bioRxiv Science⌕ Search

bioRxiv · 10.1101/2023.07.28.550952

Enhancing genetically engineered Escherichia coli bioreporters for the detection of buried TNT-based landmines

Abstract

TNT-based buried landmines are explosive devices consisting of mainly 2,4,6-trinitrotoluene (2,4,6-TNT), but also a 2,4-dinitrotoluene (2,4-DNT) impurity. It is reported that some 2,4-DNT impurity vapors slowly leak through land mine covers and into the soil above them, making this compound a potential indicator for the presence of buried landmines. There are several approaches to identify the presence of chemicals in the environment, one of which is based on a microbial detection system. This approach was previously used to address the challenge of remotely identifying buried landmines by applying genetically engineered microbial whole cell biosensors capable of sensing trace signatures of the explosive compound 2,4,6-TNT and its degradation product, 2,4-DNT. Upon exposure to 2,4-DNT, the genetically engineered bacterial reporter strains, containing a genetic fusion between a sensing element for 2,4-DNT (yqjF gene promoter) and a fluorescence/ bioluminescence reporting element (GFP/ luxCDABE genes), generate a measurable dose-dependent signal within the whole cell biosensor system. This biological component of a comprehensive remote sensing solution was significantly improved after several rounds of random mutagenesis to the yqjF promoter region using a prone-to-error PCR and selection of the best mutants. In the present project further significant improvements were obtained in signal intensity, response time, and in the limit of detection, following an additional round of random mutagenesis. Two new mutants, named A10 and C5, were identified. The new mutants present high induction levels both in terms of Ratio (sample/control) and delta (sample-control), making these strains more amenable to remote detection. To isolate a more sensitive mutant, the 3rd round of random mutagenesis library (about 1000 strains) was re-exposed to lower concentration of 2,4-DNT, but no better candidate was isolated. Possible alternative approaches to lower the detection threshold of the system, such as the use of permeability mutants, are now being tested.

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Amiel, E.. 2023-07-28. Enhancing genetically engineered Escherichia coli bioreporters for the detection of buried TNT-based landmines. https://doi.org/10.1101/2023.07.28.550952

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

SpiraMed: A Stereotactic Helix-based Therapy Delivery system for the Human Brain

Stereotactic needle-based delivery remains the standard for local administration of Advanced Therapy Medicinal Products (ATMPs) to the human brain. ATMP administration typically involves multiple trajectories, presenting cumulative risks and prolonging surgery. Reflux-prone, patchy therapy coverage compromises clinical results. We demonstrate a novel approach, deploying a helical delivery catheter via a single access trajectory per target, referred to as SpiraMed. Helix retraction is synchronised with therapy delivery, enabling comprehensive target coverage in seconds. Helix pitch and diameter can be precisely tailored to patient-specific target volume and vascular anatomy, as part of the preoperative stereotactic surgical planning process. Testing in agarose phantoms, live sheep and cadaveric human brain confirms enhanced therapy delivery volume, delivery speed and target coverage, with reductions in reflux and predicted risk of bleeding complication. SpiraMed represents a new paradigm, promising to help deliver on the transformative potential of cell and gene therapies across the spectrum of human CNS disease.

bioengineering↗

Normative Modeling of Molecular-Enriched Functional Connectivity for Detecting Deviations from Healthy Brain Aging

Inter-individual variability in adult brain aging can obscure early pathological alterations and is only partly represented by population-average or scalar brain-aging measures. We combined Receptor-Enriched Analysis of functional Connectivity by Targets (REACT) with normative modeling (NM) to derive spatially resolved, molecularly informed deviation scores for dopamine transporter (DAT)-, norepinephrine transporter (NET)-, and serotonin transporter (SERT)-enriched resting-state functional connectivity (FC). We first evaluated whether these normative models could be transferred to independently processed external data through local calibration and then explored whether the resulting deviation profiles differed with cerebral amyloid burden in cognitively unimpaired older adults. Hierarchical Bayesian regression (HBR) models with a SHASHb likelihood were estimated separately for 204 cortical molecular-enriched FC features in 4,152 healthy adults (18.0--89.8 years) from seven publicly available neuroimaging datasets and evaluated in a held-out healthy test set. Pretrained models were locally adapted to three AMYPAD-PNHS acquisition batches using cognitively unimpaired, amyloid-negative participants (global Clinical Dementia Rating [CDR] = 0; Centiloid [CL] [lt] 10). The independent primary comparison contrasted participants with intermediate amyloid burden (10 [≤] CL [lt] 30; n = 111) and amyloid-positive participants (CL [≥] 30; n = 66); secondary analyses tested linear associations with continuous CL within participants with CL [≥] 10. Of 204 normative reference models, 199 (97.5%) met the predefined diagnostic criteria; median held-out explained variance (EXPV) was 0.174. Of 612 feature-by-batch transfers, 541 (88.4%) met the strict transfer-diagnostic criteria. In the primary false discovery rate (FDR)-controlled regional analysis, DAT-enriched right caudal middle frontal cortex showed lower locally standardized deviation scores in the intermediate-amyloid-burden group than in the amyloid-positive group (adjusted difference = -0.582, q = 0.016). DAT extreme-deviation burden was also greater in the intermediate-amyloid-burden group (difference = 0.032, q = 0.044). The right frontal effect was reproduced with model-native scores across the three acquisition batches (pooled standardized effect = -0.698, q = 0.010). No NET- or SERT-enriched regional difference between these two groups survived FDR correction, and no regional or subject-level linear association with continuous CL values survived FDR correction in the CL [≥] 10 group. Normative modeling can provide spatially resolved reference distributions of molecular-enriched FC that are deployable in independently processed external data when local adaptation, calibration, and feature-level transfer diagnostics are incorporated. The AMYPAD-PNHS application identified modest, spatially selective, and molecular-system-specific categorical differences during a cognitively unimpaired stage of amyloid accumulation, while continuous analyses within CL [≥] 10 did not support a linear association. These findings support a methodological basis for distributed application of molecular-enriched normative models and motivate independent and longitudinal evaluation of their biological relevance.

bioengineering↗

AC-Diff: Anatomy-Contrast Disentangled Diffusion for Multi-Vendor Liver MRI Harmonization

Magnetic resonance imaging (MRI) exhibits substantial scanner and protocol dependent appearance variations, making harmonization challenging without distorting patient specific anatomy, particularly in abdominal imaging. We propose AC-Diff, an anatomy--contrast disentangled diffusion framework that formulates MRI harmonization as a factor-specific generative intervention. Using aligned multi-contrast supervision and cross-patient factor swapping, AC-Diff learns to separate anatomical structure from acquisition-dependent contrast. Unlike conventional image or latent diffusion, AC-Diff restricts stochastic generation to the contrast subspace while the source anatomy bypasses diffusion and is directly reused during reconstruction. Experiments on an in-house multi-vendor cohort and the external Duke Liver MRI dataset demonstrate improved target-domain alignment with strong structural preservation. In downstream liver segmentation, AC-Diff improves Dice from 0.942 for the original inputs to 0.954 for the harmonized images. These results support contrast-specific latent generation as a promising approach to anatomy-preserving MRI harmonization.

bioengineering↗