Behaviour of the Blazar CTA 102 during two giant outbursts

Blazar CTA 102 underwent exceptional optical and high-energy outbursts in 2012 and 2016-2017. We analyze its behaviour during these events, focusing on polarimetry as a tool that allows us to trace changes in the physical conditions and geometric configuration of the emission source close to the central black hole. We also use Fermi gamma-ray data in conjunction with optical photometry in an effort to localize the origin of the outbursts.AST-1615796 - Boston Universit

Multiwavelength observations of the blazar BL Lacertae: a new fast TeV γ-ray flare

Proceedings of the 35th International Cosmic Ray Conference (ICRC 2017), Busan (South Korea). Published in Proceeding of Science.Observations of fast TeV γ-ray flares from blazars reveal the extreme compactness of emitting regions in blazar jets. Combined with very-long-baseline radio interferometry measurements, they probe the structure and emission mechanism of the jet. We report on a fast TeV γ-ray flare from BL Lacertae observed by VERITAS, with a rise time of about 2.3 hours and a decay time of about 36 minutes. The peak flux at >200 GeV measured with the 4-minute binned light curve is (4.2±0.6)×10−6photonsm−2s−1, or ∼180% the Crab Nebula flux. Variability in GeV γ-ray, X-ray, and optical flux, as well as in optical and radio polarization was observed around the time of the TeV γ-ray flare. A possible superluminal knot was identified in the VLBA observations at 43 GHz. The flare constrains the size of the emitting region, and is consistent with several theoretical models with stationary shocks

Peculiarities of the Mechanism of Interactions of Catalytic Antibodies with Organophosphorus Substrates

© 2017, Pleiades Publishing, Inc. Catalytic antibodies are a promising model for creating highly specific biocatalysts with predetermined activity. However, in order to realize the directed change or improve their properties, it is necessary to understand the basics of catalysis and the specificity of interactions with substrates. In the present work, a structural and functional study of the Fab fragment of antibody A5 and a comparative analysis of its properties with antibody A17 have been carried out. These antibodies were previously selected for their ability to interact with organophosphorus compounds via covalent catalysis. It has been established that antibody A5 has exceptional specificity for phosphonate X with bimolecular reaction rate constants of 510 ± 20 and 390 ± 20 min –1 M –1 for kappa and lambda variants, respectively. 3D-Modeling of antibody A5 structure made it possible to establish that the reaction residue L-Y33 is located on the surface of the active site, in contrast to the A17 antibody, in which the reaction residue L-Y37 is located at the bottom of a deep hydrophobic pocket. To investigate a detailed mechanism of the reaction, A5 antibody mutants with replacements L-R51W and H-F100W were created, which made it possible to perform stopped-flow kinetics. Tryptophan mutants were obtained as Fab fragments in the expression system of the methylotrophic yeast species Pichia pastoris. It has been established that the effectiveness of their interaction with phosphonate X is comparable to the wild-type antibody. Using the data of the stopped-flow kinetics method, significant conformational changes were established in the phosphonate modification process. The reaction was found to proceed using the induced-fit mechanism; the kinetic parameters of the elementary stages of the process have been calculated. The results present the prospects for the further improvement of antibody-based biocatalysts

QM/MM description of newly selected catalytic bioscavengers against organophosphorus compounds revealed reactivation stimulus mediated by histidine residue in the acyl-binding loop

© 2018 Zlobin, Mokrushina, Terekhov, Zalevsky, Bobik, Stepanova, Aliseychik, Kartseva, Panteleev, Golovin, Belogurov, Gabibov and Smirnov. Butyrylcholinesterase (BChE) is considered as an efficient stoichiometric antidote against organophosphorus (OP) poisons. Recently we utilized combination of calculations and ultrahigh-throughput screening (uHTS) to select BChE variants capable of catalytic destruction of OP pesticide paraoxon. The purpose of this study was to elucidate the molecular mechanism underlying enzymatic hydrolysis of paraoxon by BChE variants using hybrid quantum mechanical/molecular mechanical (QM/MM) calculations. Detailed analysis of accomplished QM/MM runs revealed that histidine residues introduced into the acyl-binding loop are always located in close proximity with aspartate residue at position 70. Histidine residue acts as general base thus leading to attacking water molecule activation and subsequent SN2 inline hydrolysis resulting in BChE reactivation. This combination resembles canonical catalytic triad found in active centers of various proteases. Carboxyl group activates histidine residue by altering its pKa, which in turn promotes the activation of water molecule in terms of its nucleophilicity. Observed re-protonation of catalytic serine residue at position 198 from histidine residue at position 438 recovers initial configuration of the enzyme's active center, facilitating next catalytic cycle. We therefore suggest that utilization of uHTS platform in combination with deciphering of molecular mechanisms by QM/MM calculations may significantly improve our knowledge of enzyme function, propose new strategies for enzyme design and open new horizons in generation of catalytic bioscavengers against OP poisons

Microfluidic droplet platform for ultrahigh-throughput single-cell screening of biodiversity

© 2017, National Academy of Sciences. All rights reserved.Ultrahigh-throughput screening (uHTS) techniques can identify unique functionality from millions of variants. To mimic the natural selection mechanisms that occur by compartmentalization in vivo, we developed a technique based on single-cell encapsulation in droplets of a monodisperse microfluidic double water-in-oil-in-water emulsion (MDE). Biocompatible MDE enables in-droplet cultivation of different living species. The combination of droplet-generating machinery with FACS followed by next-generation sequencing and liquid chromatography-mass spectrometry analysis of the secretomes of encapsulated organisms yielded detailed genotype/phenotype descriptions. This platform was probed with uHTS for biocatalysts anchored to yeast with enrichment close to the theoretically calculated limit and cell-to-cell interactions. MDE-FACS allowed the identification of human butyrylcholinesterase mutants that undergo self-reactivation after inhibition by the organophosphorus agent paraoxon. The versatility of the platform allowed the identification of bacteria, including slow-growing oral microbiota species that suppress the growth of a common pathogen, Staphylococcus aureus, and predicted which genera were associated with inhibitory activity

MAGIC gamma-ray and multi-frequency observations of flat spectrum radio quasar PKS 1510-089 in early 2012

Aims. Amongst more than fifty blazars detected in very high energy (VHE, E> 100 GeV) γ rays, only three belong to the subclass of flat spectrum radio quasars (FSRQs). The detection of FSRQs in the VHE range is challenging, mainly because of their soft spectra in the GeV-TeV regime. MAGIC observed PKS 1510−089 (z = 0.36) starting 2012 February 3 until April 3 during a high activity state in the high energy (HE, E> 100 MeV) γ-ray band observed by AGILE and Fermi. MAGIC observations result in the detection of a source with significance of 6.0 standard deviations (σ). We study the multi-frequency behaviour of the source at the epoch of MAGIC observation, collecting quasi-simultaneous data at radio and optical (GASP-WEBT and F-Gamma collaborations, REM, Steward, Perkins, Liverpool, OVRO, and VLBA telescopes), X-ray (Swift satellite), and HE γ-ray frequencies. Methods. We study the VHE γ-ray emission, together with the multi-frequency light curves, 43 GHz radio maps, and spectral energy distribution (SED) of the source. The quasi-simultaneous multi-frequency SED from the millimetre radio band to VHE γ rays is modelled with a one-zone inverse Compton model. We study two different origins of the seed photons for the inverse Compton scattering, namely the infrared torus and a slow sheath surrounding the jet around the Very Long Baseline Array (VLBA) core. Results. We find that the VHE γ-ray emission detected from PKS 1510−089 in 2012 February-April agrees with the previous VHE observations of the source from 2009 March-April. We find no statistically significant variability during the MAGIC observations on daily, weekly, or monthly time scales, while the other two known VHE FSRQs (3C 279 and PKS 1222+216) have shown daily scale to sub-hour variability. The γ-ray SED combining AGILE, Fermi and MAGIC data joins smoothly and shows no hint of a break. The multi-frequency light curves suggest a common origin for the millimetre radio and HE γ-ray emission, and the HE γ-ray flaring starts when the new component is ejected from the 43 GHz VLBA core and the studied SED models fit the data well. However, the fast HE γ-ray variability requires that within the modelled large emitting region, more compact regions must exist. We suggest that these observed signatures would be most naturally explained by a turbulent plasma flowing at a relativistic speed down the jet and crossing a standing conical shock

Multiwavelength behaviour of the blazar OJ 248 from radio to γ -rays

© 2015 The Authors. We present an analysis of the multiwavelength behaviour of the blazar OJ 248 at z = 0.939 in the period 2006-2013. We use low-energy data (optical, near-infrared, and radio) obtained by 21 observatories participating in the Gamma-Ray Large Area Space Telescope (GLAST)-AGILE Support Program of the Whole Earth Blazar Telescope, as well as data from the Swift (optical-UV and X-rays) and Fermi (γ -rays) satellites, to study flux and spectral variability and correlations among emissions in different bands. We take into account the effect of absorption by the Damped Lyman α intervening system at z = 0.525. Two major outbursts were observed in 2006-2007 and in 2012-2013 at optical and near-IR wavelengths, while in the high-frequency radio light curves prominent radio outbursts are visible peaking at the end of 2010 and beginning of 2013, revealing a complex radio-optical correlation. Crosscorrelation analysis suggests a delay of the optical variations after the γ -ray ones of about a month, which is a peculiar behaviour in blazars. We also analyse optical polarimetric and spectroscopic data. The average polarization percentage P is less than 3 per cent, but it reaches~19 per cent during the early stage of the 2012-2013 outburst.Avague correlation of P with brightness is observed. There is no preferred electric vector polarization angle and during the outburst the linear polarization vector shows wide rotations in both directions, suggesting a complex behaviour/structure of the jet and possible turbulence. The analysis of 140 optical spectra acquired at the Steward Observatory reveals a strong Mg II broad emission line with an essentially stable flux of 6.2 × 10-15 erg cm-2 s-1 and a full width at half-maximum of 2053 km s-1