Metabolic activity of mature biofilms of Mycobacterium tuberculosis and other non-tuberculous mycobacteria

Solokhina, Anna; Bruckner, David A.; Bonkat, Gernot; Braissant, Olivier

doi:10.1038/s41598-017-10019-4

Cited by 31 publications

(32 citation statements)

References 57 publications

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“…Albeit not so numerous, the applications of ITC have gone beyond binary ligand reactions and also included the study of whole living cells. Thus, microcalorimetry studies have been applied to microorganisms (22)(23)(24) and to cells and tissues slices (25). When studying the interactions of biomolecules in solution the titration relies mainly on multiple injections of the samples, whereas with whole cells, single injections may be the method of choice (26).…”

Section: Discussionmentioning

confidence: 99%

Isothermal Microcalorimetry of Tumor Cells: Enhanced Thermogenesis by Metastatic Cells

et al. 2019

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Tumor cells exhibit rewired metabolism. We carried out comparative analyses attempting to investigate whether metabolic reprograming could be measured by isothermal microcalorimetry. Intact metastatic cell lines of tongue cell carcinoma, human and murine melanoma, lung, and breast tumors consistently released more heat than nonmetastatic cells or cells displaying lower metastatic potential. In tongue squamous carcinoma cells mitochondrial enriched extract reproduced the heat release pattern of intact cells. Cytochalasin D, an actin filament inhibitor, and suppression of metastasis marker Melanoma associated gene 10 (MAGEA10) decreased heat release. Uncoupling protein 2 was highly expressed in metastatic cells, but not in non-metastatic cells. Carnitine palmitoyl transferase-1 inhibitor, Etomoxir strongly inhibited heat release by metastatic cells, thus linking lipid metabolism to thermogenesis. We propose that heat release may be a quantifiable trait of the metastatic process.

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Section: Discussionmentioning

confidence: 99%

Isothermal Microcalorimetry of Tumor Cells: Enhanced Thermogenesis by Metastatic Cells

et al. 2019

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“…proteins (Krell, 2008). In life sciences, isothermal microcalorimetry has been used to discriminate treatments or bacteria (Solokhina et al, 2017;Butini et al, 2019) and also to estimate minimum inhibitory concentrations (MIC) of antibiotics (Tellapragda et al, 2020). Simplistic measures such as maximum heat flow, total energy release, time to peak, etc.…”

Section: Discussionmentioning

confidence: 99%

“…Bacteria residing in biofilms are thought to be phenotypically distinct from their planktonic counterparts. Many publications have identified specific biomarkers to characterize this distinction, such as differential gene expression (Folsom et al, 2010), secretion of extracellular polymers (Costerton et al, 1995;Goltermann and Tolker-Nielsen, 2017), virulence factors (Hauser, 2011), metabolism (Solokhina et al, 2017) and increased tolerance towards antibiotics and host response (Bjarnsholt et al, 2013;Ciofu and Tolker-Nielsen, 2019). However, physical aggregation (observed by microscopy) and increased antibiotic tolerance are the most dominant and consistent of these biofilm-associated phenotypes.…”

Section: Introductionmentioning

confidence: 99%

Metabolic flux fingerprinting differentiates planktonic and biofilm states ofPseudomonas aeruginosaandStaphylococcus aureus

Lichtenberg¹,

Kragh

Fritz³

et al. 2020

Preprint

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The challenges of defining the biofilm phenotype has been clear for decades. Many biomarkers for biofilm are known, but methods for identifying these are often invasive and/or complicated. These methods often rely on disrupting the biofilm matrix or examining virulence factors and compounds, which may only be expressed under certain conditions.We used microcalorimetric measurements of metabolic energy release to investigate whether unchallenged, planktonic Pseudomonas aeruginosa displayed differences in metabolism compared to surface-bound and non-attached biofilms.The pattern of energy release observed in the recorded microcalorimetric thermograms clearly depended on growth state, though the total energy expenditure was not different between growth states. To characterize these differences, we developed a classification pipeline utilizing machine learning algorithms to classify growth state, based on the observed patterns of energy release. With this approach, we could with high accuracy detect the growth form of blinded samples. To challenge the algorithm, we attempted to limit the amount of training data. By training the algorithm with only a single data point from each growth form, we obtained a mean accuracy of 90.5% using two principal components. Further validation of the classification pipeline showed that the approach was not limited to P. aeruginosa but could also be used for detection of gram-positive Staphylococcus aureus biofilm. We propose that microcalorimetric measurements, in combination with this new quantitative framework, can be used as a non-invasive biomarker to detect the presence of biofilm.These results could have a significant potential in clinical settings where the detection of biofilms in infections often means a different outcome and treatment regime for the patient.

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“…The biofilm architecture and the extracellular polymeric substance matrix limit drug diffusion, protecting individual cells. 36 Biofilm MTB differs from planktonic MTB in many ways. Biofilms are known to protect pathogens from phagocytosis and to increase tolerance to drug treatment.…”

Section: Discussionmentioning

confidence: 99%

“…However, biofilms have been shown to have higher metabolic activities than their planktonic counterparts. 36 …”

Section: Discussionmentioning

confidence: 99%

Viability, biofilm formation, and MazEF expression in drug-sensitive and drug-resistant <em>Mycobacterium tuberculosis</em> strains circulating in Xinjiang, China

Zhao¹,

Liu²,

Xie³

et al. 2018

IDR

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BackgroundTuberculosis (TB) caused by Mycobacterium tuberculosis (MTB) is one of the most common chronic infectious amphixenotic diseases worldwide. Prevention and control of TB are greatly difficult, due to the increase in drug-resistant TB, particularly multidrug-resistant TB. We speculated that there were some differences between drug-sensitive and drug-resistant MTB strains and that mazEF3,6,9 toxin–antitoxin systems (TASs) were involved in MTB viability. This study aimed to investigate differences in viability, biofilm formation, and MazEF expression between drug-sensitive and drug-resistant MTB strains circulating in Xinjiang, China, and whether mazEF3,6,9 TASs contribute to MTB viability under stress conditions.Materials and methodsGrowth profiles and biofilm-formation abilities of drug-sensitive, drug-resistant MTB strains and the control strain H37Rv were monitored. Using molecular biology experiments, the mRNA expression of the mazF3, 6, and 9 toxin genes, the mazE3, 6, and 9 antitoxin genes, and expression of the MazF9 protein were detected in the different MTB strains, H37RvΔmazEF3,6,9 mutants from the H37Rv parent strain were generated, and mutant viability was tested.ResultsEx vivo culture analyses demonstrated that drug-resistant MTB strains exhibit higher survival rates than drug-sensitive strains and the control strain H37Rv. However, there was no statistical difference in biofilm-formation ability in the drug-sensitive, drug-resistant, and H37Rv strains. mazE3,6 mRNA-expression levels were relatively reduced in the drug-sensitive and drug-resistant strains compared to H37Rv. Conversely, mazE3,9 expression was increased in drug-sensitive strains compared to drug-resistant strains. Furthermore, compared with the H37Rv strain, mazF3,6 expression was increased in drug-resistant strains, mazF9 expression was increased in drug-sensitive strains, and mazF9 exhibited reduced expression in drug-resistant strains compared with drug-sensitive strains. Protein expression of mazF9 was increased in drug-sensitive and drug-resistant strains compared to H37Rv, while drug-resistant strains exhibited reduced mazF9 expression compared to drug-sensitive strains. Compared to H37Rv, H37RvΔmazEF3,6,9-deletion mutants grew more slowly under both stress conditions, and their ability to survive in host macrophages was also weaker. Furthermore, the host macrophage-apoptosis rate was higher after infection with any of the H37RvΔmazEF3,6,9 mutants than with the H37Rv strain.ConclusionThe increased viability of MTB drug-resistant strains compared with drug-sensitive strains is likely to be related to differential MazEF mRNA and protein expression. mazEF3,6,9 TASs contribute to MTB viability under stress conditions.

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Metabolic activity of mature biofilms of Mycobacterium tuberculosis and other non-tuberculous mycobacteria

Cited by 31 publications

References 57 publications

Isothermal Microcalorimetry of Tumor Cells: Enhanced Thermogenesis by Metastatic Cells

Isothermal Microcalorimetry of Tumor Cells: Enhanced Thermogenesis by Metastatic Cells

Metabolic flux fingerprinting differentiates planktonic and biofilm states ofPseudomonas aeruginosaandStaphylococcus aureus

Viability, biofilm formation, and MazEF expression in drug-sensitive and drug-resistant <em>Mycobacterium tuberculosis</em> strains circulating in Xinjiang, China

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