Metagenomic sequencing at the epicenter of the Nigeria 2018 Lassa fever outbreak.

The 2018 Nigerian Lassa fever season saw the largest ever recorded upsurge of cases, raising concerns over the emergence of a strain with increased transmission rate. To understand the molecular epidemiology of this upsurge, we performed, for the first time at the epicenter of an unfolding outbreak, metagenomic nanopore sequencing directly from patient samples, an approach dictated by the highly variable genome of the target pathogen. Genomic data and phylogenetic reconstructions were communicated immediately to Nigerian authorities and the World Health Organization to inform the public health response. Real-time analysis of 36 genomes and subsequent confirmation using all 120 samples sequenced in the country of origin revealed extensive diversity and phylogenetic intermingling with strains from previous years, suggesting independent zoonotic transmission events and thus allaying concerns of an emergent strain or extensive human-to-human transmission.

First Complete Genome Sequences of Zika Virus Isolated from Febrile Patient Sera in Ecuador.

Here, we present the complete genome sequences of two Zika virus (ZIKV) strains, EcEs062_16 and EcEs089_16, isolated from the sera of febrile patients in Esmeraldas City, in the northern coastal province of Esmeraldas, Ecuador, in April 2016. These are the first complete ZIKV genomes to be reported from Ecuador.

Development and application of a catchment scale pesticide fate and transport model for use in drinking water risk assessment.

This paper describes the development and application of IMPT (Integrated Model for Pesticide Transport), a parameter-efficient tool for predicting diffuse-source pesticide concentrations in surface waters used for drinking water supply. The model was applied to a small UK headwater catchment with high frequency (8h) pesticide monitoring data and to five larger catchments (479-1653km(2)) with sampling approximately every 14days. Model performance was good for predictions of both flow (Nash Sutcliffe Efficiency generally >0.59 and PBIAS <10%) and pesticide concentrations, although low sampling frequency in the larger catchments is likely to mask the true episodic nature of exposure. The computational efficiency of the model, along with the fact that most of its parameters can be derived from existing national soil property data mean that it can be used to rapidly predict pesticide exposure in multiple surface water resources to support operational and strategic risk assessments.

Mastoparan causes cell permeabilisation and delayed activation of DNA synthesis in Swiss 3T3 fibroblasts.

Mastoparan, a tetradecapeptide from wasp venom, preferentially activates the heterotrimeric G proteins, Go and Gi by promoting GDP/GTP exchange. The peptide was originally identified as a potent secretagogue and has since been shown to promote DNA synthesis in Swiss 3T3 fibroblasts. Here, we have shown that mastoparan (10-20 microM), either alone or in combination with the co-mitogen insulin, had no effect on DNA synthesis when incubated with cells for 24 h. However, in the presence of insulin, the peptide evoked a small increase in DNA synthesis after incubation for 40 h. Thus, unlike other mitogens, mastoparan caused a delayed activation of DNA synthesis. At concentrations of mastoparan (15-17.5 microM) which promoted DNA synthesis, the peptide caused a rapid release of lactate dehydrogenase from the cells. These data suggest that the mitogenic action of mastoparan occurs by a mechanism distinct from that of physiological mitogens.

A novel metallo-beta-lactamase, Mbl1b, produced by the environmental bacterium Caulobacter crescentus.

Caulobacter crescentus 101123 possesses a gene (Mbl1b) encoding a metallo-beta-lactamase with 32% amino acid identity to the L1 metallo-beta-lactamase from Stenotrophomonas maltophilia. The gene was cloned into an expression vector and the enzyme, Mbl1b, was expressed in Escherichia coli. Mbl1b was purified. Catalytic properties for several antibiotics were determined. The enzyme exhibits Michaelis-Menten kinetics for imipenem, meropenem and nitrocefin but substrate inhibition kinetics with cefoxitin, cefaloridine, penicillin G and ampicillin. A homology model predicts Mbl1b has the same structural fold as other metallo-beta-lactamases with a detailed structure very similar to L1 but whereas L1 is a homotetramer, Mbl1b is monomeric. The main differences between Mbl1 and L1 are in the N-terminal region.

Recruitment & retention. A successful model in forensic psychiatric nursing.

1. The recruitment and retention of forensic psychiatric nurses in this highly competitive environment has been identified as a critical issue. 2. In response to the need to expand services, the development, implementation, and evaluation of an innovative model that has demonstrated success in the recruitment and retention of nurses for this highly specialized area of practice are described. 3. The successful recruitment and retention of forensic psychiatric nurses may be facilitated by developing and implementing strategies that integrate the goals and objectives of the organization with the needs of individual nurses.

Measurement and conceptual modelling of herbicide transport to field drains in a heavy clay soil with implications for catchment-scale water quality management.

Propyzamide and carbetamide are essential for blackgrass control in oilseed rape production. However, both of these compounds can contaminate surface waters and pose compliance problems for water utilities. The transport of propyzamide and carbetamide to an instrumented field drain in a small clay headwater tributary of the Upper Cherwell catchment was monitored over a winter season. Despite having very different sorption and dissipation properties, both herbicides were transported rapidly to the drain outlet in the first storm event after application, although carbetamide was leached more readily than propyzamide. A simple conceptual model was constructed to represent solute displacement from mobile pore water and preferential flow to drains. The model was able to reproduce the timing and magnitude of herbicide losses well, lending support to its conceptual basis. Measured losses in drainflow in the month following application were 1.1 and 8.1%, respectively, for propyzamide and carbetamide. Differences were due to a combination of differences in herbicide mobility and due to the fact that the monitoring period for carbetamide was hydrologically more active. For both compounds, losses were greater than those typically reported elsewhere for other herbicides. The data suggest that drainflow is the dominant pathway for the transfer of these herbicides to the catchment outlet, where water is abstracted for municipal supply. This imposes considerable constraints on the management options available to reduce surface water concentrations of herbicides in this catchment.

Requirement of basement membrane for the suppression of programmed cell death in mammary epithelium.

Apoptosis is an active mechanism of cell death required for normal tissue homeostasis. Cells require survival signals to avoid the engagement of apoptosis. In the mammary gland, secretory epithelial cells are removed by apoptosis during involution. This cell loss coincides with matrix metalloproteinase activation and basement membrane degradation. In this paper we describe studies that confer a new role for basement membrane in the regulation of cell phenotype. We demonstrate that the first passage epithelial cells isolated from pregnant mouse mammary gland die by apoptosis in culture, but that cell death is suppressed by basement membrane. The correct type of extracellular matrix was required, since only a basement membrane, not plastic or a collagen I matrix, lowered the rate of apoptosis. Attachment to a matrix per se was not sufficient for survival, since apoptotic cells were observed when still attached to a collagen I substratum. Experiments with individually isolated cells confirmed the requirement of basement membrane for survival, and demonstrated that survival is enhanced by cell-cell contact. A function-blocking anti-beta1 integrin antibody doubled the rate of apoptosis in single cells cultured with basement membrane, indicating that integrin-mediated signals contributed to survival. We examined the cell death-associated genes bcl-2 and bax in mammary epithelia, and found that although the expression of Bcl-2 did not correlate with cell survival, increased levels of Bax were associated with apoptosis. We propose that basement membrane provides a survival stimulus for epithelial cells in vivo, and that loss of interaction with this type of matrix acts as a control point for cell deletions that occur at specific times during development, such as in mammary gland involution.