A5041730736- Tuberculosis Research and Epidemiology
- Mycobacterium research and diagnosis
- Biochemical and Molecular Research
- Microbial Metabolic Engineering and Bioproduction
- Antibiotic Resistance in Bacteria
- Diagnosis and treatment of tuberculosis
- Mass Spectrometry Techniques and Applications
- Metabolomics and Mass Spectrometry Studies
- RNA and protein synthesis mechanisms
- Computational Drug Discovery Methods
- Advanced Proteomics Techniques and Applications
- Bioinformatics and Genomic Networks
- ATP Synthase and ATPases Research
- Bacterial Genetics and Biotechnology
- Viral gastroenteritis research and epidemiology
- Analytical Chemistry and Chromatography
- Virus-based gene therapy research
- Medical Imaging and Pathology Studies
- Escherichia coli research studies
- Heat shock proteins research
- Amoebic Infections and Treatments
- Pneumonia and Respiratory Infections
- Animal Virus Infections Studies
- Enzyme Structure and Function
- Mercury impact and mitigation studies
University of Surrey
2012-2025
University of London
2003
London School of Hygiene & Tropical Medicine
2003
Central Middlesex Hospital
2003
An impediment to the rational development of novel drugs against tuberculosis (TB) is a general paucity knowledge concerning metabolism Mycobacterium tuberculosis, particularly during infection. Constraint-based modeling provides approach investigating microbial but has not yet been applied genome-scale M. tuberculosis. GSMN-TB, metabolic model was constructed, consisting 849 unique reactions and 739 metabolites, involving 726 genes. The calibrated by growing bovis bacille Calmette Guérin in...
Whereas intracellular carbon metabolism has emerged as an attractive drug target, the sources of intracellularly replicating pathogens, such tuberculosis bacillus Mycobacterium tuberculosis, which causes long-term infections in one-third world's population, remain mostly unknown. We used a systems-based approach--(13)C-flux spectral analysis (FSA) complemented with manual analysis-to measure metabolic interaction between M. and its macrophage host cell. (13)C-FSA experimental data showed...
Mycobacterium tuberculosis requires the enzyme isocitrate lyase (ICL) for growth and virulence in vivo. The demonstration that M. also ICL survival during nutrient starvation has a role steady state glycerol limited chemostat indicates function this which extends beyond fat metabolism. As is potential drug target elucidating of importance; however, never been investigated at level vivo fluxes. Here we show deletion one two icl genes impairs replication bovis BCG slow rate carbon chemostat....
We report the development and validation of an untargeted single-cell lipidomics method based on microflow chromatography coupled to a data-dependent mass spectrometry for fragmentation-based identification lipids. Given absence lipid standards, we show how methodology should be optimized validated using dilute cell extract. The is applied pancreatic cancer macrophage extracts standards demonstrate sensitivity requirements confident assignment lipids classification type at level. then system...
Mycobacterium tuberculosis infects a third of the world's population. Primary involving active fast bacterial replication is often followed by asymptomatic latent tuberculosis, which characterised slow or non-replicating bacteria. Reactivation infection switch back to can lead post-primary transmissible tuberculosis. Mycobacterial mechanisms involved in growth switching rate provide rational targets for development new drugs against persistent mycobacterial infection. Using chemostat culture...
Abstract The approval of bedaquiline (BDQ) for the treatment tuberculosis has generated substantial interest in inhibiting energy metabolism as a therapeutic paradigm. However, it is not known precisely how BDQ triggers cell death Mycobacterium ( Mtb) . Using 13 C isotopomer analysis, we show that BDQ-treated Mtb redirects central carbon to induce metabolically vulnerable state susceptible genetic disruption glycolysis and gluconeogenesis. Metabolic flux profiles indicate dependent on ATP...
Metabolic flux is the final output of cellular regulation and has been extensively studied for carbon but much less known about nitrogen, which another important building block living organisms. For tuberculosis pathogen, this particularly in informing development effective drugs targeting pathogen's metabolism. Here we performed
The Mycobacterium tuberculosis complex includes bovine and human strains of the bacillus, including tuberculosis, bovis BCG vaccine strain. M. has evolved from a tuberculosis-like ancestor is vaccine. pathogens demonstrate distinct differences in virulence, host range metabolism, but role metabolic pathogenicity poorly understood. Systems biology approaches have been used to investigate metabolism not probe between strains. In this study genome scale networks were constructed interrogated,...
Antibiotic-tolerant bacteria, due to their unique physiology, are refractory antimicrobial killing and pose challenges for infection control. Incomplete knowledge of how bactericidal antibiotics work, limits our understanding partial resistance phenotypic tolerance in mycobacteria, a driver developing genetic resistance. Using proteomics, 13 C isotopomer analysis, biochemical assays, we investigated the physiological response M. smegmatis challenged with aminoglycoside fluoroquinolone...
ABSTRACT An experimental system of Mycobacterium tuberculosis growth in a carbon-limited chemostat has been established by the use bovis BCG as model organism. For this model, chemostats with low concentrations glycerol were used to simulate possible rates during different stages tuberculosis. A doubling time 23 h ( D = 0.03 −1 ) was adopted represent cells acute phase infection, whereas lower dilution rate equivalent 69 0.01 mycobacterial persistence. This allowed specific response cell...
Enzymes at the phosphoenolpyruvate (PEP)-pyruvate-oxaloacetate or anaplerotic (ANA) node control metabolic flux to glycolysis, gluconeogenesis, and anaplerosis. Here we used genetic, biochemical, 13C isotopomer analysis characterize role of enzymes ANA in intracellular survival world's most successful bacterial pathogen, Mycobacterium tuberculosis (Mtb). We show that each four enzymes, pyruvate carboxylase (PCA), PEP carboxykinase (PCK), malic enzyme (MEZ), phosphate dikinase (PPDK),...
Article4 May 2021Open Access Transparent process Metabolic fluxes for nutritional flexibility of Mycobacterium tuberculosis Khushboo Borah orcid.org/0000-0001-9030-2085 Department Microbial and Cellular Sciences, Faculty Health Medical University Surrey, Guildford, UK Search more papers by this author Tom A Mendum orcid.org/0000-0002-6331-2605 Nathaniel D Hawkins orcid.org/0000-0001-9806-312X Computational Analytical Rothamsted Research, Harpenden, Jane L Ward Michael H Beale Gerald...
ABSTRACT The adaptation of the tubercle bacillus to host environment is likely involve a complex set gene regulatory events and physiological switches in response environmental signals. In order deconstruct state Mycobacterium tuberculosis vivo, we used chemostat model study single aspect organism's vivo state, slow growth. bovis BCG was cultivated at high low growth rates carbon-limited chemostat, transcriptomic analysis performed identify regulation associated with results demonstrated...
Tuberculosis (TB) is caused by Mycobacterium tuberculosis (Mtb), leading to pulmonary and extrapulmonary TB, whereby Mtb disseminated many other organs tissues. Dissemination occurs early during the disease, bacteria can be found first in lymph nodes adjacent lungs then later organs, including spleen. The global gene expression response of human tissue macrophages intracellular clinical isolates has been poorly studied. Using dual RNA-seq, we have explored mRNA profiles two closely related...
New approaches are needed to control leprosy, but understanding of the biology causative agent Mycobacterium leprae remains rudimentary, principally because pathogen cannot be grown in axenic culture. Here, we applied 13C isotopomer analysis measure carbon metabolism M. its primary host cell, Schwann cell. We compared results this with those a related pathogen, tuberculosis, growing macrophage. Using glucose as tracer, show that whereas tuberculosis imports most amino acids directly from...
Metabolism underpins the pathogenic strategy of causative agent TB, Mycobacterium tuberculosis (Mtb), and therefore metabolic pathways have recently re-emerged as attractive drug targets. A powerful approach to study Mtb metabolism a whole, rather than just individual enzymatic components, is use systems biology framework, such Genome-Scale Metabolic Network (GSMN) that allows dynamic interactions all components be interrogated together. Several GSMNs networks been constructed for used...
Abstract Single cell–inductively coupled plasma–mass spectrometry (SC-ICP-MS) offers an attractive option for rapidly measuring trace metal heterogeneity at the single cell level. Chemical fixation has been previously applied to mammalian cells prior sample introduction so that they can be resuspended in a solution suitable SC-ICP-MS. However, effect of on elemental composition suspended is unknown, and robust methodologies are urgently needed community measure effects intracellular...
This work describes the development of a new approach to measure drug levels and lipid fingerprints in single living cells.
A general paucity of knowledge about the metabolic state Mycobacterium tuberculosis within host environment is a major factor impeding development novel drugs against tuberculosis. Current experimental methods do not allow direct determination global bacterial pathogen in vivo, but transcriptional activity all encoded genes has been investigated numerous microarray studies. We describe algorithm, Differential Producibility Analysis (DPA) that uses network to extract signals from...
Despite decades of research many aspects the biology Mycobacterium tuberculosis remain unclear and this is reflected in antiquated tools available to treat prevent consequently disease remains a serious public health problem. Important discoveries linking M. tuberculosis's metabolism pathogenesis have renewed interest area research. Previous experimental studies were limited analysis individual genes or enzymes whereas recent advances computational systems high throughput technologies now...
Antibiotic-tolerant bacteria, due to their unique physiology, are refractory antimicrobial killing and pose challenges for infection control. Incomplete knowledge of how bactericidal antibiotics work, limits our understanding partial resistance phenotypic tolerance in mycobacteria, a driver developing genetic resistance. Using proteomics, 13 C isotopomer analysis, biochemical assays, we investigated the physiological response M. smegmatis challenged with aminoglycoside fluoroquinolone...