Ligand-induced conformational changes in a SMALP-encapsulated GPCR.

The adenosine 2A receptor (A2AR), a G-protein-coupled receptor (GPCR), was solubilised and purified encapsulated in styrene maleic acid lipid particles (SMALPs). The purified A2AR-SMALP was associated with phospholipids characteristic of the plasma membrane of Pichia pastoris, the host used for its expression, confirming that the A2AR-SMALP encapsulated native lipids. The fluorescence spectrum of the A2AR-SMALP showed a characteristic broad emission peak at 330 nm, produced by endogenous Trp residues. The inverse agonist ZM241385 caused 30% increase in fluorescence emission, unusually accompanied by a red-shift in the emission wavelength. The emission spectrum also showed sub-peaks at 321 nm, 335 nm and 350 nm, indicating that individual Trp inhabited different environments following ZM241385 addition. There was no effect of the agonist NECA on the A2AR-SMALP fluorescence spectrum. Substitution of two Trp residues by Tyr suggested that ZM241385 affected the environment and mobility of Trp2466.48 in TM6 and Trp2687.33 at the extracellular face of TM7, causing transition to a more hydrophobic environment. The fluorescent moiety IAEDANS was site-specifically introduced at the intracellular end of TM6 (residue 2316.33) to report on the dynamic cytoplasmic face of the A2AR. The inverse agonist ZM241385 caused a concentration-dependent increase in fluorescence emission as the IAEDANS moved to a more hydrophobic environment, consistent with closing the G-protein binding crevice. NECA generated only 30% of the effect of ZM241385. This study provides insight into the SMALP environment; encapsulation supported constitutive activity of the A2AR and ZM241385-induced conformational transitions but the agonist NECA generated only small effects

G-protein-coupled receptor solubilization and purification for biophysical analysis and functional studies, in the total absence of detergent

G-protein coupled receptors (GPCRs) constitute the largest class of membrane proteins and are a major drug target. A serious obstacle to studying GPCR structure/function characteristics is the requirement to extract the receptors from their native environment in the plasma membrane, coupled with the inherent instability of GPCRs in the detergents required for their solubilization. In the present study, we report the first solubilization and purification of a functional GPCR [human adenosine A2A receptor (A2AR)], in the total absence of detergent at any stage, by exploiting spontaneous encapsulation by styrene maleic acid (SMA) co-polymer direct from the membrane into a nanoscale SMA lipid particle (SMALP). Furthermore, the A2AR-SMALP, generated from yeast (Pichia pastoris) or mammalian cells, exhibited increased thermostability (∼5°C) compared with detergent [DDM (n-dodecyl-β-D-maltopyranoside)]-solubilized A2AR controls. The A2AR-SMALP was also stable when stored for prolonged periods at 4°C and was resistant to multiple freeze-thaw cycles, in marked contrast with the detergent-solubilized receptor. These properties establish the potential for using GPCR-SMALP in receptor-based drug discovery assays. Moreover, in contrast with nanodiscs stabilized by scaffold proteins, the non-proteinaceous nature of the SMA polymer allowed unobscured biophysical characterization of the embedded receptor. Consequently, CD spectroscopy was used to relate changes in secondary structure to loss of ligand binding ([3H]ZM241385) capability. SMALP-solubilization of GPCRs, retaining the annular lipid environment, will enable a wide range of therapeutic targets to be prepared in native-like state to aid drug discovery and understanding of GPCR molecular mechanisms

Scintillation-limited photometry with the 20-cm NGTS telescopes at Paranal Observatory

Ground-based photometry of bright stars is expected to be limited by atmospheric scintillation, although in practice observations are often limited by other sources of systematic noise. We analyse 122 nights of bright star (Gmag ≲ 11.5) photometry using the 20-cm telescopes of the Next-Generation Transit Survey (NGTS) at the Paranal Observatory in Chile. We compare the noise properties to theoretical noise models and we demonstrate that NGTS photometry of bright stars is indeed limited by atmospheric scintillation. We determine a median scintillation coefficient at the Paranal Observatory of CY=1.54⁠, which is in good agreement with previous results derived from turbulence profiling measurements at the observatory. We find that separate NGTS telescopes make consistent measurements of scintillation when simultaneously monitoring the same field. Using contemporaneous meteorological data, we find that higher wind speeds at the tropopause correlate with a decrease in long-exposure (t = 10 s) scintillation. Hence, the winter months between June and August provide the best conditions for high-precision photometry of bright stars at the Paranal Observatory. This work demonstrates that NGTS photometric data, collected for searching for exoplanets, contains within it a record of the scintillation conditions at Paranal

The relative emission from chromospheres and coronae: Dependence on spectral type and age

Extreme-ultraviolet and X-ray emission from stellar coronae drives mass loss from exoplanet atmospheres, and ultraviolet emission from stellar chromospheres drives photochemistry in exoplanet atmospheres. Comparisons of the spectral energy distributions of host stars are, therefore, essential for understanding the evolution and habitability of exoplanets. The large number of stars observed with the MUSCLES, Mega-MUSCLES, and other recent Hubble Space Telescope observing programs has provided for the first time a large sample (79 stars) of reconstructed Lyα fluxes that we compare with X-ray fluxes to identify significant patterns in the relative emission from these two atmospheric regions as a function of stellar age and effective temperature. We find that as stars age on the main sequence, the emissions from their chromospheres and coronae follow a pattern in response to the amount of magnetic heating in these atmospheric layers. A single trend-line slope describes the pattern of X-ray versus Lyα emission for G and K dwarfs, but the different trend lines for M dwarf stars show that the Lyα fluxes of M stars are significantly smaller than those of warmer stars with the same X-ray flux. The X-ray and Lyα luminosities divided by the stellar bolometric luminosities show different patterns depending on stellar age. The L (Lyα)/L(bol) ratios increase smoothly to cooler stars of all ages, but the L(X)/L(bol) ratios show different trends. For older stars, the increase in coronal emission with decreasing Teff is much steeper than that of chromospheric emission. We suggest a fundamental link between atmospheric properties and trend lines relating coronal and chromospheric heating.Fil: Linsky, Jeffrey L.. State University of Colorado at Boulder; Estados UnidosFil: Wood, Brian E.. Spece Sciences División. Naval Research Laboratory; Estados UnidosFil: Youngblood, Allison. State University of Colorado at Boulder; Estados UnidosFil: Brown, Alexander. State University of Colorado at Boulder; Estados UnidosFil: Froning, Cynthia S.. University of Texas at Austin; Estados UnidosFil: France, Kevin. State University of Colorado at Boulder; Estados UnidosFil: Buccino, Andrea Paola. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Astronomía y Física del Espacio. - Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Astronomía y Física del Espacio; ArgentinaFil: Cranmer, Steven R.. State University of Colorado at Boulder; Estados UnidosFil: Mauas, Pablo Jacobo David. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Astronomía y Física del Espacio. - Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Astronomía y Física del Espacio; ArgentinaFil: Miguel, Yamila. Leiden Observatory; Países BajosFil: Sebastian Pineda, J.. State University of Colorado at Boulder; Estados UnidosFil: Rugheimer, Sarah. University of Oxford; Reino UnidoFil: Vieytes, Mariela Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Astronomía y Física del Espacio. - Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Astronomía y Física del Espacio; ArgentinaFil: Wheatley, Peter J.. University of Warwick; Reino UnidoFil: Wilson, David J.. University of Texas at Austin; Estados Unido

NGTS clusters survey -- II. White-light flares from the youngest stars in Orion

We present the detection of high energy white-light flares from pre-main sequence stars associated with the Orion complex, observed as part of the Next Generation Transit Survey (NGTS). With energies up to $5.2\times10^{35}$ erg these flares are some of the most energetic white-light flare events seen to date. We have used the NGTS observations of flaring and non-flaring stars to measure the average flare occurrence rate for 4 Myr M0-M3 stars. We have also combined our results with those from previous studies to predict average rates for flares above $1\times10^{35}$ ergs for early M stars in nearby young associations.STFC ST/M001962/1; ST/P000495/

Automatic vetting of planet candidates from ground based surveys : machine learning with NGTS

State of the art exoplanet transit surveys are producing ever increasing quantities of data. To make the best use of this resource, in detecting interesting planetary systems or in determining accurate planetary population statistics, requires new automated methods. Here we describe a machine learning algorithm that forms an integral part of the pipeline for the NGTS transit survey, demonstrating the efficacy of machine learning in selecting planetary candidates from multi-night ground based survey data. Our method uses a combination of random forests and self-organising-maps to rank planetary candidates, achieving an AUC score of 97.6% in ranking 12368 injected planets against 27496 false positives in the NGTS data. We build on past examples by using injected transit signals to form a training set, a necessary development for applying similar methods to upcoming surveys. We also make the autovet code used to implement the algorithm publicly accessible. autovet is designed to perform machine learned vetting of planetary candidates, and can utilise a variety of methods. The apparent robustness of machine learning techniques, whether on space-based or the qualitatively different ground-based data, highlights their importance to future surveys such as TESS and PLATO and the need to better understand their advantages and pitfalls in an exoplanetary context

TESS Duotransit Candidates from the Southern Ecliptic Hemisphere

Discovering transiting exoplanets with long orbital periods allows us to study warm and cool planetary systems with temperatures similar to the planets in our own Solar system. The TESS mission has photometrically surveyed the entire Southern Ecliptic Hemisphere in Cycle 1 (August 2018 - July 2019), Cycle 3 (July 2020 - June 2021) and Cycle 5 (September 2022 - September 2023). We use the observations from Cycle 1 and Cycle 3 to search for exoplanet systems that show a single transit event in each year - which we call duotransits. The periods of these planet candidates are typically in excess of 20 days, with the lower limit determined by the duration of individual TESS observations. We find 85 duotransit candidates, which span a range of host star brightnesses between 8 < $T_{mag}$ < 14, transit depths between 0.1 per cent and 1.8 per cent, and transit durations between 2 and 10 hours with the upper limit determined by our normalisation function. Of these candidates, 25 are already known, and 60 are new. We present these candidates along with the status of photometric and spectroscopic follow-up.Comment: 25 pages, 16 figures, submitted to Monthly Notices of the Royal Astronomical Societ

High C/O Chemistry and Weak Thermal Inversion in the Extremely Irradiated Atmosphere of Exoplanet WASP-12b

The carbon-to-oxygen ratio (C/O) in a planet provides critical information about its primordial origins and subsequent evolution. A primordial C/O greater than 0.8 causes a carbide-dominated interior as opposed to the silicate-dominated composition as found on Earth; the solar C/O is 0.54. Theory, shows that high C/O leads to a diversity of carbon-rich planets that can have very different interiors and atmospheres from those in the solar system. Here we report the detection of C/O greater than or equal to 1 in a planetary atmosphere. The transiting hot Jupiter WASP-12b has a dayside atmosphere depleted in water vapour and enhanced in methane by over two orders of magnitude compared to a solar-abundance chemical equilibrium model at the expected temperatures. The observed concentrations of the prominent molecules CO, CH4, and H2O are consistent with theoretical expectations for an atmosphere with the observed C/O = 1. The C/O ratios are not known for giant planets in the solar system, although they are expected to equal the solar value. If high C/O ratios are common, then extrasolar planets are likely very different in interior composition, and formed very differently, from expectations based on solar composition, potentially explaining the large diversity in observed radii. We also find that the extremely irradiated atmosphere (greater than 2500 K) of WASP-12b lacks a prominent thermal inversion, or a stratosphere, and has very efficient day-night energy circulation. The absence of a strong thermal inversion is in stark contrast to theoretical predictions for the most highly irradiated hot-Jupiter atmospheres

Physiotherapy and occupational therapy vs no therapy in mild to moderate Parkinson disease: a randomized clinical trial

IMPORTANCE It is unclear whether physiotherapy and occupational therapy are clinically effective and cost-effective in Parkinson disease (PD). OBJECTIVE To perform a large pragmatic randomized clinical trial to evaluate the clinical effectiveness of individualized physiotherapy and occupational therapy in PD. DESIGN, SETTING, AND PARTICIPANTS The PD REHAB Trial was a multicenter, open-label, parallel group, controlled efficacy trial. A total of 762 patients with mild to moderate PD were recruited from 38 sites across the United Kingdom. Recruitment took place between October 2009 and June 2012, with 15 months of follow-up. INTERVENTIONS Participants with limitations in activities of daily living (ADL) were randomized to physiotherapy and occupational therapy or no therapy. MAIN OUTCOMES AND MEASURES The primary outcome was the Nottingham Extended Activities of Daily Living (NEADL) Scale score at 3 months after randomization. Secondary outcomes were health-related quality of life (assessed by Parkinson Disease Questionnaire–39 and EuroQol-5D); adverse events; and caregiver quality of life. Outcomes were assessed before trial entry and then 3, 9, and 15 months after randomization. RESULTS Of the 762 patients included in the study (mean [SD] age, 70 [9.1] years), 381 received physiotherapy and occupational therapy and 381 received no therapy. At 3 months, there was no difference between groups in NEADL total score (difference, 0.5 points; 95%CI, −0.7 to 1.7; P = .41) or Parkinson Disease Questionnaire–39 summary index (0.007 points; 95%CI, −1.5 to 1.5; P = .99). The EuroQol-5D quotient was of borderline significance in favor of therapy (−0.03; 95%CI, −0.07 to −0.002; P = .04). The median therapist contact time was 4 visits of 58 minutes over 8 weeks. Repeated-measures analysis showed no difference in NEADL total score, but Parkinson Disease Questionnaire–39 summary index (diverging 1.6 points per annum; 95%CI, 0.47 to 2.62; P = .005) and EuroQol-5D score (0.02; 95%CI, 0.00007 to 0.03; P = .04) showed small differences in favor of therapy. There was no difference in adverse events. CONCLUSIONS AND RELEVANCE Physiotherapy and occupational therapy were not associated with immediate or medium-term clinically meaningful improvements in ADL or quality of life in mild to moderate PD. This evidence does not support the use of low-dose, patient-centered, goal-directed physiotherapy and occupational therapy in patients in the early stages of PD. Future research should explore the development and testing of more structured and intensive physical and occupational therapy programs in patients with all stages of PD