Sensitivity of Small RNA-Based Detection of Plant Viruses

Plants recognize unrelated viruses by the antiviral defense system called RNA interference (RNAi). RNAi processes double-stranded viral RNA into small RNAs (sRNAs) of 21-24 nucleotides, the reassembly of which into longer strands in silico allows virus identification by comparison with the sequences available in databases. The aim of this study was to compare the virus detection sensitivity of sRNA-based virus diagnosis with the established virus species-specific polymerase chain reaction (PCR) approach. Viruses propagated in tobacco plants included three engineered, infectious clones of Potato virus A (PVA), each carrying a different marker gene, and an infectious clone of Potato virus Y (PVY). Total RNA (containing sRNA) was isolated and subjected to reverse-transcription real-time PCR (RT-RT-PCR) and sRNA deep-sequencing at different concentrations. RNA extracted from various crop plants was included in the reactions to normalize RNA concentrations. Targeted detection of selected viruses showed a similar threshold for the sRNA and reverse-transcription quantitative PCR (RT-qPCR) analyses. The detection limit for PVY and PVA by RT-qPCR in this study was 3 and 1.5 fg of viral RNA, respectively, in 50 ng of total RNA per PCR reaction. When knowledge was available about the viruses likely present in the samples, sRNA-based virus detection was 10 times more sensitive than RT-RT-PCR. The advantage of sRNA analysis is the detection of all tested viruses without the need for virus-specific primers or probes.Peer reviewe

Impact of minority concentration on fundamental (H)D ICRF heating performance in JET-ILW

ITER will start its operation with non-activated hydrogen and helium plasmas at a reduced magnetic field of B-0 = 2.65 T. In hydrogen plasmas, the two ion cyclotron resonance frequency (ICRF) heating schemes available for central plasma heating (fundamental H majority and 2nd harmonic He-3 minority ICRF heating) are likely to suffer from relatively low RF wave absorption, as suggested by numerical modelling and confirmed by previous JET experiments conducted in conditions similar to those expected in ITER's initial phase. With He-4 plasmas, the commonly adopted fundamental H minority heating scheme will be used and its performance is expected to be much better. However, one important question that remains to be answered is whether increased levels of hydrogen (due to e. g. H pellet injection) jeopardize the high performance usually observed with this heating scheme, in particular in a full-metal environment. Recent JET experiments performed with the ITER-likewall shed some light onto this question and the main results concerning ICRF heating performance in L-mode discharges are summarized here

Molecular tools for selective recovery and detection of lignin-derived molecules

The pulp and paper industry together with lignocellulosic biofuel production provides plentiful streams of lignin and lignin-derived molecules (LDMs) that currently remain underutilized. The heterogeneity and complexity of lignin along with the lack of convenient tools significantly hamper its utilization. Selective separation of these LDMs from streams using specific tools would allow the recovery of aromatic compounds, as well as facilitate biological processes aiming at lignin valorization. To this end, here we report the isolation and characterization of single-chain variable fragment (scFv) antibodies against ferulate, coumarate, and caffeate, which are the molecular representatives of LDMs. Binders for the target LDMs were enriched by interrogating a synthetic scFv library with the phage display technique. As a result, scFv binders specific against each of the target molecules were obtained with affinities in the micromolar range. The selectivity of scFvs towards specific LDMs was proved by recovering caffeate from simulated LDM solution, Kraft lignin, and rice straw hydrolysate samples. Further proof of concept studies with model compounds demonstrated the applicability of antibody-based binders as a detection tool for monitoring microbial LDM conversion. Overall, this study demonstrates the potential of scFv binders as a specific toolset for lignin compound recovery and analysis

Fundamental cyclotron 3He minority ICRF heating experiments in H plasmas in JET in presence of the ILW

Efficient plasma heating schemes are a prerequisite for reaching fusion relevant temperatures in fusion machines. On the road to reaching ignition, non-activated scenarios - such as (3He

N=2 ICRH of H majority plasmas in JET-ILW

Heating single ion species plasmas with ICRF is a challenging task: Fundamental ion cyclotron heating (w = w(ci)) suffers from the adverse polarization of the RF electric fields near the majority cyclotron resonance while second harmonic heating (w =2w(ci)) typically requires pre-heating of the plasma ions to become efficient. Recently, w =2wci ICRF heating was tested in JET-ILW hydrogen plasmas in the absence of neutral beam injection (L-mode). Despite the lack of pre-heating, up to 6MW of ICRF power were coupled to the plasma leading to a transition to H-mode for P-ICRH>5MW in most discharges. Heating efficiencies between 0.65-0.85 were achieved as a combination of the low magnetic field adopted (enhanced finite Larmor radius effects) and the deliberate slow rise of the ICRF power, allowing time for a fast ion population to gradually build-up leading to a systematic increase of the wave absorptivity. Although fast ion tails are a common feature of harmonic ICRF heating, the N=2 majority heating features moderate tail energies (<500keV) except at very low plasma densities (n(e0)<3x10(19)/m(3)), where fast H tails in the MeV range developed and fast ion losses became significant, leading to enhanced plasma wall interaction. The main results of these experiments will be reported

Continuous mHealth Patch Monitoring for the Algorithm-Based Detection of Atrial Fibrillation: Feasibility and Diagnostic Accuracy Study

Peer reviewe

Continuous 24-h Photoplethysmogram Monitoring Enables Detection of Atrial Fibrillation

Aim: Atrial fibrillation (AF) detection is challenging because it is often asymptomatic and paroxysmal. We evaluated continuous photoplethysmogram (PPG) for signal quality and detection of AF.Methods: PPGs were recorded using a wrist-band device in 173 patients (76 AF, 97 sinus rhythm, SR) for 24 h. Simultaneously recorded 3-lead ambulatory ECG served as control. The recordings were split into 10-, 20-, 30-, and 60-min time-frames. The sensitivity, specificity, and F1-score of AF detection were evaluated for each time-frame. AF alarms were generated to simulate continuous AF monitoring. Sensitivities, specificities, and positive predictive values (PPVs) of the alarms were evaluated. User experiences of PPG and ECG recordings were assessed. The study was registered in the Clinical Trials database (NCT03507335).Results: The quality of PPG signal was better during night-time than in daytime (67.3 +/- 22.4% vs. 30.5 +/- 19.4%, p < 0.001). The 30-min time-frame yielded the highest F1-score (0.9536), identifying AF correctly in 72/76 AF patients (sensitivity 94.7%), only 3/97 SR patients receiving a false AF diagnosis (specificity 96.9%). The sensitivity and PPV of the simulated AF alarms were 78.2 and 97.2% at night, and 49.3 and 97.0% during the daytime. 82% of patients were willing to use the device at home.Conclusion: PPG wrist-band provided reliable AF identification both during daytime and night-time. The PPG data's quality was better at night. The positive user experience suggests that wearable PPG devices could be feasible for continuous rhythm monitoring.Peer reviewe

Additively manufactured hierarchical stainless steels with high strength and ductility

Many traditional approaches for strengthening steels typically come at the expense of useful ductility, a dilemma known as strength–ductility trade-off. New metallurgical processing might offer the possibility of overcoming this. Here we report that austenitic 316L stainless steels additively manufactured via a laser powder-bed-fusion technique exhibit a combination of yield strength and tensile ductility that surpasses that of conventional 316L steels. High strength is attributed to solidification-enabled cellular structures, low-angle grain boundaries, and dislocations formed during manufacturing, while high uniform elongation correlates to a steady and progressive work-hardening mechanism regulated by a hierarchically heterogeneous microstructure, with length scales spanning nearly six orders of magnitude. In addition, solute segregation along cellular walls and low-angle grain boundaries can enhance dislocation pinning and promote twinning. This work demonstrates the potential of additive manufacturing to create alloys with unique microstructures and high performance for structural applications