Bacteriophage- based tests for the detection of Mycobacterium tuberculosis in clinical specimens: a systematic review and meta- analysis

BACKGROUND: Sputum microscopy, the most important conventional test for tuberculosis, is specific in settings with high burden of tuberculosis and low prevalence of non tuberculous mycobacteria. However, the test lacks sensitivity. Although bacteriophage-based tests for tuberculosis have shown promising results, their overall accuracy has not been systematically evaluated. METHODS: We did a systematic review and meta-analysis of published studies to evaluate the accuracy of phage-based tests for the direct detection of M. tuberculosis in clinical specimens. To identify studies, we searched Medline, EMBASE, Web of science and BIOSIS, and contacted authors, experts and test manufacturers. Thirteen studies, all based on phage amplification method, met our inclusion criteria. Overall accuracy was evaluated using forest plots, summary receiver operating (SROC) curves, and subgroup analyses. RESULTS: The data suggest that phage-based assays have high specificity (range 0.83 to 1.00), but modest and variable sensitivity (range 0.21 to 0.88). The sensitivity ranged between 0.29 and 0.87 among smear-positive, and 0.13 to 0.78 among smear-negative specimens. The specificity ranged between 0.60 and 0.88 among smear-positive and 0.89 to 0.99 among smear-negative specimens. SROC analyses suggest that overall accuracy of phage-based assays is slightly higher than smear microscopy in direct head-to-head comparisons. CONCLUSION: Phage-based assays have high specificity but lower and variable sensitivity. Their performance characteristics are similar to sputum microscopy. Phage assays cannot replace conventional diagnostic tests such as microscopy and culture at this time. Further research is required to identify methods that can enhance the sensitivity of phage-based assays without compromising the high specificity

Catastrophic NAD+ Depletion in Activated T Lymphocytes through Nampt Inhibition Reduces Demyelination and Disability in EAE

Nicotinamide phosphoribosyltransferase (Nampt) inhibitors such as FK866 are potent inhibitors of NAD+ synthesis that show promise for the treatment of different forms of cancer. Based on Nampt upregulation in activated T lymphocytes and on preliminary reports of lymphopenia in FK866 treated patients, we have investigated FK866 for its capacity to interfere with T lymphocyte function and survival. Intracellular pyridine nucleotides, ATP, mitochondrial function, viability, proliferation, activation markers and cytokine secretion were assessed in resting and in activated human T lymphocytes. In addition, we used experimental autoimmune encephalomyelitis (EAE) as a model of T-cell mediated autoimmune disease to assess FK866 efficacy in vivo. We show that activated, but not resting, T lymphocytes undergo massive NAD+ depletion upon FK866-mediated Nampt inhibition. As a consequence, impaired proliferation, reduced IFN-γ and TNF-α production, and finally autophagic cell demise result. We demonstrate that upregulation of the NAD+-degrading enzyme poly-(ADP-ribose)-polymerase (PARP) by activated T cells enhances their susceptibility to NAD+ depletion. In addition, we relate defective IFN-γ and TNF-α production in response to FK866 to impaired Sirt6 activity. Finally, we show that FK866 strikingly reduces the neurological damage and the clinical manifestations of EAE. In conclusion, Nampt inhibitors (and possibly Sirt6 inhibitors) could be used to modulate T cell-mediated immune responses and thereby be beneficial in immune-mediated disorders

International Field Intercomparison of Atmospheric Mercury Measurement Methods

To determine the extent of comparability of sampling and analytical procedures for atmospheric mercury (Hg) being used by different scientific groups around the world and hence the compatibility of measurement results, the Atmospheric Environment Service (AES) co-ordinated a field intercomparison study in Windsor, Ontario, over a period of 5 days- during Sept./Oct.,1993. This study brought together 2 groups (University of Michigan Air Quality Laboratory; Chemistry Institute of GKSS) which performed conventional (manual) sample collection procedures for total gaseous mercury (TGM) and for particulate-phase mercury (PPM), followed by cold-vapor atomic fluorescence spectrophotometric (CVAFS) analysis in the respective laboratories. Two other groups (Ontario Hydro, and the Ontario Ministry of Environment & Energy) each operated a novel mercury vapor analyzer produced by Tekran Inc. of Toronto. As is the case for the manual methods, this analyzer also uses gold amalgamation and CVAFS. During the intercomparison, meteorological parameters (air temperature, barometric pressure, wind speed/direction and relative humidity) were obtained at the study site.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/43915/1/11270_2005_Article_BF01189713.pd

Cell-free biosensors for rapid detection of water contaminants

© 2020, The Author(s), under exclusive licence to Springer Nature America, Inc. Lack of access to safe drinking water is a global problem, and methods to reliably and easily detect contaminants could be transformative. We report the development of a cell-free in vitro transcription system that uses RNA Output Sensors Activated by Ligand Induction (ROSALIND) to detect contaminants in water. A combination of highly processive RNA polymerases, allosteric protein transcription factors and synthetic DNA transcription templates regulates the synthesis of a fluorescence-activating RNA aptamer. The presence of a target contaminant induces the transcription of the aptamer, and a fluorescent signal is produced. We apply ROSALIND to detect a range of water contaminants, including antibiotics, small molecules and metals. We also show that adding RNA circuitry can invert responses, reduce crosstalk and improve sensitivity without protein engineering. The ROSALIND system can be freeze-dried for easy storage and distribution, and we apply it in the field to test municipal water supplies, demonstrating its potential use for monitoring water quality

On singularities and black holes in combination-driven models of technological innovation networks

It has been suggested that innovations occur mainly by combination: the more inventions accumulate, the higher the probability that new inventions are obtained from previous designs. Additionally, it has been conjectured that the combinatorial nature of innovations naturally leads to a singularity: at some finite time, the number of innovations should diverge. Although these ideas are certainly appealing, no general models have been yet developed to test the conditions under which combinatorial technology should become explosive. Here we present a generalised model of technological evolution that takes into account two major properties: the number of previous technologies needed to create a novel one and how rapidly technology ages. Two different models of combinatorial growth are considered, involving different forms of ageing. When long-range memory is used and thus old inventions are available for novel innovations, singularities can emerge under some conditions with two phases separated by a critical boundary. If the ageing has a characteristic time scale, it is shown that no singularities will be observed. Instead, a "black hole" of old innovations appears and expands in time, making the rate of invention creation slow down into a linear regime.This work was supported by the Fundacion Botin and by the Spanish Ministry of Economy and Competitiveness, Grant FIS2013-44674-P and FEDER and by the Santa Fe Institute, where most of this work was done