DOCSLIB.ORG

Mycobacterial Glycolipids – Pathways to Synthesis and Role in Virulence

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text

Load more

Mycobacterial Glycolipids – Pathways to Synthesis and Role in Virulence by DEBASMITA SARKAR A thesis submitted to The University of Birmingham for the Degree of Doctor of Philosophy School of Biosciences College of Life and Environmental Sciences The University of Birmingham February 2012 Abstract Mycobacterial diseases are responsible for numerous deaths worldwide, the major pathogens being Mycobacterium tuberculosis and Mycobacterium leprae. Also, in recent years threats from opportunistic pathogens, such as Mycobacterium marinum and Mycobacterium kansasii have been on the rise. These mycobacteria possess a unique lipid-rich cell wall with an array of mycolic acids and species-specific antigenic glycolipids, like the lipooligosaccharides. Some of these solvent extractable lipids possess immunomodulatory properties and play an important role during infection. Lipooligosaccharides (LOS) are surface exposed, polar, antigenic glycolipids that are present in several mycobacterial species. This study used the opportunistic human pathogens M. marinum and M. kansasii as a model system to unravel the genes involved in the biosynthesis of LOSs in Mycobacterium. Using directed mutagenesis and transposon mutagenesis, mutant strains defective in various parts of the LOS biosynthetic pathway were isolated. Analysis of these strains helped in further delineating the pathway and understanding the role of LOSs in virulence. A part of this thesis focussed on studying mycolic acid processing and transport using Mycobacterium smegmatis as a surrogate system. Mycolic acids are the most distinctive components of the mycobacterial cell wall. While their biosynthesis has been studied in detail, processing and transport across the membrane is not well understood. This study attempted to explore the roles of the two putative type II mycolyltransferases MSMEG3437 and MSMEG5851 in mycolic acid processing. Additionally, the role of M. tuberculosis mmpL11 gene was probed as the Mtb-mmpL11 had been reported to be involved in virulence. A null i mutant of the M. smegmatis homologue, Ms-mmpL11 (MSMEG0241) was generated and analysed for the above study. Deletion mutant strains of the two putative mycolyltransferase II did not show any phenotype, suggesting that their roles are redundant in vivo. Although the Ms-mmpL gene was found to be non-essential, it was found to be involved in transport of free mycolic acids. ii Declaration The work presented in this thesis was carried out in the School of Biosciences at the University of Birmingham, U.K., B15 2TT during the period November 2008 to November 2011. The work in this thesis is original except where acknowledged by references. No portion of the work is being, or has been submitted for a degree, diploma or any other qualification at any other University. iii University of Birmingham Research Archive E-theses repository This unpublished thesis/dissertation is copyright of the author and/or third parties. The intellectual property rights of the author or third parties in respect of this work are as defined by The Copyright Designs and Patents Act 1988 or as modified by any successor legislation. Any use made of information contained in this thesis/dissertation must be in accordance with that legislation and must be properly acknowledged. Further distribution or reproduction in any format is prohibited without the permission of the copyright holder. iv Acknowledgement The last three years have been the most rewarding time of my life. I have learnt a lot and looking back, I did not even think I would do a PhD. This thesis would not have come to life without the encouragement of my family especially my father and my husband, help and support of my supervisors, both past and present, friends, and colleagues. I am grateful to Professor Gurdyal S. Besra, my supervisor for granting me the opportunity to pursue my PhD studies inspite of a very nervous telephone interview. I would like to express my gratitude to my supervisor, Dr. Apoorva Bhatt for all the support, guidance, patience and help with my experiments, for making my experience easier and achievable, for help with writing and giving me confidence in myself. I would like to express my gratitude to the Darwin Trust of Edinburgh, whose funding made this thesis possible. I would also like to thank Professor David E. Minnikin for his valuable comments and time. My time in the ‘Besra Lab’ has been very fulfilling both professionally and personally. It has not only been work but an opportunity to know and work with brilliant scientists and amazing friends. I would like to thank Sid and Luke, for teaching me numerous techniques. Thank you Albel for providing your patient assistance with many techniques and help with all the photography that appear in this thesis. A massive thanks to Natacha for teaching chemistry to a ‘biologist’ and for all the help and encouragement throughout. I want to thank Usha, Veemal, Amrita and Arun for all the help, support and encouragement when i needed it most. A big thank you to the wonderful friends I have made in amazing biologists - Vicky, Monika, Shipra, Jiemin, Oona, Helen, Becci, Cristian, Sarah, Georgina, Liz, Hemza, Talat, Rana, v Reenette and brilliant chemists Ting, Justyna, Peter and Petr and all members past and present. Thank you! I would like to thank Dr. David A. Lammas (Institute of Biomedical Research, University of Birmingham), for all the help with tissue culture. I would like to thank Dr. Aniek van der Woude, Dr. Astrid van der Sar, Prof. W. Bitter and their lab (VU Medical centre, The Netherlands) for the transposon mutant library (detailed in Chapter 4) and zebrafish infection experiments. Also, huge thanks to Dr. P. Ashton, Mr. N. G. May and Dr. N. Spencer for their technical superiority in performing the MALDI-MS and NMR. I acknowledge the help of friends and mentors, Dr. Lakshmi Ramachandran, Dr. J. Venkatraman, Dr. Kaveri Das, and many colleagues at Astrazeneca India, without their encouragement i would not have started to pursue my PhD. Finally, to my family for their unconditional love and support. I wish Dida, Dadu, Mesomoni were here today! Thank you Baba, Ma, Masimoni, Boro Masimoni, Mamon, Boro, Mejo and Choto Pisi, Thakuma for believing in me. Big thanks to my aunt and uncle in Blackpool for their help and support. I am thankful to my dear friends – Preeti, Sumedha, Naveen, Reena, Shubha, Shanti, Disha, Moumita, Chandrima, Moitree for being there always. I also express my thanks to the wonderful friends I have made in Birmingham, Sonali, Ravikiran, Manmeet, Bhaskar, Neelam, Janvi, Pratik, Panchi, Yogi, Indrani, Amshika, Amit, Saptarshi and Bharat Parivar members for giving me the much needed break away from lab and work. Thank you all, it has not been easy being away from home, but your support made it possible. Last but not the least I would like to acknowledge the love, support and encouragement of an amazing person - Nitin, my best friend, now my husband. This thesis would not have been possible without you. Thank you! vi This thesis is dedicated to my Family vii Table of contents Abstract ...................................................................................................................... i Declaration ............................................................................................................... iii University of Birmingham Research Archive ......................................................... iv Acknowledgement ..................................................................................................... v Table of contents .................................................................................................... viii List of Abbreviations ............................................................................................xxvi Published work associated ....................................................................................xxxi Chapter 1 General Introduction ............................................................................... 1 1.1 Mycobacterium species................................................................................... 2 1.2 Tuberculosis .................................................................................................. 4 1.3 Epidemiology ................................................................................................. 5 1.4 Immune response to Mycobacterium tuberculosis infection ......................... 7 1.5 Treatment .....................................................................................................11 1.5.1 Anti –TB drugs .................................................................................................... 11 1.5.2 Directly Observed Therapy, Short-course ............................................................. 15 1.6 Drug resistance .............................................................................................16 1.6.1 Multi drug resistant TB (MDR-TB)...................................................................... 17 1.6.2 Extensively drug resistant TB (XDR-TB) ............................................................. 19 viii 1.7 The Mycobacterium tuberculosis complex (MTBC).....................................20 1.8 Non-tuberculous mycobacteria ....................................................................22 1.9 Surrogate systems in tuberculosis research - ..............................................22 1.9.1 Corynebacterium glutamicum .............................................................................
Recommended publications
  • A Case of Miliary Tuberculosis Presenting with Whitlow of the Thumb

    A Case of Miliary Tuberculosis Presenting with Whitlow of the Thumb

    A Case of Miliary Tuberculosis Presenting with Whitlow of the Thumb Romaric Larcher, Albert Sotto, Jean-Marc Mauboussin, Jean-Philippe Lavigne, François-Xavier Blanc, Didier Laureillard To cite this version: Romaric Larcher, Albert Sotto, Jean-Marc Mauboussin, Jean-Philippe Lavigne, François-Xavier Blanc, et al.. A Case of Miliary Tuberculosis Presenting with Whitlow of the Thumb. Acta Dermato- Venereologica, Society for Publication of Acta Dermato-Venereologica, 2016, 96 (4), pp.560 - 561. 10.2340/00015555-2285. hal-01909474 HAL Id: hal-01909474 https://hal.archives-ouvertes.fr/hal-01909474 Submitted on 25 May 2021 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 - NonCommercial| 4.0 International License Acta Derm Venereol 2016; 96: 560–561 SHORT COMMUNICATION A Case of Miliary Tuberculosis Presenting with Whitlow of the Thumb Romaric Larcher1, Albert Sotto1*, Jean-Marc Mauboussin1, Jean-Philippe Lavigne2, François-Xavier Blanc3 and Didier Laureillard1 1Infectious Disease Department, 2Department of Microbiology, University Hospital Caremeau, Place du Professeur Robert Debré, FR-0029 Nîmes Cedex 09, and 3L’Institut du Thorax, Respiratory Medicine Department, University Hospital, Nantes, France. *E-mail: [email protected] Accepted Nov 10, 2015; Epub ahead of print Nov 11, 2015 Tuberculosis remains a major public health concern, accounting for millions of cases and deaths worldwide.
  • S1 Sulfate Reducing Bacteria and Mycobacteria Dominate the Biofilm

    S1 Sulfate Reducing Bacteria and Mycobacteria Dominate the Biofilm

    Sulfate Reducing Bacteria and Mycobacteria Dominate the Biofilm Communities in a Chloraminated Drinking Water Distribution System C. Kimloi Gomez-Smith 1,2 , Timothy M. LaPara 1, 3, Raymond M. Hozalski 1,3* 1Department of Civil, Environmental, and Geo- Engineering, University of Minnesota, Minneapolis, Minnesota 55455 United States 2Water Resources Sciences Graduate Program, University of Minnesota, St. Paul, Minnesota 55108, United States 3BioTechnology Institute, University of Minnesota, St. Paul, Minnesota 55108, United States Pages: 9 Figures: 2 Tables: 3 Inquiries to: Raymond M. Hozalski, Department of Civil, Environmental, and Geo- Engineering, 500 Pillsbury Drive SE, Minneapolis, MN 554555, Tel: (612) 626-9650. Fax: (612) 626-7750. E-mail: [email protected] S1 Table S1. Reference sequences used in the newly created alignment and taxonomy databases for hsp65 Illumina sequencing. Sequences were obtained from the National Center for Biotechnology Information Genbank database. Accession Accession Organism name Organism name Number Number Arthrobacter ureafaciens DQ007457 Mycobacterium koreense JF271827 Corynebacterium afermentans EF107157 Mycobacterium kubicae AY373458 Mycobacterium abscessus JX154122 Mycobacterium kumamotonense JX154126 Mycobacterium aemonae AM902964 Mycobacterium kyorinense JN974461 Mycobacterium africanum AF547803 Mycobacterium lacticola HM030495 Mycobacterium agri AY438080 Mycobacterium lacticola HM030495 Mycobacterium aichiense AJ310218 Mycobacterium lacus AY438090 Mycobacterium aichiense AF547804 Mycobacterium
  • Successful Drug Discovery Informed by Actinobacterial Systematics

    Successful Drug Discovery Informed by Actinobacterial Systematics

    Successful Drug Discovery Informed by Actinobacterial Systematics Verrucosispora HPLC-DAD analysis of culture filtrate Structures of Abyssomicins Biological activity T DAD1, 7.382 (196 mAU,Up2) of 002-0101.D V. maris AB-18-032 mAU CH3 CH3 T extract H3C H3C Antibacterial activity (MIC): S. leeuwenhoekii C34 maris AB-18-032 175 mAU DAD1 A, Sig=210,10 150 C DAD1 B, Sig=230,10 O O DAD1 C, Sig=260,20 125 7 7 500 Rt 7.4 min DAD1 D, Sig=280,20 O O O O Growth inhibition of Gram-positive bacteria DAD1 , Sig=310,20 100 Abyssomicins DAD1 F, Sig=360,40 C 75 DAD1 G, Sig=435,40 Staphylococcus aureus (MRSA) 4 µg/ml DAD1 H, Sig=500,40 50 400 O O 25 O O Staphylococcus aureus (iVRSA) 13 µg/ml 0 CH CH3 300 400 500 nm 3 DAD1, 7.446 (300 mAU,Dn1) of 002-0101.D 300 mAU Mode of action: C HO atrop-C HO 250 atrop-C CH3 CH3 CH3 CH3 200 H C H C H C inhibitior of pABA biosynthesis 200 Rt 7.5 min H3C 3 3 3 Proximicin A Proximicin 150 HO O HO O O O O O O O O O A 100 O covalent binding to Cys263 of PabB 100 N 50 O O HO O O Sea of Japan B O O N O O (4-amino-4-deoxychorismate synthase) by 0 CH CH3 CH3 CH3 3 300 400 500 nm HO HO HO HO Michael addition -289 m 0 B D G H 2 4 6 8 10 12 14 16 min Newcastle Michael Goodfellow, School of Biology, University Newcastle University, Newcastle upon Tyne Atacama Desert In This Talk I will Consider: • Actinobacteria as a key group in the search for new therapeutic drugs.
  • Phylogenomic Analysis of the Species of the Mycobacterium Tuberculosis

    Phylogenomic Analysis of the Species of the Mycobacterium Tuberculosis

    Phylogenomic analysis of the species of the Mycobacterium tuberculosis complex demonstrates that Mycobacterium africanum, Mycobacterium bovis, Mycobacterium caprae, Mycobacterium microti and Mycobacterium pinnipedii are later heterotypic synonyms of Mycobacterium tuberculosis Marco Riojas, Katya Mcgough, Cristin Rider-Riojas, Nalin Rastogi, Manzour Hernando Hazbón To cite this version: Marco Riojas, Katya Mcgough, Cristin Rider-Riojas, Nalin Rastogi, Manzour Hernando Hazbón. Phy- logenomic analysis of the species of the Mycobacterium tuberculosis complex demonstrates that My- cobacterium africanum, Mycobacterium bovis, Mycobacterium caprae, Mycobacterium microti and Mycobacterium pinnipedii are later heterotypic synonyms of Mycobacterium tuberculosis. Inter- national Journal of Systematic and Evolutionary Microbiology, Microbiology Society, 2018, 68 (1), pp.324-332. 10.1099/ijsem.0.002507. pasteur-01986654 HAL Id: pasteur-01986654 https://hal-riip.archives-ouvertes.fr/pasteur-01986654 Submitted on 18 Jan 2019 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. RESEARCH ARTICLE Riojas et al., Int J Syst Evol Microbiol 2018;68:324–332 DOI 10.1099/ijsem.0.002507 Phylogenomic analysis of the species of the Mycobacterium tuberculosis complex demonstrates that Mycobacterium africanum, Mycobacterium bovis, Mycobacterium caprae, Mycobacterium microti and Mycobacterium pinnipedii are later heterotypic synonyms of Mycobacterium tuberculosis Marco A.
  • Numerical Analysis of Rapidly Growing, Scotochromogenic Mycobacteria, Including Mycobacterium O Buense Sp

    Numerical Analysis of Rapidly Growing, Scotochromogenic Mycobacteria, Including Mycobacterium O Buense Sp

    INTERNATIONALJOURNAL OF SYSTEMATICBACTERIOLOGY, July 1981, p. 263-275 Vol. 31, No. 3 0020-7713/81/030263-13$02.00/0 Numerical Analysis of Rapidly Growing, Scotochromogenic Mycobacteria, Including Mycobacterium o buense sp. nov., norn. rev., Mycobacterium rhodesiae sp. nov., nom. rev., Mycobacterium aichiense sp. nov., norn. rev., Mycobacterium chubuense sp. nov., norn. rev., and Mycobacterium tokaiense sp. nov., nom. rev. MICHIO TSUKAMURA, SHOJI MIZUNO, AND SUM10 TSUKAMURA The National Chubu Hospital, Obu, Aichi, Japan 474 We performed numerical analyses of 155 strains of rapidly growing, scotochrom- ogenic mycobacteria by two different methods; in one method we used 104 characters, and in the other we used 84 characters. The following taxa appeared as distinct clusters: Myco bacterium thermoresistibile, Myco bacterium flavescens, Mycobacterium duvalii, Mycobacterium phlei, “Mycobacterium o buense,” My- co bacterium parafortuitum, Mycobacterium vaccae, Mycobacterium sphagni, “Mycobacterium aichiense,” “Mycobacterium rhodesiae,” Mycobacterium neoaurum, “Mycobacterium chubuense,” “Mycobacterium tokaiense,” and My- cobacterzum komossense (names in quotation marks are not on the Approved Lists of Bacterial Names). M. flavescens strains were divided into two subgroups, one consisting of strains isolated in Japan and the other consisting of strains isolated in Rhodesia and strains received from the American Type Culture Collection, including the type strain of M. flavescens (ATCC 14474). We found that there are many species of rapidly growing, scotochromogenic mycobacteria, and we believe that new species should be recognized and named on the basis of at least three strains. The following species appeared to be distinct from all presently named species: “Mycobacteriumgallinarum,” “Mycobacterium armen- tun,” “Mycobacterium pelpallidurn,” and “Mycobacterium taurus.” However, each of these species was proposed on the basis of only one or two strains.
  • Role of Immunotherapy in the Treatment of Tuberculosis

    Role of Immunotherapy in the Treatment of Tuberculosis

    ROLE OF IMMUNOTHERAPY IN THE TREATMENT OF TUBERCULOSIS MURTHY, K.J.R. VIJAYA LAKSHMI, V. and Singh, S. Bhagwan Mahavir Medical Research Centre, 10-1-1, Mahavir Marg, Hyderabad - 500 029. India. ABSTRACT Tuberculosis is caused by Mycobacterium tuberculosis, an intracellular pathogen residing in macrophages. Cell mediated immune (CMI) and delayed type of hypersensitive (DTH) responses play a pivotal role in providing protection to the host. The most important cell is the CD4 T lymphocyte, which is divided into TH1 and TH2 subsets depending on the type of cytokines produced. TH1 cells produce the cytokines interferon-gamma and interleukin-2, which are important for activa- tion of antimycobacterial activities and essential for the DTH response. Grange opines that the immune response in an individual with tuberculous infection gets locked in' to one or other pattern of response viz. TH1 or TH2 response, the latter response leading to tissue damage and progression of disease. Stanford and co-workers conducted several studies on the effectiveness of Mycobacterium vaccae, as an immunotherapeutic agent for tuberculosis. It is non-pathogenic in humans and is thought to be a powerful TH1 adjuvant. A series of small studies pointed that M. vaccae has a beneficial effect and there is enough evidence now to show that its use as an immunotherapeutic agent, as an adjunct to chemotherapy in the treatment of tuberculosis especially at a time when drug resistance is rampant, appears promising. KEY WORDS : Tuberculosis, Drug-Resistance, Immunotherapy, T Cell Responses. ROLE OF IMMUNOTHERAPY IN THE cytokines secreted by the TH1 cell are interferon- TREATMENT OF TUBERCULOSIS gamma (IFN-~,) and interleukin-2 (IL-2).
  • Opportunist Mycobacteria

    Opportunist Mycobacteria

    Thorax: first published as 10.1136/thx.44.6.449 on 1 June 1989. Downloaded from Thorax 1989;44:449454 Editorial Treatment of pulmonary disease caused by opportunist mycobacteria During the early 1950s it was recognised that recommended that surgical treatment for opportunist mycobacteria other than Mycobacterium tuberculosis mycobacterial infection should be given to those could cause pulmonary disease in man.' Over 30 years patients who are suitable surgical candidates.'7 The later there is no general agreement about the treatment failure of chemotherapy was often attributed to drug of patients with these mycobacterial infections. The resistance,'2 18 but the importance of prolonging the greatest controversy surrounds the treatment of in- duration of chemotherapy in opportunist mycobac- fection caused by the M aviwn-intracellulare- terial disease beyond that which would normally be scrofulaceum complex (MAIS), for which various required in tuberculosis was not appreciated. In treatments have been advocated, including chemo- several surgically treated series preoperative chemo- therapy with three23 or more' drugs or, alternatively, therapy was given on average for only four to seven surgical resection of the affected lung.78 Although the months,'3 19-22 some patients receiving as little as eight treatment of infection caused by M kansasii is less weeks of treatment before surgery.2324 In contrast, controversial there is no uniform approach to treat- chemotherapy alone, with isoniazid, p-aminosalicyclic ment. Disease caused by M xenopi has been described acid, and streptomycin for 24 months, produced by some as easy to treat with chemotherapy,9 whereas successful results in 80-100% of patients with M others have found the response to drug treatment to be kansasii infection despite reports of in vitro resistance unpredictable.'' " to these agents.2125 This diversity ofopinion and approach to treatment has arisen for two reasons.
  • A Simple and Efficient Multiplex PCR Assay for the Identification of Mycobacteriumgenus and Mycobacterium Tuberculosis Complex T

    A Simple and Efficient Multiplex PCR Assay for the Identification of Mycobacteriumgenus and Mycobacterium Tuberculosis Complex T

    http://dx.doi.org/10.3349/ymj.2013.54.5.1220 Original Article pISSN: 0513-5796, eISSN: 1976-2437 Yonsei Med J 54(5):1220-1226, 2013 A Simple and Efficient Multiplex PCR Assay for the Identification ofMycobacterium Genus and Mycobacterium tuberculosis Complex to the Species Level Yeun Kim,1 Yeonim Choi,1 Bo-Young Jeon,1 Hyunwoo Jin,1,2 Sang-Nae Cho,3 and Hyeyoung Lee1 1Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, Wonju; 2Department of Clinical Laboratory Science, College of Health Sciences, Catholic University of Pusan, Busan; 3Department of Microbiology, Yonsei University College of Medicine, Seoul, Korea. Received: September 19, 2012 Purpose: The Mycobacterium tuberculosis complex comprises M. tuberculosis, Revised: October 25, 2012 M. bovis, M. bovis bacillus Calmette-Guérin (BCG) and M. africanum, and causes Accepted: October 29, 2012 tuberculosis in humans and animals. Identification of Mycobacterium spp. and M. Corresponding author: Dr. Hyeyoung Lee, tuberculosis complex to the species level is important for practical use in microbi- Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, ological laboratories, in addition to optimal treatment and public health. Materials 1 Yonseidae-gil, Wonju 220-710, Korea. and Methods: A novel multiplex PCR assay targeting a conserved rpoB sequence Tel: 82-33-760-2740, Fax: 82-33-760-2561 in Mycobacteria spp., as well as regions of difference (RD) 1 and RD8, was devel- E-mail: [email protected] oped and evaluated using 37 reference strains and 178 clinical isolates. Results: All mycobacterial strains produced a 518-bp product (rpoB), while other bacteria ∙ The authors have no financial conflicts of produced no product.
  • Nontuberculous Mycobacterial Species and Mycobacterium Tuberculosis Complex Coinfection in Patients with Pulmonary Tuberculosis in Dr

    Nontuberculous Mycobacterial Species and Mycobacterium Tuberculosis Complex Coinfection in Patients with Pulmonary Tuberculosis in Dr

    [Downloaded free from http://www.ijmyco.org on Saturday, September 28, 2019, IP: 210.57.215.50] Original Research Article Nontuberculous Mycobacterial Species and Mycobacterium Tuberculosis Complex Coinfection in Patients with Pulmonary Tuberculosis in Dr. Soetomo Hospital, Surabaya, Indonesia Ni Made Mertaniasih1,2, Deby Kusumaningrum1,2, Eko Budi Koendhori1,2, Soedarsono3, Tutik Kusmiati3, Desak Nyoman Surya Suameitria Dewi2 Departments of 1Clinical Microbiology and 3Pulmonology, Faculty of Medicine, Airlangga University, Dr. Soetomo Hospital, Surabaya 60131, 2Institute of Tropical Disease, Airlangga University, Surabaya 60115, Indonesia Abstract Objective/Background: The aim of this study was to analyze the detection of nontuberculous mycobacterial (NTM) species derived from sputum specimens of pulmonary tuberculosis (TB) suspects. Increasing prevalence and incidence of pulmonary infection by NTM species have widely been reported in several countries with geographical variation. Materials and Methods: Between January 2014 and September 2015, sputum specimens from chronic pulmonary TB suspect patients were analyzed. Laboratory examination of mycobacteria was conducted in the TB laboratory, Department of Clinical Microbiology, Dr. Soetomo Hospital, Surabaya. Detection and identification of mycobacteria were performed by the standard culture method using the BACTEC MGIT 960 system (BD) and Lowenstein–Jensen medium. Identification of positive Mycobacterium tuberculosis complex (MTBC) was based on positive acid-fast bacilli microscopic smear, positive niacin accumulation, and positive TB Ag MPT 64 test results (SD Bioline). If the growth of positive cultures and acid-fast bacilli microscopic smear was positive, but niacin accumulation and TB Ag MPT 64 (SD Bioline) results were negative, then the isolates were categorized as NTM species. MTBC isolates were also tested for their sensitivity toward first-line anti-TB drugs, using isoniazid, rifampin, ethambutol, and streptomycin.
  • Mycobacterium Goodii Endocarditis Following Mitral Valve Ring Annuloplasty Rohan B

    Mycobacterium Goodii Endocarditis Following Mitral Valve Ring Annuloplasty Rohan B

    Parikh and Grant Ann Clin Microbiol Antimicrob (2017) 16:14 DOI 10.1186/s12941-017-0190-4 Annals of Clinical Microbiology and Antimicrobials CASE REPORT Open Access Mycobacterium goodii endocarditis following mitral valve ring annuloplasty Rohan B. Parikh1 and Matthew Grant2* Abstract Background: Mycobacterium goodii is an infrequent human pathogen which has been implicated in prosthesis related infections and penetrating injuries. It is often initially misidentified as a gram-positive rod by clinical microbio- logic laboratories and should be considered in the differential diagnosis. Case presentation: We describe here the second reported case of M. goodii endocarditis. Species level identification was performed by 16S rDNA (ribosomal deoxyribonucleic acid) gene sequencing. The patient was successfully treated with mitral valve replacement and a prolonged combination of ciprofloxacin and trimethoprim/sulfamethoxazole. Conclusion: Confirmation of the diagnosis utilizing molecular techniques and drug susceptibility testing allowed for successful treatment of this prosthetic infection. Keywords: Mycobacterium goodii, Endocarditis, Gene sequencing, Prostheses related infections Background appreciated at the apex, and a drain was in place for a Mycobacterium goodii is a rapidly growing non-tubercu- groin seroma related to recent left heart catheterization. lous mycobacterium (NTM) belonging to the Mycobac- He had an unsteady gait and exhibited mild left lower terium smegmatis [1] group. Its importance has become extremity weakness (4/5). His brain magnetic resonance increasingly appreciated as a pathogen over the last imaging showed multiple ring-enhancing lesions in the 20 years, with a predilection towards infecting tissues at pons and posterior fossa suggestive of septic emboli. the site of penetrating injuries. Antibacterial treatment Transthoracic echocardiography showed moderate strategies against this pathogen are diverse but reported mitral regurgitation without any vegetation.
  • Immunotherapy of TB…

    Immunotherapy of TB…

    Results from Phase III, placebo-controlled, 2:1 randomized, double-blind trial of tableted TB vaccine (V7) containing 10 μg of heat-killed Mycobacterium vaccae administered daily for one month Correspondence: Aldar Bourinbaiar * Tel: +976 95130306; +1 301 476-0930 * e-mail: [email protected] Tuberculosis • 33% of people carry TB bacteria = 2.5 billion • Every second, a person becomes ill with TB • Every year 10 mln people develop TB and 2 mln die • Drug-resistant TB poised to become global pandemic • Less than 3% of drug-resistant TB is treated today Th-1 cells = IFN-ɣ, IL-2, GM-CSF, IFN-ɑ, TNF-ɑ, IL-12, IL-18 Th-2 cells = IL-4, IL-5, IL-13 Th-17 cells = IL-6, IL-17, IL-22, TNF-ɑ Treg cells = IL-10, TGF-ɓ B cells = IFN-ɑ, IL-1ɓ, IL-12 Monocytes = IL-8, IL-18, TNF-ɑ State-of-the-art: immunology of TB – a great deal of information gathered but… can't see the forest through the trees When this vast knowledge was applied to immunotherapy of TB…. it failed… It also resulted in negative attitude toward immunotherapy • IL-2 (increased bacterial load) • IL-12 (no effect) • GM-CSF (clearance not confirmed) • IFN-ɣ (irreproducible, no effect) • IFN-ɑ (negative outcome) • anti-TNF-ɑ (negative outcome) • Thalidomide (negative outcome) • Corticosteroids (irreproducible, negative effect) Paradoxes of Tuberculosis • 1/3 of world population (~2.5bln) have latent M. tuberculosis • Yet only about 10 Million people (0.004%) develop TB • M. tuberculosis is in symbiotic relationship with the host • In some cases it’s even beneficial (asthma, cancer) • M.
  • Antibitoic Treatment for Tuberculosis Induces a Profound Dysbiosis of the Gut Microbiome That Persists Long After Therapy Is Completed

    Antibitoic Treatment for Tuberculosis Induces a Profound Dysbiosis of the Gut Microbiome That Persists Long After Therapy Is Completed

    ANTIBITOIC TREATMENT FOR TUBERCULOSIS INDUCES A PROFOUND DYSBIOSIS OF THE GUT MICROBIOME THAT PERSISTS LONG AFTER THERAPY IS COMPLETED A Thesis Presented to the Faculty of the Weill Cornell Graduate School of Medical Sciences in Partial Fulfillment of the Requirements for the Degree of Masters of Science by Matthew F. Wipperman May 2017 © 2017 Matthew F. Wipperman ABSTRACT Mycobacterium tuberculosis, the cause of Tuberculosis (TB), infects one third of the world’s population and causes substantial mortality worldwide. In its shortest format, treatment of drug sensitive TB requires six months of multidrug therapy with a mixture of broad spectrum and mycobacterial specific antibiotics, and treatment of multidrug resistant TB is much longer. The widespread use of this regimen worldwide makes this one the largest exposures of humans to antimicrobials, yet the effects of antimycobacterial agents on intestinal microbiome composition and long term stability are unknown. We compared the microbiome composition, assessed by both 16S rDNA and metagenomic DNA sequencing, of Haitian TB cases during antimycobacterial treatment and following cure by 6 months of TB therapy. TB treatment does not perturb overall diversity, but nonetheless dramatically depletes multiple immunologically significant commensal bacteria. The perturbation by TB therapy lasts at least 1.5 years after completion of treatment, indicating that the effects of TB treatment are long lasting and perhaps permanent. These results demonstrate that TB treatment has dramatic and durable effects on the intestinal microbiome and highlight unexpected extreme consequences of treatment for the world’s most common infection on human ecology. BIOGRAPHICAL SKETCH NAME POSITION TITLE Wipperman, Matthew Frederick Postdoctoral Researcher at eRA COMMONS USER NAME Memorial Sloan Kettering Cancer Center MFWIPPERMAN DEGREE INSTITUTION AND (if MM/YY FIELD OF STUDY LOCATION applicable) Franklin & Marshall College B.A.