Generation of Thymidine Kinase-Deficient Mutants of Infectious Laryngotracheitis Virus

AbstractCurrent vaccines for the avian respiratory disease infectious laryngotracheitis consist of naturally attenuated strains of the causative agent—the herpesvirus infectious laryngotracheitis virus (ILTV). Due to the dissemination of these viruses from vaccinated chickens as well as their possible reversion to more pathogenic forms, the use of genetically engineered viral vaccines lacking virulence factors while retaining antigenicity is being considered. Since the thymidine kinase (TK) activity of herpesviruses has been associated with virulence, inactivation of the encoding gene in the ILTV genome should attenuate the virus. Moreover, by analogy to other TK- herpesviruses, the ability of such ILTV mutants to induce a protective response in chickens should not be compromised. Therefore, the deliberate genetic alteration of ILTV was attempted. In order to prevent reversion and also to enable identification of the modified virus, a "marker" transcriptional unit (Escherichia coli lacZ gene fused to a SV-40 3′-polyadenylation signal sequence and regulated by the pseudorabies virus gX gene promoter) was inserted via homologous recombination at one of two loci within the ILTV TK gene. Recombinant viruses were identified and plaque-purified on the basis of their ability to produce β-galactosidase. Retention of the foreign DNA at the predicted sites in the genomes of the recombinant ILTV was verified by Southern hybridization. Since their replication was unaffected by the thymine analog 1-(2-fluoro-2-deoxy-β-d-arabinofuranosyl)-5-methyluracil, the recombinants appeared to have a TK- phenotype. Despite this apparent deficiency, prior inoculation of either recombinant virus into chickens afforded the birds protection against a lethal challenge of virulent ILTV. Moreover, the degree of respiratory distress in the chickens vaccinated with the recombinants was relatively mild compared to the severe reaction in birds receiving the parental virus. Thus, ILTV can be genetically attenuated without an accompanying loss of immunogenicity

The composition of cosmic rays near the Bend (10 to the 15th power eV) from a study of muons in air showers at sea level

The distribution of muons near shower cores was studied at sea level at Fermilab using the E594 neutrino detector to sample the muon with E testing 3 GeV. These data are compared with detailed Monte Carlo simulations to derive conclusions about the composition of cosmic rays near the bend in the all particle spectrum. Monte Carlo simulations generating extensive air showers (EAS) with primary energy in excess of 50 TeV are described. Each shower record contains details of the electron lateral distribution and the muon and hadron lateral distributions as a function of energy, at the observation level of 100g/cm. The number of detected electrons and muons in each case was determined by a Poisson fluctuation of the number incident. The resultant predicted distribution of muons, electrons, the rate events are compared to those observed. Preliminary results on the rate favor a heavy primary dominated cosmic ray spectrum in energy range 50 to 1000 TeV

Study of muons near shower cores at sea level using the E594 neutrino detector

The E594 neutrino detector has been used to study the lateral distribution of muons of energy 3 GeV near shower cores. The detector consists of a 340 ton fine grain calorimeter with 400,000 cells of flash chamber and dimensions of 3.7 m x 20 m x 3.7 m (height). The average density in the calorimeter is 1.4 gm/sq cm, and the average Z is 21. The detector was triggered by four 0.6 sq m scintillators placed immediately on the top of the calorimeter. The trigger required at least two of these four counters. The accompanying extensive air showers (EAS) was sampled by 14 scintillation counters located up to 15 m from the calorimeter. Several off line cuts have been applied to the data. Demanding five particles in at least two of the trigger detectors, a total of 20 particles in all of them together, and an arrival angle for the shower 450 deg reduced the data sample to 11053 events. Of these in 4869 cases, a computer algorithm found at least three muons in the calorimeter

Improved Boundary Layer Depth Retrievals from MPLNET

Continuous lidar observations of the planetary boundary layer (PBL) depth have been made at the Micropulse Lidar Network (MPLNET) site in Greenbelt, MD since April 2001. However, because of issues with the operational PBL depth algorithm, the data is not reliable for determining seasonal and diurnal trends. Therefore, an improved PBL depth algorithm has been developed which uses a combination of the wavelet technique and image processing. The new algorithm is less susceptible to contamination by clouds and residual layers, and in general, produces lower PBL depths. A 2010 comparison shows the operational algorithm overestimates the daily mean PBL depth when compared to the improved algorithm (1.85 and 1.07 km, respectively). The improved MPLNET PBL depths are validated using radiosonde comparisons which suggests the algorithm performs well to determine the depth of a fully developed PBL. A comparison with the Goddard Earth Observing System-version 5 (GEOS-5) model suggests that the model may underestimate the maximum daytime PBL depth by 410 m during the spring and summer. The best agreement between MPLNET and GEOS-5 occurred during the fall and they diered the most in the winter

A simple method for simulating drought effects on plants

Drought is expected to increase in frequency and severity in many regions in the future, so it is important to improve our understanding of how drought affects plant functional traits and ecological interactions. Imposing experimental water deficits is key to gaining this understanding, but has been hindered by logistic difficulties in maintaining consistently low water availability for plants. Here, we describe a simple method for applying soil water deficits to potted plants in glasshouse experiments. We modified an existing method (the “Snow and Tingey system”) in order to apply a gradual, moderate water deficit to 50 plant species of different life forms (grasses, vines, shrubs, trees). The method requires less maintenance and manual handling compared to other water deficit methods, so it can be used for extended periods of time and is relatively inexpensive to implement. With only a few modifications, it is possible to easily establish and maintain soil water deficits of differing intensity and duration, as well as to incorporate interacting stress factors. We tested this method by measuring physiological responses to an applied water deficit in a subset of 11 tree/shrub species with a wide range of drought tolerances and water-use strategies. For this subgroup of species, stomatal conductance was 2–17 times lower in droughted plants than controls, although only half of the species (5 out of 11) experienced midday leaf water potentials that exceeded their turgor loss (i.e., wilting) point. Leaf temperatures were up to 8°C higher in droughted plants than controls, indicating that droughted plants are at greater risk of thermal damage, relative to unstressed plants. The largest leaf temperature differences (between droughted and well-watered plants) were in species with high rates of water loss. Rapid osmotic adjustment was observed in leaves of five species when drought stress was combined with an experimental heatwave. These results highlight the potential value of further ecological and physiological experiments utilizing this simple water deficit method to study plant responses to drought stress

Acute Liver Failure Secondary to Niacin Toxicity

A 17-year-old male was transferred to the pediatric intensive care unit for evaluation of acute liver failure. He was recently released from an alcohol treatment center with acute onset of chest pain. Cardiac workup was negative but he was found to have abnormal coagulation studies and elevated liver transaminases. Other evaluations included a normal toxicology screen and negative acetaminophen level. Autoimmune and infectious workups were normal providing no identifiable cause of his acute liver failure. He initially denied any ingestions or illicit drug use but on further query he admitted taking niacin in an attempt to obscure the results of an upcoming drug test. Niacin has been touted on the Internet as an aid to help pass urine drug tests though there is no evidence to support this practice. Niacin toxicity has been associated with serious multisystem organ failure and fulminant hepatic failure requiring liver transplantation. Pediatric providers should be aware of the risks associated with niacin toxicity and other experimental medical therapies that may be described on the Internet or other nonreputable sources

Observation of TeV Gamma Rays from the Crab Nebula with Milagro Using a New Background Rejection Technique

The recent advances in TeV gamma-ray astronomy are largely the result of the ability to differentiate between extensive air showers generated by gamma rays and hadronic cosmic rays. Air Cherenkov telescopes have developed and perfected the "imaging" technique over the past several decades. However until now no background rejection method has been successfully used in an air shower array to detect a source of TeV gamma rays. We report on a method to differentiate hadronic air showers from electromagnetic air showers in the Milagro gamma ray observatory, based on the ability to detect the energetic particles in an extensive air shower. The technique is used to detect TeV emission from the Crab nebula. The flux from the Crab is estimated to be 2.68(+-0.42stat +- 1.4sys) x10^{-7} (E/1TeV)^{-2.59} m^{-2} s^{-1} TeV^{-1}, where the spectral index is assumed to be as given by the HEGRA collaboration.Comment: 22 pages, 11 figures, submitted to Astrophysical Journa

Extreme heat increases stomatal conductance and drought-induced mortality risk in vulnerable plant species

Tree mortality during global-change-type drought is usually attributed to xylem dysfunction, but as climate change increases the frequency of extreme heat events, it is necessary to better understand the interactive role of heat stress. We hypothesized that some drought-stressed plants paradoxically open stomata in heatwaves to prevent leaves from critically overheating. We experimentally imposed heat (>40°C) and drought stress onto 20 broadleaf evergreen tree/shrub species in a glasshouse study. Most well-watered plants avoided lethal overheating, but drought exacerbated thermal damage during heatwaves. Thermal safety margins (TSM) quantifying the difference between leaf surface temperature and leaf critical temperature, where photosynthesis is disrupted, identified species vulnerability to heatwaves. Several mechanisms contributed to high heat tolerance and avoidance of damaging leaf temperatures—small leaf size, low leaf osmotic potential, high leaf mass per area (i.e., thick, dense leaves), high transpirational capacity, and access to water. Water-stressed plants had smaller TSM, greater crown dieback, and a fundamentally different stomatal heatwave response relative to well-watered plants. On average, well-watered plants closed stomata and decreased stomatal conductance (gs) during the heatwave, but droughted plants did not. Plant species with low gs, either due to isohydric stomatal behavior under water deficit or inherently low transpirational capacity, opened stomata and increased gs under high temperatures. The current paradigm maintains that stomata close before hydraulic thresholds are surpassed, but our results suggest that isohydric species may dramatically increase gs (over sixfold increases) even past their leaf turgor loss point. By actively increasing water loss at high temperatures, plants can be driven toward mortality thresholds more rapidly than has been previously recognized. The inclusion of TSM and responses to heat stress could improve our ability to predict the vulnerability of different tree species to future droughts

Calibration of the Milagro Cosmic Ray Telescope

The Milagro detector is an air shower array which uses the water Cherenkov technique and is capable of continuously monitoring the sky at energies near 1 TeV. The detector consists of 20000 metric tons of pure water instrumented with 723 photo-multiplier tubes (PMTs). The PMTs are arranged in a two-layer structure on a lattice of 3 m spacing covering 5000 $m^2$ area. The direction of the shower is determined from the relative timing of the PMT signals, necessitating a common time reference and amplitude slewing corrections to improve the time resolution. The calibration system to provide these consists of a pulsed laser driving 30 diffusing light sources deployed in the pond to allow cross-calibration of the PMTs. The system is capable of calibrating times and the pulse-heights from the PMTs using the time-over-threshold technique. The absolute energy scale is provided using single muons passing through the detector. The description of the calibration system of the Milagro detector and its prototype Milagrito will be presented.Comment: 4 pages, submitted to the XXVI International Cosmic Ray Conferenc