Search bioRxivSearch

bioRxiv · 10.1101/2020.05.29.122994

Development and on-site evaluation of an easy-to-perform and low-cost food pathogen diagnostic workflow for low-resource communities

Abstract

Food-borne diseases are a leading cause of illness and death in many developing countries and thus, there is a real need to develop affordable and practical technologies that can help improve food safety in these countries. The ability to efficiently identify food pathogens is essential to allow national regulatory authorities to monitor food quality and implement safety protocols. In this study, we have developed a simple, low-cost ($0.76 (USD)) complete food pathogen diagnostic workflow ideally suited for deployment in low-resource environments that uses a simple four step process (sample enrichment, cell lysis, DNA amplification, and naked-eye readout). The minimal number of steps and equipment involved in our diagnostic workflow, as well as the simplicity of the yes/no flocculation readout, allows non-technical personnel to perform and interpret the assay. To evaluate the systems performance, we tested the entire system on fresh produce samples collected from local farms and markets in Cambodia for the presence of the E. coli O157 O-antigen polymerase, wzy. Although this was a proof-of-concept study, our system successfully revealed a clear correlation between the origin and condition of the produce collected and their likelihood of contamination. In conclusion, we believe that our easy-to-perform diagnostic system can have a significant impact on improving food quality and human health if adopted by regulatory authorities in developing countries due to the assays simplicity, affordability, and adaptability.

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Mason, M. G., Botella, J. R.. 2020-05-30. Development and on-site evaluation of an easy-to-perform and low-cost food pathogen diagnostic workflow for low-resource communities. https://doi.org/10.1101/2020.05.29.122994

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

KEEP EXPLORING

Related preprints

Trans-branching of polyubiquitin chains orchestrates the DNA replication stress response

Polyubiquitin chain geometry dictates functional consequences of ubiquitylation. Although branched polyubiquitin chains are abundant in cells, little is known about their functions. Here we show that branching on the DNA replication factor PCNA, mediated by the ubiquitin-conjugating enzyme UBE2K and involving lysines 63 and 48 of ubiquitin, orchestrates the sequence of events in response to replication stress. By inducing VCP-dependent extraction of PCNA from chromatin, branching promotes re-priming of stalled forks and necessitates a BRCA1-dependent pathway of daughter-strand gap repair. Our study identifies hyper-accumulation of daughter-strand gaps as the mechanistic basis underlying the toxicity of inhibitors of the PCNA-specific isopeptidase, USP1, in BRCA1-deficient cells. Moreover, an unexpected preference of UBE2K to operate in trans suggests a general timing mechanism to organize hierarchies amongst ubiquitin signals.

molecular biology

Impaired proteostasis is an early feature of the diabetic heart in humans and mice

Diabetes and obesity increase cardiac lipid levels leading to cardiomyopathy and heart failure. We hypothesized that intermittent fasting would reduce cardiac lipid levels. Surprisingly, intermittent fasting increased myocardial triglyceride content, but rescued mortality and attenuated cardiomyopathy in mice overexpressing cardiomyocyte acyl-CoA synthetase 1 (MHC-ACSL1). Lipid overload caused cardiomyocyte accumulation of polyubiquitinated protein aggregates containing desmin, a scaffolding intermediate filament protein, which intermittent fasting prevented. Furthermore, intermittent fasting reversed elevated myocardial C16:0 ceramide content, and knockdown of ceramide synthase CerS5 and CerS6 reduced palmitate-induced protein aggregation, highlighting a role for C16:0 ceramides in this pathology. Conversely, impairing aggrephagy with cardiomyocyte-specific p62 ablation induced heart failure in mice fed a high-fat diet, with paradoxically reduced cardiac lipid content. Crucially, non-failing diabetic human hearts also exhibited protein aggregate pathology. Taken together, these results demonstrate that impaired proteostasis characterizes cardiomyopathy from cardiac lipid overload and identify a promising new therapeutic target for this condition.

molecular biology

Spatial profiling and neurovascular communication in the developing and adolescent cortex following prenatal alcohol exposure

Fetal alcohol spectrum disorders (FASD) constitute a wide range of developmental, cognitive, and behavioral impairments caused by prenatal alcohol exposure (PAE). Although neuronal and vascular consequences of PAE have been studied, how alcohol affects the cerebrovasculature within the framework of the neurovascular unit (NVU) across development remains poorly understood. At minimum, the NVU comprises neurons, astrocyte endfeet, and endothelial cells (ECs), which coordinate to maintain brain homeostasis. Here, we used the NanoString Digital Spatial Profiling platform to characterize spatial transcriptomic data from neurons, astrocytes, and ECs from PAE and saccharin (SAC) control cortices at embryonic day 18 (E18) and postnatal day 28 (P28). Differentially expressed genes were then used for Ingenuity Pathway Analysis (IPA) to identify altered biological pathways and perform comparison analyses across developmental time points, while CellChat was used to infer cell cell communication networks. We uncovered thousands of differentially expressed genes and numerous altered pathways and biological processes in PAE cortices across development. Both IPA and CellChat analyses implicated dysregulation of vascular and extracellular matrix (ECM) remodeling, cell adhesion, and neuroinflammatory signaling. CellChat further predicted the loss of several key bidirectional relationships and altered ligand-receptor interactions among neurovascular cell types at E18 and P28. Overall, these findings identify PAE associated alterations in neurovascular gene expression and intercellular signaling across development, providing potential mechanisms by which PAE may disrupt neurodevelopment.

molecular biology