8 Conclusion ........................................................................................................... 95 Chapter 4.Structure Activity Studies and Structural Elaborations of the 1,2,3-Triazole Ring 97 4.1 Introduction.......................................................................................................... 97 4.2 Plan
Load more

Recommended publications
  • Structure-Based Design, Synthesis, and Biological Evaluation Of
    Structure-Based Design, Synthesis, and Biological Evaluation of Triazole-Based smHDAC8 Inhibitors Dmitrii Kalinin, Sunit Jana, Maxim Pfafenrot, Alokta Chakrabarti, Jelena Melesina, Tajith Shaik, Julien Lancelot, Raymond Pierce, Wolfgang Sippl, Christophe Romier, et al. To cite this version: Dmitrii Kalinin, Sunit Jana, Maxim Pfafenrot, Alokta Chakrabarti, Jelena Melesina, et al.. Structure-Based Design, Synthesis, and Biological Evaluation of Triazole-Based smHDAC8 Inhibitors. ChemMedChem, Wiley-VCH Verlag, 2019, 15 (7), pp.571-584. 10.1002/cmdc.201900583. hal- 02999295 HAL Id: hal-02999295 https://hal.archives-ouvertes.fr/hal-02999295 Submitted on 23 Nov 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. Distributed under a Creative Commons Attribution| 4.0 International License Full Papers ChemMedChem doi.org/10.1002/cmdc.201900583 1 2 3 Structure-Based Design, Synthesis, and Biological 4 5 Evaluation of Triazole-Based smHDAC8 Inhibitors 6 [a, b, c, d] [c, e] [c] [f] 7 Dmitrii V. Kalinin, Sunit K. Jana, Maxim Pfafenrot, Alokta Chakrabarti, [g]
    [Show full text]
  • WO 2015/179249 Al 26 November 2015 (26.11.2015) P O P C T
    (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (10) International Publication Number (43) International Publication Date WO 2015/179249 Al 26 November 2015 (26.11.2015) P O P C T (51) International Patent Classification: (81) Designated States (unless otherwise indicated, for every C12N 15/11 (2006.01) A61K 38/08 (2006.01) kind of national protection available): AE, AG, AL, AM, C12N 15/00 (2006.01) AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DK, DM, (21) Number: International Application DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, PCT/US2015/031213 HN, HR, HU, ID, IL, IN, IR, IS, JP, KE, KG, KN, KP, KR, (22) International Filing Date: KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, 15 May 2015 (15.05.2015) MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, (25) Filing Language: English SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, (26) Publication Language: English TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW. (30) Priority Data: (84) Designated States (unless otherwise indicated, for every 62/000,43 1 19 May 2014 (19.05.2014) US kind of regional protection available): ARIPO (BW, GH, 62/129,746 6 March 2015 (06.03.2015) US GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, (72) Inventors; and TJ, TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, (71) Applicants : GELLER, Bruce, L.
    [Show full text]
  • Quantitative Analysis of Spectinomycin and Lincomycin in Poultry Eggs by Accelerated Solvent Extraction Coupled with Gas Chromatography Tandem Mass Spectrometry
    foods Article Quantitative Analysis of Spectinomycin and Lincomycin in Poultry Eggs by Accelerated Solvent Extraction Coupled with Gas Chromatography Tandem Mass Spectrometry Bo Wang 1,2, Yajuan Wang 2,3, Xing Xie 4, Zhixiang Diao 2,3, Kaizhou Xie 2,3,*, Genxi Zhang 2,3 , Tao Zhang 2,3 and Guojun Dai 2,3 1 College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China; [email protected] 2 Joint International Research Laboratory of Agriculture & Agri-Product Safety, Yangzhou University, Yangzhou 225009, China; [email protected] (Y.W.); [email protected] (Z.D.); [email protected] (G.Z.); [email protected] (T.Z.); [email protected] (G.D.) 3 College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China 4 Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture, Nanjing 210014, China; [email protected] * Correspondence: [email protected]; Tel.: +86-139-5275-0925 Received: 30 March 2020; Accepted: 13 May 2020; Published: 18 May 2020 Abstract: A method based on accelerated solvent extraction (ASE) coupled with gas chromatography tandem mass spectrometry (GC-MS/MS) was developed for the quantitative analysis of spectinomycin and lincomycin in poultry egg (whole egg, albumen and yolk) samples. In this work, the samples were extracted and purified using an ASE350 instrument and solid-phase extraction (SPE) cartridges, and the parameters of the ASE method were experimentally optimized. The appropriate SPE cartridges were selected, and the conditions for the derivatization reaction were optimized. After derivatization, the poultry egg (whole egg, albumen and yolk) samples were analyzed by GC-MS/MS.
    [Show full text]
  • Oxazolidinones for TB
    Oxazolidinones for TB: Current Status and Future Prospects 12th International Workshop on Clinical Pharmacology of Tuberculosis Drugs London, UK 10 September 2019 Lawrence Geiter, PhD Disclosures • Currently contract consultant with LegoChem Biosciences, Inc., Daejeon, Korea (LCB01-0371/delpazolid) • Previously employed with Otsuka Pharmaceutical Development and Commercialization, Inc. (delamanid, OPC-167832, LAM assay) What are Oxazolidinones • A family of antimicrobials mostly targeting an early step in protein synthesis • Cycloserine technically oxazolidinone but 2-oxazolidinone different MOA and chemical properties • New generation oxazolidinones bind to both 50S subunit and 30S subunit • Linezolid (Zyvox) and Tedizolid (Sivestro) approved for drug resistant skin infections and community acquired pneumonia Cycloserine • Activity against TB demonstrated in non- clinical and clinical studies • Mitochondrial toxicity >21 days limits use in TB treatment Linezolid Developing Oxazolidinones for TB Compound Generic Brand Sponsor Development Status TB Code- Activity/Trials PNU-100766 Linezolid Zyvox Pfizer Multiple regimen Yes/Yes TR-201 Tedizolid Sivextro Merck Pre-clinical efficacy Yes/No PNU-100480 Sutezolid Pfizer Multiple regimen studies Sequella Yes/Yes (PanACEA) TB Alliance LCB01-0371 Delpazolid LegoChem Bio EBA trial recruitment completed Yes/Yes TBI-223 - Global Alliance SAD trial launched Yes/Yes AZD5847 Posizolid AstraZenica Completed EBA Yes/No RX-1741 Radezolid Melinta IND for vaginal infections ?/No RBX-7644 Ranbezolid Rabbaxy None found ?/No MRX-4/MRX-1 Contezolid MicuRx Skin infections Yes/No U-100592 Eperezolid ? No clinical trials ?/No PK of Oxazolidinones in Development for TB Steady State PK Parameters Parameter Linezolid 600 Delpazolid 800 mg QD2 mg QD3 Cmax (mg/L) 17.8 8.9 Cmin (mg/L) 2.43 0.1 Tmax (h) 0.87 0.5 T1/2 (h) 3.54 1.7 AUC0-24 (µg*h/mL) 84.5 20.1 1 MIC90 (µg/mL) 0.25 0.5 References 1.
    [Show full text]
  • Triazole Fungicide Residues and Their Inhibitory Effect on Some Trichothecenes Mycotoxin Excretion in Wheat Grains
    molecules Article Triazole Fungicide Residues and Their Inhibitory Effect on Some Trichothecenes Mycotoxin Excretion in Wheat Grains Tamer M. A. Thabit 1,2, Eman M. Abdelkareem 3,* , Nahla A. Bouqellah 4 and Shokr A. Shokr 1 1 Central Agricultural Pesticides Laboratory (CAPL), Agricultural Research Center (ARC), Giza 12611, Egypt; [email protected] (T.M.A.T.); [email protected] (S.A.S.) 2 Saudi Arabia Grains Organization (SAGO), Riyadh 11471, Saudi Arabia 3 Plant Pathology Research Institute, Agricultural Research Center (ARC), Giza 12619, Egypt 4 Biology Department, Collage of Science, Taibah University, Al-Madinah Al-Munawarah 344, Saudi Arabia; [email protected] * Correspondence: [email protected] Abstract: Wheat is one of the global strategic crops and ranks third in terms of cereals production. Wheat crops are exposed to many fungal infections during their cultivation stages, some of which have the ability to secrete a number of toxic secondary metabolites that threaten the quality of the grains, consumer health, producer economics, and global trade exchange. Fifty-four random samples were collected from wheat which originated from different countries. The samples included 14 types of soft wheat to study the extent of their contamination with deoxynivalenol (DON) and T-2 toxin by auto-ELISA technology and r-biopharm microtiter plate. All samples were contaminated with −1 DON toxin except one sample, and the values ranged between 40.7 and 1018.8 µg/kg . The highest contamination rates were in Lithuanian wheat and the lowest was in Indian wheat. Meanwhile, the Citation: Thabit, T.M.A.; highest average level of T-2 toxin contamination was in Lithuanian wheat grains with 377.4 µg/kg−1, Abdelkareem, E.M.; Bouqellah, N.A.; and the lowest average was 115.3 µg/kg−1 in Polish wheat.
    [Show full text]
  • Dissertation
    DISSERTATION FOLATE PATHWAY INHIBITOR RESISTANCE MECHANISMS IN BURKHOLDERIA PSEUDOMALLEI Submitted by Nicole L. Podnecky Department of Microbiology, Immunology and Pathology In partial fulfillment of the requirements For the Degree of Doctor of Philosophy Colorado State University Fort Collins, Colorado Fall 2013 Doctoral Committee: Advisor: Herbert P. Schweizer Steven W. Dow Richard A. Slayden Laurie A. Stargell ABSTRACT FOLATE PATHWAY INHIBITOR RESISTANCE MECHANISMS IN BURKHOLDERIA PSEUDOMALLEI Antimicrobials are invaluable tools used to facilitate the treatment of infectious diseases. Their use has saved millions of lives since their introduction in the early 1900’s. Unfortunately, due to the increased incidence and dispersal of antimicrobial resistance determinants, many of these drugs are no longer efficacious. This greatly limits the options available for treatment of serious bacterial infections, including melioidosis, which is caused by Burkholderia pseudomallei, a Gram-negative saprophyte. This organism is intrinsically resistant to many antimicrobials. Additionally, there have been reports of B. pseudomallei isolates resistant to several of the antimicrobials currently used for treatment, including the trimethoprim and sulfamethoxazole combination, co-trimoxazole. The overarching goal of this project was to identify and characterize mechanisms of trimethoprim and sulfamethoxazole resistance in clinical and environmental isolates, as well as in laboratory induced mutants. Prior to these studies, very little work has been done to identify and characterize the mechanisms by which B. pseudomallei strains are or could become resistant to folate-pathway inhibitors, specifically trimethoprim and sulfamethoxazole. During the initial phases of these studies, we determined the antimicrobial susceptibilities of a large collection of clinical and environmental isolates from Thailand and Australia (n = 65).
    [Show full text]
  • 1,4-Disubstituted-1,2,3-Triazole Compounds Induce Ultrastructural Alterations in Leishmania Amazonensis Promastigote
    International Journal of Molecular Sciences Article 1,4-Disubstituted-1,2,3-Triazole Compounds Induce Ultrastructural Alterations in Leishmania amazonensis Promastigote: An in Vitro Antileishmanial and in Silico Pharmacokinetic Study Fernando Almeida-Souza 1,2,* , Verônica Diniz da Silva 3,4, Gabriel Xavier Silva 5, Noemi Nosomi Taniwaki 6, Daiana de Jesus Hardoim 2, Camilla Djenne Buarque 3, 1, , 2, Ana Lucia Abreu-Silva * y and Kátia da Silva Calabrese y 1 Pós-graduação em Ciência Animal, Universidade Estadual do Maranhão, São Luís 65055-310, Brazil 2 Laboratório de Imunomodulação e Protozoologia, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro 21040-900, Brazil; [email protected] (D.d.J.H.); calabrese@ioc.fiocruz.br (K.d.S.C.) 3 Laboratório de Síntese Orgânica, Pontifícia Universidade Católica, Rio de Janeiro 22451-900, Brazil; [email protected] (V.D.d.S.); [email protected] (C.D.B.) 4 Faculdade de Ciência e Tecnologia, Universidade Nova de Lisboa, 2825-149 Caparica, Portugal 5 Rede Nordeste de Biotecnologia, Universidade Federal do Maranhão, São Luís 65080-805, Brazil; [email protected] 6 Núcleo de Microscopia Eletrônica, Instituto Adolfo Lutz, São Paulo 01246-000, Brazil; [email protected] * Correspondence: [email protected] (F.A.-S.); [email protected] (A.L.A.-S.) These authors equally contributed to this work. y Received: 26 June 2020; Accepted: 14 July 2020; Published: 18 September 2020 Abstract: The current standard treatment for leishmaniasis has remained the same for over 100 years, despite inducing several adverse effects and increasing cases of resistance. In this study we evaluated the in vitro antileishmanial activity of 1,4-disubstituted-1,2,3 triazole compounds and carried out in silico predictive study of their pharmacokinetic and toxicity properties.
    [Show full text]
  • (12) Patent Application Publication (10) Pub. No.: US 2012/0115729 A1 Qin Et Al
    US 201201.15729A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2012/0115729 A1 Qin et al. (43) Pub. Date: May 10, 2012 (54) PROCESS FOR FORMING FILMS, FIBERS, Publication Classification AND BEADS FROM CHITNOUS BOMASS (51) Int. Cl (75) Inventors: Ying Qin, Tuscaloosa, AL (US); AOIN 25/00 (2006.01) Robin D. Rogers, Tuscaloosa, AL A6II 47/36 (2006.01) AL(US); (US) Daniel T. Daly, Tuscaloosa, tish 9.8 (2006.01)C (52) U.S. Cl. ............ 504/358:536/20: 514/777; 426/658 (73) Assignee: THE BOARD OF TRUSTEES OF THE UNIVERSITY OF 57 ABSTRACT ALABAMA, Tuscaloosa, AL (US) (57) Disclosed is a process for forming films, fibers, and beads (21) Appl. No.: 13/375,245 comprising a chitinous mass, for example, chitin, chitosan obtained from one or more biomasses. The disclosed process (22) PCT Filed: Jun. 1, 2010 can be used to prepare films, fibers, and beads comprising only polymers, i.e., chitin, obtained from a suitable biomass, (86). PCT No.: PCT/US 10/36904 or the films, fibers, and beads can comprise a mixture of polymers obtained from a suitable biomass and a naturally S3712). (4) (c)(1), Date: Jan. 26, 2012 occurring and/or synthetic polymer. Disclosed herein are the (2), (4) Date: an. AO. films, fibers, and beads obtained from the disclosed process. O O This Abstract is presented solely to aid in searching the sub Related U.S. Application Data ject matter disclosed herein and is not intended to define, (60)60) Provisional applicationpp No. 61/182,833,sy- - - s filed on Jun.
    [Show full text]
  • Molecular and Immunological Sd0100025 Characterization of Mycobacteria Associated with Bovine Farcy
    MOLECULAR AND IMMUNOLOGICAL SD0100025 CHARACTERIZATION OF MYCOBACTERIA ASSOCIATED WITH BOVINE FARCY. Thesis submitted in accordance with requirements of the University of Khartoum for the Degree of Doctor of Philosophy (Ph.D). By Victor Loku Kwajok B.V.Med. 1978, University of Cairo/Egypt. M.V.Sc.1992, University of Khartoum/Sudan. Supervisor: Dr. Maowia M. Mukhtar. Institute of Endemic Diseases, University of Khartoum. Department of Preventive Medicine and Veterinary Public Health. Faculty of Veterinary Science, University of Khartoum. 2000. 32/27 SOME PAGES ARE MISSING IN THE ORIGINAL DOCUMENT LIST OF CONTENTS DEDICATION. * ii v ACKNOWLEDGEMENTS vi ABBREVIATIONS viii ABSTRACT. xi CHAPTER ONE. REVIEW OF LITERATURE. 1. General Introduction. 1. 1.1. Molecular Systematics of genus Mycobacterium. 2. 1.2. Molecular taxonomy of M. farcinogenese and M. senegalense 11. 1.3. Immunology of bovine farcy agents. 36. CHAPTER TWO. MATERIALS AND METHODOLOGY 41. Isolation, identification and characterization of M. farcinogenes. 2.1 Phenotypic characterization. 41. 2.1.1. Morphological and biochemical tests. 2.1.2. Degradation tests. 2.1.3. Rapid fluorogenic enzyme tests. 2.1.4. Nutritional tests. 2.1.5. Physiological tests. 2.2. Molecular characterization. 52. 2.2.1. DNA extraction and purification 2.2.2. PCR amplification and application. 2.2.3. DNA sequencing of 16SrDNA. 2.2.4. PCR-based restriction fragment length polymorphism. 2.3 Imrmmological analyses of bovine farcy agents. 71. 2.3.1 .EL1SA technique for sera diagnosis. 2.3.2 Animal pathogenicity Tests. 2.3.3 Protein antigen profiles determination using. CHAPTER THREE: RESULTS. 78. CHAPTER. FOUR: DISCUSSION. 124. REFERENCES. 133.
    [Show full text]
  • Antibacterial Drugs
    Antibacterial drugs Iwona Smolarek, MD, PhD Department of Clinical Pharmacology The antibacterial drugs destroy or slow down the growth of bacteria They are widely used in medicine and dentistry. They are administered generally and locally, in prophylaxis and treatment. ANTIBIOTICS antimicrobial agents produced by microorganisms synthetic drugs designed based upon the structure of molecules produced in nature by microorganisms CHEMOTHERAPEUTIC AGENTS antimicrobial agents synthesized in the laboratory Colloquially – all antibacterial agents are named antibiotics Antibiotics classification and mechanism of action Antibiotics classification bacteriostatic – slow growth or reproduction of bacteria (macrolides, lincosamides, tetracyclines, sulfonamides and chloramphenicol) bactericidal - kill bacteria (β-lactams, vancomycin, daptomycin, fluoroquinolones, metronidazole, co-trimoxazole) The classification is in part arbitrary because most bacteriostatic drugs are bactericidal at high concentrations, under certain incubation conditions in vitro, and against some bacteria. Β - lactams inhibit bacterial growth by interfering with bacterial cell wall synthesis - bactericidal this group of antibiotic include some groups such as penicillins, cephalosporins, monobactams, carbapenems, and β – lactamase inhibitors Penicillins - classification The penicillin group include: prototype: penicillin G, penicillin V penicillinase – resistant: nafcillin, methilcillin, oxacillin, cloxacillin, dicloxacillin extendend – spectrum: ampicillin, amoxicillin,
    [Show full text]
  • Antibiotic Sensitivity of Bacterial Pathogens Isolated from Bovine Mastitis Milk
    Content of Research Report A. Project title: Antibiotic Sensitivity of Bacterial Pathogens Isolated From Bovine Mastitis Milk B. Abstract: Antibiotic sensitivity of bacteria isolated from bovine milk samples was investigated. The 18 antibiotics that were evaluated (e.g., penicillin, novobiocin, gentamicin) are commonly used to treat various diseases in cattle, including mastitis, an inflammation of the udder of dairy cows. A common concern in using antibiotics is the increase in drug resistance with time. This project studies if antibiotic resistance is a threat to consumers of raw milk products and if these antibiotics are still effective against mastitis pathogens. The study included isolating and culturing bacteria from quarter milk samples (n=205) collected from mastitic dairy cows from farms in Chino and Ontario, CA. The isolated bacteria were tested for sensitivity to antibiotics using the Kirby Bauer disk diffusion method. The prevalence (%) of resistance to the individual antibiotics was reported. Resistance to penicillin was 45% which may support previous data on penicillin-resistant bacteria, especially Staphylococcus and Streptococcus. Resistance rates (%) for oxytetracycline (26.8%) and tetracycline (22.9%) were low compared to previous studies but a trend was seen in our results that may support concerns of emerging resistance to tetracyclines in both gram- positive and gram-negative bacteria. Similarly, 31.9% of bacterial isolates showed resistance to erythromycin which is at least 30% less than in reported literature concerning emerging resistance to macrolides. More numbers (%) that should be noted are cefazolin (26.3%), ampicillin (29.7%), novobiocin (33.0%), polymyxin B (31.5%), and resistance ranging from 9­ 18% for the other antibiotics.
    [Show full text]
  • Unexpected Drug Residuals in Human Milk in Ankara, Capital of Turkey Ayşe Meltem Ergen1 and Sıddıka Songül Yalçın2*
    Ergen and Yalçın BMC Pregnancy and Childbirth (2019) 19:348 https://doi.org/10.1186/s12884-019-2506-1 RESEARCH ARTICLE Open Access Unexpected drug residuals in human milk in Ankara, capital of Turkey Ayşe Meltem Ergen1 and Sıddıka Songül Yalçın2* Abstract Background: Breast milk is a natural and unique nutrient for optimum growth and development of the newborn. The aim of this study was to investigate the presence of unpredictable drug residues in mothers’ milk and the relationship between drug residues and maternal-infant characteristics. Methods: In a descriptive study, breastfed infants under 3 months of age and their mothers who applied for child health monitoring were enrolled for the study. Information forms were completed for maternal-infant characteristics, breastfeeding problems, crying and sleep characteristics of infants. Maternal and infant anthropometric measurements and maternal milk sample were taken. Edinburgh Postpartum Depression Scale was applied to mothers. RANDOX Infiniplex kit for milk was used for residual analysis. Results: Overall, 90 volunteer mothers and their breastfed infants were taken into the study and the mean age of the mothers and their infants was 31.5 ± 4.2 years and 57.8 ± 18.1 days, respectively. Anti-inflammatory drug residues in breast milk were detected in 30.0% of mothers and all had tolfenamic acid. Overall, 94.4% had quinolone, 93.3% beta-lactam, 31.1% aminoglycoside and 13.3% polymycin residues. Drugs used during pregnancy or lactation period were not affected by the presence of residues. Edinburgh postpartum depression scores of mothers and crying and sleeping problems of infants were similar in cases with and without drug residues in breast milk.
    [Show full text]