Production of recombinant disulfide-rich venom peptides for structural and functional analysis via expression in the periplasm of E. coli

Abstract Disulfide-rich peptides are the dominant component of most animal venoms. These peptides have received much attention as leads for the development of novel therapeutic agents and bioinsecticides because they target a wide range of neuronal receptors and ion channels with a high degree of potency and selectivity. In addition, their rigid disulfide framework makes them particularly well suited for addressing the crucial issue of in vivo stability. Structural and functional characterization of these peptides necessitates the development of a robust, reliable expression system that maintains their native disulfide framework. The bacterium Escherichia coli has long been used for economical production of recombinant proteins. However, the expression of functional disulfide-rich proteins in the reducing environment of the E. coli cytoplasm presents a significant challenge. Thus, we present here an optimised protocol for the expression of disulfide-rich venom peptides in the periplasm of E. coli, which is where the endogenous machinery for production of disulfide-bonds is located. The parameters that have been investigated include choice of media, induction conditions, lysis methods, methods of fusion protein and peptide purification, and sample preparation for NMR studies. After each section a recommendation is made for conditions to use. We demonstrate the use of this method for the production of venom peptides ranging in size from 2 to 8 kDa and containing 2-6 disulfide bonds

Terahertz time-domain spectroscopy of electromagnons in multiferroic perovskite manganites

Recent spectroscopic studies at terahertz frequencies for a variety of multiferroics endowed with both ferroelectric and magnetic orders have revealed the possible emergence of a new collective excitation, frequently referred to as electromagnon. It is magnetic origin, but becomes active in response to the electric field component of light. Here we give an overview on our recent advance in the terahertz time-domain spectroscopy of electromagnons or electric-dipole active magnetic resonances, focused on perovskite manganites--$R$MnO$_3$ ($R$ denotes rare-earth ions). The respective electric and magnetic contributions to the observed magnetic resonance are firmly identified by the measurements of the light-polarization dependence using a complete set of the crystal orientations. We extract general optical features in a variety of the spin ordered phases, including the $A$-type antiferromagnetic, collinear spin ordered, and ferroelectric $bc$ and $ab$ spiral spin ordered phases, which are realized by tuning the chemical composition of $R$, temperature, and external magnetic field. In addition to the antiferromagnetic resonances of Mn ions driven by the magnetic field component of light, we clarify that the electromagnon appears only for light polarized along the a-axis even in the collinear spin ordered phase and grows in intensity with evolution of the spiral spin order, but independent of the direction of the spiral spin plane ($bc$ or $ab$) or equivalently the direction of the ferroelectric polarization $P_{\rm s}$ ($P_{\rm s}\| c$ or $P_{\rm s}\| a$). A possible origin of the observed magnetic resonances at terahertz frequencies is discussed by comparing the systematic experimental data presented here with theoretical considerations based on Heisenberg model.Comment: 19 pages including 15 figures and 2 tables; Invited Paper in Special Issue on Terahertz Wave Photonics in J. Opt. Soc. Am. B (Accepted for publication

New features in the phase diagram of TbMnO3_3

The (H,T)-phase diagram of the multiferroic perovskite TbMnO$_3$ was studied by high-resolution thermal expansion $\alpha(T)$ and magnetostriction $\Delta L(H)/L$ measurements. Below $T_{N}\simeq 42$ K, TbMnO$_3$ shows antiferromagnetic order, which changes at $T_{FE}\simeq 28$ K where simultaneously a spontaneous polarization $P||c$ develops. Sufficiently high magnetic fields applied along $a$ or $b$ induce a polarization flop to $P||a$. We find that all of these transitions are strongly coupled to the lattice parameters. Thus, our data allow for a precise determination of the phase boundaries and also yield information about their uniaxial pressure dependencies. The strongly hysteretic phase boundary to the ferroelectric phase with $P||a$ is derived in detail. Contrary to previous reports, we find that even in high magnetic fields there are no direct transitions from this phase to the paraelectric phase. We also determine the various phase boundaries in the low-temperature region related to complex reordering transitions of the Tb moments.Comment: 17 pages including 9 figure

Far-infrared optical excitations in multiferroic TbMnO_3

We provide a detailed study of the reflectivity of multiferroic TbMnO_3 for wave numbers from 40 cm^{-1} to 1000 cm^{-1} and temperatures 5 K < T < 300 K. Excitations are studied for polarization directions E || a, the polarization where electromagnons are observed, and for E || c, the direction of the spontaneous polarization in this material. The temperature dependencies of eigenfrequencies, damping constants and polar strengths of all modes are studied and analyzed. For E || a and below the spiral ordering temperature of about 27 K we observe a transfer of optical weight from phonon excitations to electromagnons, which mainly involves low-frequency phonons. For E || c an unusual increase of the total polar strength and hence of the dielectric constant is observed indicating significant transfer of dynamic charge probably within manganese-oxygen bonds on decreasing temperatures.Comment: 8 pages, 7 figures, accepted for submission in European Physical Journal

Cement degradation in CO2 storage sites: a review on potential applications of nanomaterials

© 2018 The Author(s) Carbon capture and sequestration (CCS) has been employed to reduce global warming, which is one of the critical environmental issues gained the attention of scientific and industrial communities worldwide. Once implemented successfully, CCS can store at least 5 billion tons of CO2per year as an effective and technologically safe method. However, there have been a few issues raised in recent years, indicating the potential leakages paths created during and after injection. One of the major issues might be the chemical interaction of supercritical CO2with the cement, which may lead to the partial or total loss of the cement sheath. There have been many approaches presented to improve the physical and mechanical properties of the cement against CO2attack such as changing the water-to-cement ratio, employing pozzolanic materials, and considering non-Portland cements. However, a limited success has been reported to the application of these approaches once implemented in a real-field condition. To date, only a few studies reported the application of nanoparticles as sophisticated additives which can reinforce oil well cements. This paper provides a review on the possible application of nanomaterials in the cement industry where physical and mechanical characteristics of the cement can be modified to have a better resistance against corrosive environments such as CO2storage sites. The results obtained indicated that adding 0.5 wt% of Carbon NanoTubes (CNTs) and NanoGlass Flakes (NGFs) can reinforce the thermal stability and coating characteristics of the cement which are required to increase the chance of survival in a CO2sequestrated site. Nanosilica can also be a good choice and added to the cement by as much as 3.0 wt% to improve pozzolanic reactivity and thermal stability as per the reports of recent studies

Overview of the 2010 Carbonaceous Aerosols and Radiative Effects Study (CARES)

Substantial uncertainties still exist in the scientific understanding of the possible interactions between urban and natural (biogenic) emissions in the production and transformation of atmospheric aerosol and the resulting impact on climate change. The US Department of Energy (DOE) Atmospheric Radiation Measurement (ARM) program's Carbonaceous Aerosol and Radiative Effects Study (CARES) carried out in June 2010 in Central Valley, California, was a comprehensive effort designed to improve this understanding. The primary objective of the field study was to investigate the evolution of secondary organic and black carbon aerosols and their climate-related properties in the Sacramento urban plume as it was routinely transported into the forested Sierra Nevada foothills area. Urban aerosols and trace gases experienced significant physical and chemical transformations as they mixed with the reactive biogenic hydrocarbons emitted from the forest. Two heavily-instrumented ground sites – one within the Sacramento urban area and another about 40 km to the northeast in the foothills area – were set up to characterize the evolution of meteorological variables, trace gases, aerosol precursors, aerosol size, composition, and climate-related properties in freshly polluted and "aged" urban air. On selected days, the DOE G-1 aircraft was deployed to make similar measurements upwind and across the evolving Sacramento plume in the morning and again in the afternoon. The NASA B-200 aircraft, carrying remote sensing instruments, was also deployed to characterize the vertical and horizontal distribution of aerosols and aerosol optical properties within and around the plume. This overview provides: (a) the scientific background and motivation for the study, (b) the operational and logistical information pertinent to the execution of the study, (c) an overview of key observations and initial findings from the aircraft and ground-based sampling platforms, and (d) a roadmap of planned data analyses and focused modeling efforts that will facilitate the integration of new knowledge into improved representations of key aerosol processes and properties in climate models.United States. Dept. of Energy. Atmospheric System Research Program (Contract DE-AC06-76RLO 1830)United States. National Oceanic and Atmospheric AdministrationUnited States. National Aeronautics and Space Administration. HQ Science Mission Directorate Radiation Sciences ProgramUnited States. National Aeronautics and Space Administration. CALIPSO ProgramUnited States. Dept. of Energy. Atmospheric Radiation Measurement Program (Interagency Agreement No. DE-AI02-05ER63985

Influence of fly ash blending on hydration and physical behavior of Belite-Alite-Ye'elimite cements

A cement powder, composed of belite, alite and ye’elimite, was blended with 0, 15 and 30 wt% of fly ash and the resulting lended cements were further characterized. During hydration, the presence of fly ash caused the partial inhibition of both AFt degradation and belite reactivity, even after 180 days. The compressive strength of the corresponding mortars increased by increasing the fly ash content (68, 73 and 82 MPa for mortars with 0, 15 and 30 wt% of fly ash, respectively, at 180 curing days), mainly due to the diminishing porosity and pore size values. Although pozzolanic reaction has not been directly proved there are indirect evidences.This work is part of the Ph.D. of D. Londono-Zuluaga funded by Beca Colciencias 646—Doctorado en el exterior and Enlaza Mundos 2013 program grant. Cement and Building materials group (CEMATCO) from National University of Colombia is acknowledged for providing the calorimetric measurements. Funding from Spanish MINECO BIA2017-82391-R and I3 (IEDI-2016-0079) grants, co-funded by FEDER, are acknowledged

Geographical and temporal distribution of SARS-CoV-2 clades in the WHO European Region, January to June 2020

We show the distribution of SARS-CoV-2 genetic clades over time and between countries and outline potential genomic surveillance objectives. We applied three available genomic nomenclature systems for SARS-CoV-2 to all sequence data from the WHO European Region available during the COVID-19 pandemic until 10 July 2020. We highlight the importance of real-time sequencing and data dissemination in a pandemic situation. We provide a comparison of the nomenclatures and lay a foundation for future European genomic surveillance of SARS-CoV-2.Peer reviewe