LplA1-dependent utilization of host lipoyl peptides enables Listeria cytosolic growth and virulence

The bacterial pathogen Listeria monocytogenes replicates within the cytosol of mammalian cells. Mechanisms by which the bacterium exploits the host cytosolic environment for essential nutrients are poorly defined. L. monocytogenes is a lipoate auxotroph and must scavenge this critical cofactor, using lipoate ligases to facilitate attachment of the lipoyl moiety to metabolic enzyme complexes. Although the L. monocytogenes genome encodes two putative lipoate ligases, LplA1 and LplA2, intracellular replication and virulence require only LplA1. Here we show that LplA1 enables utilization of host-derived lipoyl peptides by L. monocytogenes . LplA1 is dispensable for growth in the presence of free lipoate, but necessary for growth on low concentrations of mammalian lipoyl peptides. Furthermore, we demonstrate that the intracellular growth defect of the δ lplA1 mutant is rescued by addition of exogenous lipoic acid to host cells, suggesting that L. monocytogenes dependence on LplA1 is dictated by limiting concentrations of available host lipoyl substrates. Thus, the ability of L. monocytogenes and other intracellular pathogens to efficiently use host lipoyl peptides as a source of lipoate may be a requisite adaptation for life within the mammalian cell.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/72528/1/MMI+5956+Supp.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/72528/2/j.1365-2958.2007.05956.x.pd

Temperature dependence of single particle excitations in a S=1 chain: exact diagonalization calculations compared to neutron scattering experiments

Exact diagonalization calculations of finite antiferromagnetic spin-1 Heisenberg chains at finite temperatures are presented and compared to a recent inelastic neutron scattering experiment for temperatures T up to 7.5 times the intrachain exchange constant J. The calculations show that the excitations at the antiferromagnetic point q=1 and at q=0.5 remain resonant up to at least T=2J, confirming the recent experimental observation of resonant high-temperature domain wall excitations. The predicted first and second moments are in good agreement with experiment, except at temperatures where three-dimensional spin correlations are most important. The ratio of the structure factors at q=1 and at q=0.5 is well predicted for the paramagnetic infinite-temperature limit. For T=2J, however, we found that the experimentally observed intensity is considerably less than predicted. This suggests that domain wall excitations on different chains interact up to temperatures of the order of the spin band width.Comment: 9 pages revtex, submitted to PR

Reverse-Engineering a Transcriptional Enhancer: A Case Study in Drosophila

Abstract Enhancers, or cis-regulatory elements, are the principal determinants of spatiotemporal patterning of gene expression. For reasons of clinical and research utility, it is desirable to build customized enhancers that drive novel gene expression patterns, but currently, we largely rely on “found” genomic elements. Synthetic enhancers, assembled from transcription factor binding sites taken from natural signal-regulated enhancers, generally fail to behave like their wild-type counterparts when placed in transgenic animals, suggesting that important aspects of enhancer function are still unexplored. As a step toward the creation of a truly synthetic regulatory element, we have undertaken an extensive structure–function study of an enhancer of the Drosophila decapentaplegic (dpp) gene that drives expression in the developing visceral mesoderm (VM). Although considerable past efforts have been made to dissect the dppVM enhancer, transgenic experiments presented here indicate that its activity cannot be explained by the known regulators alone. dppVM contains multiple, previously uncharacterized, regulatory sites, some of which exhibit functional redundancy. The results presented here suggest that even the best-studied enhancers must be further dissected before they can be fully understood, and before faithful synthetic elements based on them can be created. Implications for developmental genetics, mathematical modeling, and therapeutic applications are discussed.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/63213/1/ten.tea.2008.0074.pd

Neighbourhood prevalence-to-notification ratios for adult bacteriologically-confirmed tuberculosis reveals hotspots of underdiagnosis in Blantyre, Malawi

Local information is needed to guide targeted interventions for respiratory infections such as tuberculosis (TB). Case notification rates (CNRs) are readily available, but systematically underestimate true disease burden in neighbourhoods with high diagnostic access barriers. We explored a novel approach, adjusting CNRs for under-notification (P:N ratio) using neighbourhood-level predictors of TB prevalence-to-notification ratios. We analysed data from 1) a citywide routine TB surveillance system including geolocation, confirmatory mycobacteriology, and clinical and demographic characteristics of all registering TB patients in Blantyre, Malawi during 2015-19, and 2) an adult TB prevalence survey done in 2019. In the prevalence survey, consenting adults from randomly selected households in 72 neighbourhoods had symptom-plus-chest X-ray screening, confirmed with sputum smear microscopy, Xpert MTB/Rif and culture. Bayesian multilevel models were used to estimate adjusted neighbourhood prevalence-to-notification ratios, based on summarised posterior draws from fitted adult bacteriologically-confirmed TB CNRs and prevalence. From 2015-19, adult bacteriologically-confirmed CNRs were 131 (479/371,834), 134 (539/415,226), 114 (519/463,707), 56 (283/517,860) and 46 (258/578,377) per 100,000 adults per annum, and 2019 bacteriologically-confirmed prevalence was 215 (29/13,490) per 100,000 adults. Lower educational achievement by household head and neighbourhood distance to TB clinic was negatively associated with CNRs. The mean neighbourhood P:N ratio was 4.49 (95% credible interval [CrI]: 0.98-11.91), consistent with underdiagnosis of TB, and was most pronounced in informal peri-urban neighbourhoods. Here we have demonstrated a method for the identification of neighbourhoods with high levels of under-diagnosis of TB without the requirement for a prevalence survey; this is important since prevalence surveys are expensive and logistically challenging. If confirmed, this approach may support more efficient and effective targeting of intensified TB and HIV case-finding interventions aiming to accelerate elimination of urban TB

The Mw 5.1, 9 August 2020, Sparta Earthquake, North Carolina: The First Documented Seismic Surface Rupture in the Eastern United States

At 8:07 a.m. EDT on 9 Aug. 2020 a Mw 5.1 earthquake located ~3 km south of Sparta, North Carolina, USA, shook much of the eastern United States, producing the first documented surface rupture due to faulting east of the New Madrid seismic zone. The co-seismic surface rupture was identified along a 2-km-long traceable zone of predominantly reverse displacement, with folding and flexure generating a scarp averaging 8–10-cm-high with a maximum observed height of ~25 cm. Widespread deformation south of the main surface rupture includes cm-dm–long and mm-cm–wide fissures. Two trenches excavated across the surface rupture reveal that this earthquake propagated to the surface along a preexisting structure in the shallow bedrock, which had not been previously identified as an active fault. Surface ruptures by faulting are rarely reported for M <6 earthquakes, and hence the Sparta earthquake provides an opportunity to improve seismic hazard knowledge associated with these moderate events. Furthermore, this earthquake occurred in a very low strain rate intraplate setting, where earthquake surface deformation, regardless of magnitude, is sparse in time and rare to observe and characterize

Effects of N-acetyl-cysteine on endothelial function and inflammation in patients with type 2 diabetes mellitus

Endothelial dysfunction has been associated with premature vascular disease. There is increasing data that N-acetyl-cysteine (NAC) may prevent or improve endothelial dysfunction. The aim of this study was to assess the effects of NAC on endothelial function in patients with type 2 diabetes mellitus, a population at high risk for endothelial dysfunction. Twenty-four patients with diabetes mellitus were assigned randomly to initial therapy with either 900 mg NAC or placebo twice daily in a double-blind, cross-over study design. Flowmediated vasodilation (FMD) of the brachial artery was assessed at baseline, after four weeks of therapy, after a four-week wash-out period, and after another four weeks on the opposite treatment. Plasma and red blood cell glutathione levels and high-sensitivity C-reactive protein (CRP) were measured at all four visits. At baseline, FMD was moderately impaired (3.7±2.9%). There was no significant change in FMD after four weeks of NAC therapy as compared to placebo (0.1±3.6% vs. 1.2±4.2%). Similarly, there was no significant change in glutathione levels. However, median CRP decreased from 2.35 to 2.14 mg/L during NAC therapy (p=0.04), while it increased from 2.24 to 2.65 mg/L with placebo. No side effects were noted during the treatment period. In this double-blind, randomized cross-over study, four weeks of oral NAC therapy failed to improve endothelial dysfunction in patients with diabetes mellitus. However, NAC therapy decreased CRP levels, suggesting that this compound may have some efficacy in reducing systemic inflammation

The transfer of diatoms from freshwater to footwear materials: An experimental study assessing transfer, persistence, and extraction methods for forensic reconstruction

In recent years there has been growing interest in environmental forms of trace evidence, and ecological trace evidence collected from footwear has proved valuable within casework. Simultaneously, there has been growing awareness of the need for empirical experimentation to underpin forensic inferences. Diatoms are unicellular algae, and each cell (or ‘frustule’) consists of two valves which are made of silica, a robust material that favours their preservation both in sediments and within forensic scenarios. A series of experiments were carried out to investigate the transfer and persistence of diatoms upon common footwear materials, a recipient surface that has historically been overlooked by studies of persistence. The effectiveness of two novel extraction techniques (jet rinsing, and heating and agitation with distilled water) was compared to the established extraction technique of hydrogen peroxide digestion, for a suite of five common footwear materials: canvas, leather, and ‘suede’ (representing upper materials), and rubber and polyurethane (representing sole materials). It was observed that the novel extraction technique of heating and agitation with distilled water did not extract fewer diatom valves, or cause increased fragmentation of valves, when compared to peroxide digestion, suggesting that the method may be viable where potentially hazardous chemical reactions may be encountered with the peroxide digestion method. Valves could be extracted from all five footwear materials after 3 min of immersion, and more valves were extracted from the rougher, woven upper materials than the smoother sole materials. Canvas yielded the most valves (a mean of 2511/cm2) and polyurethane the fewest (a mean of 15/cm2). The persistence of diatoms on the three upper materials was addressed with a preliminary pilot investigation, with ten intervals sampled between 0 and 168 h. Valves were seen to persist in detectable quantities after 168 h on all three upper materials. However, some samples produced slides with no valves, and the earliest time after which no diatom valves were found was 4 h after the transfer. Analysis of the particle size distributions over time, by image analysis, suggests that the retention of diatoms may be size-selective; after 168 h, no particles larger than 200 μm2 could be found on the samples of canvas, and > 95% of the particles on the samples of suede were less than or equal to 200 μm2. A pilot investigation into the effects of immersion interval was carried out upon samples of canvas. Greater numbers of valves were extracted from the samples with longer immersion intervals, but even after 30 s, > 500 valves could be recovered per cm2, suggesting that footwear may be sampled for diatoms even if the contact with a water body may have been brief. These findings indicate that, if the variability within and between experimental runs can be addressed, there is significant potential for diatoms to be incorporated into the trace analysis of footwear and assist forensic reconstructions

Middle to late Pleistocene palaeoecological reconstructions and palaeotemperature estimates for cold/cool stage deposits at Whittlesey, eastern England

Fossiliferous beds in a complex sequence of late Middle to Late Pleistocene deposits at Whittlesey, eastern England, provided a rare opportunity for a multidisciplinary study of the palaeoecology of cool/cold stage deposits from different glacial stages. The fossiliferous sediments investigated form part of the River Nene 1st Terrace. Three of the four fossil assemblages investigated pre-date the last interglacial stage (Ipswichian/Eemian/marine oxygen isotope stage (MIS) 5e), whereas the other dates to part of the MIS 3 interstadial complex (Middle Devensian/Weichselian). Pollen, plant macrofossil, molluscan, coleopteran, ostracod, foraminifera and vertebrate data are available to a greater or lesser extent for each cool/cold stage assemblage, and they broadly present the same ecological picture for each one: a continuum from low-energy permanent to non-permanent aquatic habitats through marshland with associated waterside taxa, together with flood influxes of fluvial, riparian and ruderal taxa. Although each fossil assemblage records cool/cold climatic conditions, to a greater or lesser extent, these conditions are more apparent in the insect and ostracod faunas. In comparison with results published for the Last Glacial Maximum (LGM) stadial in The Netherlands, palaeotemperature estimates based on ranges of mutual agreement between independent coleopteran and ostracod methods for the three pre-Ipswichian/Eemian assemblages indicate minimum mean July air temperatures that are from +1° to +3 °C warmer, but January values that embrace the −8 °C estimate for the LGM. There is, however, a disparity between the coleopteran and ostracod palaeotemperature estimates for the Middle Devensian/Weichselian fossil assemblage, which are based on two different sample stratigraphic levels; the lower, coleopteran assemblage is indicative of very cool, continental climates, whereas the stratigraphically slightly higher ostracod assemblage suggests a climatic amelioration. Lack of numerical age-estimates prevents a robust stratigraphical interpretation, but the youngest pre-Ipswichian/Eemian fossil assemblage could date to the MIS 7–6 transition, at a time when cooling possibly preceded glacially driven sea-level fall. It is apparent from the rich coleopteran data that some continental cold-indicator taxa also appeared in pre-Ipswichian/Eemian cold stages and therefore assignment of continental cold-indicator taxa to particular Devensian/Weichselian intervals should be undertaken with care