PFKP, STMN2, CFP, KIAA1324 and TRHDE were common targets and were also differentially expressed in TDP-43-immunonegative neurons. DEG enrichment analysis revealed diverse biological pathways including immune and synaptic functions. Common DEU events in human datasets included well-known targets POLDIP3 and STMN2, and novel targets EXD3, MMAB, DLG5 and GOSR2. Our interactive database (https://www.scotterlab.auckland.ac.nz/research-themes/tdp43-lof-db/) allows further exploration of TDP-43 DEG and DEU targets. Together, these data identify TDP-43 targets that can be exploited therapeutically or used to validate loss-of-function processes.TDP-43 proteinopathy is the major pathology in amyotrophic lateral sclerosis (ALS) and tau-negative frontotemporal dementia (FTD). Mounting evidence implicates loss of normal TDP-43 RNA-processing function as a key pathomechanism. However, the RNA targets of TDP-43 differ by report, and have never been formally collated or compared between models and disease, hampering understanding of TDP-43 function. Here, we conducted re-analysis and meta-analysis of publicly available RNA-sequencing datasets from six TDP-43-knockdown models, and TDP-43-immunonegative neuronal nuclei from ALS/FTD brain, to identify differentially expressed genes (DEGs) and differential exon usage (DEU) events. There was little overlap in DEGs between knockdown models, but This article has an associated First Person interview with the first author of the paper. Summary: A meta-analysis and interactive database of TDP-43 target genes and splicing events identified in publicly available RNA-sequencing datasets from six TDP-43 knockdown models and TDP-43-immunonegative neurons in amyotrophic lateral sclerosis/frontotemporal dementia. TARDBP) is a predominantly nuclear DNA- and RNA-binding protein first discovered to bind to the trans-active response element in the human immunodeficiency virus (HIV)-1 sequence  that almost all inherited ALS including minantly ; (2) thaminantly ; and (3)minantly . Recogniminantly . Nuclearminantly  Fig. 1)TARDBP-muminantly . Howeverminantly . These iminantly . Nuclearminantly , so lossminantly  minantly . Notablyminantly . Thus, lRANBP1 and POLDIP3, are clearly reproducible in multiple studies ; GSE116456 . An additional RNA-seq dataset was identified in which neuronal nuclei had been sorted from ALS/FTD tissue according to nuclear TDP-43 immunoreactivity [GSE126543 ]; however, only six of these met quality control thresholds and were fully processed . Of thesTARDBP/Tardbp was examined in HeLa, SH-SY5Y, ihMN, mouse striatum, mouse mammary gland and rat astrocytes .There were three upregulated differentially expressed genes (DEGs) and five downregulated DEGs shared by all human-derived TDP-43 knockdown models, and nine upregulated and three downregulated DEGs shared by all rodent-derived TDP-43 knockdown models B. There KIAA1324, CFP and ITGA4. The shared downregulated DEGs were TRHDE, PFKP and MASP2 <0.05] in each model are listed in Tables S2 and S3 (only genes with log2FC between −0.5 and >0.5), or can be explored interactively at https://www.scotterlab.auckland.ac.nz/research-themes/tdp43-lof-db/ , filtering by model system or gene of interest. Overall, DEGs were highly variable between datasets, but the sharing of DEGs between model systems validates them as conserved TDP-43 targets, and sharing of DEGs with ALS/FTD TDP neg neuronal nuclei validates those DEGs as being disease relevant.The upregulated DEGs shared by the largest number of TDP-43 knockdown models and with ALS/FTD TDP neg neuronal nuclei were nd MASP2 D. DEGs [Table S4.Having identified certain genes with conserved patterns of regulation by TDP-43 between cell types and species, we examined the effect of TDP-43 knockdown on biological processes (BPs), molecular functions (MFs) and cellular components (CCs) using gene ontology (GO) enrichment analysis. The top three significant terms for BPs, MFs and CCs for both upregulated genes , left anTwo patterns emerged after redundancy was eliminated from the GO terms: many terms were related to immune response  using the blastn algorithm, to examine sequence homology of DEU loci between human and rodent datasets. Only two genes showed a DEU event that was shared between ALS/FTD TDP neg neuronal nuclei and a rodent dataset, and that also showed sequence homology of the region between human and mouse: STMN2 and POLDIP3 , the 3′ UTR  and a lncRNA (TBL1XR1-AS1)  . We consTARDBP mRNA targets drives pathogenesis and phenotype remains unclear. Central to this is the need to identify the complement of TARDBP mRNA targets in distinct cell types and model systems, and to verify which of these targets is regulated by TDP-43 in human ALS/FTD neurons. We approached this problem by assessing TARDBP mRNA targets in human and rodent models, in neuronal and non-neuronal cell types, and identifying shared and unique targets and biological pathways under TDP-43 control. TDP-43 regulates certain common targets across myriad cell types, implying that TDP-43 proteinopathy in non-neuronal cells initiates processes that at least partially overlap with those occurring in neurons. Notably, TDP-43-dependent changes identified in multiple model systems were validated to change in human ALS/FTD neuronal nuclei with TDP-43 nuclear depletion, supporting TDP-43 loss of function as being pathomechanistic.As the common neuropathological feature in ALS with or without FTD, aggregated TDP-43 and its acquired toxic functions represent tractable molecular targets for therapy. Yet, increasing evidence suggests that successful therapies will also require rescue of normal physiological functions of TDP-43 that are compromised in disease. Loss of gene expression and splicing regulatory function are now widely accepted features of ALS with TDP-43 proteinopathy, but how the specific complement of PFKP mRNA has been identified as a TDP-43 loss-of-function marker by several studies in addition to those in our meta-analysis . This cryptic exon may therefore be preferable to overall PFKP transcript levels as a TDP-43 loss-of-function marker.Certain transcripts were robustly regulated by TDP-43 in multiple datasets and in human ALS/FTD neurons with nuclear TDP-43 loss. Decreased analysis . PFKP en miR-520 . Yet, decontrols . Howeverdirectly , and TAReraction . Indeed,STMN2 expression following TDP-43 depletion is associated with the emergence of a cryptic exon after exon 1, which introduces a premature polyadenylation site  is an M1 family metallopeptidase enriched in brain regions  and endosome/lysosome-associated apoptosis and autophagy regulator (ELAPOR1) in humans (Elapor1 or insulin inhibitory receptor (Iir) in mice  the cell types in which the transcript is expressed, and whether those cell types are subject to TDP-43 dysfunction in the tissue sampled; (2) whether the transcript is subject to secondary regulation, aside from regulation by TDP-43; and (3) whether regulation by TDP-43 is direct or indirect via other mediators also subject to change. Overall, the ability of DEGs to report upon TDP-43 function is likely to differ across different transcriptomic profiles .POLDIP3). The skipping of exon 3 in POLDIP3 is a frequently reproduced marker in TDP-43 knockdown studies  of ALS/FTD tissue, in conjunction with cell-type-specific immunohistochemical markers, to identify additional cell types with TDP-43 loss of function. This panel of DEU events could also act as primers/probes for quantitative RT-PCR to assess the fidelity of ALS models or the restoration of TDP-43 function by therapeutics.Half of the DEU events common to all human datasets were modifiers of 3′ UTR usage. The 3′ UTR is an important regulatory region, of mRNA stability , localizPOLDIP3)  (GOSR2) (IL18BP)  (EXD3) (USP31)  , mitocho NDUFA5) , endosom (GOSR2) , immune (IL18BP) , microtu3, DLG5) , exonucl) (EXD3)  and deub (USP31) . How theSeveral of our data suggest our DESeq2 and DEXSeq analyses to be conservative methods for identifying DEGs and DEU events, and comparative studies of DEG packages have also demonstrated DESeq2 to err on the conservative side . It is eSTMN2 that has attracted significant attention in ALS/FTD research . The search was performed using the keyword ‘TDP-43’, and the results were filtered by setting Entry Type as ‘Series’ to capture all potential samples that belonged to a common study, and setting Study Type as ‘Expression profiling by high throughput sequencing’. Inclusion criteria for re-analyzing these datasets were as follows: (1) raw RNA-seq data available; (2) at least three TDP-43 knockdown samples and two appropriate control samples; (3) experimental depletion of TDP-43. In addition, an RNA-seq dataset was identified in which neuronal nuclei had been sorted from ALS/FTD tissue according to nuclear TDP-43 immunoreactivity; either TDP pos  or TDP neg (no nuclear TDP-43) (A repository search was conducted in September 2020 to identify TDP-43 knockdown studies with available RNA-seq data using the Gene Expression Omnibus (GEO) from the NCBI ( TDP-43)  Fig. 2)http://nchttp://www.ncbi.nlm.nih.gov/sra), and quality control was applied using FastQC v0.11.9 software (https://www.bioinformatics.babraham.ac.uk/projects/fastqc/). Details of each dataset including study design, GEO accession number and library information are supplied in Table S1. Code is deposited at https://github.com/mcao051/TDP_LOF. Raw data that still contained adapter sequence content were trimmed with Trimmomatic v0.39  was used for GO enrichment analysis using genes with Padj<0.05 and log2FC of either <−0.5 or >0.5. The genes detected after low-count filtering for each dataset were used as the background gene list. Redundant GO terms were eliminated using the ReViGO tool available at http://revigo.irb.hr/.The data processing pipeline is shown in ic v0.39 . If feweic v0.39 . Referenic v0.39  to perfoic v0.39  and DEU ic v0.39 . DEU is ic v0.39 . DESeq2 https://www.scotterlab.auckland.ac.nz/research-themes/tdp43-lof-db/. They enable the reader to visualize and interact with all differential gene expression and exon usage analyses described herein, including genes of interest not highlighted in our study.Interactive graphical reports were generated from the packages Glimma  and DEXShttps://ggplot2.tidyverse.org/), ggvenn (https://CRAN.R-project.org/package=ggvenn), the plotCounts function from within DESeq2 (https://www.r-project.org/), Prism 9.0 software  and Integrative Genomics Viewer (Data were visualized using R software with ggplot2 (n DESeq2 , DEXSeq s Viewer . Adobe P


An optical fiber Fabry–Perot interferometer (FPI) is constructed for relative humidity measurement by fusion splicing a short hollow core fiber (HCF) to the end of a single-mode fiber and coating the tip of the HCF with a layer of gelatin. The thickness of the gelatin film changes with ambient humidity level and modulates cavity length of the FPI. Humidity measurement is therefore realized by measuring the wavelength shift of the interreference fringe. RH sensitivity of 0.192 nm/%RH is achieved within a measurement range of 20–80%RH. Dynamic measurement shows a response and recovery time of 240 and 350 ms, respectively. Sensor performance testing shows good repeatability and stability at room temperature but also reveals slight dependence of the RH sensitivity on environmental temperature. Therefore, a fiber Bragg grating is cascaded to the FPI sensing probe to monitor temperature simultaneously with temperature sensitivity of 10 pm/°C. Humidity measurement is widely needed in the many fields such as food, medicines, storage and environmental monitoring. High-performance humidity sensors with wide measurement range, high accuracy and fast response speed are desired. Since conventional humidity sensors based on electronic technologies are hardly used for long-distance measurement or in strong electromagnetic-interference environments, optical fiber humidity sensors have attracted a growing number of research interests in recent years because of their long-distance signal transmission capability and immunity to electromagnetic interference.An optical fiber humidity sensor is usually composed of an optical fiber structure interacting with a humidity-sensitive material. The optical fiber structure can be a fiber grating ,2,3,4, fIn this paper, an optical fiber interferometric humidity sensor by using a short hollow core fiber (HCF) interacting with a gelatin film is demonstrated. It is constructed by fusion splicing a short length of HCF to the end of a single-mode fiber (SMF) and coating the tip of the HCF with a layer of gelatin to form a Fabry–Perot interferometer (FPI). Humidity changes the thickness of the gelatin film that modulates the interference cavity length and hence the dip wavelength of the interference spectrum of the FPI in turn. A relative humidity (RH) measurement with a sensitivity of 0.192 nm/%RH is therefore achieved within a measurement range of 20–80%RH and with the response and recovery times of 240 and 350 ms, respectively. It is found that the RH sensitivity shows a slight dependence on environmental temperature. A fiber Bragg grating (FBG) is therefore cascaded in the FPI sensing probe to monitor temperature changes simultaneously.A schematic diagram of the proposed optical fiber FPI humidity sensor probe is shown in The working principle for humidity measurement is based on the humidity-induced thickness change in the gelatin film that changes the cavity length of the FPI formed by the two reflective surfaces, as shown in Based on the theory of optical interference, intensity of the reflected light of the FPI probe can be derived by using the two-beam approximation and expressed as :(1)I=I0[When the external humidity level increases, the gelatin film will absorb more water molecules from the surrounding air and expand in volume. Its thickness will be increased accordingly, which will reduce cavity length of the FPI and shift the dip wavelengths of the interference spectrum to the short wavelength direction. When the external humidity level decreases, the gelatin film will release water molecules to the surrounding air and shrink in volume. Its thickness will be decreased and the cavity length of the FPI will become larger to make the dip wavelengths of the interference spectrum move to longer wavelengths. The changes in dip wavelengths of the FPI can be expressed by:The experimental setup of the humidity measurement is shown in Three FPI probes with different HCF lengths of 73, 110 and 220 μm were fabricated, and their interference spectra are shown in The humidity level was increased at a step of 5%RH within the range of 20 to 80 %RH. We tested the responses of the FPI probe in both ascending and descending orders of RH level. The measured wavelength shifts in the FPI probe against RH in both orders as shown in −2, indicating quite good repeatability for the proposed FPI humidity probe. The minor errors among different measurements are most possibly caused by the nonuniform RH distribution and unstable airflows in the THCC.To further study the repetitive characteristics of the FPI humidity probe, two more iterations of repetitive testing were carried out. The measured results of dip wavelength shift against RH level in both ascending and descending orders are shown in To study the effect of environmental temperature on the response of the FPI humidity probe, the testing was carried out under two more different temperatures of 15 and 25 °C. The measured results, together with that measured at 20 °C, were shown in Then, the stability test of the FPI probe against time was carried out at three different RH levels of 20, 40 and 80%RH under a fixed temperature of 20 °C. The dip wavelength was measured every 5 min within a period of 30 min, and the results are shown in The dynamic response of the FPI probe to humidity change was then carried out by changing the humidity level from 50 to 80%RH quickly and then changing it back after 10 s. We set the OSA to automatic acquisition mode to record the wavelength shift of interference spectrum of the FPI probe at a high speed. The measured results, as shown in The fast response and recovery time may be attributed to the compact structure of the FPI probe and the small thickness of the gelatin film, which allow the fast diffusion of water vapor molecules. Based on the fast response time, the FPI probe provides a near-real-time humidity measurement technique which can be used in areas such as human breathing monitoring for the improvement of healthcare, as the response time of current commercial humidity sensors is in the order of several seconds.2 = 0.99723. Since the FBG is not sensitive to RH variations, it can provide the accurate information of environmental temperature for reference when the FPI humidity probe works.Finally, we tested the sensitivity of the FBG to temperature within a temperature range of 10 to 50 °C. The measured wavelength of the FBG’s reflection peak against temperature are shown in To better evaluate the proposed FPI humidity sensor, we compare the key performances including sensitivity, measurement range and dynamic response time of our sensor, with other reported ones also based on optical fiber interferometers. The results, as shown in An optical fiber FPI-based humidity sensor has been demonstrated by fusion splicing a short HCF with a single-mode fiber and coating the tip of the HCF with a layer of gelatin. The working principle is based on the humidity-induced thickness change in the gelatin film that changes the cavity length of the FPI. A humidity measurement with a high sensitivity of 0.192 nm/%RH and a wide measurement range of 20–80%RH has been realized by measuring the wavelength shift of the interreference fringe. Good repeatability and stability have been obtained, and the response and recovery times are only 240 and 350 ms, respectively. In addition, simultaneous measurement of temperature with a sensitivity of 10 pm/°C has been achieved by cascading an FBG sensor in the FPI probe.


P-type ternary switch devices are crucial elements for the practical implementation of complementary ternary circuits. This report demonstrates a p-type ternary device showing three distinct electrical output states with controllable threshold voltage values using a dual-channel dinaphthothieno-thiophene–graphene barristor structure. To obtain transfer characteristics with distinctively separated ternary states, novel structures called contact-resistive and contact-doping layers were developed. The feasibility of a complementary standard ternary inverter design around 1 V was demonstrated using the experimentally calibrated ternary device model. MVL provides significant efficiency gains in terms of the number of transistors and the interconnection length required to perform equivalent functions designed with binary logic. In particular, the ternary logic that is composed of three logic states—0, 1, and 2—has several advantages over Boolean logic design using the conventional complementary metal–oxide–semiconductor (CMOS) logic, with the possibility of the lowest power consumption among MVLs9.Recently, multi-valued logic (MVL) technology has attracted interest as an alternative architecture to address the rapid increase in energy consumption required for massive data processingVth) in the form of integrated circuits8. More recently, a quantum dot gate FET was also proposed for ternary logic operation, but the process of fabricating two layers of well-aligned quantum dots is difficult to control and further scalability can be a serious challenge because the size of the quantum dots and the separation distance between them cannot be modulated flexibly9. Ternary devices using transition metal dichalcogenide (TMDC) heterojunctions have also been proposed to utilize the negative differential resistance or negative differential transconductance13. Shim et al. reported a ternary device based on a BP/ReS2 heterojunction forming a broken-gap band structure10. Nourbakhsh et al. demonstrated a MoS2/WSe2 ternary device with band-to-band tunneling mechanism11. Although these devices show promising functional feasibility, large-scale device integration remains quite challenging for TMDC materials. The wafer scale growths of TMDCs above oxides such as crystalline SiO2 or sapphire have been recently researched, but the direct growth on the oxide when the device structure exists is not desirable because of high growth temperatures over 800 °C15.Owing to the apparent merit of architectural simplicity, various kinds of ternary devices have been investigated for many decades. Among them, carbon nanotube field-effect transistors (CNTFETs) have been the most widely investigated, but all studies using CNTFETs have been theoretical because it is difficult to arrange various types of CNTs with different threshold voltages , where external doping processes have been used in graphene channels to customize various PN junctions in the channel region21. However, the low noise margins because of the low on/off current ratios of GFETs, which are less than ~ 10, constitute a serious drawback in ternary circuits.Graphene FETs are also used for ternary logic applications because of their high mobility, linearly controllable Fermi levels, and large area growth via thermal chemical vapor deposition (TCVD)n-type ternary graphene barristor by forming dual junctions with different Vth22. The graphene barristor, composed of graphene–semiconductor heterojunction, is the Schottky barrier triode modulating by control of the Fermi level of graphene using an electrical gate field. For implementing a ternary logic device, a graphene barristor is a better option, because of its linear drivability of current and large on/off ratio than GFETs. However, the benefits of ternary logic could not be fully accomplished by the resistive-load approach; a p-type counterpart ternary device is necessary to overcome this limit. Unfortunately, there are not many candidate materials for the p-type semiconductor that can be deposited at low temperatures, without damaging the graphene channel at low temperatures. However, several methods of fabricating p-type “binary” graphene barristors have been reported using pentacene and dinaphthothieno-thiophene (DNTT), despite the relatively low field-effect mobility ~ 2 cm2/Vs26. In addition, DNTT showed better air stability than pentacene28. Thus, it is worthwhile to pursue a p-type ternary graphene barristor utilizing the works on air-stable DNTT–graphene barristors.Kim et al. demonstrated an p-type ternary graphene barristor using a DNTT–graphene heterojunction. Multi-Vthp-type DNTT–graphene barristors with dual-channel structures were successfully demonstrated by modulating the carrier transport through the DNTT and metal contact interface. Subsequently, the functionalities and benefits of a standard ternary inverter (STI) were examined using the complementary ternary graphene barristors (ZnO–graphene barristor for an n-type device and DNTT–graphene barristor for a p-type device).In this study, we investigated a dual-channel p-type graphene barristor with the DNTT–graphene heterojunction device. In this study, a buried gate structure was adopted to control the Fermi level of graphene more stably  metals were deposited using an e-beam evaporator to fill the oxide trench. The buried gate pattern was formed in the trench region by applying a combination of lift-off and chemical–mechanical polishing processes. The buried gate structure was chosen because it can provide a more uniform electric field in the channel region than the bottom gate structure that is commonly employed in the early stages of graphene device studies29. This step was followed by atomic layer deposition of 30 nm of Al2O3 gate dielectric at 200 °C and annealed at 300 °C in a high vacuum (~ 10−6 Torr) for 1 h to improve the quality of the gate dielectric  for certain devices, whereas for other devices, a contact-doping layer (CDL) was formed with 10 nm of a 7:1 co-evaporation layer of DNTT and 2,3,5,6-tetrafluoro-7,7,8,8-tetracyanoquinodimethane  deposited on the drain region using a shadow mask , which is a ask Fig. f. Upon cn-doped graphene channels were 5.1 and 5.3, respectively, which confirmed that the graphene channel was a monolayer. Subsequently, the undoped graphene channel was slightly p-doped with impurity charges at the graphene–Al2O3 interface and the redshifts of the G and 2D peaks indicated that the PEI doping process successfully changed the doping state of the graphene channel to n-type31.During device fabrication, the quality of the graphene channel was examined using Raman spectroscopy, as shown in Fig. The electrical properties of the fabricated devices were characterized using a semiconductor parameter analyzer (Keithley 4200) at room temperature.p-type graphene barristor, where the arrow indicates the direction of carrier flow. The DNTT partially overlaps the graphene channel, and the Schottky junction is formed at the DNTT–graphene interface. The flux of charge carriers through the DNTT–graphene Schottky barrier can be modulated using the buried gate bias (Vg), which modulates the Fermi level of graphene and thereby adjusts the Schottky barrier height (SBH).Figure p-type graphene barristors are shown in Fig. Vg is low (more positive Vg), the graphene barristor is turned off because the SBH is too high to allow the flow of hole carriers, as shown in Fig. Vg increases (more negative Vg), the graphene barristor is turned on as the SBH is reduced, as illustrated in Fig. Vth. Therefore, the surface of the graphene channel region was treated with PEI solutions of various concentrations, viz., 0 (no doping), 0.0025, 0.005, and 0.01 wt%. As the concentration of the PEI solution increases, Vth of the graphene barristor shifts toward negative Vg; Vth is − 6.8 V in the case of 0.01 wt% PEI concentration. The n-type dopant, PEI, induces additional electron charges in graphene, which shifts the Fermi level of graphene upward and increases the SBH. The effects of graphene doping are schematically illustrated by comparing the band diagrams of undoped and n-doped graphene barristors in Fig. Vg ranging from − 15 to 15 V, the extracted SBHs are modulated from 0.19 to 0.34 V for the undoped device and from 0.21 to 0.39 V for the n-doped device, indicating that the Vth shift of the graphene devices is caused by the change in the SBH due to chemical doping. , as shown in Fig. p-type ternary device is presented in Fig. 2, which is sufficient to divide the current level between ternary states. This is the first demonstration of a p-type ternary logic switch with a saturated intermediate state. Figure Id–Vg of the p-type ternary device on a linear scale. The saturation of the intermediate current is distinctly observed even on this scale, which is important for the ternary logic gate circuit. Another positive result of this study is that the electrical characteristics of the p-type ternary device can be maintained for more than 45 days, confirming that this device is air-stable , and the other was the  channel , as show2O3). In principle, further performance enhancement is quite feasible because there are several structural parameters, such as the electrical thickness of the gate dielectric, thickness and doping state of the DNTT, and area of the Schottky junction, that can be optimized further. Because the experimental optimization requires a rather extensive material-based study, theoretical analysis of the future scalability and performance was performed based on the fitting result shown in Fig. The performance of the ternary device is not yet competitive with CMOS technology on a similar scale. The operation voltage range is quite large, and the subthreshold swings remain very high due to the thickness of the gate dielectric (30 nm of Alp-type ternary device was well-fitted by a capacitor-based ternary device model (Supplementary information, Fig. The electrical operation of the n-type dual-channel ternary graphene barristor, the ZnO–graphene barristor model reported in the literature22 was used (Supplementary information, Fig. Then, a complementary STI was designed using the device model developed for 1 V operation, as shown in Fig. SPICE for Vdd = 1 ± 0.1 V. Well-behaving STI transfer characteristics were obtained around Vdd = 1 V. For STI, four noise margins were defined, as demonstrated in Fig. dd, in which the minimum noise margin was 71 mV at Vdd = 1.06 V. The very narrow peak region in Fig. n-type and p-type ternary switches are necessary. The gains of the two-state transitions were 2.3 and 2.8, respectively, for Vdd = 1.06 V (Fig. 7. However, the large static power dissipation in the intermediate state due to the half turn-on of both n- and p-type ternary devices remains an issue for improvement for low-power systems (Supplementary information, Fig. Figure 6 V Fig. d. As a rp-type ternary device is the last essential element for the operation of complementary ternary logic circuits. By combining the demonstration of this device with the recent progress related to n-type ternary devices, reasonable projections of the performance and functionality of a complementary ternary inverter were achieved, and the feasibility of low Vdd operation at 1 V was confirmed. Although further optimization to scale down the device is necessary, this report confirms the achievement of a major milestone in ternary logic technology for extremely low-power computing.A fully functional Supplementary Information.


We report an ultra-broadband and compact TM-pass polarizer based on graphene-buried polymer waveguides. The characteristic parameters of the polarizer were carefully designed and optimized. The standard microfabrication processes were employed to fabricate the device. The presented polarizers exhibit high polarization-dependent transmission imposing a TE mode cutoff while leaving the TM mode almost unaffected. We experimentally demonstrated the polarizer that has an ultra-high extinction ratio of more than 22.9 dB and 41.9 dB for the monolayer graphene film placed on the surface of core layer and buried in the center of core layer, respectively, and as low insertion loss as ~4.0 dB for the TM mode with the bandwidth over 110 nm. The presented polarizer has the advantages of high extinction ratio, ultra-broadband, low cost, and easy integration with other polymer-based planar lightwave devices. Polarization state control of the light has important applications in optical communications and photonic integrated circuits, especially for the systems with only one operating polarization . OpticalHere, we report an ultra-broadband and compact TM-pass polarizer based on graphene-buried polymer waveguides. The polarization mechanism is mainly due to the differential attenuation of the two polarization modes in the presented hybrid waveguides. The change of the modal characteristics for the TE mode is more than that of the TM mode, which is derived from the fact that the electric field of the TE mode interacts with graphene more effectively than that of the TM mode. Moreover, thanks to the flexible fabrication process of the polymer waveguide, the graphene film can be easily placed anywhere inside the waveguide by using the standard microfabrication process. The presented polarizers have the advantages of low insertion loss, high extinction ratio, and easy integration with other polymer-based photonic integrated devices.λ0 is 1550 nm. The relations between the waveguide core width a and mode effective indices effN of the strip waveguide without a graphene layer is shown in b = 0.8a. In order to realize single-mode transmission in the waveguide, the core width a should be selected in the range of 2~7 µm, meanwhile, the core thickness b was chosen in the range of 1.6~5.6 µm. The simulated intensity patterns of the fundamental mode with the optimized core dimension for TE and TM polarizations are shown in Polarizers with the graphene layer buried in different locations are designed to examine the effects of the buried location on their loss performance. The schematic structures of the demonstrated two types of polarizers are shown in MPA) can be calculated with the imaginary part of the effective refractive index according to the following formula [effN) is the imaginary part of the effective mode index of the waveguide. The modal loss induced by the graphene was affected by the core dimension, which is mainly due to the effect of the core dimension on the modal field intensity distributions in the waveguide. As shown in The complex effective indices of the hybrid polymer waveguide embedded with graphene layer were calculated based on the interface model by using the finite-element method with the commercial full-vector mode solver COMSOL. In the simulation, we have established triangle meshes with a maximum mesh length of 500 nm and applied perfect matched layers as the boundary condition, which can help us get an accurate numerical result since electromagnetic waves are perfectly absorbed in the boundary without reflections. The modal loss MPA=The designed polarizers were fabricated by using standard microfabrication techniques, wherein polymer materials EpoCore and EpoClad  were chosen as the core and cladding materials, respectively. To characterize the performance of the fabricated polarizers based on the graphene-buried polymer waveguides, we used the polarization optics to control and monitor the polarization states of the input light and the output light. The input signal light from a semiconductor tunable laser  is coupled directly into the input port of the polarizer through a lensed single-mode fiber (SMF) with a mode-field diameter of 2.5 µm. The output light beam from the polarizer can be collected with a lens, which images the output near-field patterns, and observed by an infrared charge-coupled device (CCD) camera. The images can be displayed on a video monitor during the testing process. After finding the output pattern, the output light was coupled into an optical power meter through a SMF to measure the output optical power from the polarizer and calculate the loss induced by graphene film and the insertion loss of the polarizer. By comparing the losses measured for the graphene-buried waveguide and the reference waveguide without graphene film, the losses induced only by the graphene films can be obtained. The variation of graphene-induced losses of the polarizers with the operating wavelength is indicated in In fact, we should get a larger extinction ratio, however, this is limited by the measuring system rather than the graphene layer. Therefore, we dissociated the samples and further experimentally measured the loss performance of the polarizers embedded with a shorter length of graphene film. As shown in n is the number of graphene layers and t is the thickness of EpoCore between graphene sheets, where the t is 100 nm in our design. The variation of the graphene-induced loss of the polarizer with the numbers of graphene layers is shown in Moreover, we further theoretically verified the effect of the number of the buried graphene layers on the fundamental mode losses. In conclusion, to develop the polymer integrated waveguide polarizer in the optical communication system applications, we have proposed and experimentally demonstrated an ultra-broadband TM-pass polarizer based on polymer waveguide embedded with graphene layer. Polarizers with the graphene film buried in different locations are designed and fabricated to demonstrate the effects of the buried locations as well as the layers of the graphene on their loss performance. Our typical fabricated polarizer presents an ultra-high extinction ratio of more than 22.9 dB and 41.9 dB for the monolayer graphene placed on the surface of core layer and buried in the center of core layer, respectively, and as low insertion loss as ~4.0 dB for the TM mode with the bandwidth over 110 nm at telecommunication wavelengths. Furthermore, we can increase the graphene layers buried in the center of core to develop a polarizer with compact size and higher extinction ratio. The proposed polymer waveguide polarizers with high extinction ratio, low insertion loss, ultra-broadband, and easy fabrication process have potential applications for on-chip photonic integrated circuits.


This review article looks at the use of lead-free halide perovskite materials for humidity sensing. A complete description of the underlying mechanisms and charge transport characteristics that are necessary for a thorough comprehension of the sensing performance will be provided. An overview of considerations and potential recommendations for the creation of new lead-free perovskite nanostructure-based sensors is presented.Over the past decade, perovskite-based nanomaterials have gained notoriety within the scientific community and have been used for a variety of viable applications. The unique structural properties of these materials, namely good direct bandgap, low density of defects, large absorption coefficient, high sensitivity, long charge carrier lifetime, good selectivity, acceptable stability at room temperature, and good diffusion length have prompted researchers to explore their potential applications in photovoltaics, light-emitting devices, transistors, sensors, and other areas. Perovskite-based devices have shown very excellent sensing performances to numerous chemical and biological compounds in both solid and liquid mediums. When used in sensing devices, Perovskite nanomaterials are for the most part able to detect O Double perovskite is the resultant substance, having the formula A2B’B”X6 and the cubic space group Fm3 m . A. A3) typup Fm3 m . The geoup Fm3 m . The thee formed . Lead-fr2BiAgCl6, Cs2BiAgBr6, Cs2SbAgCl6, and Cs2SbAgBr6  octahedra, even though researchers would want to name them, perovskites. Due to the isolated [BX6] octahedra in A2BX6 compounds, the optoelectronic properties of A2BX6-type perovskites deviate from those of 3D ABX3  perovskites. A2SnI6  [2TiBr6 [2BX6 compounds [Previous research has shown that tetravalent B(IV) replacements may be used to replace Pb in lead halide perovskites. To accommodate these heterovalent replacements, the chemical formula must be changed to Aeen them ,74,75,76 Cs, MA) ,78 and C [2TiBr6  have beeompounds ,77,78,792BiAgBr6 and Cs2PdBr6 are both recently developed lead-free perovskite with good photoelectric properties and high stability. Zhan Yiqiang et al. and Ye et al. both found that these materials reported humidity-dependent electrical properties and good stability and therefore giving them excellent humidity sensitivity [Cssitivity ,38. By aThere are two types of humidity measurements, i.e., absolute humidity, and relative humidity related to qualitative assessment. The proportion of vapor (moisture) in a unit volume of air, independent of temperature, is referred to as absolute humidity. The proportion of water vapor present in a unit volume of air is also referred to as relative humidity, but this time in connection to the temperature of the air.Most humidity sensors work by measuring the change in capacitance or resistance of a certain conductive substance as a function of relative humidity. Several fundamental parameters determine the good functioning of a humidity sensor. The proper operation of a humidity sensor is determined by several essential parameters. The key parameters to consider when it comes to evaluating the functioning of a humidity sensor are sensitivity, accuracy, and response/recovery time.Humidity sensors are categorized into several classes of sensors. These groups include electrical, mechanical, optical, and integrated sensors. Electrical sensors are types of sensors whose functioning is based on impedance and capacitance , strain The general humidity sensing mechanism of ceramic oxides or perovskites as a whole relies on the superficial water vapor absorption from chemisorption, physisorption, and capillary condensation process as can be seen in 2BiAgBr6 humidity sensor. When there are no water molecules adsorbed on the film surface, the conductivity comes from the carriers inside the p-type Cs2BiAgBr6 material, which may principally be the Ag vacancy [+ protons are transported to nearby water molecules, H3O+ are generated. In the presence of external electric fields, the protons can then be moved from one site to another across energy barriers. As a result, when the humidity level is low, the sensor’s electrical conductivity rises.In general, the adsorption of water molecules on the sensor surface during the humidity sensing process is divided into different phases, chemisorption, and physisorption ,83, as i vacancy . Water m vacancy . Meanwhi vacancy ,85, in a2BiAgBr6 thin film  on the flat (or porous) surface, these different processes can be involved in explaining the resistance or capacitance changes. Under applied voltage, singly bonded water vapor molecules, for example, become mobile and capable of forming dipoles and electrolyte layers, leading to an enhanced dielectric constant and bulk conductivity. As a result, the surface water protonation, as well as protonic conduction mechanisms, might cause a minor fluctuation in conductivity with humidity adsorption.2AgBiX6 and Cs2InBr5H2O  in their stable state ranges from t = 0.8 to t = 1 [3 configuration, inorganic perovskites based on Cesium exhibit low t-factor values, indicating poor phase stability. As can be observed from One of the most important ways in which scientists have been able to improve not just the structural characteristics of perovskite materials, but also their optical characteristics, has been to modify their compositional structure. These modifications have proven to have an impact on the material’s performance and stability in device applications. The capacity that perovskite materials have, to tune their bandgap by using mixed-cation or mixed-halide compositions, enables their optical absorption to be expanded spanning a broader spectrum of wavelengths ,89. The to t = 1 ,90,91. WHumidity sensors typically use two methods to measure humidity. One of those is resistive sensing, in which water molecules affect the bulk material’s resistance due to chemisorption or physisorption. The second is capacitive sensing, in which the sensor’s capacitance changes when it interacts with water vapor . The typThe various perovskite materials utilized in the production of the capacitive and resistive sensors reported in Typically, noble precious metal electrodes are fabricated on a glass or ceramic substrate via thick film printing techniques. Ref.  or thin-The key active materials used for the fabrication of humidity sensors include metal oxides, with perovskites being a subpart , carbon Studies conducted on polymeric humidity sensors have progressed and been incorporated into the industry over the past four decades. The majority of these sensors are made from porous polymer thin films  and emplOrganic polymer thin film humidity sensors with their applications, on the other hand, have a lower degree of satisfaction and significance when compared to metal-oxide thick or thin-film ceramic sensors , yet, thElectroceramic materials having single/polycrystalline structures, as single species or composites, could be great contenders for humidity sensing applications. The use of these innovative materials with novel humidity sensing properties is being investigated to overcome some of the limitations of many of the conventional materials, such as insufficient sensitivity or selectivity, low catalytic grade, insufficiency of cavities, surface degradation due to harsh contaminants in extreme conditions, and failure to operate in extremely dry or moistened environments. The addition of suitable nanomaterials with variable particle sizes/morphologies, hybridization of materials by replacement or doping of new atoms in the lattice, and particle size reduction to sub-nano scales can all be used to overcome most of the flaws.2O3) [With techniques such as thick film screen printing, porous ceramic or nanorod-based ceramic humidity sensors have also been manufactured and produced ,130, whe2O3) . In this2O4-TiO2 [2O4-LiZnVO4 [2-K2Ti6O3 [l, Fe)2O4-TiO2 [2O4 [2 [2O4-TiO2 porous ceramic humidity sensor for microwave oven applications [Ionic and electrical conduction sensors, as indicated previously, are two types of resistive sensors. Surface chemisorption and physisorption are used by ionic conduction types of humidity sensors to measure ambient air relative humidity, and they include MgCr2O4-TiO2 , ZnCr2O4-LiZnVO4 , TiO2-K2-K2Ti6O3 , . [3 [3 [3 [The research and analysis of perovskite films and bulk materials’ humidity sensing behavior and morphological structure continue to generate unique preliminary and novel findings ,141,142.oxygen). , and forgen). [3 , ZnSnO3 ). [3 [3 , and NaT[3 [3 [3 , were cl3 nanocrystalline films with great sensitivity, strong linearity with impedance values ranging in more than three times the normal magnitude, limited hysteresis, and quick response time at 100 Hz, AC 1 V from 33 to 95 percent relative humidity. The complex impedance was also used to investigate the humidity sensing mechanism. The findings suggested that NaTaO3 may be used in humidity sensors. In the example of ZnSnO3, Bauskar et al. [3 cubic crystallites produced using a hydrothermal technique to create a quick and stable humidity sensor. Linearity, rapid response and recovery behavior, hysteresis within 3.5 percent, outstanding repeatability, stability, and a wide range of operation (11–97% RH) were all excellent humidity sensing properties of the sensor. Finally, Mahmoud et al. [3 powder for moisture sensing measurements. The solid-state reaction approach was used to make perovskite PbTiO3 (PT) powder and ceramics in a cost-effective manner. It made it possible to create a perovskite tetragonal single phase. At an ambient temperature, however, pure PbTiO3 did not demonstrate a satisfactory reaction to relative humidity.Zhang et al.  were ablr et al.  conducteation 11–% RH were3NH3PbI3-xClx films and investigated its sensitivity in RH ranges of 32 to 97 percent with a recovery time of 74 s. Xu et al. [3NH3PbBr3 humidity sensor with a quicker response time (250 s) and quicker recovery (30–70 s) in a broader range of humidity detection (7–98%). A CH3NH3PbI3-xClx-based humidity sensor with a reaction time of 21 s, quicker than the quickest commonly available psychrometer on the market, was recently produced by Ren et al. [2+, such as double, triple and vacancy ordered perovskites, have gotten a lot of interest because of their flexibility in photoelectric devices. The produced metal-based halide perovskites such as (NH4)3Bi2I9 [2AgBiBr6 [3Bi2Br9 [2PdBr6 [2InBr5·H2O [3+, Pd2+, In+3 with a comparable electron configuration to Pb2+.Hu et al.  developeu et al.  have devn et al.  with vern et al. ,148,149.4)3Bi2I9 , Cs2AgBi2AgBiBr6 , and Cs3[3Bi2Br9 , Cs2PdBr [2PdBr6  and Cs2InBr5·H2O  provide Capacitive humidity sensors are often designed with a layered structure with two electrode interfaces, or as an interdigitated structure with comb electrodes, analogous to resistive RH sensors, with the insulating polymer film sandwiched in between ,127. SevPolymeric capacitive humidity sensors have been widely employed in the industrial and automation industries due to their simplicity in terms of coating, mass production, long-term stability, and a wide variety of potential sensing polymers such as polyimides ,155. On-Variations in the sensing film dielectric constant as a result of water absorption have a big impact on the capacitive RH sensors’ architecture, which alters the overall capacitance of the system. Differences in capacitance can be used to detect humidity changes because polymer dielectrics’ physical properties, such as permittivity, alter proportionally with the high dipole moments of water molecules. At ambient temperature, capacitive polymer sensors have a relative dielectric permittivity of roughly 5, whereas pure water has a permittivity of about 80. The dielectric permittivity increases the magnitude of the adsorption of water vapor by polymers (78.54), resulting in a sensitive linear change in capacitance. Polyimides (PI) and cellulose acetates are examples of such polymers, with relative dielectric permittivity values in the range from 3 to 6.Based on established observations, the porosity of the humidity-sensitive polymer sheet may be changed to boost sensor responses. In 1983, Delapierre et al.  proposedYang et al.  made a cBi et al.  develope2O3) with a microporous structure is the best material for these types of sensors [2O3 with a small pore radius is extremely prone to very low humidity levels, according to the electron tunneling mechanism effect inside the compacted immobile layers of water [2O3 (amorphous) and Al2O3 (corundum) are often used; however, the former is somewhat more sensitive than the latter due to its higher porosity [2O3 humidity sensors for their low cost and convenience of usage. As per the electron tunneling mechanism effect within the condensed immobile layers of water, Al2O3 with a tiny pore radius is particularly sensitive to very low humidity levels. Aluminum oxide (Al2O3) with a porous structure is the most suited material for these types of sensors. Only the two phases γ-Al2O3 (amorphous) and α-Al2O3 (corundum) are typically utilized in humidity sensing applications; however, the former is more sensitive than the latter due to its greater porosity [2O3 humidity sensors. McGhee et al. [Aluminum oxide  shells were placed over hydrothermally generated zinc oxide (ZnO) nanorod cores. Per the morphological study, the initial zinc oxide nanorods were coated by anatase titanium oxide shells as a second layer. The ZnO/TiO2 nanocomposite (ZTNA) sensors have substantially better sensitivity at 95 percent RH than separate ZnO and TiO2 sensors . Moreover, at ambient temperature, the capacitance of the core/shell arrays varied from 101 to 106 pF throughout the whole humidity range of 11 to 95 percent RH [On glass substrates, Gu et al. built ZnO/TiOrcent RH . The design configurations of impedance- (resistive) and capacitive-based humidity sensors that have proven to be the best suited and most prevalent in advanced applications such as laboratory research or automated industries were examined. An interesting fact to note was that resistive sensors operate on the same principle as capacitive sensors, measuring electrical changes to provide a relative humidity value. Although resistive sensors use hygroscopic materials similar to capacitive sensors, the humidity measurement processes in these systems differ. The distinction made was that the resistance change in the material is measured instead of the capacitance, and capacitive measurement, which is based on electrical capacitance, is used by a capacitive humidity sensor. In comparison to resistive measurements, capacitive measurements are widely utilized in the literature because they are more resilient and stable with temperature and humidity changes. Although both capacitive and resistive sensors have the same goal, which is that of measuring humidity, their methodologies are different. Capacitive sensors have also proven to be more accurate and stable than resistive sensors while giving reproducible findings, making them more ideal for medical applications where precision is critical. However, they are more expensive than resistive sensors. The less expensive resistive sensors are more commonly utilized in circumstances where frequent measurements are required but precision is not required.The success of these configurations moreover is linked to their ability to address common demands such as streamlined construction, free selection of sensing materials from several accessible kinds, cost, circuit adaptability, ease of manufacture, and measurement setup. Due to advantages such as low cost, proper gasketing, design flexibility, and quick deposition rate, film-based humidity sensors are more frequently employed. Semiconducting metal oxide and metal oxide/polymer-based sensors, which are primarily manufactured using thick film and thin film deposition techniques, stand out among the various types of humidity sensors due to their wide range of sensitive element options, post-processing capabilities, and superior response characteristics. 2BX6 or Cs2B”B’X6 and Cs2InX5.H2O, should be prioritized as a viable alternative to the endless difficulties that plagued other metal halides and oxides perovskites with lower stabilities and more efficient performances. Their nanoscale qualities may boost performance, having a significant impact on accuracy, reliability, and cost. In the future, nanocrystalline lead-free perovskite composites with ceramics, polymers, or ceramics/polymers, which are among the most promising materials for humidity sensors, may yield significant improvements in terms of humidity sensing performances.Exploration of the synthesis and use of lead-free metal halide perovskites, such as Cs


Spinal cord injuries and neurodegenerative diseases, including Parkinson’s, Alzheimer’s, and traumatic brain injuries, remain challenging to treat. Nowadays, neural stem cell therapies excite high expectations within academia. The increasing demand for innovative solutions in regenerative medicine has drawn considerable attention to graphene materials. Due to unique properties, carbon materials are increasingly used as cellular scaffolds. They provide a biological microenvironment supporting cell adhesion and proliferation. The topography and mechanical properties of the graphene culture surface influence the forces exerted by the cells on their extracellular matrix. Which consequently affects the cell proliferation and differentiation. As a result, material properties such as stiffness, elasticity and mechanical strength play an important role in stem cells’ growth and life. The ink unification process is crucial while the layer homogeneity is essential for obtaining suitable surface for specific cell growth. Different ink unification processes were tested to achieve appropriate layer homogeneity and resistivity to successfully applied the GNPs layers in neural cell electrostimulation. The GNP coatings were then used to electrostimulate mouse NE-4C neural stem cells. In this study, the authors investigated how the stimulation voltage amplitude’s value affects cell behaviour, particularly the number of cells. Sinusoidal alternating current was used for stimulation. Three different values of stimulation voltage amplitude were investigated: 5, 10, and 15 V. It was noticed that a lower stimulation voltage amplitude had the most favourable effect on the stem cell count. Stem cell therapies are gaining popularity due to their versatility, self-renewal, and multipotency . NervousThe ideal material for neuronal stimulation should be characterised by low electrical impedance, sizeable electroactive surface area, and appropriate biomimetics . Hence, Creating a scaffold and a microenvironment similar to the ECM enables cells to adhere, proliferate and differentiate appropriately , which aThe existing solutions do not achieve the desired reproducible therapeutic effect, and there is a significant discrepancy between scientific reports . SimultaModifying the particle surface by attaching surfactants is a standard procedure to improve the homogeneity of solutions and counteract the agglomeration of hydrophobic particles. The homogeneous dispersion of graphene particles in solution and their efficient distribution in the polymer matrix can be achieved using ultrasound . The larDue to the unique electrical properties of graphene, it is possible to combine two types of stimulation: material and electrical . The eluin vitro and in vivo, studying the degree of particle accumulation in tissues or blood vessels  and a Sonics Vibra Cell VC505 sonotrode homogeniser (maximum power 500 W and 20 kHz frequency). The baseline sonication process using the homogeniser lasted 1 min, while sonication using the scrubber lasted 30, 60, 90, or 120 min . The beaAfter combining the components with the pre-prepared base, the mix was sonicated again. In the case of inks homogenised using a sonotrode, the re-sonication time was 1 min, while the sonication time for inks in an ultrasonic washer was 15 min.® polyamide film substrate (from Dupont™) with screen-printed silver contacts and polystyrene 6-well cell culture plates from Nest Scientific Biotechnology. The printed silver contacts allowed for a measurement of the resistivity of the layers. The dimensions of the tested layers were 24 mm × 24 mm in width and length. As in the previous study, coatings were applied via spraying. A manual Infinity Airbrush CR Plus Solo spray gun from Harder & Steenbeck was used. For Kapton film substrates, the produced layers were dried in a laboratory dryer at 120°C for 60 min . A UNI-T laboratory multimeter, model UT804, was used to measure the resistance of the fabricated coatings. The thickness of the layers was measured with the DektakXT Bruker BNS stylus profilometer. Microscopic observations of layers were made using a Hitachi SU8230 scanning electron microscope (SEM) with an accelerating voltage of 5.0 kV. Images were taken with SEM at magnifications of 1 × 100 and 1 × 500.RRID: CVCL_B063) were used to perform electrostimulation on graphene substrates. NE-4C neuroepithelial cell lines were obtained from brain follicles of 9-day-old mouse embryos. In this study, neural stem cells from 8 to 10 passages were used. The cells medium and culture substrate had been prepared, as described in detail in previous work (cm2), which were then incubated at 37°C in an atmosphere of 5% CO2. Cells from the control group and stimulated with an electric field were cultured for 3 days.Mouse NE-4C neural stem cells of neuroectodermal origin  with a SE secondary electron detector at an accelerating voltage of 5 kV. Specimens are observed in high vacuum and high resolution. Images were taken with an SEM microscope at 1 × 1,000 magnification.Inadequate ink viscosity results in uncontrolled spreading over the coated surface, resulting in a layer of non-uniform thickness. The composite’s viscosity depends on several factors, including the content and type of functional phase, the matrix material used and the solvent. Inappropriate viscosity degrades the mechanical and electrical properties of the graphene layer. Previous studies showed the particle size’s effect on solution viscosity and the effect of type and surfactant content on the rheological properties of inks. It also demonstrates that depending on the surfactant content, the viscosity values of the inks were in the range of 1.3–2.0 mPas. The viscosity dependence on shear rate for graphene inks with different sonication times and methods are shown in The viscosity curves of all the tested inks have the desired shape for spray-coating techniques, characteristic of shear-thinning non-Newtonian fluids. The curves represent a decrease in viscosity values as the shear rate increases, results from the gradual stacking of functional phase particles along the flow line. The viscosity decreases until no further increase in system ordering is possible – which corresponds to the Newtonian behaviour of systems at high shear rates . FurtherGraphene coatings, made using inks with different sonication times, were applied to flexible Kapton film substrates to determine the layer micro-geometry. The example images of graphene coatings on polystyrene substrates made using inks sonicated with a sonotrode are presented in Electrical conductivity is a crucial factor in using graphene substrates in the electrostimulation of nerve cells. As-low-as-possible resistance of conductive culture substrates is sought to increase the efficiency of stimulation and allow the use of a lower current signal to reduce potential cell weakness caused by the flow of an extensively strong electrical signal . AchieviTo determine resistivity, the thickness of layers was measured using a profilometer. The average thickness of the coatings was 37.46 μm.The measurements show a significant scattering of resistivity values for different sonication times. As the sonication time increases in the ultrasonic bath, a visible rise in coating resistance is present. The highest resistance values were obtained for inks sonicated for 90 and 120 min using the ultrasonic bath. On average, the resistance values of the coatings made with the inks sonicated for 120 min were twice as high as the resistance values achieved for the inks sonicated with the sonotrode. The lowest resistance values were obtained for the coatings made with the inks sonicated using a sonotrode and ultrasonic bath for 30 min; therefore, both were used for further electrostimulation studies.cm2) is also included in the graph (red column – seeded cells).To evaluate the effect of electrostimulation on NE-4C cells, the cell counts were examined on substrates without graphene coating, on graphene substrates without stimulation and with stimulation . To faciFirstly, it can be seen that the use of graphene substrates significantly increases cell counts. The use of graphene substrates alone increases cell counts more than threefold. When applying graphene substrates, cell counts approximating those on polystyrene control substrate without graphene coating were obtained. A high increase in cell number is observable for the graphene substrate prepared with ink sonicated using a sonotrode.For both types of substrates, the highest population size was obtained for cells stimulated with the lowest voltage amplitude of 5 V. More than the sevenfold increase of the cell count (compared to seeded cells) was obtained using 5 V electrostimulation and graphene substrate prepared using the ink sonicated in the ultrasonic bath. In the case of the other substrate, an almost fourfold increase in cell count was obtained. Slightly lower cell counts were obtained with a stimulation voltage of 10 V. The weakest stimulation effects, for both types of substrates, were obtained using the highest voltage amplitude of 15 V. The 15 V sonotrode substrate was the only one in which the decrease in cell count was observed. In the 15 V US bath substrate, the cell count only slightly higher than the number of seeded cells was observed.Observations of selected culture substrates were also made using SEM microscopy to assess the effect of electrostimulation. The cell count study and their microscopic observations confirmed the positive effect of graphene substrates and electrostimulation on neural stem cell behaviour. Using graphene substrates increases cell counts by 300% (without electrostimulation) and over 700% by using 5 V stimulation. Graphene layers improve the shape and adhesion of cells to the substrate. It is clear that lower values of stimulation voltage amplitude improve the cell behaviour. The highest cell count for both types of substrates was obtained when electrostimulation with a voltage amplitude of 5 V was applied. Therefore, these substrates were selected for further observations using SEM microscopy.The literature analysis of graphene surface application evidences a high demand for innovative solutions in the field of neural tissue engineering. The production of a biocompatible layer supporting the development of nerve cells, promoting their adhesion and proliferation will allow for significant improvements in the treatment of many such neurodegenerative diseases as neural tube defects, spinal cord injuries, or neuropathies. Achieving the desired homogeneity of the produced composite is crucial for the substrate’s conductivity, which makes them useful in the electrostimulation of nerve cells. The properties of graphene layers are also affected by the composite fabrication process itself particularly the time and method of solution homogenisation.The studies showed a significant influence of the graphene ink preparation method on the properties of the graphene coatings. Here, two alternative ultrasound sonication methods were used, which allowed for high dispersion of graphene particles, resulting in homogeneous, highly conductive graphene layers successfully used in the electrostimulation of neural stem cells.While comparable ink viscosity values decreased with the increasing homogenisation time. Many factors can influence the rheological properties of fluids, including the particle’s shape and size, the molecular weight of the functional phase or particle size distribution in solution . The visDespite the lack of differences in ink viscosity, apparent differences were observed in the conductivity of layers produced at different sonication times. As the viscosity decreased (with longer sonication times), the conductivity of the layer decreased. This is presumably related to the degree of graphene particle dispersion in the produced inks. The sonication time probably affected the fragmentation of graphene nanoplatelets and also changed the degree of the graphene particles’ surroundings by the surfactant, thus influencing the formation of particle agglomerates. According to Nevertheless, each manufactured ink fulfils the rheological requirements enabling its application in the spray coating technology. The sonication time and method did not cause significant discrepancies in the viscosity values of the inks. It can be concluded that both sonication methods are effective ways to homogenise solutions. The relationship between viscosity and conductivity of the layers requires further investigation and additional studies – including the size of graphene nanoplatelets obtained in ink after the sonication process.Observations of the micro and macro geometries of the substrates conclude that both sonication methods can be successfully used in the homogenisation of graphene inks. However, the taken photographs do not allow an assessment of the effect of sonication time on the dimensions of the graphene particles themselves. Particle dimensions are an essential parameter for coating applications in tissue engineering, as they could strongly influence the occurrence of cytotoxic phenomena. Therefore, in future work, it is necessary to perform SEM photography with higher accuracy to measure the size of the flakes.–3 Ω⋅m and 2,154 ⋅ 10–3Ω ⋅m, respectively, for sonotrode and ultrasonic bath. The results show that a short, 1-min sonication process using a homogeniser or extending the process to 30 min when using an ultrasonic bath is optimal for the accurate combination of surfactant particles with graphene flakes. The study showed that the obtained resistance values strongly depend on the graphene ink’s method and sonication time. With an extension of the sonication time, apparent increase in the resistance of the coatings can be seen. This phenomenon may be due to the excessive fragmentation of graphene nanoplatelets, as flakes with smaller dimensions are weaker conductors.The lowest resistivity values were obtained for the coatings made with the ink sonicated using a sonotrode for 1 min and an ultrasonic bath for 30 min. The obtained resistivity of the films were 1,623 ⋅ 10Through the successful use of a sonotrode, the efficiency of the fabrication process of graphene heterophasic inks was increased by shortening the sonication time from 90 to 2 min while increasing the conductivity of the layers. Comparing the fabrication times of the inks made in our previous work (90 + 15 min sonication) , one canUpon analysing the results, graphene substrates and electrostimulation significantly increase the cell counts. The simulations carried out in this work, using different stimulating voltages with amplitudes, showed over a 700%-cell-count increase when 5 V was applied. The produced substrates improved cell adhesion and morphology, as shown in the SEM photographs in the section below. The fabricated graphene substrates had a beneficial effect not only on cell adhesion but also on cell proliferation. Using graphene substrates enabled to increase cell count by 300% (without electrostimulation).The occurrence of functional groups on graphene surfaces could influence the types of cell surface receptors and proteins, such as the cellular adhesion molecules (CAM), which cells use to bind to graphene surfaces . Lee et Cell adhesion plays a significant role in regulating such processes as growth, proliferation and migration. The increase in the number of cells on graphene illustrates a positive effect on cell adhesion, also confirmed by For both types of substrates, a decrease in cell count is observed as the stimulation voltage amplitude increases. This observation indicates that the stimulation voltage amplitude of 15 V is too high and probably hinders the cells from adhering correctly to the substrate, thus preventing their proliferation, which is also stated by In the study, the 5 V voltage amplitude was used. However, the literature reports confirm the efficiency of even lower stimulating voltages . This isIn conclusion, we can state that using graphene substrates as cellular scaffolds enhance and improve neural tissue regeneration protocols. The preliminary data showed the efficiency of the proposed simulation method and the beneficial effects of graphene substrates and electrical stimulation. We propose an alternative tissue stimulation method, which could be a starting point for new, innovative treatment routes. The high homogeneity and conductivity of the layers allowed them to be effectively used in the electrostimulation of neuronal stem cells. The increased cell count of cells cultured of the graphene substrates indicates that our substrates provide an environment suitable for cells’ development, mainly supporting cell adhesion, which enables their proper growth and proliferation. Graphene substrates show significant potential in cellular scaffolds and regenerative medicine, which in the future may contribute to the development of effective clinical therapy for peripheral nerve injuries.The original contributions presented in this study are included in the article/supplementary material, further inquiries can be directed to the corresponding author.ŁD-S: conceptualisation and methodology. WS: writing—first draft, experiments—materials preparation and spray coating, and figures preparation. AG: experiments—cell culture. JK: formal analysis. WS and JK: writing—review and editing. ŁD-S and MJ: supervision and funding acquisition. All authors contributed to the article and approved the submitted version.


In the design, graphene is served as excellent carrier to improve the catalysis of Mn3(PO4)2 nanoparticles; and DNA adsorbed on graphene acts as medium to assist the growth of Mn3(PO4)2 on graphene. The fabricated graphene/DNA/Mn3(PO4)2 composites exhibit excellently electrochemical activity, significantly decrease the response time and increase the sensitivity of the sensor towards O2˙−. The successful detection of O2˙− released from cancer cell demonstrated its potential applications in biology and medicine.In this work, we successfully fabricate a rapid, sensitive sensor for the detection of superoxide anions O 2˙− based on graphene/DNA/Mn3(PO4)2 biomimetic enzyme.In this work, we successfully fabricate a rapid, sensitive sensor for the detection of superoxide anions O Under normal conditions, O2˙− is highly reactive and unstable and its metabolism is a rapid and spontaneous process. Overproduction of O2˙− can cause various disease, such as aging, asthma, ulcer disease, cancer, atherosclerosis, neurodegenerative diseases and other diseases.1–4 Thus the detection of O2˙− is very important in various biological systems. Many methods have been developed for the detection of O2˙− including chemiluminescent, spectrophotometric, fluorometric, and electrochemical technique, etc.5–8 Among all these techniques, electrochemical method receives most attention due to its advantages of real-time assay, high sensitivity and selectivity. And most of the reported electrochemical methods for O2˙− detection are based on enzyme catalysts such as cytochrome c and superoxide dismutase (SOD).2,9–11 Though enzyme improves the assay performance of sensors, but its high cost and poor long-term stability increase difficulty of direct monitoring of O2˙− in biological samples. Therefore, it is a serious challenge for us to establish a fast, reliable, and sensitive non-enzyme approach for O2˙− monitoring in physiological and pathological processes.Superoxide anions 2) has the ability to catalyze the dismutation of O2˙− compared with free Mn2+ ion that only stoichiometrically reacts with O2˙−.13 Therefore Mn3(PO4)2 is usually selected as a substitute for enzyme to detect O2˙−.14 Mao et al. synthesize SiO2-Mn3(PO4)2 nanoparticles derived from phytic acid and utilize for O2˙− detection.15 Lan and co-workers deposit Mn3(PO4)2 nanoparticles on carbon nanomaterials directly for the fabrication of O2˙− sensor.16 These works verify the catalysis of Mn-superoxide dismutase mimics towards O2˙−. But the synthesis and property of nanoparticles need further study to improve the catalytic performance of sensor. In this work, we synthesize Mn3(PO4)2 nanoparticles by DNA induction, which results in even dispersion and excellent catalytic activity towards O2˙− disproportionation reaction.As early as 1982, manganese was reported possessing an effective catalytic effect 17–20 However, the two dimensional structure is easy to aggregate due to the single-atom-thickness, and hydrophobic aromatic structure. To solve this problem, we functionalize graphene with deoxyribonucleic acid (DNA) which can be adsorbed on graphene through π–π stacking and don't destroy the intact structure of the carbon materials.20 Under the induction of ssDNA, Mn3(PO4)2 nanoparticles was evenly deposited on the surface of graphene. The obtained graphene/DNA/Mn3(PO4)2 nanocomposites displayed significant biomimetic enzyme activity, rapid and sensitive response towards O2˙−. This approach holds a great promise for broad applications in biomedical research and clinical test.Graphene is an increasingly important nanomaterials due to its excellent electronic conductivity, good stability, and promising catalytic performance in sensing.4), potassium phosphate tribasic (K3PO4) and Nafion were purchased from East Sichuan Chemical Industry (Group) Co., Ltd.  and used as received. Graphene was obtained from Sinocarbon Materials Technology Co., Ltd., China. Zymosan A  SOD, DNA , and potassium superoxide were purchased from Sigma-Aldrich and used without further purification. The O2˙− solutions were prepared by dissolving KO2 in PBS solution . The concentration of O2˙− was determined by the reduction of ferri cytochrome c spectrophotometrically.21 All the other solutions were prepared using deionized water (18 MΩ cm) and were degassed with high purity nitrogen before experiments. All electrochemical experiments were carried out at room temperature.MCF-7 was bought from Cell bank of the representative culture preservation committee of the Chinese Academy of Sciences, China. Manganese sulfate (MnSO3(PO4)2 were characterized by transmission electron microscopy . Fourier transform infrared spectroscopy (FTIR) spectra were determined using a Thermo Nicolet 6700 FTIR spectrometer. Electrochemical performances were characterized using a CHI 660 system . Graphene/DNA/Mn3(PO4)2 modified glassy carbon electrode (graphene/DNA/Mn3(PO4)2/GCE) served as working electrodes, platinum sheet as counter electrode and Ag/AgCl (in 3 M KCl) acted as reference electrode. All O2˙− measurements were performed in 10 mL 0.01 M PBS solution (pH = 7.4).Scanning electron microscopy (SEM) images were taken by JSM-6510LV, Japan. Field emission scanning electron microscopy (FESEM) images were taken by JSM-7800F, Japan. X-Ray diffraction (XRD) measurements were performed on a Shimadzu diffractometer  operating in reflection mode with Cu Kα radiation at a step size of 0.06 per second. The nanosheets of graphene/DNA/Mn3(PO4)2 nanosheets were prepared by the previously reported method.22 Thirty milligrams of DNA was dissolved in 30 mL de-ionized water under stirring, after which the solution was annealed at 95 °C for 15 min to produce single-stranded DNA (ssDNA). Then the obtained ssDNA was mixed with 15 mL of 1 mg mL−1 graphene by mildly sonicated at 4 °C for 2 h, thereafter filtration and washing were performed to remove excess ssDNA. Subsequently, the mixture was dispersed into 0.1 M MnSO4 solution, then 0.1 M K3PO4 solution were dropwisely added under stirring and kept at room temperature for 30 min. Then graphene/DNA/Mn3(PO4)2 nanosheets were obtained by centrifugation at 8000 rpm for 10 minutes and washing with deionized water for 3 times.Graphene/DNA/Mnd = 3 mm) was polished with alumina slurry to a smooth and bright surface, washed by sonication for 30 s and dried under nitrogen. Then 5 μL of the obtained graphene/DNA/Mn3(PO4)2 suspension was dropped on the electrode surface and dried at room temperature. Finally, 5% Nafion solution was coated to stabilize the fabricated electrode.Glassy carbon electrode  at 37 °C. The cells were cultured in Dulbecco's Modified Eagle's Medium (DMEM)  supplemented with 1 mol L−1 glutamine, 50 U mL−1 penicillin/streptomycin and 10% heat inactivated fetal bovine serum.14 For the detection of O2˙− released from living cell, the incubation solution was removed and washed with PBS solution (pH 7.4) for three times. Before electrochemical measurement, zymosan (Zym) was added to motivate cells generation of O2˙−. Amperometric response was recorded by CHI-660B electrochemical station at applied potential of 700 mV (versus Hg/Hg2Cl2).MCF-7 cells were cultured in a humidified incubator (95% air with 5% CO3(PO4)2 enzyme mimics by growth of Mn3(PO4)2 on graphene under the induction of DNA. As we all know, single-strand DNA (ssDNA) can be adsorbed on graphene by π–π interaction.23,24 Here ssDNA was prepared by annealing double-strand DNA (dsDNA) and applied for graphene modification. In the presence of Mn2+, the divalent cations accumulate along the ssDNA backbone by the electrostatic interaction, which facilitate the formation of Mn3(PO4)2 crystal nucleus on graphene and growth of Mn3(PO4)2 nanoparticles upon the addition of negatively charged PO43− groups.3(PO4)2 nanosheets is characterized with scanning electron microscope (SEM). 4 and K3PO4, typical wrinkle of graphene can be clearly observed and no evident Mn3(PO4)2 nanoparticles is found in 3(PO4)2 nanoparticles on graphene surface without DNA. However, when ssDNA is modified on graphene (3(PO4)2 nanoparticles can be evidently observed on the surface of graphene. The phenomena reveal the crucial role of ssDNA on the growth of Mn3(PO4)2 on graphene. The TEM images in 3(PO4)2 nanoparticles due to the overlapping of particles. In order to verify the results, energy disperse spectrometer (EDS) was performed to explore chemical composition of the materials. The EDS results in Fig. S23(PO4)2 nanoparticles. The results further confirm the formation of well-defined thin-layer structure of the nanocomposites of Mn3(PO4)2 nanoparticles on graphene surface.The morphology of the as-prepared graphene/DNA/Mngraphene , even-di3(PO4)2 nanosheets. As shown in −1 can be assigned to the aliphatic carboxylic acid salts, the peaks at 996.0 cm−1 and 3196.3 cm−1 were attributed to inorganic phosphates. The results suggest that DNA and Mn3(PO4)2 have been successfully modified on the graphene. In addition, the X-ray diffraction pattern (XRD) of graphene/Mn3(PO4)2 in 3(PO4)2. The results further confirm that Mn3(PO4)2 nanoparticles are successfully modified on the surface of graphene. Then the zeta potential at various stages of the synthesis were measured to further characterize the formation of the nanocomposites. The results in Fig. S13(PO4)2 was deposited, the zeta potential increase to −15.6 mV, indicating the formation of graphene/DNA/Mn3(PO4)2 nanocomposites.Then FTIR spectra are determined to further verify the formation of graphene/DNA/Mn2˙− and the mixture of O2˙− and SOD (red column) in PBS. When graphene/DNA/Mn3(PO4)2 nanosheets were modified on GCE, the obtained modified electrode displayed evident redox peaks around 0.7 V and 0.5 V in the absence of O2˙− 2 nanosheets towards O2˙−. These results verify the biomimetic enzyme activity of graphene/DNA/Mn3(PO4)2 to catalyze the dismutation of O2˙−.253(PO4)2 nanosheets, graphene/Mn3(PO4)2 were synthesized by direct depositing Mn3(PO4)2 nanoparticles on graphene in the absence of DNA, DNA/Mn3(PO4)2 nanoparticles were by prepared depositing Mn3(PO4)2 on dsDNA template and graphene/DNA/Mn2+ was synthesized by adsorbing DNA on graphene and subsequent depositing Mn2+ on DNA. Electrochemical performance of these materials were explored with electrodes modified with the above three nanomaterials (3(PO4)2 (graphene/Mn3(PO4)2/GCE) possesses excellent electrical conductivity but the current response towards O2˙− is weak, verifying the good conductivity of graphene and poor modification of Mn3(PO4)2 (3(PO4)2/DNA and graphene/DNA/Mn2+ display evident current response to O2˙−, but the current response is lower than that on graphene/DNA/Mn3(PO4)2/GCE (as shown in 3(PO4)2 nanosheets. In addition, in comparison with the Mn3(PO4)2/DNA/GCE (3(PO4)2/GCE 2 . Similar/DNA/GCE , we can O4)2/GCE  displays3(PO4)2 nanoparticles on graphene can be ascribed to two factors. First, the π–π interaction between DNA and graphene induces ordered assembly of DNA on the surface of graphene, which is a critical factor to the synthesis of Mn3(PO4)2 nanoparticles. Next, Mn2+ is adsorbed on DNA backbone to assist the nucleation of Mn3(PO4)2 nanoparticles. In addition, in comparison with other conventional method, the DNA induced synthesis is easier to control the morphology and dispersion of products by adjusting temperature, pH, or DNA concentration.The reason for DNA facilitating the growth of Mn3(PO4)2/GCE towards O2˙− in 10 mM PBS (pH = 7.4). 2˙− concentration from 5 nM to 400 nM. From the results in 2˙− concentrations in the range of 5 nM to 400 nM with a regression equation expressed as I (μA) = 0.00354 c (nM) + 0.1504 (R2 = 0.999). The detection limit of the assay is calculated to be about 1.67 nM (S/N = 3) with a sensitivity of 3.54 μA μM−1, which is much more superior to most of previous reports.14,23,26–28 The response time of the electrode to O2˙− is as short as 5 s, which is an advantage for the detection of O2˙− due to its short lifetime. These results demonstrate that the present sensor may meet the requirement of O2˙− detection in normal physiological conditions.29Chronoamperometry is utilized to investigate the electrochemical response of graphene/DNA/Mn+, Na+, Cl−, NO3−, SO42−, hydrogen peroxide (H2O2), glucose (Glu), glutathione (GSH) uric acid (UA) and ascorbic acid (AA). Among these interferences, H2O2, UA and AA possess good electrochemical activity and perplex the detection of O2˙− due to their wide existence in biological systems. As shown in 2˙−, the presence of 10 μM K+, Na+, Cl−, SO42−, NO3−, Glu. GSH, AA, UA and H2O2 did not cause any noticeable current response. The result suggests that the proposed sensor possesses excellent specificity to O2˙− detection in biological systems. In addition, the reproducibility and stability of the electrode were explored. The electrochemical response of the sensor was performed using five different electrodes fabricated at the same time, the standard derivations of current response was lower than 5.2%. When the electrode was stored at 4 °C for one month, the current response towards O2˙− decrease 6.8% of original value  of zymosan , a significant current response was observed (corresponding to O2˙− oxidation). According to the calibration curve in 2˙− released from MCF-7 cells is calculated to be around 156.38 nM, 315.71 nM and 631.81 nM, respectively 2 nanosheets is successfully developed. The synthesized nanosheets display strong electrocatalytic activity towards O2˙− with high sensitivity, excellent selectivity and fast current response. The successful determination of O2˙− released from cancer cells demonstrates the great potential for application in biological system. This work provides a new method for the fabrication of biomimetic enzyme sensor and a great promise for biosensing and biomedical application.A novel OThere are no conflicts to declareRA-008-C7RA12962A-s001


Chronic cough is experienced by most patients with idiopathic pulmonary fibrosis (IPF). It is often the first symptom and is associated with reduced quality of life, increased rates of depression and anxiety, more severe physiological impairment, and disease progression. Although not fully understood, recent gains in understanding the pathophysiology of chronic cough in IPF have been made. The pathophysiology is likely multifactorial and includes alterations in mucous production and clearance, architectural distortion, and increased cough reflex sensitivity, suggesting a role for targeted therapies and multidisciplinary treatment. Modifiable comorbidities can also induce cough in patients with IPF. There is a renewed emphasis on measuring cough in IPF, with clinical trials of novel and repurposed therapies for chronic cough emerging in this population. This review provides an update on the clinical characteristics, pathophysiology, and measurement of chronic cough in patients with IPF and summarizes recent developments in non-pharmacological and pharmacological therapies. Idiopathic pulmonary fibrosis (IPF) is a chronic fibrosing idiopathic interstitial pneumonia, most prevalent after the 6th decade of life. It is associated with significant morbidity and mortality, with progressive worsening of respiratory symptoms over time. Cough is a common symptom, often occurring years prior to diagnosis, contributing to poor prognosis and reduced quality of life. The severity of cough encompasses its frequency, intensity, and impact to patients' lives . Cough fCough and dyspnoea are the two most common symptoms described in patients with IPF, with up to 80% experiencing chronic cough . Cough iCough is more prevalent in patients with advanced IPF . IncreasCough due to IPF is typically described as dry or non-productive and refractory to antitussive therapy. A minority of patients have a chesty cough from the onset of disease. The severity of cough in IPF is greater than in other types of interstitial lung disease (ILD) –8. The iPatients identify cough as a key symptom associated with physical and emotional distress, with a more limited appreciation by health care professionals . Physicavia rapidly adapting receptors, slowly adapting stretch receptors, and c-fibers in the afferent pathway that conduct mechanoreceptor and chemoreceptor stimuli to the medullary control center by branches of the vagus nerve  gene, the most well-validated genetic risk factor for IPF –35, incrIPF-related comorbidities and other health conditions can be important contributors to cough in this population and are therefore key targets for treatment. The role of GOR in IPF remains unsettled, with inconclusive results on therapeutic effects of antacid medication and anti-reflux surgery on progression-free survival, quality of life, and all-cause mortality in this population , 52. As The prevalence of GOR is higher in IPF than in other chronic lung diseases , 54, 55 P and mast cell-tryptase are both elevated in the sputum of patients with chronic cough and comorbid GOR, which inversely correlate with cough threshold  on lung function testing may prompt further investigation with computed tomography of the chest to assess for acute exacerbation, infection, or disease progression. Red flag symptoms for lung cancer include haemoptysis, dysphonia or dysphagia, and weight loss. Relevant comorbidities  and the Leicester cough monitor are two validated monitors , 71, whiPatient reported outcomes (PROs) have gained importance in recognition of their value to capture patients' perceptions of their symptoms and health status. They are measured using standardized questionnaires, which may be generic, disease-specific, or symptom-specific. PROMs for cough have been developed in other conditions such as lung cancer, while no cough-specific instruments have been developed and validated prospectively in the IPF population. The currently used cough questionnaires were developed in patients with chronic cough and other respiratory conditions such as chronic obstructive pulmonary disease. Instead of comprehensively reporting on all PROMs for cough that have been used in IPF, this review focuses on selected commonly used cough-specific instruments, including the VAS, LCQ , Cough QVAS subjectively assesses cough severity and is popular due to its simplicity. Patients are asked to place a vertical mark on a 0–100-mm linear scale reflecting their cough intensity or frequency, with higher scores indicating greater severity. It can be used to assess both the status and changes in cough severity. There is limited robust data on its reliability and validity in specific respiratory conditions, including asthma and chronic obstructive pulmonary disease . The VASThe LCQ is the most widely used validated cough instrument in clinical and research settings. Its advantages are that it is relatively easy to administer, translated into many languages, and used in many cultures. It is a self-administered questionnaire consisting of 19 items covering three health domains  using a 7-point Likert response scale for each item. The total LCQ score ranges from 3 to 21, with lower scores indicating greater impairment in health status because of cough. All domains of the LCQ inversely correlate with objective daytime cough frequency  in patieThe CQLQ consists of 28 items and six health domains . Each item is rated using a 4-point Likert scale. The CQLQ has good internal consistency in IPF but this does not apply to all domains . InternaThe CASA-Q comprises four domains: symptom and impact for cough and sputum. The score for each domain ranges from 0 to 100, and lower scores indicate worse symptoms. It has been used in landmark trials for antifibrotic therapies in IPF , 92 withClinical trial design in IPF has historically been hampered by a lack of consensus on appropriate outcome measures to reliably assess treatment response . For exaThere is no therapy for cough in IPF that has proved effective in clinical trials. Aside from treatment of relevant comorbidities, several therapies have been tried for chronic cough in IPF and may be useful in some individuals, which include both non-pharmacological and pharmacological interventions . It is sWhile the evidence is scarce in patients with IPF, multimodality speech therapy, an established therapy for unexplained chronic cough, may be a suitable treatment. Speech therapy has been reported to decrease cough frequency and improve quality of life in chronic cough . The PhyThe outlook for patients with IPF changed with the availability of two antifibrotic drug therapies; nintedanib and pirfenidone. Nintedanib alters cell signaling mediated by tyrosine kinases resulting in the inhibition of mechanisms fundamental to the development of fibrosis . PirfeniStudies evaluating antitussive therapies in IPF are limited. Opioids have been shown to decrease cough frequency and improve cough-related quality of life in patients with chronic cough in a study of slow-release oral morphine at a dose of 5 mg twice daily . Common via high-efficiency nebuliser reduced mean daytime cough frequency by 31% at 14 days in patients with IPF and was generally well-tolerated (In a randomized placebo-controlled trial of 24 patients with IPF, low-dose thalidomide was shown to improve cough severity and respiratory-related quality of life measured by the CQLQ and VAS scores . Howeverolerated . This beolerated . An oldeolerated . Howeverolerated , and itsolerated . Furtherolerated .Cough is a common and often under-recognized symptom of IPF, which is associated with disease progression. Identified as a central symptom by patients and their caregivers, cough impacts health-related quality of life. Careful and timely clinical evaluation and assessment using validated tools that objectively and subjectively measure cough in this specific patient population are recommended. Currently available measurement tools for chronic cough have been suboptimally validated in patients with IPF. The development of disease-specific PROMs for cough in the IPF population is required to accurately and reliably show the difference between competing therapies. Further clinical research into effective non-pharmacological and pharmacological interventions for cough is vital to improve symptom management and health outcomes for patients with this progressive disease. In light of the limited proven efficacy of single therapeutic agents for cough in IPF, future clinical trials may consider multimodal interventions or combination drug therapy in this patient population.YK and JM conceived the presented idea. JM performed the literature search and wrote the manuscript with support and critical feedback from YK. Critical feedback was also provided by NG and AH. All authors contributed to the final manuscript.YK reports grants and personal fees from Boehringer Ingelheim, personal fees from Roche, other from Air Liquide Healthcare, outside the submitted work. NG reports personal fees from Boehringer Ingelheim, Roche, AstraZenaca, and Novartis, other from Air Liquide Healthcare, outside the submitted work. AH reports non-financial support from BOC Ltd Australia and Air Liquide Healthcare, outside the submitted work. The remaining author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.