Expression of Chlamydomonas reinhardtii periplasmic carbonic anhydrase in Escherichia coli and transgenic tobacco plants

Carbonic anhydrase (CA) catalyzes the reversible hydration of CO2. The periplasmic CA gene CAH1 of Chlamydomonas reinhardtii codes for a highly processed secreted glycoprotein. The primary translation product of the CAH1 gene is targeted to the algae\u27s ER, where it is proteolytically processed to yield two different subunits, glycosylated, assembled into an active heterotetramer, and secreted. Expression of CAH1 in Escherichia coli and tobacco plants were effected in this study. For bacterial expression, two constructs, one with and another without, the algal target leader sequence were used. Similarly, after replacing the algal\u27s target leader sequence with that from tobacco anionic peroxidase, expression of this gene in transgenic tobacco plants was also investigated;No expression was detected in bacteria from the construct with the algal leader. The gene was however efficiently expressed in the cytoplasm of E. coli from the construct without the leader sequence. The expressed protein which accumulated to very high levels, was confirmed to be periplasmic CA 1 (peri-CA1) based on the expected size and by cross-reaction with polyclonal antibody. The overexpressed polypeptide was present in inclusion bodies and no CA activity was detected in cell homogenates. The band was excised from polyacrylamide gel, and after some processing, used to raise polyclonal antibodies in rabbits;Of several transgenic plants screened by immunoblotting and found to express peri-CA1, a single plant, TL1, with a high level of expression was chosen for further analysis. SDS-PAGE gels of the purified protein from this tobacco plant, showed that it migrated as a series of discrete bands (two large and one small) with slightly faster mobility than the comparable bands in the purified algal protein. The expressed protein in the transgenic plant was active, and staining with thymol and sulfuric acid confirmed that it was also glycosylated. Analysis of vacuum infiltrates from leaves expressing peri-CA1 showed that it was enriched in the intercellular fluid. The sensitivity of the enzyme to sulfonamide inhibitors was similar to that of the native algal enzyme. These results suggest that the post translational processing of Chlamydomonas peri-CA1 is largely conserved in a higher plant

Inert shell coating for enhanced laser refrigeration of nanoparticles: application in levitated optomechanics

We report on a study exploring the design of nanoparticles that can enhance their laser refrigeration efficiency for applications in levitated optomechanics. In particular, we developed lanthanide-doped nanocrystals with an inert shell coating and compared their performance with bare nanocrystals. While optically levitated, we studied the refrigeration of both types of nanoparticles while varying the pressure. We found that the core-shell design shows an improvement in the minimum final temperature: a fourth of the core-shell nanoparticles showed a significant cooling compared to almost none of the bare nanoparticles. Furthermore, we measured a core-shell nanoparticle cooling down to a temperature of 147 K at 26 mbar in the underdamped regime. Our study is a first step towards engineering nanoparticles that are suitable for achieving absolute (centre-of-mass and internal temperature) cooling in levitation, opening new avenues for force sensing and the realization of macroscopic quantum superpositions.Comment: Any comments are welcome

Hexabromocyclododecane and hexachlorocyclohexane: How lessons learnt have led to improved regulation

This is the author's accepted manuscript. The final published article is available from the link below. Copyright @ 2014 Taylor & Francis.The use of chemicals by society has many benefits but contamination of the environment is an unintended consequence. One example is the organochlorine compound hexachlorocyclohexane (HCH). During the 1980s, when HCH was banned in many countries, the brominated flame retardant, hexabromocyclododecane (HBCD), found increasing use. The persistent, bioaccumulative, and toxic characteristics of HBCD are, 30 years later, likely to warrant global action on production and use under the Stockholm Convention on persistent organic pollutants. Historical lessons have taught us that we need to control the use of chemicals and programs are in place worldwide in an attempt to do so.Tertiary Education Trust Fund, Nigeri

Guaranteeing Canadian lamb meat quality using near-infrared spectroscopy on intact rack

Lamb racks from commercial carcasses were scanned using near-infrared spectroscopy. The prediction accuracies (R 2) for meat quality traits were assessed. Prediction accuracy ranged between 0.40 and 0.94. When predicted values were used to classify meat based on quality, 88.7%–95.2% of samples were correctly classified as quality guaranteed

Combination of mesoscale and synoptic mechanisms for triggering an isolated thunderstorm: Observational case study of CSIP IOP 1

Copyright @ 2007 AMSAn isolated thunderstorm formed in the southern United Kingdom on 15 June 2005 and moved through the area where a large number of observational instruments were deployed as part of the Convective Storm Initiation Project. Earlier, a convergence line had formed downstream of Devon in the southwest of the United Kingdom in a southwesterly airflow, along which a series of light showers had formed. The depth of these showers was limited by a capping inversion, or lid, at around 2.5 km. The deep thunderstorm convection developed from one of these showers when the convection broke through the lid and ascended up to the next inversion, associated with a tropopause fold at around 6 km. A series of clear-air reflectivity RHIs are used to map the height of the capping inversion and its lifting resulting from the ascent along the convergence line. The origins of the lid are tracked back to some descent from the midtroposphere along dry adiabats. The strength of the lid was weaker along a northwest-to-southeast-oriented region located behind an overrunning upper cold front. The transition from shallow to deep convection occurred where this region with a weaker lid intersected the region with a raised lid, oriented southwest to northeast, downstream of Devon. A very high resolution forecast model that is being developed by the Met Office predicted the isolated thunderstorm successfully. This success depended on the accurate representation of the following two scales: the synoptic-scale and the surface-forced mesoscale convergence line. The interaction between these scales localized the convection sufficiently in space and time for the initiation and subsequent development to be highly predictable despite the relatively poor representation in the model of processes at the cloud scale.This work was funded from the Natural Environment Research Council and the Met Office National Meteorology Programme

The surprising role of orography in the initiation of an isolated thunderstorm in southern England

Many factors, both mesoscale and larger scale, often come together in order for a particular convective initiation to take place. The authors describe a modeling study of a case from the Convective Storms Initiation Project (CSIP) in which a single thunderstorm formed behind a front in the southern United Kingdom. The key features of the case were a tongue of low-level high θw air associated with a forward-sloping split front (overrunning lower θw air above), a convergence line, and a “lid” of high static stability air, which the shower was initially constrained below but later broke through. In this paper, the authors analyze the initiation of the storm, which can be traced back to a region of high ground (Dartmoor) at around 0700 UTC, in more detail using model sensitivity studies with the Met Office Unified Model (MetUM). It is established that the convergence line was initially caused by roughness effects but had a significant thermal component later. Dartmoor had a key role in the development of the thunderstorm. A period of asymmetric flow over the high ground, with stronger low-level descent in the lee, led to a hole in a layer of low-level clouds downstream. The surface solar heating through this hole, in combination with the tongue of low-level high θw air associated with the front, caused the shower to initiate with sufficient lifting to enable it later to break through the lid

Ram-pressure stripped radio tail and two ULXs in the spiral galaxy HCG 97b

We report LOFAR and VLA detections of extended radio emission in the spiral galaxy HCG 97b, hosted by an X-ray bright galaxy group. The extended radio emission detected at 144 MHz, 1.4 GHz and 4.8 GHz is elongated along the optical disk and has a tail that extends 27 kpc in projection towards the centre of the group at GHz frequencies or 60 kpc at 144 MHz. Chandra X-ray data show two off-nuclear ultra-luminous X-ray sources (ULXs) with the more distant one being a suitable candidate for an accreting intermediate-mass black hole (IMBH) embedded in an environment with an increased density of molecular gas. Given the observed morphology in optical, CO, and radio continuum, we propose that the galaxy is undergoing ram-pressure stripping and the relativistic plasma accelerated in star-forming regions is transported from the galactic disc by galaxy-intragroup medium interaction. Finally, we also demonstrate that the formation of the radio tail could, in principle, be the result of putative IMBH-induced activity, which could facilitate the stripping or inject the radio plasma via jets.Comment: 15 pages, 11 figures, submitted to MNRAS, comments are welcom

Statistical study of linear magnetic hole structures near Earth

The Magnetospheric Multiscale mission (MMS1) data for 8 months in the winter periods of 2017-2018 and 2018-2019, when MMS had its apogee in the upstream solar wind of the Earth's bow shock, are used to study linear magnetic holes (LMHs). These LMHs are characterized by a magnetic depression of more than 50 % and a rotation of the background magnetic field of less then 10g . A total of 406 LMHs are found and, based on their magnetoplasma characteristics, are split into three categories: cold (increase in density, little change in ion temperature), hot (increase in ion temperature, decrease in density) and sign change (at least one magnetic field component changes sign). The occurrence rate of LMHs is 2.3 per day. All LMHs are basically in pressure balance with the ambient plasma. Most of the linear magnetic holes are found in ambient plasmas that are stable against the mirror-mode generation, but only half of the holes are mirror-mode-stable inside