An underlying clock in the extreme flip-flop state transitions of the black hole transient Swift J1658.2-4242

Aims: Flip-flops are top-hat-like X-ray flux variations which occur in some transient accreting black hole binary systems and feature simultaneous changes in the spectral hardness and the Power Density Spectrum (PDS). They occur at a crucial time in the evolution of these systems, when the accretion disk emission starts to dominate over coronal emission. Flip-flops have only rarely been observed and are poorly understood. Methods: We detect 15 flip-flops in the 2018 outburst of Swift J1658.2-4242, in observations by XMM-Newton, NuSTAR, Astrosat, Swift, Insight-HXMT, INTEGRAL, and ATCA. We analyse their light curves, search for periodicities, compute their PDS, and fit their X-ray spectra, to investigate the source behaviour during flip-flop transitions, and how the interval featuring flip-flops differs from the rest of the outburst. Results: The flip-flops of Swift J1658.2-4242 are of an extreme variety, exhibiting flux differences of up to 77% within ~100s, much larger than has been seen so far. We observe radical changes in the PDS simultaneous with the sharp flux variations, featuring transitions between the Quasi-Periodic Oscillation types C and A, which have never been observed before. Changes to the PDS are delayed, but more rapid than changes in the light curve. Flip-flops occur in two intervals, separated by two weeks in which these phenomena were not seen. Transitions between the two flip-flop states occurred at random integer multiples of a fundamental period, of 2.761ks in the first interval, and 2.61ks in the second. Spectral analysis reveals the high and low flux flip-flop states to be similar, but distinct from intervals lacking flip-flops. A change in the inner temperature of the accretion disk is responsible for most of the flux difference in the flip-flops. We highlight the importance of correcting for the influence of the dust scattering halo on the X-ray spectra.Comment: 30 pages, 28 figures, accepted for publication in Astronomy & Astrophysic

0312: Characterization of human valvular interstitial cells isolated from normal and fibrocalcified aortic valves

PurposeAortic Valve Stenosis (AVS) affects 2% to 6% of population over 65 years in industrialized countries. This atherosclerosis-like pathology involves Valve Interstitial Cell (VIC) proliferation and commitment to osteoblast- like cells. This prevalent cell type of aortic valve presents five identifiable phenotypes: embryonic progenitor endothelial/mesenchymal cells, progenitor, quiescent, activated and osteoblastic VICs. To study the pathophysiology of AVS, their in vitro cultures are frequently used. Our purpose is to characterize VICs isolated from normal and fibrocalcified human aortic valves and analyze their in vitro behavior.MethodsWe collected 5 normal and 5 fibrocalcified human aortic valves. VICs were isolated by collagenase digestion. Characterization is assessed at different passages (2 to 5) by immunofluorescence. Analyzed markers consist of progenitor cell markers (SSEA4, ABCG2, CD90, NG2 and OsteoBlast CaDHerin (OB-CDH)), fibroblast markers (vimentin and HSP47) and smooth muscle cell (SMC) marker (α-actin). By blue trypan and MTS, we compared the viability and proliferation of VICs in standard and starvation medium at 48 hours.ResultsIndependently of their origin, VICs express all progenitor cell markers. Fibroblasts markers are expressed twice more by pathological VICs and four times more for SMC marker. In standard medium, VICs viability is similar (96,7±2,4% vs 96,4±2,3% ; normal vs pathological ± SEM). Pathological VICs proliferate more than normal VICs (2,2±0,7 vs 1,6±0,4 ; OD/OD control). In starvation medium, viability is significantly reduced for pathological VICs (89,6±7,9% vs 76,5±5,3%) but still proliferate in opposition with normal VICs (1,7±0,6 vs 1,2±0,3).ConclusionAll VICs phenotypes are found in vitro with no culture selection but in different ratios according to their origin. These new data in VICs isolated from normal or pathological human aortic valves allow us to approve their use in vitro

A jet model for the fast IR variability of the black hole X-ray binary GX 339-4

Using the simultaneous Infra-Red (IR) and X-ray light curves obtained by Kalamkar et al., we perform a Fourier analysis of the IR/X-ray timing correlations of the black hole X-ray binary (BHB) GX 339-4. The resulting IR vs X-ray Fourier coherence and lag spectra are similar to those obtained in previous studies of GX 339-4 using optical light curves. In particular, above 1 Hz, the lag spectrum features an approximately constant IR lag of about 100 ms. We model simultaneously the radio to IR Spectral Energy Distribution (SED), the IR Power Spectral Density (PSD), and the coherence and lag spectra using the jet internal shock model ISHEM assuming that the fluctuations of the jet Lorentz factor are driven by the accretion flow. It turns out that most of the spectral and timing features, including the 100-ms lag, are remarkably well-reproduced by this model. The 100-ms time-scale is then associated with the travel time from the accretion flow to the IR emitting zone. Our exploration of the parameter space favours a jet which is at most mildly relativistic (¯ < 3), and a linear and positive relation between the jet Lorentz factor and X-ray light curve i.e. (t) − 1∝LX(t). The presence of a strong Low-Frequency Quasi-Periodic Oscillation (LFQPO) in the IR light curve could be caused by jet precession driven by Lense–Thirring precession of the jet-emitting accretion flow. Our simulations confirm that this mechanism can produce an IR LFQPO similar to that observed in GX 339-4

Monalysin, a Novel ß-Pore-Forming Toxin from the Drosophila Pathogen Pseudomonas entomophila, Contributes to Host Intestinal Damage and Lethality

Pseudomonas entomophila is an entomopathogenic bacterium that infects and kills Drosophila. P. entomophila pathogenicity is linked to its ability to cause irreversible damages to the Drosophila gut, preventing epithelium renewal and repair. Here we report the identification of a novel pore-forming toxin (PFT), Monalysin, which contributes to the virulence of P. entomophila against Drosophila. Our data show that Monalysin requires N-terminal cleavage to become fully active, forms oligomers in vitro, and induces pore-formation in artificial lipid membranes. The prediction of the secondary structure of the membrane-spanning domain indicates that Monalysin is a PFT of the ß-type. The expression of Monalysin is regulated by both the GacS/GacA two-component system and the Pvf regulator, two signaling systems that control P. entomophila pathogenicity. In addition, AprA, a metallo-protease secreted by P. entomophila, can induce the rapid cleavage of pro-Monalysin into its active form. Reduced cell death is observed upon infection with a mutant deficient in Monalysin production showing that Monalysin plays a role in P. entomophila ability to induce intestinal cell damages, which is consistent with its activity as a PFT. Our study together with the well-established action of Bacillus thuringiensis Cry toxins suggests that production of PFTs is a common strategy of entomopathogens to disrupt insect gut homeostasis

Transapical mitral valve implantation for treatment of symptomatic mitral valve disease: a real-world multicentre experience.

AIMS Transcatheter mitral valve implantation (TMVI) is a new treatment option for patients with symptomatic mitral valve (MV) disease. Real-world data have not yet been reported. This study aimed to assess procedural and 30-day outcomes of TMVI in a real-world patient cohort. METHOD AND RESULTS All consecutive patients undergoing implantation of a transapically delivered self-expanding valve at 26 European centres from January 2020 to April 2021 were included in this retrospective observational registry. Among 108 surgical high-risk patients included (43% female, mean age 75 ± 7 years, mean STS-PROM 7.2 ± 5.3%), 25% was treated for an off-label indication (e.g. previous MV intervention or surgery, mitral stenosis, mitral annular calcification). Patients were highly symptomatic (New York Heart Association [NYHA] functional class III/IV in 86%) and mitral regurgitation (MR) was graded 3+/4+ in 95% (38% primary, 37% secondary, and 25% mixed aetiology). Technical success rate was 96%, and MR reduction to ≤1+ was achieved in all patients with successful implantation. There were two procedural deaths and 30-day all-cause mortality was 12%. At early clinical follow-up, MR reduction was sustained and there were significant reductions of pulmonary pressure (systolic pulmonary artery pressure 52 vs. 42 mmHg, p < 0.001), and tricuspid regurgitation severity (p = 0.013). Heart failure symptoms improved significantly (73% in NYHA class I/II, p < 0.001). Procedural success rate according to MVARC criteria was 80% and was not different in patients treated for an off-label indication (74% vs. 81% for off- vs. on-label, p = 0.41). CONCLUSION In a real-world patient population, TMVI has a high technical and procedural success rate with efficient and durable MR reduction and symptomatic improvement

Small-scale, semi-automated purification of eukaryotic proteins for structure determination

A simple approach that allows cost-effective automated purification of recombinant proteins in levels sufficient for functional characterization or structural studies is described. Studies with four human stem cell proteins, an engineered version of green fluorescent protein, and other proteins are included. The method combines an expression vector (pVP62K) that provides in vivo cleavage of an initial fusion protein, a factorial designed auto-induction medium that improves the performance of small-scale production, and rapid, automated metal affinity purification of His8-tagged proteins. For initial small-scale production screening, single colony transformants were grown overnight in 0.4 ml of auto-induction medium, produced proteins were purified using the Promega Maxwell 16, and purification results were analyzed by Caliper LC90 capillary electrophoresis. The yield of purified [U-15N]-His8-Tcl-1 was 7.5 μg/ml of culture medium, of purified [U-15N]-His8-GFP was 68 μg/ml, and of purified selenomethione-labeled AIA–GFP (His8 removed by treatment with TEV protease) was 172 μg/ml. The yield information obtained from a successful automated purification from 0.4 ml was used to inform the decision to scale-up for a second meso-scale (10–50 ml) cell growth and automated purification. 1H–15N NMR HSQC spectra of His8-Tcl-1 and of His8-GFP prepared from 50 ml cultures showed excellent chemical shift dispersion, consistent with well folded states in solution suitable for structure determination. Moreover, AIA–GFP obtained by proteolytic removal of the His8 tag was subjected to crystallization screening, and yielded crystals under several conditions. Single crystals were subsequently produced and optimized by the hanging drop method. The structure was solved by molecular replacement at a resolution of 1.7 Å. This approach provides an efficient way to carry out several key target screening steps that are essential for successful operation of proteomics pipelines with eukaryotic proteins: examination of total expression, determination of proteolysis of fusion tags, quantification of the yield of purified protein, and suitability for structure determination

Obesity Paradox in the Clinical Significance of Effective Prosthetic Orifice Area after Aortic Valve Replacement.

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Matter ejections behind the highs and lows of the transitional millisecond pulsar PSR J1023+0038

Transitional millisecond pulsars are an emerging class of sources that link low-mass X-ray binaries to millisecond radio pulsars in binary systems. These pulsars alternate between a radio pulsar state and an active low-luminosity X-ray disc state. During the active state, these sources exhibit two distinct emission modes (high and low) that alternate unpredictably, abruptly, and incessantly. X-ray to optical pulsations are observed only during the high mode. The root cause of this puzzling behaviour remains elusive. This paper presents the results of the most extensive multi-wavelength campaign ever conducted on the transitional pulsar prototype, PSR J1023+0038, covering from the radio to X-rays. The campaign was carried out over two nights in June 2021 and involved 12 different telescopes and instruments, including XMM-Newton, HST, VLT/FORS2 (in polarimetric mode), ALMA, VLA, and FAST. By modelling the broadband spectral energy distributions in both emission modes, we show that the mode switches are caused by changes in the innermost region of the accretion disc. These changes trigger the emission of discrete mass ejections, which occur on top of a compact jet, as testified by the detection of at least one short-duration millimetre flare with ALMA at the high-to-low mode switch. The pulsar is subsequently re-enshrouded, completing our picture of the mode switches.F.C.Z. is supported by a Ramón y Cajal fellowship (grant agreement RYC2021-030888-I) and Catalan grant SGR-Cat 2021 (PI: Graber). F.C.Z. and D.F.T. are supported by the program Unidad de Excelencia María de Maeztu CEX2020-001058-M. F.C.Z., S.C.a., P.D.’A., F.A., P.C., D.d.M. and A.P. acknowledge financial support from INAF-Fundamental research astrophysics project “Uncovering the optical beat of the fastest magnetised neutron stars” (FANS; PI: AP). S.C.a., S.C.o. and P.D.’A. acknowledge support from ASI grant I/004/11/5. G.B. acknowledges support from the PID2020-117710GB-I00 grant funded by MCIN/ AEI /10.13039/501100011033. A.M.Z. is supported by PRIN-MIUR 2017 UnIAM (Unifying Isolated and Accreting Magnetars, PI S. Mereghetti). X.H. is supported by the National Natural Science Foundation of China through grant No. 12041303. J.L. is supported by the National Natural Science Foundation of China through grant No.12273038. D.d.M. and A.P. acknowledge financial support from the Italian Space Agency (ASI) and National Institute for Astrophysics (INAF) under agreements ASI-INAF I/037/12/0 and ASI-INAF n.2017- 14-H.0, from INAF ‘Sostegno alla ricerca scientifica main streams dell‘INAF’, Presidential Decree 43/2018 and from INAF ‘SKA/CTA projects’, Presidential Decree 70/2016. D.M.R. and D.M.B. acknowledge the support of the NYU Abu Dhabi Research Enhancement Fund under grant RE124. D.F.T. is supported by the grants PID2021-124581OB-I00 funded by MCIN/AEI/10.13039/501100011033, 2021SGR00426 of the Generalitat de Catalunya and by MCIN with funding from European Union NextGeneration EU (PRTR-C17.I1). M.M. acknowledges the research programme Athena with project number 184.034.002, which is (partly) financed by the Dutch Research Council (NWO) and thanks the Team Meeting at the International Space Science Institute (Bern) for fruitful discussions. F.M.V. acknowledges support from the Spanish Ministry of Science for the grant PID2020-120323GB-I00 and for the FJC2020-043334-I grant financed by MCIN/AEI/10.13039/501100011033 and Next Generation EU/PRTR. This work was also partially supported by the COST Action ‘PHAROS’ (CA 16124).With funding from the Spanish government through the "Severo Ochoa Centre of Excellence" accreditation (CEX2020-001058-M).Peer reviewe