A rapid, antibiotic susceptibility test for multidrug-resistant, Gram-negative bacterial uropathogens using the biochemical assay, DETECT.

The increasing prevalence of extended spectrum ß-lactamases (ESBLs) and plasmid-mediated AmpC (pAmpC) ß-lactamases among Enterobacterales threatens our ability to treat urinary tract infections (UTIs). These organisms are resistant to most ß-lactam antibiotics and are frequently multidrug-resistant (MDR). Consequently, they are often resistant to antibiotics used to empirically treat UTIs. The lack of rapid diagnostic and antibiotic susceptibility tests (AST) makes clinical management of UTIs caused by such organisms difficult, as standard culture and susceptibility assays require several days. We have adapted a biochemical detection assay, termed dual-enzyme trigger-enabled cascade technology (DETECT) for rapid detection of resistance (time-to-result of 3 h) to other antibiotics commonly used in treatment of UTIs. DETECT is activated by the presence of CTX-M and pAmpC ß-lactamases. In this proof-of-concept study, the adapted DETECT assay (AST-DETECT) has been performed on pure-cultures of Klebsiella pneumoniae and Escherichia coli (48 isolates) expressing ESBL or pAmpC ß-lactamases to perform AST for ciprofloxacin (sensitivity 96.9%, specificity 100%, accuracy 97.9%) nitrofurantoin (sensitivity 95.7%, specificity 91.7%, accuracy 94%) and trimethoprim/sulfamethoxazole (sensitivity 83.3%, specificity 100%, accuracy 89.4%). These results suggest that AST-DETECT may be adapted as a potential diagnostic platform to rapidly detect multidrug-resistant E. coli and K. pneumoniae that cause UTI.

Author Correction: L1-associated genomic regions are deleted in somatic cells of the healthy human brain.

In the version of this article initially published, NIH grant U01 MH106882 to F.H.G. was missing from the Acknowledgments. The error has been corrected in the HTML and PDF versions of the article.

Acetylcholinesterase activity and time after a peak pesticide-use period among Ecuadorian children.

PURPOSE: Mother's Day (May) is a holiday with substantial demand for flowers, associated with heightened flower production and escalated pesticide use. The effect of spray seasons on pesticide exposures of children living in agricultural communities but who do not work in agriculture is poorly understood. In this study, we estimated the association of time after Mother's Day harvest with children's acetylcholinesterase (AChE) activity. AChE is a physiological marker of organophosphate/carbamate pesticide exposures that may take up to 3 months to normalize after its inhibition. METHODS: We examined 308 children, aged 4-9 years, in Ecuadorian agricultural communities during a low flower-production season but within 63-100 days (mean: 81.5 days, SD: 10.9) after Mother's Day harvest. We quantified AChE activity (mean: 3.14 U/mL, SD: 0.49) from a single finger-stick sample. RESULTS: We observed positive linear associations between time after the harvest and AChE among participants living near plantations. The associations were strongest among participants living within 233 m [(0.15 U/mL (95% CI 0.02, 0.28)], slightly weaker among participants living within 234-532 m [0.11 U/mL (0.00, 0.23)], and not associated among participants at greater distances. Similar findings were observed across categories of areas of flower plantations within 500 m of homes. CONCLUSIONS: These cross-sectional findings suggest that a peak pesticide-use period can decrease AChE activity of children living near plantations. These seasonal pesticide exposures could induce short- and long-term developmental alterations in children. Studies assessing exposures at multiple times in relation to pesticide spray seasons among children who do not work in agriculture are needed.

L1-associated genomic regions are deleted in somatic cells of the healthy human brain.

The healthy human brain is a mosaic of varied genomes. Long interspersed element-1 (LINE-1 or L1) retrotransposition is known to create mosaicism by inserting L1 sequences into new locations of somatic cell genomes. Using a machine learning-based, single-cell sequencing approach, we discovered that somatic L1-associated variants (SLAVs) are composed of two classes: L1 retrotransposition insertions and retrotransposition-independent L1-associated variants. We demonstrate that a subset of SLAVs comprises somatic deletions generated by L1 endonuclease cutting activity. Retrotransposition-independent rearrangements in inherited L1s resulted in the deletion of proximal genomic regions. These rearrangements were resolved by microhomology-mediated repair, which suggests that L1-associated genomic regions are hotspots for somatic copy number variants in the brain and therefore a heritable genetic contributor to somatic mosaicism. We demonstrate that SLAVs are present in crucial neural genes, such as DLG2 (also called PSD93), and affect 44-63% of cells of the cells in the healthy brain.