Laboratory procedures manual for the firefly luciferase assay for adenosine triphosphate (ATP)

A manual on the procedures and instruments developed for the adenosine triphosphate (ATP) luciferase assay is presented. Data cover, laboratory maintenance, maintenance of bacterial cultures, bacteria measurement, reagents, luciferase procedures, and determination of microbal susceptibility to antibiotics

The mysterious optical afterglow spectrum of GRB140506A at z=0.889

Context. Gamma-ray burst (GRBs) afterglows probe sightlines to star-forming regions in distant star-forming galaxies. Here we present a study of the peculiar afterglow spectrum of the z = 0.889 Swift GRB 140506A. Aims. Our aim is to understand the origin of the very unusual properties of the absorption along the line-of-sight. Methods. We analyse spectroscopic observations obtained with the X-shooter spectrograph mounted on the ESO/VLT at two epochs 8.8 h and 33 h after the burst as well as imaging from the GROND instrument. We also present imaging and spectroscopy of the host galaxy obtained with the Magellan telescope. Results. The underlying afterglow appears to be a typical afterglow of a long-duration GRB. However, the material along the line-of- sight has imprinted very unusual features on the spectrum. Firstly, there is a very broad and strong flux drop below 8000 AA (4000 AA in the rest frame), which seems to be variable between the two spectroscopic epochs. We can reproduce the flux-drops both as a giant 2175 AA extinction bump and as an effect of multiple scattering on dust grains in a dense environment. Secondly, we detect absorption lines from excited H i and He i. We also detect molecular absorption from CH+ . Conclusions. We interpret the unusual properties of these spectra as reflecting the presence of three distinct regions along the line-of-sight: the excited He i absorption originates from an H ii-region, whereas the Balmer absorption must originate from an associated photodissociation region. The strong metal line and molecular absorption and the dust extinction must originate from a third, cooler region along the line-of-sight. The presence of (at least) three separate regions is reflected in the fact that the different absorption components have different velocities relative to the systemic redshift of the host galaxy.Comment: 8 pages, 4 figures. Accepted for publications in A&

The evolution of superluminous supernova LSQ14mo and its interacting host galaxy system

We present and analyse an extensive dataset of the superluminous supernova (SLSN) LSQ14mo (z = 0.256), consisting of a multi-colour light curve from-30 d to +70 d in the rest-frame (relative to maximum light) and a series of six spectra from PESSTO covering-7 d to +50 d. This is among the densest spectroscopic coverage, and best-constrained rising light curve, for a fast-declining hydrogen-poor SLSN. The bolometric light curve can be reproduced with a millisecond magnetar model with 4 M⊙ ejecta mass, and the temperature and velocity evolution is also suggestive of a magnetar as the power source. Spectral modelling indicates that the SN ejected 6 M⊙ of CO-rich material with a kinetic energy of 7 × 1051 erg, and suggests a partially thermalised additional source of luminosity between-2 d and +22 d. This may be due to interaction with a shell of material originating from pre-explosion mass loss. We further present a detailed analysis of the host galaxy system of LSQ14mo. PESSTO and GROND imaging show three spatially resolved bright regions, and we used the VLT and FORS2 to obtain a deep (five-hour exposure) spectra of the SN position and the three star-forming regions, which are at a similar redshift. The FORS2 spectrum at + 300 days shows no trace of SN emission lines and we place limits on the strength of [O i] from comparisons with other Ic supernovae. The deep spectra provides a unique chance to investigate spatial variations in the host star-formation activity and metallicity. The specific star-formation rate is similar in all three components,as is the presence of a young stellar population. However, the position of LSQ14mo exhibits a lower metallicity, with 12 + log (O/H) = 8.2 in both the R23 and N2 scales (corresponding to 0.3 Z⊙). We propose that the three bright regions in the host system are interacting, which could induce gas flows triggering star formation in low-metallicity regions. © ESO, 2017

The optical/NIR afterglow of GRB 111209A: Complex yet not unprecedented

Context. Afterglows of gamma-ray bursts (GRBs) are simple in the most basic model, but can show many complex features. The ultra-long duration GRB 111209A, one of the longest GRBs ever detected, also has the best-monitored afterglow in this rare class of GRBs. Aims. We want to address the question whether GRB 111209A was a special event beyond its extreme duration alone, and whether it is a classical GRB or another kind of high-energy transient. The afterglow may yield significant clues. Methods. We present afterglow photometry obtained in seven bands with the GROND imager as well as in further seven bands with the Ultraviolet/Optical Telescope (UVOT) on-board the Neil Gehrels Swift Observatory. The light curve is analysed by multi-band modelling and joint fitting with power-laws and broken power-laws, and we use the contemporaneous GROND data to study the evolution of the spectral energy distribution. We compare the optical afterglow to a large ensemble we have analysed in earlier works, and especially to that of another ultra-long event, GRB 130925A. We furthermore undertake a photometric study of the host galaxy. Results. We find a strong, chromatic rebrightening event at ≈0.8 days after the GRB, during which the spectral slope becomes redder. After this, the light curve decays achromatically, with evidence for a break at about 9 days after the trigger. The afterglow luminosity is found to not be exceptional. We find that a double-jet model is able to explain the chromatic rebrightening. The afterglow features have been detected in other events and are not unique. Conclusions. The duration aside, the GRB prompt emission and afterglow parameters of GRB 111209A are in agreement with the known distributions for these parameters. While the central engine of this event may differ from that of classical GRBs, there are multiple lines of evidence pointing to GRB 111209A resulting from the core-collapse of a massive star with a stripped envelope

IκB kinase 2 determines oligodendrocyte loss by non-cell-autonomous activation of NF-κB in the central nervous system

The IκB kinase complex induces nuclear factor kappa B activation and has recently been recognized as a key player of autoimmunity in the central nervous system. Notably, IκB kinase/nuclear factor kappa B signalling regulates peripheral myelin formation by Schwann cells, however, its role in myelin formation in the central nervous system during health and disease is largely unknown. Surprisingly, we found that brain-specific IκB kinase 2 expression is dispensable for proper myelin assembly and repair in the central nervous system, but instead plays a fundamental role for the loss of myelin in the cuprizone model. During toxic demyelination, inhibition of nuclear factor kappa B activation by conditional ablation of IκB kinase 2 resulted in strong preservation of central nervous system myelin, reduced expression of proinflammatory mediators and a significantly attenuated glial response. Importantly, IκB kinase 2 depletion in astrocytes, but not in oligodendrocytes, was sufficient to protect mice from myelin loss. Our results reveal a crucial role of glial cell-specific IκB kinase 2/nuclear factor kappa B signalling for oligodendrocyte damage during toxic demyelination. Thus, therapies targeting IκB kinase 2 function in non-neuronal cells may represent a promising strategy for the treatment of distinct demyelinating central nervous system disease

The evolution of superluminous supernova LSQ14mo and its interacting host galaxy system

We present and analyse an extensive dataset of the superluminous supernova (SLSN) LSQ14mo (z = 0.256), consisting of a multi-colour light curve from-30 d to +70 d in the rest-frame (relative to maximum light) and a series of six spectra from PESSTO covering-7 d to +50 d. This is among the densest spectroscopic coverage, and best-constrained rising light curve, for a fast-declining hydrogen-poor SLSN. The bolometric light curve can be reproduced with a millisecond magnetar model with 4 M⊙ ejecta mass, and the temperature and velocity evolution is also suggestive of a magnetar as the power source. Spectral modelling indicates that the SN ejected 6 M⊙ of CO-rich material with a kinetic energy of 7 × 1051 erg, and suggests a partially thermalised additional source of luminosity between-2 d and +22 d. This may be due to interaction with a shell of material originating from pre-explosion mass loss. We further present a detailed analysis of the host galaxy system of LSQ14mo. PESSTO and GROND imaging show three spatially resolved bright regions, and we used the VLT and FORS2 to obtain a deep (five-hour exposure) spectra of the SN position and the three star-forming regions, which are at a similar redshift. The FORS2 spectrum at + 300 days shows no trace of SN emission lines and we place limits on the strength of [O i] from comparisons with other Ic supernovae. The deep spectra provides a unique chance to investigate spatial variations in the host star-formation activity and metallicity. The specific star-formation rate is similar in all three components,as is the presence of a young stellar population. However, the position of LSQ14mo exhibits a lower metallicity, with 12 + log (O/H) = 8.2 in both the R23 and N2 scales (corresponding to 0.3 Z⊙). We propose that the three bright regions in the host system are interacting, which could induce gas flows triggering star formation in low-metallicity regions. © ESO, 2017

Selective inhibition of extra-synaptic α5-GABA_A receptors by S44819, a new therapeutic agent

In the mammalian central nervous system (CNS) GABAA receptors (GABAARs) mediate neuronal inhibition and are important therapeutic targets. GABAARs are composed of 5 subunits, drawn from 19 proteins, underpinning expression of 20-30 GABAAR subtypes. In the CNS these isoforms are heterogeneously expressed and exhibit distinct physiological and pharmacological properties. We report the discovery of S44819, a novel tricyclic oxazolo-2,3-benzodiazepine-derivative, that selectively inhibits α5-subunit-containing GABAARs (α5-GABAARs). Current α5-GABAAR inhibitors bind to the “benzodiazepine site”. However, in HEK293 cells expressing recombinant α5-GABAARs, S44819 had no effect on 3H-flumazenil binding, but displaced the GABAAR agonist 3H-muscimol and competitively inhibited the GABA-induced responses. Importantly, we reveal that the α5-subunit selectivity is uniquely governed by amino acid residues within the α-subunit F-loop, a region associated with GABA binding. In mouse hippocampal CA1 neurons, S44819 enhanced long-term potentiation (LTP), blocked a tonic current mediated by extrasynaptic α5-GABAARs, but had no effect on synaptic GABAARs. In mouse thalamic neurons, S44819 had no effect on the tonic current mediated by δ-GABAARs, or on synaptic (α1β2γ2) GABAARs. In rats, S44819 enhanced object recognition memory and reversed scopolamine-induced impairment of working memory in the eight-arm radial maze. In conclusion, S44819 is a first in class compound that uniquely acts as a potent, competitive, selective antagonist of recombinant and native α5-GABAARs. Consequently, S44819 enhances hippocampal synaptic plasticity and exhibits pro-cognitive efficacy. Given this profile, S44819 may improve cognitive function in neurodegenerative disorders and facilitate post-stroke recovery

Control of Neural Daughter Cell Proliferation by Multi-level Notch/Su(H)/E(spl)-HLH Signaling

The Notch pathway controls proliferation during development and in adulthood, and is frequently affected in many disorders. However, the genetic sensitivity and multi-layered transcriptional properties of the Notch pathway has made its molecular decoding challenging. Here, we address the complexity of Notch signaling with respect to proliferation, using the developing Drosophila CNS as model. We find that a Notch/Su(H)/E(spl)-HLH cascade specifically controls daughter, but not progenitor proliferation. Additionally, we find that different E(spl)-HLH genes are required in different neuroblast lineages. The Notch/Su(H)/E(spl)-HLH cascade alters daughter proliferation by regulating four key cell cycle factors: Cyclin E, String/Cdc25, E2f and Dacapo (mammalian p21CIP1/p27KIP1/p57Kip2). ChIP and DamID analysis of Su(H) and E(spl)-HLH indicates direct transcriptional regulation of the cell cycle genes, and of the Notch pathway itself. These results point to a multi-level signaling model and may help shed light on the dichotomous proliferative role of Notch signaling in many other systems