Free-standing kinked nanowire transistor probes for targeted intracellular recording in three dimensions

Recording intracellular bioelectrical signals is central to understanding the fundamental behaviour of cells and cell-networks in, for example, neural and cardiac systems1–4. The standard tool for intracellular recording, the patch-clamp micropipette5 is widely applied, yet remains limited in terms of reducing the tip size, the ability to reuse the pipette5, and ion exchange with the cytoplasm6. Recent efforts have been directed towards developing new chip-based tools1–4,7–13, including micro-to-nanoscale metal pillars7–9, transistor-based kinked nanowire10,11 and nanotube devices12,13. These nanoscale tools are interesting with respect to chip-based multiplexing, but, to date, preclude targeted recording from specific cell regions and/or subcellular structures. Here we overcome this limitation in a general manner by fabricating free-standing probes where a kinked silicon nanowire with encoded field-effect transistor detector serves as the tip end. These probes can be manipulated in three dimensions (3D) within a standard microscope to target specific cells/cell regions, and record stable full-amplitude intracellular action potentials from different targeted cells without the need to clean or change the tip. Simultaneous measurements from the same cell made with free-standing nanowire and patch-clamp probes show that the same action potential amplitude and temporal properties are recorded without corrections to the raw nanowire signal. In addition, we demonstrate real-time monitoring of changes in the action potential as different ion-channel blockers are applied to cells, and multiplexed recording from cells by independent manipulation of two free-standing nanowire probes

Fully Convolutional Network Ensembles for White Matter Hyperintensities Segmentation in MR Images

White matter hyperintensities (WMH) are commonly found in the brains of healthy elderly individuals and have been associated with various neurological and geriatric disorders. In this paper, we present a study using deep fully convolutional network and ensemble models to automatically detect such WMH using fluid attenuation inversion recovery (FLAIR) and T1 magnetic resonance (MR) scans. The algorithm was evaluated and ranked 1 st in the WMH Segmentation Challenge at MICCAI 2017. In the evaluation stage, the implementation of the algorithm was submitted to the challenge organizers, who then independently tested it on a hidden set of 110 cases from 5 scanners. Averaged dice score, precision and robust Hausdorff distance obtained on held-out test datasets were 80%, 84% and 6.30mm respectively. These were the highest achieved in the challenge, suggesting the proposed method is the state-of-the-art. In this paper, we provide detailed descriptions and quantitative analysis on key components of the system. Furthermore, a study of cross-scanner evaluation is presented to discuss how the combination of modalities and data augmentation affect the generalization capability of the system. The adaptability of the system to different scanners and protocols is also investigated. A quantitative study is further presented to test the effect of ensemble size. Additionally, software and models of our method are made publicly available. The effectiveness and generalization capability of the proposed system show its potential for real-world clinical practice.Comment: final version in NeuroImag

Intracellular Recordings of Action Potentials by an Extracellular Nanoscale Field-Effect Transistor

The ability to make electrical measurements inside cells has led to many important advances in electrophysiology. The patch clamp technique, in which a glass micropipette filled with electrolyte is inserted into a cell, offers both high signal-to-noise ratio and temporal resolution. Ideally, the micropipette should be as small as possible to increase the spatial resolution and reduce the invasiveness of the measurement, but the overall performance of the technique depends on the impedance of the interface between the micropipette and the cell interior, which limits how small the micropipette can be. Techniques that involve inserting metal or carbon microelectrodes into cells are subject to similar constraints. Field-effect transistors (FETs) can also record electric potentials inside cells, and because their performance does not depend on impedance, they can be made much smaller than micropipettes and microelectrodes. Moreover, FET arrays are better suited for multiplexed measurements. Previously, we have demonstrated FET-based intracellular recording with kinked nanowire structures, but the kink configuration and device design places limits on the probe size and the potential for multiplexing. Here, we report a new approach in which a $SiO_2$ nanotube is synthetically integrated on top of a nanoscale FET. This nanotube penetrates the cell membrane, bringing the cell cytosol into contact with the FET, which is then able to record the intracellular transmembrane potential. Simulations show that the bandwidth of this branched intracellular nanotube FET (BIT-FET) is high enough for it to record fast action potentials even when the nanotube diameter is decreased to 3 nm, a length scale well below that accessible with other methods. Studies of cardiomyocyte cells demonstrate that when phospholipid-modified BIT-FETs are brought close to cells, the nanotubes can spontaneously penetrate the cell membrane to allow the full-amplitude intracellular action potential to be recorded, thus showing that a stable and tight seal forms between the nanotube and cell membrane. We also show that multiple BIT-FETs can record multiplexed intracellular signals from both single cells and networks of cells.Chemistry and Chemical BiologyEngineering and Applied SciencesPhysic

Broadband and continuous wave pumped second-harmonic generation from microfiber coated with layered GaSe crystal

The conversion-efficiency for second-harmonic (SH) in optical fibers is significantly limited by extremely weak second-order nonlinearity of fused silica, and pulse pump lasers with high peak power are widely employed. Here, we propose a simple strategy to efficiently realize the broadband and continuous wave (CW) pumped SH, by transferring a crystalline GaSe coating onto a microfiber with phase-matching diameter. In the experiment, high efficiency up to 0.08 %W-1mm-1 is reached for a C-band pump laser. The high enough efficiency not only guarantees SH at a single frequency pumped by a CW laser, but also multi-frequencies mixing supported by three CW light sources. Moreover, broadband SH spectrum is also achieved under the pump of a superluminescent light-emitting diode source with a 79.3 nm bandwidth. The proposed scheme provides a beneficial method to the enhancement of various nonlinear parameter processes, development of quasi-monochromatic or broadband CW light sources at new wavelength regions

PPAR-γ in Macrophages Limits Pulmonary Inflammation and Promotes Host Recovery Following Respiratory Viral Infection

Alveolar macrophages (AM) play pivotal roles in modulating host defense, pulmonary inflammation, and tissue injury following respiratory viral infections. However, the transcriptional regulation of AM function during respiratory viral infections is still largely undefined. Here we have screened the expression of 84 transcription factors in AM in response to influenza A virus (IAV) infection. We found that the transcription factor PPAR-γ was downregulated following IAV infection in AM through type I interferon (IFN)-dependent signaling. PPAR-γ expression in AM was critical for the suppression of exaggerated antiviral and inflammatory responses of AM following IAV and respiratory syncytial virus (RSV) infections. Myeloid PPAR-γ deficiency resulted in enhanced host morbidity and increased pulmonary inflammation following both IAV and RSV infections, suggesting that macrophage PPAR-γ is vital for restricting severe host disease development. Using approaches to selectively deplete recruiting monocytes, we demonstrate that PPAR-γ expression in resident AM is likely important in regulating host disease development. Furthermore, we show that PPAR-γ was critical for the expression of wound healing genes in AM. As such, myeloid PPAR-γ deficiency resulted in impaired inflammation resolution and defective tissue repair following IAV infection. Our data suggest a critical role of PPAR-γ expression in lung macrophages in the modulation of pulmonary inflammation, the development of acute host diseases, and the proper restoration of tissue homeostasis following respiratory viral infections.IMPORTANCE Respiratory viral infections, like IAV and respiratory syncytial virus (RSV) infections, impose great challenges to public health. Alveolar macrophages (AM) are lung-resident immune cells that play important roles in protecting the host against IAV and RSV infections. However, the underlying molecular mechanisms by which AM modulate host inflammation, disease development, and tissue recovery are not very well understood. Here we identify that PPAR-γ expression in AM is crucial to suppress pulmonary inflammation and diseases and to promote fast host recovery from IAV and RSV infections. Our data suggest that targeting macrophage PPAR-γ may be a promising therapeutic option in the future to suppress acute inflammation and simultaneously promote recovery from severe diseases associated with respiratory viral infections

Ecological vulnerability assessment of coral islands and reefs in the South China Sea based on remote sensing and reanalysis data

Coral reefs are ecosystems that are highly vulnerable to external environmental impacts, including changes associated with ocean acidification and global warming. Assessing the vulnerability of coral reef growth environments over large areas of the sea is a difficult and complex process, as it is influenced by many variables. There are few studies on environmental vulnerability assessment of coral islands and reefs in the South China Sea. It is therefore particularly important to understand the environmental sensitivity of corals and how coral communities respond to changes in climate-related environmental variables. In this study, indicators were selected mainly from natural environmental factors that hinder the development of coral reefs. The sea surface temperature (SST), sea surface salinity (SSS), wind velocity (WV) and direction, sea level height (SL), ocean currents (OC), and chlorophyll concentration (Chl) of coral reefs in South China Sea Island were integrated to calculate the coral reef environmental vulnerability region. In a GIS environment, Spatial Principal Component Analysis (SPCA) was used to develop sensitivity models and evaluate the ecological vulnerability of coral reefs. Based on the Environmental vulnerability indicator (EVI) values, the study area was classified as 5 grades of ecological vulnerability: Potential (0.000–0.577), Light (0.577–0.780), Medium (0.780–0.886), Heavy (0.886–0.993) and Very Heavy (0.993–1.131). Sensitivity models identified regional gradients of environmental stress and found that some coral reefs in western Malaysia and southwestern Philippines have higher vulnerability. Meanwhile, the study found that the reefs of Paracel Islands and Macclesfield Bank areas of medium vulnerability. Future use of high-precision data from long time series will allow better estimates of site-specific vulnerability and allow for the precise establishment of marine protected areas so that the ecological diversity of coral reefs can be sustained

Real-world therapy with pembrolizumab: outcomes and surrogate endpoints for predicting survival in advanced melanoma patients in Germany

Knowledge on the real-world characteristics and outcomes of pembrolizumab-treated advanced melanoma patients in Germany and on the value of different real-world endpoints as surrogates for overall survival (OS) is limited. A sample of 664 pembrolizumab-treated patients with advanced melanoma from the German registry ADOReg was used. We examined OS, real-world progression-free survival (rwPFS), real-world time to next treatment (rwTtNT), and real-world time on treatment (rwToT). Spearman’s rank and iterative multiple imputation (IMI)-based correlation coefficients were computed between the OS and the rwPFS, rwTtNT, and rwToT and reported for the first line of therapy and the overall sample. The median OS was 30.5 (95%CI 25.0–35.4) months, the rwPFS was 3.9 months (95%CI 3.5–4.9), the rwTtNT was 10.7 months (95%CI 9.0–12.9), and the rwToT was 6.2 months (95%CI 5.1–6.8). The rwTtNT showed the highest correlation with the OS based on the IMI (rIMI = 0.83), Spearman rank correlations (rs = 0.74), followed by the rwToT (rIMI = 0.74 and rs = 0.65) and rwPFS (rIMI = 0.69 and rs = 0.56). The estimates for the outcomes and correlations were similar for the overall sample and those in first-line therapy. The median OS was higher compared to recent real-world studies, supporting the effectiveness of pembrolizumab in regular clinical practice. The rwTtNT may be a valuable OS surrogate, considering the highest correlation was observed with the OS among the investigated real-world endpoints

Adjuvant pembrolizumab versus placebo in resected high-risk stage II melanoma: Health-related quality of life from the randomized phase 3 KEYNOTE-716 study

Background: Adjuvant pembrolizumab significantly improved recurrence-free survival (RFS) versus placebo in resected stage IIB and IIC melanoma in the phase 3 KEYNOTE-716 study. Health-related quality of life (HRQoL) results are reported. Methods: Patients were randomly assigned 1:1 to pembrolizumab 200 mg (2 mg/kg, patients ≥ 12 to \u3c 18 years) Q3W or placebo for ≤ 17 cycles or until disease recurrence, unacceptable toxicity, or withdrawal. Change from baseline in EORTC QLQ-C30 global health status (GHS)/quality of life (QoL) was a prespecified exploratory end point. Change in EORTC QLQ-C30 functioning, symptom, and single-item scales, and EQ-5D-5L visual analog scale (VAS) were also summarized. Primary analyses were performed at week 48 to ensure adequate completion/compliance. The HRQoL population comprised patients who received ≥ 1 dose of treatment and completed ≥ 1 assessment. Results: The HRQoL population included 969 patients (pembrolizumab, n = 483; placebo, n = 486). Compliance at week 48 was ≥ 80 % for both instruments. EORTC QLQ-C30 GHS/QoL, physical functioning, role functioning, and EQ-5D-5L VAS scores were stable from baseline to week 48 in both arms, with no clinically meaningful decline observed. Scores did not differ significantly between pembrolizumab and placebo. EORTC QLQ-C30 GHS/QoL, physical functioning, role functioning, and EQ-5D-5L VAS scores remained stable through week 96 in both arms. Conclusions: HRQoL was stable with adjuvant pembrolizumab, with no clinically meaningful decline observed. Change from baseline in HRQoL was similar between arms. These results, in conjunction with the improved RFS and manageable safety previously reported, support the use of adjuvant pembrolizumab for high-risk stage II melanoma