Influence of blood collection methods and long-term plasma storage on quorum-sensing peptide stability

Finding adequate biomarkers for rapid and accurate disease detection, prognosis, and therapy is increasingly important. Quorum-sensing peptides are herein a new emerging group, produced by bacteria, fungi, protozoa, and viruses, with blood being the most straightforward sample type to detect/quantitate them. However, detailed information about suitable blood sample collection methods and storage conditions for measuring these quorum-sensing peptides hampers further clinical research and development. Here, we first tested the time-dependent stability of a set of chemically diverse quorum-sensing peptides, spiked in blood at different temperatures (4, 21, and 37 °C) in four different ethylenediamine tetraacetic acid (EDTA)-containing plasma tubes (with different protein-stabilizing additives) over a period of up to 7.5 h. Next, we determined the storage stability of these quorum-sensing peptides in plasma at different temperatures (4, −35, and −80 °C). UPLC/MS–MS was used to selectively detect and quantify the spiked quorum-sensing peptides. The results of this study indicate that a cost-effective tube, designed for traditional proteomics and stored at 4 °C, is the preferred collection condition when quorum-sensing peptides need to be detected/quantified in human plasma. When the tubes are handled at room temperature (21 °C), a more specialized tube is required. Long-term storage of plasma samples, even under low-temperature conditions (−80 °C), indicates rapid degradation of certain quorum-sensing peptides

High-order harmonic transient grating spectroscopy of SF6 molecular vibrations

special issue : Ultrafast electron and molecular dynamicsInternational audienceStrong field transient grating spectroscopy has shown to be a very versatile tool in time-resolved molecular spectroscopy. Here we use this technique to investigate the high-order harmonic generation from SF6 molecules vibrationally excited by impulsive stimulated Raman scattering. Transient grating spectroscopy enables us to reveal clear modulations of the harmonic emission. This heterodyne detection shows that the harmonic emission generated between 14 to 26 eV is mainly sensitive to two among the three active Raman modes in SF6, i.e. the strongest and fully symmetric nu 1-A1g mode (774 cm-1, 43 fs) and the slowest mode nu5-T2g (524 cm-1, 63 fs). A time-frequency analysis of the harmonic emission reveals additional dynamics: the strength and central frequency of the nu 1 mode oscillate with a frequency of 52 cm-1 (640 fs). This could be a signature of the vibration of dimers in the generating medium. Harmonic 11 shows a remarkable behavior, oscillating in opposite phase, both on the fast (774 cm-1) and slow (52 cm-1) timescales, which indicates a strong modulation of the recombination matrix element as a function of the nuclear geometry. These results demonstrate that the high sensitivity of high-order harmonic generation to molecularvibrations, associated to the high sensitivity of transient grating spectroscopy, make their combination a unique tool to probe vibrational dynamics

Extracorporeal Membrane Oxygenation for Severe Acute Respiratory Distress Syndrome associated with COVID-19: An Emulated Target Trial Analysis.

RATIONALE: Whether COVID patients may benefit from extracorporeal membrane oxygenation (ECMO) compared with conventional invasive mechanical ventilation (IMV) remains unknown. OBJECTIVES: To estimate the effect of ECMO on 90-Day mortality vs IMV only Methods: Among 4,244 critically ill adult patients with COVID-19 included in a multicenter cohort study, we emulated a target trial comparing the treatment strategies of initiating ECMO vs. no ECMO within 7 days of IMV in patients with severe acute respiratory distress syndrome (PaO2/FiO2 <80 or PaCO2 ≥60 mmHg). We controlled for confounding using a multivariable Cox model based on predefined variables. MAIN RESULTS: 1,235 patients met the full eligibility criteria for the emulated trial, among whom 164 patients initiated ECMO. The ECMO strategy had a higher survival probability at Day-7 from the onset of eligibility criteria (87% vs 83%, risk difference: 4%, 95% CI 0;9%) which decreased during follow-up (survival at Day-90: 63% vs 65%, risk difference: -2%, 95% CI -10;5%). However, ECMO was associated with higher survival when performed in high-volume ECMO centers or in regions where a specific ECMO network organization was set up to handle high demand, and when initiated within the first 4 days of MV and in profoundly hypoxemic patients. CONCLUSIONS: In an emulated trial based on a nationwide COVID-19 cohort, we found differential survival over time of an ECMO compared with a no-ECMO strategy. However, ECMO was consistently associated with better outcomes when performed in high-volume centers and in regions with ECMO capacities specifically organized to handle high demand. This article is open access and distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives License 4.0 (http://creativecommons.org/licenses/by-nc-nd/4.0/)

Biostability Assessment of Flexible Parylene C-based Implantable Sensor in Wireless Chronic Neural Recording

International audienceThe stability of polymer-based sensors in a biological environment remains a challenge, as delamination and swelling often compromise mechanical and electrical capability. We have developed a neural implant based on Parylene C, a biocompatible flexible polymer, with PEDOT-nanostructured gold patterns to record the brain electrical activity. Here, we show first evidence of device biostability through in vitro soaking tests in artificial brain environment and in vivo recording in mice. Our results indicate that after over the six months trial, more than 75% of the in vitro electrodes have stable impedance, and the implanted sensors in mice were able to accurately record signals from mice hippocampi. None of the implants presented with signs of Parylene degradation or metal corrosion. Overall, the devices are promising candidates for reliable, chronically implanted sensors in the biomedical field

In vitro and in vivo biostability assessment of chronically-implanted Parylene C neural sensors

International audienceParylene C has rapidly gained attention as a exible biomaterial for a new generation of chronic neural probes. However, polymeric material failure in the form of delamination, swelling or tearing, often compromises device biostability in the long term. This work constitutes a rst step towards lifetime assessment of Parylene C implanted devices. We have conceived a Parylene C-based neural probe with PEDOT-nanostructured gold electrodes for the recording of brain activity. The material response to its biological environment was studied through in vitro soaking tests and in vivo wireless recordings in mice brain, both carried out for up to 6 months. Impedance monitoring and SEM images indicate that over the length of this trial, none of the implants presented with apparent signs of material degradation. Packaging reliability was a predominant factor in device failure, with a certain number of faulty connection appearing over time. This parameter aside, all soaked devices were stable in Articial Cerebro-Spinal Fluid, with impedances within 10% of their initial value after 6 months at 37°C. Besides, at least 70% of the implanted device were able to accurately record wirelessly high amplitude hippocampal Local Field Potentials from freely-moving mice, with steady Signal-to-Noise Ratio. In other terms, Parylene C implantable sensors responded minimally to articial and actual physiological conditions during a period of 6 months, which makes them promising candidates for reliable, chronically implanted sensors in the biomedical eld

Postcompression of high-energy terawatt-level femtosecond pulses and application to high-order harmonic generation

We perform a postcompression of high-energy pulses by using optical-field ionization of low pressure helium gas in a guided geometry. We apply this approach to a terawatt (TW) chirped-pulse-amplification-based Ti:sapphire laser chain and show that spectral broadening can be controlled with both the input pulse energy and gas pressure. Under optimized conditions, we generate 10 fs pulses at the TW level directly under vacuum and demonstrate a high stability of the postcompressed pulse duration. These high-energy postcompressed pulses are thereafter used to perform high harmonic generation in a loose focusing geometry. The extreme ultraviolet (XUV) beam is characterized both spatially and spectrally on a single shot basis, and structured continuous XUV spectra are observed

Peptidomics : LC-MS operational parameters do matter

The sensitive and specific detection of peptides at low levels in biofluids is critical to increase the lab-to-human translation of peptidomic research. An interesting group of peptides with increasing evidence for involvement in human diseases are quorum sensing peptides. To obtain more reliable conclusions on peptide measurands in biofluids, a selection of often neglected parts of the analytical process using LC-MS were investigated, with novel approaches recommended for each part. Quorum sensing peptides were used as the main model-peptides. The peptidomic parts investigated and discussed here are: 1. Sample collection: obtaining and storing biofluids such as plasma/serum, feces or saliva from mice and humans up to the laboratory handling (sample preparation) should ensure no degradation/metabolization of the peptide measurand (yielding false negatives) or of proteins (yielding false positives). 2. Sample preparation: to remove interfering compounds as well as to release peptides from the adsorbing matrix-components and to preconcentrate them; this is a crucial step which should assure analytical stability and adsorption-minimization. 3. Chromatography: not only the separation power and orthogonality of the complementary systems is critical, but a careful characterisation of the gradient-system, such as the starting conditions, need to be addressed as well because of worsened detection limits (yielding false negatives) or carry-over (false positives). 4. MS detection: operational parameters such as duty cycle characteristics applied are critical in obtaining low-level, reliable results. Our work addresses aQbD-approached solutions to these challenges, encompassing sample stabilization measures, a suitable peptide anti-adsorption tool, judicious choice of injection solvent versus gradient system and optimal duty cycle parameters. Our recommendations will improve the peptidomics bio-analytics of not only quorum sensing peptides, but can also be of value for other measurands at low concentrations

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Combined high-harmonic interferometries for vectorial spectroscopy

We present a new method to characterize transverse vectorial light produced by high-harmonic generation (HHG). The incoherent sum of the two components of the electric field is measured using a bi-dimensional transient grating while one of the components is simultaneously characterized using two-source interferometry. The combination of these two interferometric setups enables the amplitude and phase measurement of the two vectorial components of the extreme ultraviolet radiation. We demonstrate the potential of this technique in the case of HHG in aligned nitrogen, revealing the vectorial properties of harmonics 9–17 of a Ti:sapphire laser

Quantum-path resolved attosecond high-harmonic spectroscopy

International audienceStrong-field ionization of molecules releases electrons which can be accelerated and driven back to recombine with their parent ion, emitting high-order harmonics. This ionization also initiates attosecond electronic and vibrational dynamics in the ion, evolving during the electron travel in the continuum. Revealing this subcycle dynamics from the emitted radiation usually requires advanced theoretical modeling. We show that this can be avoided by resolving the emission from two families of electronic quantum paths in the generation process. The corresponding electrons have the same kinetic energy, and thus the same structural sensitivity, but differ by the travel time between ionization and recombination—the pump-probe delay in this attosecond self-probing scheme. We measure the harmonic amplitude and phase in aligned CO$_2$ and N$_2$ molecules and observe a strong influence of laser-induced dynamics on two characteristic spectroscopic features: a shape resonance and multichannel interference. This quantum-path resolved spectroscopy thus opens wide prospects for the investigation of ultrafast ionic dynamics, such as charge migration