Long-term Recovery from Acute Cold Shock in Caenorhabditis Elegans

Background Animals are exposed to a wide range of environmental stresses that can cause potentially fatal cellular damage. The ability to survive the period of stress as well as to repair any damage incurred is essential for fitness. Exposure to 2 °C for 24 h or longer is rapidly fatal to the nematode Caenorhabditis elegans, but the process of recovery from a shorter, initially non-lethal, cold shock is poorly understood. Results We report that cold shock of less than 12-hour duration does not initially kill C. elegans, but these worms experience a progression of devastating phenotypes over the next 96 h that correlate with their eventual fate: successful recovery from the cold shock and survival, or failure to recover and death. Cold-shocked worms experience a marked loss of pigmentation, decrease in the size of their intestine and gonads, and disruption to the vulva. Those worms who will successfully recover from the cold shock regain their pigmentation and much of the integrity of their intestine and gonads. Those who will die do so with a distinct phenotype from worms dying during or immediately following cold shock, suggesting independent mechanisms. Worms lacking the G-protein coupled receptor FSHR-1 are resistant to acute death from longer cold shocks, and are more successful in their recovery from shorter sub-lethal cold shocks. Conclusions We have defined two distinct phases of death associated with cold shock and described a progression of phenotypes that accompanies the course of recovery from that cold shock. The G-protein coupled receptor FSHR-1 antagonizes these novel processes of damage and recovery

The Conserved G-Protein Coupled Receptor FSHR-1 Regulates Protective Host Responses to Infection and Oxidative Stress

The innate immune system’s ability to sense an infection is critical so that it can rapidly respond if pathogenic microorganisms threaten the host, but otherwise maintain a quiescent baseline state to avoid causing damage to the host or to commensal microorganisms. One important mechanism for discriminating between pathogenic and non-pathogenic bacteria is the recognition of cellular damage caused by a pathogen during the course of infection. InCaenorhabditis elegans, the conserved G-protein coupled receptor FSHR-1 is an important constituent of the innate immune response. FSHR-1 activates the expression of antimicrobial infection response genes in infected worms and delays accumulation of the ingested pathogenPseudomonas aeruginosa. FSHR-1 is central not only to the worm’s survival of infection by multiple pathogens, but also to the worm’s survival of xenobiotic cadmium and oxidative stresses. Infected worms produce reactive oxygen species to fight off the pathogens; FSHR-1 is required at the site of infection for the expression of detoxifying genes that protect the host from collateral damage caused by this defense response. Finally, the FSHR-1 pathway is important for the ability of worms to discriminate pathogenic from benign bacteria and subsequently initiate an aversive learning program that promotes selective pathogen avoidance

Development of a low-maintenance measurement approach to continuously estimate methane emissions: a case study

The chemical breakdown of organic matter in landfills represents a significant source of methane gas (CH4). Current estimates suggest that landfills are responsible for between 3% and 19% of global anthropogenic emissions. The net CH4 emissions resulting from biogeochemical processes and their modulation by microbes in landfills are poorly constrained by imprecise knowledge of environmental constraints. The uncertainty in absolute CH4 emissions from landfills is therefore considerable. This study investigates a new method to estimate the temporal variability of CH4 emissions using meteorological and CH4 concentration measurements downwind of a landfill site in Suffolk, UK from July to September 2014, taking advantage of the statistics that such a measurement approach offers versus shorter-term, but more complex and instantaneously accurate, flux snapshots. Methane emissions were calculated from CH4 concentrations measured 700 m from the perimeter of the landfill with observed concentrations ranging from background to 46.4 ppm. Using an atmospheric dispersion model, we estimate a mean emission flux of 709 μg m−2 s−1 over this period, with a maximum value of 6.21 mg m−2 s−1, reflecting the wide natural variability in biogeochemical and other environmental controls on net site emission. The emissions calculated suggest that meteorological conditions have an influence on the magnitude of CH4 emissions. We also investigate the factors responsible for the large variability observed in the estimated CH4 emissions, and suggest that the largest component arises from uncertainty in the spatial distribution of CH4 emissions within the landfill area. The results determined using the low-maintenance approach discussed in this paper suggest that a network of cheaper, less precise CH4 sensors could be used to measure a continuous CH4 emission time series from a landfill site, something that is not practical using far-field approaches such as tracer release methods. Even though there are limitations to the approach described here, this easy, low-maintenance, low-cost method could be used by landfill operators to estimate time-averaged CH4 emissions and their impact downwind by simultaneously monitoring plume advection and CH4 concentrations

HST ultraviolet spectral energy distributions for three ultraluminous infrared galaxies

We present HST Faint Object Camera ultraviolet (230 nm and 140 nm) images of three ultraluminous infrared galaxies (ULIG: L_ir > 10^12 L_sun) selected from the IRAS Revised Bright Galaxy Sample. The purpose is to estimate spectral energy distributions (SEDs) to facilitate the identification of similar objects at high redshift in deep optical, infrared, and submm surveys. All three galaxies (VII Zw031 = IRAS F12112+0305, and IRAS F22491-1808) were well detected at 230 nm. Two of the three were marginally detected at 140 nm. The fluxes, together with ground-based optical and infrared photometry, are used to compute SEDs over a wide wavelength range. The measured SEDs drop from the optical to the ultraviolet, but the magnitude of the drop ranges from a factor of ~3 in IRAS F22491-1808 to a factor of ~100 in VIIZw031. This is most likely due to different internal extinctions. Such an interpretation is also suggested by extrapolating to ultraviolet wavelengths the optical internal extinction measured in VIIZw031. K-corrections are calculated to determine the colors of the sample galaxies as seen at high redshifts. Galaxies like VIIZw031 have very low observed rest-frame UV fluxes which means that such galaxies at high redshift will be extremely red or even missing in optical surveys. On the other hand, galaxies like IRAS F12112+0305 and IRAS F22491-1808, if seen at high redshift, would be sufficiently blue that they would not easily be distinguished from normal field galaxies, and therefore, identified as ULIGs. The implication is then that submillimeter surveys may be the only means of properly identifying the majority of ULIGs at high redshift.Comment: AJ in press, TeX, 23 pages, 7 tab, 17 figs available also (at higher resolution) from http://www.ast.cam.ac.uk~trentham/ufigs.htm

Treatment of calibration uncertainty in multi-baseline cross-correlation searches for gravitational waves

Uncertainty in the calibration of gravitational wave (GW) detector data leads to systematic errors, which must be accounted for in setting limits on the strength of GW signals. When cross-correlation measurements are made using data from a pair of instruments, as in searches for a stochastic GW background, the calibration uncertainties of the individual instruments can be combined into an uncertainty associated with the pair. With the advent of multi-baseline GW observation (e.g., networks consisting of multiple detectors such as the LIGO observatories and Virgo), a more sophisticated treatment is called for. We have described how the correlations between calibration factors associated with different pairs can be taken into account by marginalizing over the uncertainty associated with each instrument

Enhanced Sample Multiplexing of Tissues Using Combined Precursor Isotopic Labeling and Isobaric Tagging (cPILOT)

There is an increasing demand to analyze many biological samples for disease understanding and biomarker discovery. Quantitative proteomics strategies that allow simultaneous measurement of multiple samples have become widespread and greatly reduce experimental costs and times. Our laboratory developed a technique called combined precursor isotopic labeling and isobaric tagging (cPILOT), which enhances sample multiplexing of traditional isotopic labeling or isobaric tagging approaches. Global cPILOT can be applied to samples originating from cells, tissues, bodily fluids, or whole organisms and gives information on relative protein abundances across different sample conditions. cPILOT works by 1) using low pH buffer conditions to selectively dimethylate peptide N-termini and 2) using high pH buffer conditions to label primary amines of lysine residues with commercially-available isobaric reagents (see Table of Materials/Reagents). The degree of sample multiplexing available is dependent on the number of precursor labels used and the isobaric tagging reagent. Here, we present a 12-plex analysis using light and heavy dimethylation combined with six-plex isobaric reagents to analyze 12 samples from mouse tissues in a single analysis. Enhanced multiplexing is helpful for reducing experimental time and cost and more importantly, allowing comparison across many sample conditions (biological replicates, disease stage, drug treatments, genotypes, or longitudinal time-points) with less experimental bias and error. In this work, the global cPILOT approach is used to analyze brain, heart, and liver tissues across biological replicates from an Alzheimer's disease mouse model and wild-type controls. Global cPILOT can be applied to study other biological processes and adapted to increase sample multiplexing to greater than 20 samples