Ferrous Campylobacter jejuni truncated hemoglobin P displays an extremely high reactivity for cyanide - A comparative study

Campylobacter jejuni hosts two hemoglobins (Hbs). The Camplylobacter jejuni single-domain Hb (called Cgb) is homologous to the globin domain of flavohemoglobin, and it has been proposed to protect the bacterium against nitrosative stress. The second Hb is called Ctb (hereafter Cj-trHbP), belongs to truncated Hb group III, and has been hypothesized to be involved in O 2 chemistry. Here, the kinetics and thermodynamics of cyanide binding to ferric and ferrous Cj-trHbP [Cj-trHbP(III) and Cj-trHbP(II), respectively] are reported and analyzed in parallel with those of related heme proteins, with particular reference to those from Mycobacterium tuberculosis. The affinity of cyanide for Cj-trHbP(II) is higher than that reported for any known (in)vertebrate globin by more than three orders of magnitude (K = 1.2 × 10-6 m). This can be fully attributed to the highest (ever observed for a ferrous Hb) cyanide-binding association rate constant (kon = 3.3 × 103 m-1·s-1), even though the binding process displays a rate-limiting step (kmax = 9.1 s -1). Cj-trHbP(III) shows a very high affinity for cyanide (L = 5.8 × 10-9 m); however, cyanide association kinetics are independent of cyanide concentration, displaying a rate-limiting step (l max = 2.0 × 10-3 s-1). Values of the first-order rate constant for cyanide dissociation from Cj-trHbP(II)-cyanide and Cj-trHbP(III)-cyanide (koff =5.0 × 10-3 s -1 and loff ≥ 1 × 10-4 s-1, respectively) are similar to those reported for (in)vertebrate globins. The very high affinity of cyanide for Cj-trHbP(II), reminiscent of that of horseradish peroxidase(II), suggests that this globin may participate in cyanide detoxification. © 2008 The Authors

Determinants of Oral Corticosteroid Responsiveness in Wheezing Asthmatic Youth (DOORWAY): Protocol for a prospective multicentre cohort study of children with acute moderate-to-severe asthma exacerbations

Introduction: Oral corticosteroids are the cornerstone of acute asthma management in the emergency department. Recent evidence has raised doubts about the efficacy of this treatment in preschool-aged children with viral-induced wheezing and in smoking adults. The aims of the study were to: (1) document the magnitude of response to oral corticosteroids in children presenting to the emergency department with moderate or severe asthma; (2) quantify potential determinants of response to corticosteroids and (3) explore the role of gene polymorphisms associated with the responsiveness to corticosteroids. Methods and analysis: The design is a prospective cohort study of 1008 children aged 1-17 years meeting a strict definition of asthma and presenting with a clinical score of ≥4 on the validated Pediatric Respiratory Assessment Measure. All children will receive standardised severity-specific treatment with prednisone/prednisolone and cointerventions (salbutamol with/without ipratropium bromide). Determinants, namely viral aetiology, environmental tobacco smoke and single nucleotide polymorphism, will be objectively documented. The primary efficacy endpoint is the failure of emergency department (ED) management within 72 h of the ED visit. Secondary endpoints include other measures of asthma severity and time to recovery within 7 days of the index visit. The study has 80% power for detecting a risk difference of 7.5% associated with each determinant from a baseline risk of 21%, at an α of 0.05. Ethics and dissemination: Ethical approval has been obtained from all participating institutions. An impaired response to systemic steroids in certain subgroups will challenge the current standard of practice and call for the immediate search for better approaches. A potential host-environment interaction will broaden our understanding of corticosteroid responsiveness in children. Documentation of similar effectiveness of corticosteroids across determinants will provide the needed reassurance regarding current treatment recommendations. Results: Results will be disseminated at international conferences and manuscripts targeted at emergency physicians, paediatricians, geneticists and respirologists. Trial registration number: This study is registered at Clinicaltrials.gov (NCT02013076)

Pulsed Laser System to Simulate Effects of Cosmic Rays in Semiconductor Devices

Spaceflight system electronic devices must survive a wide range of radiation environments with various particle types including energetic protons, electrons, gamma rays, x-rays, and heavy ions. High-energy charged particles such as heavy ions can pass straight through a semiconductor material and interact with a charge-sensitive region, generating a significant amount of charge (electron-hole pairs) along their tracks. These excess charges can damage the device, and the response can range from temporary perturbations to permanent changes in the state or performance. These phenomena are called single event effects (SEE). Before application in flight systems, electronic parts need to be qualified and tested for performance and radiation sensitivity. Typically, their susceptibility to SEE is tested by exposure to an ion beam from a particle accelerator. At such facilities, the device under test (DUT) is irradiated with large beams so there is no fine resolution to investigate particular regions of sensitivity on the parts. While it is the most reliable approach for radiation qualification, these evaluations are time consuming and costly. There is always a need for new cost-efficient strategies to complement accelerator testing: pulsed lasers provide such a solution. Pulsed laser light can be utilized to simulate heavy ion effects with the advantage of being able to localize the sensitive region of an integrated circuit. Generally, a focused laser beam of approximately picosecond pulse duration is used to generate carrier density in the semiconductor device. During irradiation, the laser pulse is absorbed by the electronic medium with a wavelength selected accordingly by the user, and the laser energy can ionize and simulate SEE as would occur in space. With a tightly focused near infrared (NIR) laser beam, the beam waist of about a micrometer can be achieved, and additional scanning techniques are able to yield submicron resolution. This feature allows mapping of all of the sensitive regions of the studied device with fine resolution, unlike heavy ion experiments. The problematic regions can be precisely identified, and it provides a considerable amount of information about the circuit. In addition, the system allows flexibility for testing the device in different configurations in situ

A Sea Turtle Population Assessment for Florida\u27s Big Bend, Northeastern Gulf of Mexico

Coastal waters of Florida’s Big Bend, Gulf of Mexico (GOM) once supported one of the largest sea turtle fisheries in the United States. To fill an information gap in this region on abundance and distribution of sea turtles, we used vessel—based distance sampling and active capture methods to characterize current foraging aggregations near the St. Martins Marsh Aquatic Preserve. Over 10 sampling periods between 2012—2018, we completed 513 km of transects and recorded 819 turtles among 4 species—green turtle (Chelonia mydas, n = 624), Kemp’s ridley (Lepidochelys kempii, n = 147), loggerhead (Caretta caretta, n = 47), and a single hawksbill (Eretmochelys imbricata). Turtle densities in 4 study plots within the 200 km2 study site ranged from 57—221 immature green turtles/km2, 16—56 immature Kemp’s ridleys/km2, and 1—14 juvenile—to—adult loggerheads/km2. Of 200 green turtles captured, 67.5% showed skin tumors consistent with fibropapillomatosis, a frequency similar to that from urbanized estuaries of Florida’s east coast. The largest green turtles (\u3e 60 cm straight standard carapace length), abundant in the southern portion of our study area, are of note because this size class is uncommonly recorded within US territorial waters. Analyses of green turtle mtDNA haplotypes found contributions from rookeries in the western GOM, Mexican Caribbean, and Costa Rica. Although Big Bend protected areas were principally designed to conserve marine and coastal habitats, these regulatory zones have also effectively encompassed a hotspot for foraging sea turtles

GAGA Factor Maintains Nucleosome-Free Regions and Has a Role in RNA Polymerase II Recruitment to Promoters

Previous studies have shown that GAGA Factor (GAF) is enriched on promoters with paused RNA Polymerase II (Pol II), but its genome-wide function and mechanism of action remain largely uncharacterized. We assayed the levels of transcriptionally-engaged polymerase using global run-on sequencing (GRO-seq) in control and GAF-RNAi Drosophila S2 cells and found promoter-proximal polymerase was significantly reduced on a large subset of paused promoters where GAF occupancy was reduced by knock down. These promoters show a dramatic increase in nucleosome occupancy upon GAF depletion. These results, in conjunction with previous studies showing that GAF directly interacts with nucleosome remodelers, strongly support a model where GAF directs nucleosome displacement at the promoter and thereby allows the entry Pol II to the promoter and pause sites. This action of GAF on nucleosomes is at least partially independent of paused Pol II because intergenic GAF binding sites with little or no Pol II also show GAF-dependent nucleosome displacement. In addition, the insulator factor BEAF, the BEAF-interacting protein Chriz, and the transcription factor M1BP are strikingly enriched on those GAF-associated genes where pausing is unaffected by knock down, suggesting insulators or the alternative promoter-associated factor M1BP protect a subset of GAF-bound paused genes from GAF knock-down effects. Thus, GAF binding at promoters can lead to the local displacement of nucleosomes, but this activity can be restricted or compensated for when insulator protein or M1BP complexes also reside at GAF bound promoters

KINEROS2-AGWA: Model Use, Calibration, and Validation

KINEROS (KINematic runoff and EROSion) originated in the 1960s as a distributed event-based model that conceptualizes a watershed as a cascade of overland flow model elements that flow into trapezoidal channel model elements. KINEROS was one of the first widely available watershed models that interactively coupled a finite difference approximation of the kinematic overland flow equations to a physically based infiltration model. Development and improvement of KINEROS continued from the 1960s on a variety of projects for a range of purposes, which has resulted in a suite of KINEROS-based modeling tools. This article focuses on KINEROS2 (K2), a spatially distributed, event-based watershed rainfall-runoff and erosion model, and the companion ArcGIS-based Automated Geospatial Watershed Assessment (AGWA) tool. AGWA automates the time-consuming tasks of watershed delineation into distributed model elements and initial parameterization of these elements using commonly available, national GIS data layers. A variety of approaches have been used to calibrate and validate K2 successfully across a relatively broad range of applications (e.g., urbanization, pre- and post-fire, hillslope erosion, erosion from roads, runoff and recharge, and manure transport). The case studies presented in this article (1) compare lumped to stepwise calibration and validation of runoff and sediment at plot, hillslope, and small watershed scales; and (2) demonstrate an uncalibrated application to address relative change in watershed response to wildfire

Destruction of the Mott Insulating Ground State of Ca_2RuO_4 by a Structural Transition

We report a first-order phase transition at T_M=357 K in single crystal Ca_2RuO_4, an isomorph to the superconductor Sr_2RuO_4. The discontinuous decrease in electrical resistivity signals the near destruction of the Mott insulating phase and is triggered by a structural transition from the low temperature orthorhombic to a high temperature tetragonal phase. The magnetic susceptibility, which is temperature dependent but not Curie-like decreases abruptly at TM and becomes less temperature dependent. Unlike most insulator to metal transitions, the system is not magnetically ordered in either phase, though the Mott insulator phase is antiferromagnetic below T_N=110 K.Comment: Accepted for publication in Phys. Rev. B (Rapid Communications

Adipocyte ACLY Facilitates Dietary Carbohydrate Handling to Maintain Metabolic Homeostasis in Females

Sugars and refined carbohydrates are major components of the modern diet. ATP-citrate lyase (ACLY) is upregulated in adipocytes in response to carbohydrate consumption and generates acetyl-coenzyme A (CoA) for both lipid synthesis and acetylation reactions. Here, we investigate the role of ACLY in the metabolic and transcriptional responses to carbohydrates in adipocytes and unexpectedly uncover a sexually dimorphic function in maintaining systemic metabolic homeostasis. When fed a high-sucrose diet, Acly(FAT-/-) females exhibit a lipodystrophy-like phenotype, with minimal fat accumulation, insulin resistance, and hepatic lipid accumulation, whereas Acly(FAT-/-) males have only mild metabolic phenotypes. We find that ACLY is crucial for nutrient-dependent carbohydrate response element-binding protein (ChREBP) activation in adipocytes and plays a key role, particularly in females, in the storage of newly synthesized fatty acids in adipose tissue. The data indicate that adipocyte ACLY is important in females for the systemic handling of dietary carbohydrates and for the preservation of metabolic homeostasis

Nucleon-induced reactions at intermediate energies: New data at 96 MeV and theoretical status

Double-differential cross sections for light charged particle production (up to A=4) were measured in 96 MeV neutron-induced reactions, at TSL laboratory cyclotron in Uppsala (Sweden). Measurements for three targets, Fe, Pb, and U, were performed using two independent devices, SCANDAL and MEDLEY. The data were recorded with low energy thresholds and for a wide angular range (20-160 degrees). The normalization procedure used to extract the cross sections is based on the np elastic scattering reaction that we measured and for which we present experimental results. A good control of the systematic uncertainties affecting the results is achieved. Calculations using the exciton model are reported. Two different theoretical approches proposed to improve its predictive power regarding the complex particle emission are tested. The capabilities of each approach is illustrated by comparison with the 96 MeV data that we measured, and with other experimental results available in the literature.Comment: 21 pages, 28 figure