Terahertz modulators with high tunability of both intensity and phase are essential for effective control of electromagnetic properties. Due to the underlying physics behind existing approaches there is still a lack of broadband devices able to achieve deep modulation. Here, we demonstrate the effect of tunable Brewster angle controlled by graphene, and develop a highly-tunable solid-state graphene/quartz modulator based on this mechanism. The Brewster angle of the device can be tuned by varying the conductivity of the graphene through an electrical gate. In this way, we achieve near perfect intensity modulation with spectrally flat modulation depth of 99.3 to 99.9 percent and phase tunability of up to 140 degree in the frequency range from 0.5 to 1.6 THz. Different from using electromagnetic resonance effects , this principle ensures that our device can operate in ultra-broadband. Thus it is an effective principle for terahertz modulation. Low-dimensional materials show promise for applications in imaging, spectroscopy and ultra-broadband communications. Here, the authors report an effect of Brewster angle control at graphene-quartz interface for applications in terahertz modulation over a broadband range from 0.5 to 1.6 THz. These applications drive the need for THz modulators with high modulation depth, broad operation bandwidth, and high modulation speed5. Manipulating THz light to achieve both high modulation depth and broadband response in a device is a great challenge. One major limitation of the state-of-the-art THz modulator is the narrow bandwidth; current technologies rely primarily on narrow band resonance effects13  for enhancing the interaction between the THz wave and the tunable conductivity materials. Semiconductor hybrid structures are developed, but these architectures require the assistance of high-power optical pumping. Another issue is the low modulation speed of the non-solid-state devices, such as the modulators based on ion-gel gate or liquid crystal.Terahertz technology shows promise for many applications including imaging, spectroscopy, and communications19. It has attracted great attention in the THz region because of its tunable conductivity, typically from comparatively high-resistance state to semi-metal state23. The first graphene THz modulator is demonstrated by Sensale-Rodriguez et al.24 with an ultra-broad operation band and a modulation depth of 15% using monolayer graphene. To improve the modulation depth, complex optical resonance structures 25–27, semiconductor hybridization29, and ion gel31, are assembled to work together. However, these structures could not overcome the above-mentioned problems. Therefore, there is still a lack of an effective fundamental approach to achieving a broadband device with deep and wideband modulation range.Graphene is an ideal material for photoelectronic devices due to its unique propertiesp-polarization light experiences zero reflection when it is incident to a medium at the Brewster angle and a π-phase shift occurs to the reflected light when the incident angle jumps across the Brewster angle. Furthermore, the Brewster angle can be tuned by varying the surface conductivity of the medium33. Based on this fundamental principle, we experimentally employ an atomic layer of graphene to tune the surface conductivity and demonstrate that the Brewster angle of a graphene–quartz device can be tuned from 65 to 71° by varying the conductivity of graphene. By properly selecting the incident angle, the device can work as a THz-intensity modulator with spectrally flat modulation depth of 99.3–99.9% in a wide range from 0.5 to 1.6 THz, and operate as a phase modulator with tunability higher than 140°. Compared to the high-performance THz modulators based on light-trapping effects, this principle ensures that the device can operate as an ultra-broadband spectrally flat terahertz modulator. Actually, the reported bandwidth in this work is mainly limited by the effective bandwidth of the THz system. An even broader bandwidth can be theoretically extrapolated from the operation principle. Our device presents a THz modulation based on a graphene-controlled Brewster angle device and achieves record performance at room temperature.Here, as a proof of concept, we demonstrate a solid-state modulator with near-perfect tunability, ultra-broad operation bandwidth, and fast modulation speed based on tunable Brewster reflection at graphene/quartz surface. It is well known that σg is placed on the dielectric silica substrate. Light with p-polarization is incident from air onto the graphene/silica medium. According to Maxwell’s equations, together with the boundary conditions . The insertion loss of this device, We first present the simulation results based on the physical model in Fig. ero Fig.  and the ero Fig. , which imentclass2pt{minim2O3/TiOx sandwich structure is constructed for a solid-state electrical gate to tune the conductivity of graphene (details of the fabrication can be seen in the Methods section). A 50-nm Al2O3 layer is used as a dielectric gate, which has a negligible effect on the light reflection. The back gate electrode is made of 10-nm thickness of a TiOx (sheet resistance ~2000 Ω) film. This thin and high-resistance film ensures the transparency for the THz light. The transparency of TiOx is verified by THz transmission spectrum and the loss of TiOx is also discussed in Supplementary Figure The real device in this work is schematically shown in Fig. 35. However, these methods are hard to active tuning of Brewster angle. To change the Brewster angle, the unit structure or periodic parameter for the metamaterial or metasurface should be changed. Besides, Brewster angle of metamaterials/metasurface is always frequency-dependent, due to their resonance nature. As far as we know, this work is the first time to realize an active tunable Brewster angle and achieve broadband response.In order to obtain the Brewster angle of the device, we measured the reflected THz pulse with an incident angle of 55–75°, as well as gate voltage from −12 to 12 V. Three typically reflected THz pulses in the time domain are shown in Fig. 38–40. This experimental insertion loss is mainly due to the poor quality of commercial graphene and the fabrication process.From the result in Fig. In order to further clarify this phenomenon, we simulated the electric field distribution at the frequency of 0.8 THz, when graphene’s conductivity is 0.5 and 3.5 mS, corresponding to the gate voltage of 14 and −12 V in the experiment. As shown in Fig. 62341–43. More importantly, owing to the solid-state-based modulations, our device provides a high modulation speed, as shown in Fig. x. Here, we take a resistance of 250 Ω for the graphene and 1000 Ω for the TiOx, which are calculated by the half of averaged square resistance over the gate voltage range. A capacitance (C) of ~140 nF is calculated by adopting a 50-nm Al2O3 and an active device (graphene) area of 1.0 × 1.0 cm. Therefore, the calculated RC time constant is 1.75 × 10–4 S, corresponding to a modulation speed of ~6 kHz, which agrees well with the experimental result. Higher modulation speed (shorter RC time constant) can be realized through replacing TiOx by a higher-conductivity material  and reducing the device size. For instance, by utilizing graphene to replace TiOx and reducing the device size to ~1.0 × 1.0 mm (about two times the wavelength of 0.8 THz), the modulation speed will be ~2.4 MHz, which is comparable with other solid-state THz modulators in previous reports27.The performance of the device, both of the modulation depth and operation band, is much higher than that of previous resultsVg = 16 V to Vg = −12 V, as shown in Fig. According to the physical model, a broadband phase modulation with a deep modulation range can be achieved by utilizing the phase-jump property across the Brewster angle. Here, we chose the incident angle of 68° to discuss this issue. The reflected THz time-domain signal changes from an asymmetry “W” shape at the gate voltages of 16 V to an asymmetry “M” shape at the gate voltages of –12 V, which indicates that there should be a large phase shift from 2O3/TiOx sandwich structure on a quartz substrate. The conductivity of the graphene can be electrically controlled to generate a variable Brewster angle. Based on this device, an ultra-broadband THz-intensity modulator with a modulation depth larger than 99.3% and an ultra-broadband-phase modulator with a tunability higher than 140° are demonstrated. Our work presents graphene-controlled Brewster angle device for ultra-broadband THz intensity and phase modulation at room temperature. The promising performance of our device also suggests that there exists a great potential to use graphene in deliberate architecture for active THz devices.In this work, THz modulation-based graphene-controlled Brewster angle device is experimentally demonstrated for the first time. The device is fabricated using a graphene/Alx through annealing at 150 °C. After oxidization, the conductivity of this film was about a reciprocal of sheet resistance of 2000 Ω, which was measured using a four-probe method. Then, a layer of Al2O3 (50 nm) was deposited by atomic layer deposition onto TiOx as a dielectric gate. Afterward, graphene (~1 × 1 cm) was transferred onto the as-prepared quartz/TiOx/Al2O3 substrate. Subsequently, two electrodes (Cr/Au) were fabricated by thermal deposition. One was on graphene and the other one was on TiOx for the gate voltage. The device is schematically shown in Fig. A Ti film of 8 nm was deposited on a quartz substrate (diameter of 2 cm) for the bottom-gate electrode. In order to avoid further reducing the conductivity of Ti, the Ti film was oxidized into TiOA THz-TDS system from Menlo Systems (TERA-K15) was used for the terahertz reflection measurements. A reflection guide was used for manual adjustment of the reflection angle, ranging from a minimal reflection angle of 55 to 75°.Supplementary InformationPeer Review File


Umingmakstrongylus pallikuukensis and Varestrongylus eleguneniensis are two potentially pathogenic lungworms of caribou and muskoxen in the Canadian Arctic. These parasites are currently undergoing northward range expansion at differential rates. It is hypothesized that their invasion and spread to the Canadian Arctic Archipelago are in part driven by climate warming. However, very little is known regarding their physiological ecology, limiting our ability to parameterize ecological models to test these hypotheses and make meaningful predictions. In this study, the developmental parameters of V. eleguneniensis inside a gastropod intermediate host were determined and freezing survival of U. pallikuukensis and V. eleguneniensis were compared.Deroceras laeve, were collected from their natural habitat and experimentally infected with first-stage larvae (L1) of V. eleguneniensis. Development of L1 to third-stage larvae (L3) in D. laeve was studied at constant temperature treatments from 8.5 to 24 °C. To determine freezing survival, freshly collected L1 of both parasite species were held in water at subzero temperatures from -10 to -80 °C, and the number of L1 surviving were counted at 2, 7, 30, 90 and 180 days.Slug intermediate hosts, 0) below which the larvae of V. eleguneniensis did not develop into L3 was 9.54 °C and the degree-days required for development (DD) was 171.25. Both U. pallikuukensis and V. eleguneniensis showed remarkable freeze tolerance: more than 80% of L1 survived across all temperatures and durations. Larval survival decreased with freezing duration but did not differ between the two species.The lower threshold temperature (TU. pallikuukensis and V. eleguneniensis have high freezing survival that allows them to survive severe Arctic winters. The higher T0 and DD of V. eleguneniensis compared to U. pallikuukensis may contribute to the comparatively slower range expansion of the former. Our study advances knowledge of Arctic parasitology and provides ecological and physiological data that can be useful for parameterizing ecological models.Both  The extreme climate of the Arctic makes it one of the most challenging environments for living organisms. Winter temperatures can drop to -50 °C in many parts of the Arctic, and sub-zero temperatures last for nearly two-thirds of the year. The summers are short, cool, and dry, providing a narrow developmental window for ectotherms, including parasites, and a short growing season for endotherms and plants –3. DespiIn the animal kingdom, nematodes are considered the most diverse and successful organisms for their ability to adapt to diverse habitats and survive extreme environmental conditions , 10. NemUmingmakstrongylus pallikuukensis Hoberg, Polley, Gunn & Nishi, 1995 and Varestrongylus eleguneniensis Verocai, Kutz, Simard & Hoberg, 2014, are parasites of Arctic ungulates in areas of the Canadian Arctic mainland and Victoria Island in the Arctic Archipelago [Umingmakstrongylus pallikuukensis is a host specialist and only infects muskoxen (Ovibos moschatus) [V. eleguneniensis is a multi-host parasite that infects muskoxen, caribou (Rangifer tarandus) and moose  [0; lower threshold temperature), larval development inside the intermediate host is minimal, and development to L3 does not take place [0, L1 develop to L3 after accumulating a certain amount of heat, quantified as development degree-days (DD) [Two lung nematodes, hipelago . Umingmaschatus) , 23, whes alces) , 24. Theke place , 26. Aboays (DD) . The L3 ays (DD) . The L3 ays (DD) . The devays (DD) . HoweverUmingmakstrongylus pallikuukensis and V. eleguneniensis co-occur in muskoxen of the western Canadian Arctic and, until recently, were limited to the North American mainland and had not been found in the Arctic Archipelago. However, in 2008, U. pallikuukensis, and in 2010, V. eleguneniensis, were reported for the first time on southern Victoria Island, Nunavut, Canada [U. pallikuukensis establishing at higher latitudes prior to V. eleguneniensis ., Canada  Fig. 1)Umingmaks, Canada , 19. SinU. pallikuukensis in its intermediate host, but similar information for V. eleguneniensis is lacking. Few previous studies have investigated the freezing survival of L1 of protostrongylids [One hypothesis for the differential range expansion of the two parasites is different species-specific thermal requirements and tolerances for the development and survival of their larval stages. Indeed, understanding the developmental responses to temperature in intermediate hosts and freeze tolerance of L1 in the environment is vital for understanding the ecology and transmission dynamics of protostrongylids in general. Kutz et al.  studied ongylids –32, and V. eleguneniensis and U. pallikuukensis. Specifically, we investigated the temperature-dependent development of V. eleguneniensis in a gastropod intermediate host, the meadow slug Deroceras laeve , and compared the freezing survival of L1 of U. pallikuukensis and V. eleguneniensis. This study is essential to our understanding of the thermal ecology of these two emerging parasites in the Canadian Arctic. The resulting data on thermal tolerances provide essential parameter estimates for parasite distribution and transmission models [The objectives of this study were to determine the temperature requirements and tolerances of n models  and willU. pallikuukensis and V. eleguneniensis. First, temperature-dependent development of V. eleguneniensis in D. laeve was determined following the methodology used for U. pallikuukensis by Kutz et al. [Two sets of experiments were carried out to determine the impact of temperature on development and survival of z et al. . SecondlD. laeve, were collected from their natural habitat near Sheep River Provincial Park  and Sundre , Alberta, Canada. Refuge traps comprised of dampened cardboard, masonite board, and fabric mats, often baited with grape juice, were used for collecting slugs near creeks and wetlands [V. eleguneniensis were isolated from the feces obtained from wild muskoxen in northern Quebec  using the modified beaker Baermann technique [V. eleguneniensis L1 [The gastropod intermediate hosts, wetlands , 35. Thewetlands  and tranwetlands  at 8 ± 2echnique . The fecensis L1 , 38. Theensis L1  and PCR.The experimental infection was performed as previously described by Hobert et al.  and KutzV. eleguneniensis [For each trial, three slugs were haphazardly selected at designated days post infection and digested in a pepsin hydrochloride solution , 25. Theeniensis , and knoeniensis , 38. Theeniensis . After dVarestrongylus eleguneniensis L1 were obtained from the fresh feces of a captive muskox that was experimentally infected with the larvae obtained from wild muskoxen from northern Quebec . Umingmakstrongylus pallikuukensis L1 were obtained from fresh fecal samples collected from wild muskoxen near Norman Wells, Northwest Territories . Within 24 h of collection, fecal samples were transported to the lab in whirl packs  maintained at 4 ± 1 °C (temperature monitored by LogTag® temperature recorder) and stored at 4 ± 1.5 °C until processed. For both parasite species, L1 were extracted using the Baermann method [n method  within on method , 39.U. pallikuukensis and V. eleguneniensis was studied under four  and three  sub-zero temperatures, respectively , and degree-days for development (DD), were estimated by fitting the daily development rate D is the number of days from infection to the first appearance of intermediate third-stage larvae (iL3) and T is temperature. The parameters were estimated by linear regression of development rate (1/D) over temperature (T), where slope equals 1/DD and the intercept equals T0/DD [0, and (iii) we wanted to compare to other protostrongylids, especially with lungworm U. pallikuukensis, and linear models were used to derive the parameters for these protostrongylids.All analyses were conducted using R statistical software . The thels T0/DD , 42. We U. pallikuukensis and V. eleguneniensis. The response was the proportion of individuals in each well surviving until inspection at 2, 7, 30, 90 or 180 days freezing duration, calculated as the surviving L1s divided by the initial number of viable parasites in each well. Fixed effects included parasite species, temperature, freezing duration, and the interaction between temperature and species, representing the differential effect of freezing temperature on the two species. The plate was initially included as a random effect, but the variance among the plates was small, and the inclusion of the random effect did not change the parameter estimates for the fixed effects, so the effect of the plates was ignored. Ten different models were fitted representing different combinations of the four fixed effects. The models were compared using Akaike Information Criterion (AIC). There was a high degree of model and parameter uncertainty, and so we report the model-averaged predictions for survival over the top five models which comprised more than 90% of the cumulative Akaike weight [Binomial generalized linear models  were fitted to investigate the effect of temperature and freezing duration on the survival of e weight , using te weight .Varestrongylus eleguneniensis larvae successfully developed from L1 to L3 at temperatures between 12.5 to 24 °C  = 1297, P < 0.0001), with R2 of 0.99. From equation 0 = 9.54 °C  and 74 dpi (two slugs). Emergence from all four slugs continued to day 83, and although no emergence was detected on subsequent observations, L3 were found inside all of the slugs on digestion after they died at days 85 and 87, respectively.U. pallikuukensis and V. eleguneniensis had high freezing survival at all temperatures and all durations  for V. eleguneniensis.L1 of both ons Fig. . There wons Fig. . Temperaons Fig. . Survivaons Fig. . Despiteons Fig. , within ons Fig. , suggestlar Fig. . The modV. eleguneniensis inside the intermediate host, D. laeve, was positively related to temperature, and the lower threshold temperature (T0) and the thermal constant required for development into infective stage (DD) were within the range determined for other northern protostrongylids  at -80 °C, frozen for a similar time. Developmental thresholds and freezing survival of more temperate species seem to be comparatively lower. For example: Muellerius capillaris, a temperate protostrongylid, has a considerably lower T0 (4.2 °C) and is less tolerant to freezing despite being relatively closely phylogenetically related to U. pallikuukensis [Parelaphostrongylus tenuis, another temperate species, also has a lower freezing tolerance than it northern counterparts - approximately 70% survival of L1 when frozen in feces at -15 to -20 °C for 182 days [P. tenuis L1 survive a constant temperature of -14 °C in the lab when frozen in fecal pellets for four months. The variation in developmental and survival parameters among protostrongylid species is most likely a result of biological differences between both parasite and host species that are shaped by various ecological and evolutionary processes [0 for northern protostrongylids is an ecological adaptation to the Arctic conditions  [0 ensures that L1 enter the developmental phase only when the temperature is consistently warm, thereby increasing the chances of successful development to L3 in a single season. This further prevents mortality of developing larvae  and the intermediate hosts themselves over the winter [The developmental parameters and freeze tolerance capabilities among protostrongylids with Arctic and sub-Arctic distributions are quite comparable - developmental thresholds lie around Tfreezing –32. Withte hosts , 48. Basangiferi . The litangiferi  reportedalvorsen  did not uukensis . Parelap182 days  found throcesses . For ins period) . Higher e winter .U. pallikuukensis and V. eleguneniensis warrants further explorations on mechanisms of freeze resistance. For other nematodes, several biochemical and physiological mechanisms to survive freezing have been identified [U. pallikuukensis and V. eleguneniensis L1, but because the survival was very high in water, they may employ a freeze-tolerance strategy and survive inoculative freezing, as described for other nematodes [The extreme freeze tolerance of entified , 52–54. entified , 55, 56.entified . We do nematodes , 58.The Arctic continues to warm at an unprecedented rate, driving the emergence and spread of pathogens , 59, 60


The LC wireless passive humidity sensor was optimized by performing HFSS simulations and fabricated via a screen-printing technique. The MoS2 nanoflakes were characterized by laser scanning confocal microcopy, scanning electron microscope, and X-ray diffraction. The measurements show the sensor can operate stably for a long time with a hysteresis of 4% RH (relative humidity) in 10–95% RH. At low humidity environment (10–60% RH), the sensitivity of the as-prepared humidity sensor is 2.79 kHz/% RH, and a sensitivity of 76.04 kHz/% RH was realized in a high humidity environment (60–95% RH). In this regard, the sensing mechanism was discussed in the scope of proton transfer theory. The test results also indicate that the response time and recovery time of the prepared sensor are 10 s, 15 s, respectively and between 15~40 °C the sensitivity of sensor was not temperature-dependent in the range of 10~80% RH. In addition, the sensor shows less sensitivity to temperature in the 15–25 °C range at 90% RH. All of these experimental results show that the prepared LC wireless passive humidity sensor can stably monitor the rapidly changing humidity in a sealed and narrow environment for a long time.In this study, an LC wireless passive humidity sensor based on MoS Humidity sensors have been widely used in industrial, agricultural, medical, food package and household life ,2,3,4,5.2 have been widely studied and demonstrated their gas sensing capability [2 being as a gas sensing material is gaining more attention due to the unique properties including tunable ban gap, lower intrinsic background carrier density, etc. Moreover, owing to the special molecular structure, there exist many dangling bonds on the edge of the MoS2 structure and these provide a large amount active sites interacting with gas molecules, which implies MoS2 is a promising material for humidity sensing [2 have not been reported.Owing to the high surface area to volume ratio, 2D materials such as graphene, MoSpability ,28. Howepability . On the  sensing . Neverth2 nanoflakes. With the help of High Frequency Structure Simulator (HFSS) software, the structure and distance–S11 curve of the as-designed humidity sensor was optimized, and the sensor circuit was fixed to the alumina substrate by screen printing technology which is simple to operate and low cost. By dint of laser scanning confocal microcopy (LSCM) we investigated the morphology and topography of the as–sprayed MoS2 film and MoS2 nanoflakes were characterized by a scanning electron microscope (SEM) and X-ray diffraction (XRD). Then the experimental and simulation results were compared and the prepared sensor was tested over a wide humidity range at room temperature. Moreover, other sensing performances such as hysteresis, the response and recovery time of as-prepared humidity sensor were investigated.In this work, we studied a LC wireless passive humidity based on MoSsL, and variable sC, resonates based on Equation (1):The principle circuit of the designed LC wireless passive sensor is shown in f, which is the resonant frequency (RP) of the sensor resonant circuit, is uniquely determined by the variable C. The structure of the proposed humidity sensor is shown as 2 nanoflakes as humidity sensing film. Here, the capacitance of the MoS2-covered interdigital electrode is calculated using Equation (2) as follows:N is the number of interdigitated electrodes, l, g, h are the length, width, thickness of a interdigitated electrodes, and ε, which changes as the MoS2 nanoflakes absorbs or desorbs the moisture in ambient environment, is the relative dielectric permittivity of the MoS2-moisture mixture. cD and outD are the width and the length of inductance. As the capacitance the humidity sensor circuit resonates, the sweep source connecting with an antenna such as network analyzer with a certain frequency range can detect the resonance frequency of humidity sensor circuit, thereby realizing wireless measurement of humidity parameter.Therefore, −3 and 3960 kg/m2, while the bulk conductivity and mass density of the Ag conductor was 6.1 × 107 Siemens/m and 10,500 kg/m2. The dielectric constant of MoS2 was set to be 12.33. In the analysis, we primarily probed the S11 values of sensor in the coupling distance range of 0–30 mm with the increment of 5 mm, and the derived results shown in 11 value decreases as distance increases in 0–10 mm, and then dramatically creeps with increasing distances in 10–25 mm. The S11 value at the distance of 10 mm was higher than at any other distances and its value is up to −12.77 dB which indicates a robust signal level at this position. In this regard, we assume the preferable coupling distance between humidity sensor and antenna was 10 mm owing to the high S11 value. Further, the electric field distribution of the sensor at the distance of 10 mm is shown in 11-f resonance curve of as-designed humidity sensor at 10 mm is shown in 11 value is up to −12.77 dB.Then, the proposed humidity sensor structure was optimized by High Frequency Structure Simulator (HFSS) software. 2 humidity sensor using screen printing technology which has the advantages of simple operation, high flexibility, and low cost, etc. when compared with other microelectronic processing methods. CN33-398 Ag conductor  with a lower resistance was selected as the inductor and the interdigitated electrodes series material. MoS2 nanoflakes (particle diameter: 1.5 μm) were supplied by XFNANO Company, . First, to remove the impurities in the MoS2 nanoflakes. MoS2 nanoflakes (2.0 g) were dispersed in 200 mL deionized water and then magnetically stirred for 10 min, and then the MoS2 nanoflakes were collected by vacuum filtration. After repeating the above process three times, the as-prepared MoS2 nanoflakes were dried at 90 °C and later dissolved with 200 mL of ethanol for further use. As 2 nanoflakes solution was evenly sprayed onto the surface of IDEs at room temperature as the humidity sensitive layer of the sensor. After that, the as-sprayed MoS2 nanoflakes were dried at 90 °C for 5 min to evaporate the solvent.In this work, we fabricated the proposed MoSUnder the same conditions, we fabricated four sensors and the frequency test results of these four sensors at 10% RH and 95% RH of room temperature are presented in 2 humidity sensor. By dint of laser scanning confocal microcopy (LSCM) we investigated the morphology and topography of the as-sprayed MoS2 film and 2 nanoflakes were sprayed evenly on the IDEs and the thickness of the MoS2 thin film was about 32.07 μm. Further, the MoS2 nanoflakes were characterized by the SEM, and the results are shown in 2 nanoflakes is uniform and it has a large surface area to volume ratio. Moreover, owing to the random sequence of nanoflakes, large amount cracks can be absorbed in the SEM image, which may be beneficial for humidity capture. 2 sample prepared by the hydrothermal method. The diffraction peaks of MoS2 nanosheets are observed at 2θ of 14.4°, 33.1°, 39.7° and 58.5°, which were attributed with the MoS2 nanocrystals of (002), (100), (103), and (110) plane. Moreover, the high and sharp diffraction peak (002) of the as-prepared MoS2 samples indicates the sample has good crystallinity and a layered structure.11 data of the humidity sensor in the experimental chamber. According to the simulation results, the frequency sweeping range was set between 150–200 MHz. Note all the tests were processed at room temperature.The sensing performance tests of as-fabricated sensor were arranged in the experimental equipment shown in 11-f curve of the humidity sensor under 10% RH was extracted as shown in 11-f of the as-prepared humidity sensor in a fluctuating humidity environment ranging from 10% RH to 95% RH in increments of 5% RH every 5 min and the S11-f curve of the as-prepared sensor is given as 11 and frequency decrease as humidity variess. Moreover, as the S11 value remained below −6.7 dB all the time, this indicates that the humidity sensor shows a robust signal level from 10–95% RH. Further, by repeating the test procedures in 10–95% RH five times, we obtained the f-humidity curve with error bars as First, the as-prepared sensor was placed in the test chamber and the S2 nanoflakes and the large amount of cracks induced by the random sequence of nanoflakes, water molecules in the ambient can be absorbed by MoS2 nanoflakes. Because of the active defects caused by the Mo atoms on the edge of nanoflakes, the absorbed water molecules were then captured by the defects in the form of Mo-O bonding and ionized by the electric field E between IDEs and dissociated into protons (H+) and hydroxyl ions (OH−) accumulating and mainly distributing on the surface [2 nanoflakes. Due to the low humidity level, the water molecule adsorption on the surface was discontinuous and thus the protons transfer was confined and only happened in the discontinuous water clusters of the surface. Consequently, the dielectric constant shift weakly and the capacitance of IDEs was perturbed in a small range, which account for the low sensitivity in 10–60% RH. At high humidity range (60–95% RH), As shown in 2 and further dissociated into a large amount of protons (H+) and hydroxyl ions (OH−). In this way, proton transfer occurred in the bulk volume of the MoS2 nanoflakes and became continuous in the condensed water. As a result, the dielectric constant shifts acutely and then the sensitivity was significantly improved compared with the sensitivity in a lower humidity range. Also, as the hydrated water dielectric is 2.2 in the low humidity range, the hydrated water dielectric in the high humidity range is 78, which also gave more capacitance improvement of the humidity sensor in the high humidity range (60–95% RH) [As  surface . Then, t–95% RH) ,33.2 nanoflakes are chemically stable to water molecules and the as-prepared humidity sensor has good stability. Also, the hysteresis results shown in Later, we investigated the response and recovery time of the prepared sensor, and frequency-time curve obtained by the experiment is shown in 2 nanoflake wireless passive humidity sensor. The sensor coil is bonded to the alumina ceramic substrate by screen printing technology, which is easy to perform and facile. The sensor structure was optimized by HFSS software simulation and experimental tests show that the prepared LC wireless passive humidity sensor can operate stably at a humidity range of 10~95% RH and has the advantages of high sensitivity and small hysteresis. Under low and high humidity, the sensitivities are respectively 2.79 kHz/% RH and 76.04 kHz/% RH and the response and recovery time of as-prepared sensor is about 10 s and 15 s. In addition, the hysteresis measurements show the sensor has a hysteresis value of 4% RH. Further, it was found that in the 15–40 °C below 80% RH, the temperature has little effect on the sensitivity of the humidity sensor. In summary, the experimental results show that the proposed LC wireless passive humidity sensor can monitor the rapidly changing humidity in a tight, sealed environment.This paper presented a MoS


Scientific and engineering progress associated with increased interest in healthcare monitoring, therapy, and human-machine interfaces has rapidly accelerated the development of bio-integrated multifunctional devices. Recently, compensation for the cons of existing materials on electronics for health care systems has been provided by carbon-based nanomaterials. Due to their excellent mechanical and electrical properties, these materials provide benefits such as improved flexibility and stretchability for conformal integration with the soft, curvilinear surfaces of human tissues or organs, while maintaining their own unique functions. This review summarizes the most recent advanced biomedical devices and technologies based on two most popular carbon based materials, carbon nanotubes (CNTs) and graphene. In the beginning, we discuss the biocompatibility of CNTs and graphene by examining their cytotoxicity and/or detrimental effects on the human body for application to bioelectronics. Then, we scrutinize the various types of flexible and/or stretchable substrates that are integrated with CNTs and graphene for the construction of high-quality active electrode arrays and sensors. The convergence of these carbon-based materials and bioelectronics ensures scalability and cooperativity in various fields. Finally, future works with challenges are presented in bio-integrated electronic applications with these carbon-based materials. Over the past few decades, commercial nanoscale electronic devices, based on semiconductor wafers composed of single-crystal inorganic materials, have achieved high performance with a significant development of fabrication processes. Despite these technological advances, devices with bulky, rigid, and planar materials have limitations for use as flexible or stretchable electronics ,2,3 becaUsing solely biocompatible materials is one of the most significant undertakings in biomedical systems, especially for epidermal electronics ,11 and iOver the last few decades, carbon-based materials such as pyrolytic carbon (PC) and diamond-like carbon (DLC) have been used in the biomedical field. PC has been used in biomedical implants and coating materials, especially in the production of heart valve prostheses . Most PCCNTs have always been controversial in terms of biocompatibility. These conflicting arguments are influenced by the effects of the impurities in CNTs, the length of CNTs, surface chemistry, dispersion interaction between various factors, and experimental variables, including different doses of CNTs, cell population, and evaluation systems . CNTs arThe chemical composition and crystalline structure of both graphene and CNTs are similar. However, some properties of these two materials are different, because the two-dimensional (2D) structure of graphene is fundamentally different from the cylindrical structure of CNT. These materials have different mechanisms of interaction with cell systems, resulting in definite differences in the toxicity of living cells between graphene and CNT. Therefore, the biocompatibility of graphene should be carefully examined separately from CNTs. Recent studies on dispersed graphene flakes using a liquid-phase exfoliation (LPE)  of graphCNTs are considered promising materials for flexible neural interfaces because of their high conductivity and mechanical strength. The electrophysiological signals such as electroencephalogram (EEG) or electrocorticogram (ECoG) are directly affected by the impedance of the electrodes, which lead to the extremely low current injected into the electrodes . High imCNT-based microelectrodes have much higher surface areas than those of flat surface electrodes in the same planar surface as a result of the unique one-dimensional (1D) geometry of CNTs , which gSimilar to CNTs, PEDOT, well known as a representative conducting polymer, has also been regarded as a biocompatible sensing material that provides improved selectivity and enables further surface modification . The eleElectrophysiological signals recorded using novel microelectrodes provide an excellent temporal resolution, but the optical imaging of neural circuits must be followed to achieve both high temporal and spatial resolution ,105,106.Here, we introduce recent advances in transparent and flexible microelectrode arrays, based on CNTs and graphene for simultaneous electrical and optical interaction with brain tissues. Graphene is another candidate for a transparent flexible electrodes arrays because of its ultrathin geometry with a unique two-dimensional structure and mechanical flexibility. Kuzum et al. presented a transparent, flexible neural interface for simultaneous ECoG and calcium imaging, by transferring CVD-grown graphene electrodes on a polyimide substrate a 38]. T. T38]. TThe development of transparent graphene electrode arrays enables the optogenetic stimulation of the underlying brain tissue, as in the case of CNTs, as previously described. Park et al. developed a fully transparent and flexible graphene-based carbon-layered electrode array (CLEAR) by coating the graphene electrodes with Parylene C films e 112]. . 112]. TMicroelectrodes are capable of not only recording the neural signals, but also exciting or inhibiting the action potentials by applying an electrical stimulation to neurons . As the 2/V·s) is much lower than that of a perfectly aligned SWCNT in one direction (~1000 cm2/V·s) . . 161]. TThis review covers some of the latest bioelectronics based on carbon nanotubes and graphene, the most representative carbon nanomaterials. In addition to their high electrical conductivity and optical transparency, excellent mechanical properties that are not found in conventional metal or semiconducting materials have allowed them to be extensively studied as wearable or implantable electronics. However, even though those physical properties of CNTs and graphene are outstanding, they cannot be integrated into the human body if their definite biocompatibility is not established. CNTs and graphene show varying biocompatibility depending on their concentration, method of synthesis, functionalization, and the type of cells to which they are applied. Although most studies using carbon nanomaterials in mammalian cells have shown biocompatibility, careful consideration of the application types is needed, as there are some studies that show toxicity to certain tissues, such as lung and skin. We have introduced biocompatible and flexible CNT/graphene-based bioelectronics that can be applied in epidermal or implantable electronics in two categories, according to their application, namely, microelectrodes and sensors.The surface coating of traditional microelectrodes with CNTs or composites of CNTs and other conductive materials, including PEDOT and gold, remarkably increases the surface roughness, thereby resulting in a high SNR. Furthermore, the high optical transmittance of graphene and CNT thin films enables optical imaging and optogenetic stimulation, at the same time as electrical recording, offering high temporal and spatial resolution. CNTs and graphene with tunable bandgaps are suitable for sensors that detect biomolecules, such as proteins and nucleic acids, with a high sensitivity due to their high mobility and surface area. Recent studies have developed a novel human–machine interface that converts human facial expressions and movements into electrical signals using carbon-based piezoresistive, capacitive, and piezoelectric strain sensors. The integration of various electrochemical sensors and microneedles for drug delivery presents a new scheme that enables the diagnosis and treatment of hyperglycemia with a thin and transparent graphene-based patch.These technological advances in carbon-based materials have contributed greatly to the realization of neural mapping, diagnosis and treatment of various diseases, and to the human–machine interface. Nevertheless, there are still unsolved issues that must be addressed to make these applications even better. Although CNTs and graphene have electrochemical properties that surpass other conventional electrodes for neural interfaces, there are still size-related problems in measuring the activity of all single neurons. To be integrated with a highly stretchable substrate for a more conformal interface with the human body, improved manufacturing technologies, such as low-temperature processes or transfer processes, are required. Furthermore, although many examples of implantable devices have been introduced, there is still a lack of research on devices for long-term usage in the biological environments. For example, CNTs and graphene can be oxidized and peeled off from the electrode surfaces under the electrical stimulation . Establi


Idiopathic pulmonary fibrosis (IPF) is a progressive fibrosing interstitial lung disease (ILD). In this review, we describe the central role of high-resolution computed tomography (HRCT) in the diagnosis of IPF and discuss how communication between pulmonologists and radiologists might be improved to make the interpretation of HRCT scans more effective. Clinical information is important in the interpretation of HRCT scans, as the likelihood that specific radiologic features reflect IPF is not absolute, but dependent on the clinical context. In cases where the clinical context or HRCT pattern are inconclusive, multidisciplinary discussion (MDD) between a pulmonologist and radiologist  experienced in the differential diagnosis of ILD is necessary to establish a diagnosis. While it can be challenging to convene a face-to-face meeting, MDD can be conducted virtually or by telephone to enable each specialty group to contribute. To make the MDD most effective, it is important that relevant clinical information  is shared with all parties in advance. A common lexicon to describe HRCT features observed in ILD can also help improve the effectiveness of MDD. A working diagnosis may be made in patients who do not fulfill all the diagnostic criteria for any specific type of ILD, but this diagnosis should be reviewed at regular intervals, with repeat of clinical, radiological, and laboratory assessments as appropriate, as new information pertinent to the patient’s diagnosis may become available. Idiopathic pulmonary fibrosis (IPF) is a chronic and ultimately fatal interstitial lung disease (ILD) characterized by progressive fibrosis and loss of lung function . IPF is Prompt diagnosis of IPF is important to ensure that patients receive appropriate care and support and have the opportunity to receive anti-fibrotic therapy and be evaluated for lung transplantation. Accurate diagnosis of IPF is critical, as other forms of ILD that have similar clinical presentations to IPF require different treatment strategies –11. ImagCentral to the diagnosis of IPF is the performance and interpretation of an HRCT scan . A surgiCurrent international guidelines for the diagnosis of IPF include recommendations for the optimal HRCT technique for evaluation of ILD . These iMDD involving a radiologist, a pulmonologist and, when necessary, a pathologist, with experience in the differential diagnosis of ILD is the gold standard in the diagnosis of IPF , 12, 13 A white paper recently published by the Fleischner Society  proposedWhile it can be challenging to convene a multidisciplinary team face-to-face, MDD can be conducted virtually or by telephone to enable each specialty group to contribute. To make the MDD most effective, comprehensive clinical and HRCT data should be made available in advance of the MDD so that all parties can consider the case in advance. A common understanding among all parties of the terms used to describe HRCT features observed in ILD  can alsoClinical information is key to the interpretation of HRCT scans, as the likelihood that specific radiologic features reflect IPF is not absolute, but dependent on the clinical context. Indeed, making a diagnosis of IPF specifically requires the exclusion of known causes of ILD, including autoimmune diseases, exposure to potential inducers of chronic hypersensitivity pneumonitis, occupational exposures, and the use of certain drugs . To assiCO] with a normal ratio of forced expiratory volume in one second [FEV1] to FVC) [The typical presentation of IPF is a male ex-smoker over the age of 50 years who presents with chronic exertional dyspnea and cough and has “Velcro”-like bibasilar inspiratory crackles on auscultation of the chest , 30, 31. to FVC) , 33. How to FVC)  and that to FVC) .A family history of pulmonary fibrosis should prompt consideration of familial interstitial pneumonia, although this is very rare . SeveralCO of 50% predicted and FVC of 65% predicted). An HRCT scan showed definite features of UIP  and had a significant exposure history for parakeets, dogs, and guinea pigs. She also had a history of asthma and arthritis. PFTs showed a DLCO of 60% predicted and FVC of 70% predicted. Laboratory findings were mildly positive for antinuclear antibody and C-reactive protein. An HRCT scan was inconsistent with UIP and most suggestive of a non-specific interstitial pneumonia pattern (Fig. A 67-year-old woman presented with chronic cough, fatigue, and mild basilar crackles on chest auscultation. She was a current smoker with a 20 pack-year history. Exposure history was significant for cats. She had a history of gastroesophageal reflux disease, hypertension, and migraines and comorbidities including back pain, and intermittent constipation and diarrhea. PFTs revealed a DLern Fig. . HoweverCentral to the diagnosis of ILD is an HRCT scan performed and interpreted by an expert radiologist. However, the likelihood that specific radiologic features reflect IPF is dependent on the clinical context. Although both radiologists and pulmonologists can identify findings that suggest an ILD, neither can make a differential diagnosis without input from the other. In cases where the diagnosis is not clear based on the patient’s radiological and clinical features, MDD is required to agree on the most appropriate diagnosis. For some patients, a working diagnosis is appropriate, but this should be reviewed at regular intervals as new information pertinent to the patient’s diagnosis may become available over time. Diagnostic guidelines for IPF will be revised in future to reflect our growing knowledge of the clinical course of patients with specific clinical and radiological features, as well as new information on molecular signatures that predict UIP.


Sporadic inclusion body myositis (sIBM) is the most commonly acquired myopathy in middle-aged and elderly people. The muscle histology is characterized by both inflammation and degeneration, including sarcoplasmic aggregation of TDP-43. Cylindromatosis (CYLD) is a deubiquitinating enzyme that targets Lys63-linked ubiquitin chains and negatively regulates signal transduction pathways, such as NF-κB signalling pathways. We examined localization of CYLD as well as phosphorylated TDP-43, phosphorylated p62, and Lys63-linked ubiquitin in muscle tissues of sIBM patients and muscle-specific wild-type TDP-43 transgenic (TDP-43 TG) mice. We investigated whether overexpression of CYLD can affect muscle toxicity in the cell models treated by endoplasmic reticulum (ER) stress inducers tunicamycin and thapsigargin. CYLD expressed with phosphorylated TDP-43, phosphorylated p62, and Lys63-linked ubiquitin in the nuclear and perinuclear regions of muscle fibres of wild-type TDP-43 TG mice and the degenerative myofibres of sIBM patients with rimmed vacuoles and endomysial cellular infiltration. Although expression levels of CYLD decreased and cell viability was reduced in cells treated with ER stress inducers, wild-type CYLD, but not the catalytic mutant, substantially improved cell viability based on the deubiquitinase activity. Dysregulation of CYLD may reinforce myodegeneration in the pathophysiology of sIBM by attenuating autophagic clearance of protein aggregates. Regulating CYLD in muscle fibres might serve as a novel therapeutic strategy for sIBM treatment. Immunosuppressive treatments have been disappointing, and many patients become wheelchair-bound 7–10 years after onset of the disease4.Sporadic inclusion body myositis (sIBM) is the most commonly acquired myopathy in middle-aged and elderly people in Western countries and possibly also in Japan6. Recent histological investigations have shown that p62/SQSTM1 and TDP-43 accumulate frequently within damaged myofibres of sIBM tissues9. Nakano et al. showed that p62, lysine (Lys) 63-linked ubiquitin, and LC3 were collaboratively involved in selective autophagy against ubiquitinated protein aggregates in sIBM-affected tissues10. During this process, compromised binding of the p62-ubiquitinated protein complex to LC3 terminates the autophagic process at the initial stage, and then causes the formation of p62-oligomer aggregates with Lys63-ubiquitinated proteins.The muscle histology of sIBM is characterized by inflammatory features such as mononuclear cells infiltrating the endomysium and non-necrotic myofibres, which has resulted in its classification as an autoimmune inflammatory disease like dermatomyositis and polymyositis; and degenerative features including formation of rimmed vacuoles and abnormal sarcoplasmic aggregation of proteins12. Activation of nuclear factor κB (NF-κB) has been shown to be an inflammatory response to the onset of sIBM14. Importantly, ER stress, induced by tunicamycin or thapsigargin treatment, resulted in NF-κB activation followed by induction of the myostatin precursor protein in a culture model of human muscle, mimicking the inflammatory and degenerative aspects of sIBM14.Endoplasmic reticulum (ER) stress and the unfolded protein response (UPR) have been identified in the muscle fibres of sIBM tissuesCYLD gene15. CYLD is a deubiquitinating enzyme that targets Lys63-linked ubiquitin chains and negatively regulates signal transduction factors and pathways, including transforming growth factor-β (TGF-β) and NF-κB signalling pathways16. So far, CYLD has been found to regulate various biological pathways: including cell proliferation, survival, and migration; immune responses; osteoclastogenesis; and spermatogenesis17.Cylindromatosis (CYLD) was initially identified in the diagnosis of familial cylindromatosis, which is caused by mutations to the tumour suppressor 9. OPTN has been shown to regulate NF-κB activation by mediating the interaction of CYLD with the ubiquitinated receptor interacting protein (RIP) in order to facilitate the deubiquitination of RIP18. Therefore, our study aimed to investigate whether dysregulation of CYLD is involved in sIBM by comparing muscle tissues of sIBM patients and muscle-specific wild-type TDP-43 transgenic (TDP-43 TG) mice, as well as myogenic cell lines treated with the ER stress inducers, tunicamycin and thapsigargin.We previously reported that optineurin (OPTN) is involved in the pathophysiological mechanisms of skeletal muscular degeneration in cooperation with TDP-43 in sIBMWe first compared the localization of CYLD in healthy control samples, control disease samples, and muscle samples from patients with sIBM. Our control disease samples consisted of patients with dermatomyositis, polymyositis, and progressive muscular atrophy. The immunohistochemical study revealed nuclear staining for CYLD in all samples Table , Fig. 1.We next examined localization of CYLD in muscle tissues of TDP-43 TG mice. As shown in Fig. We then investigated localization between pathogenic TDP-43 and CYLD in muscle specimens from patients with sIBM and TDP-43 TG mice. Phosphorylated TDP-43 (p-TDP-43) colocalized with CYLD mainly in the nuclear and perinuclear areas Fig. . In contPhosphorylated p62 (p-p62) is a putative marker for selective autophagy in the pathogenesis of sIBM. Thus, we next examined whether p-p62 was associated with CYLD in muscle tissues of patients with sIBM and TDP-43 TG mice. P-p62 formed punctate aggregates in the sarcoplasm of sIBM tissues Fig. . Most ofBecause Lys63-linked ubiquitin is involved in the selective autophagy of sIBM, we then investigated whether Lys63-linked ubiquitin cooperated with CYLD in the nuclear and perinuclear areas of sIBM and TDP-43 TG mice. Confocal microscopic analysis demonstrated frequent colocalization of Lys63-linked ubiquitin and CYLD in the nuclear and perinuclear areas of muscle fibres from sIBM-affected tissues Fig.  and TDP-20, although we failed to detect CYLD expression in these samples.Then, we conducted proteomic analysis to identify the components of degenerating myofibres from sIBM patients. We dissected the myofibres with rimmed vacuoles using a laser microdissector, digested the samples with trypsin, and analysed them with LC-MS/MS. The analysis identified expression of 26, 17, 49, and 73 peptides in the sarcoplasm with rimmed vacuoles of sIBM patients 5, 6, 7, and 9, respectively  with thapsigargin (0–2 µM). Of note, expression levels of CYLD decreased   Fig.  but CYLDµM) Fig. . OverexpWe have demonstrated that CYLD localized in the nuclear and perinuclear regions of degenerative myofibres with rimmed vacuoles in sIBM-affected tissues. CYLD showed localization associated with p-TDP-43, p-p62, and Lys63-linked ubiquitin in the myofibres of sIBM patients and wild-type TDP-43 mice. Although ER stress inducers reduced the expression levels of CYLD and decreased cell viability, wild-type CYLD substantially improved cell viability, possibly by reducing the misfolded protein load.21. In cardiovascular research, CYLD has been shown to interact with p62 directly and CYLD can directly inactivate HDAC6, thereby controlling autophagy22. Thus, it has been suggested that CYLD can regulate the autophagic pathway that clears damaged protein aggregates and reduces the chance of lesion formation by its ability to control the cell cycle, inflammation, and autophagic factors in vascular diseases21. Consistent with these findings, our observation that CYLD overexpression reduced expression levels of BiP, an ER stress marker, suggests that CYLD promotes autophagic clearance of misfolded proteins resulting from ER stress inducers in cell models. Because insoluble protein cannot be detected by our proteomic methods, CYLD might be aggregated and insolubilized with TDP-43, phosphorylated-TDP-43, phosphor-p62 and Lys63-linked ubiquitin, which could explain why we failed to detect CYLD expression in our proteomic samples from degenerating myofibres of sIBM patients.CYLD localizes primarily in the cytoplasm and perinuclear region of various cell types23. The inflammatory changes, specifically those that mimic polymyositis, are more often observed at the early stages of the disease, whereas the degenerative changes are seen more commonly in the advanced stages24. Delineating the relationship between inflammatory and degenerative changes in sIBM is a prime concern in the field of clinical myology. Recently, two independent groups identified the target antigen of sIBM autoantibodies as cytosolic 5ʹ-nucleotidase 1 A (cN1A) in the plasma and serum samples of patients with sIBM26. Although the pathogenic role of the anti-cN1A autoantibodies has not been precisely determined, our previous experiments revealed p62-positive sarcoplasmic aggregates in the myofibres of mice injected with anti-cN1A-positive sIBM IgG in an in vivo passive immunization model27. Thus, both inflammation and degeneration may provoke a vicious circle, reinforcing the myopathic process in sIBM. Although we have not shown direct evidence for this, reduced expression of CYLD may fail to negatively regulate NF-κB signalling pathways, thus promoting an inflammatory mechanism in sIBM pathogenesis.There is some contention as to whether sIBM is primarily an inflammatory or a degenerative myopathy28. Necroptosis is involved in various pathological conditions, such as ischemia-reperfusion injury in the heart and brain, and injury-induced inflammatory diseases and neurodegenerative diseases28. Recently, CYLD has been identified as an essential factor for tumour necrosis factor-induced necroptosis30. Although it remains unclear whether necroptosis is involved in the pathogenesis of sIBM, the possibility that reduced expression of CYLD prevents necroptosis should be investigated.Necroptosis is a novel form of programmed cell death that is characterized by necrosis-like morphological changes, and is triggered by RIP kinases RIPK-1 and RIPK3, and pseudokinase mixed lineage kinase domain-like (MLKL)31. Second, CYLD may be another unfortunate bystander in the underlying pathology of IBM, because many proteins are accumulated in the myofibres of sIBM6 tissues, and as a result it is somewhat difficult to identify which are primary or secondary events.The present study has a few limitations. First, the wild-type TDP-43 transgenic mouse model bears little resemblance to sIBM, such as TDP-43-positive inclusions; however, the model lacked inflammatory cell infiltrates as well as the presence of rimmed vacuolesIn conclusion, dysregulation of CYLD may reinforce myodegeneration in the pathophysiology of sIBM by attenuating autophagic clearance. Further investigation is required for a better understanding of the role of CYLD in the pathogenesis of sIBM. Regulating CYLD in myofibres might be a novel therapeutic strategy in treating sIBM.32. All muscle samples were first examined by routine histological techniques for diagnostic purposes. Fresh frozen tissues were kept at −80 °C. The study was approved by the Ethics Committee of the Kumamoto University Hospital, Japan. All methods were carried out in accordance with the relevant guidelines and regulations.We investigated muscle biopsy samples from 11 patients with sIBM, 3 with polymyositis, 3 with dermatomyositis, 3 with neurogenic atrophy, and 3 healthy controls  or RD cells (rhabdomyosarcoma) were purchased from Riken Bioresource Center . The pcDNA3-FLAG-CYLD vector was provided by Dr. Satoru Shinriki and Dr. Hirofumi Jono . The fragment of catalytic mutant CYLD (CYLD 1–932) was generated by PCR, and subcloned into the pcDNA3 vector. Cells were transfected with plasmids using Lipofectamine 2000 . Twenty-four hours after transfection, cells were shifted to fresh medium supplemented with foetal calf serum (10% vol/vol) containing thapsigargin  or 5, 15, and 45 μM of tunicamycin (Sigma-Aldrich) for 24 h.The muscle specimens were fixed with paraformaldehyde (4%), and blocked with normal donkey serum (5%)/Triton-X in phosphate-buffered saline (0.1%). The primary antibodies were as follows: rabbit anti-CYLD ; rabbit anti-CYLD (H-419) ; mouse anti-CD4 ; mouse anti-CD8 ; rabbit anti-TDP-43 ; rabbit anti-OPTN ; mouse anti-phosphorylated TDP-43  ; mouse p-p62/SQSTM1 (Ser351) ; and mouse anti-polyubiquitin (Lys63-linkage-specific) . Immunolabeled proteins were visualized by the avidin-biotin-peroxidase complex method with a Vectastain Elite ABC Kit , or anti-mouse Ig antibody-conjugated Alexa Fluor 488 and anti-rabbit Ig antibody-conjugated Alexa Fluor 594 . Nuclei were stained with 4’,6-diamidino-2-phenylindole , and observed with confocal microscopy .31. In brief, muscle biopsy specimens from sIBM patients  were dried overnight. Myofibres with rimmed vacuoles  were collected using the LMD7000 system . Collected tissue samples in Tris (10 mM), EDTA (1 mM), and Zwittergent (0.002%) were heated at 98 °C for 90 min, sonicated for 90 min, and digested with 1.5 μl trypsin  overnight at 37 °C. Digested peptides were analysed using nanoflow reversed-phase LC–MS/MS .The procedure for proteomic analysis was described previouslyCells were solubilized with radioimmunoprecipitation assay (RIPA) buffer (Tris-HCl (50 mM), pH 8.0; NaCl (150 mM); NP-40 (1%); sodium deoxycholate (0.5%); and SDS (0.1%)) and protease inhibitor cocktail . The primary antibodies were as follows: rabbit anti-CYLD , mouse anti-GADD153 (B-3) , anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) , mouse anti-BiP/GRP78 , and mouse anti-β-actin antibody . After incubation with the primary antibodies for 16 h at 4 °C, the membranes were reacted with the corresponding horseradish peroxidase (HRP)-conjugated secondary antibodies  for 1 h. Bands were detected by ECL prime western blotting detection system  with ImageQuant LAS-4000 mini EPUV . Grouping of blots were cropped from different parts of the same gel, and intensity of bands was quantified by ImageJ 1.50i software .Cells transfected with CYLD or pcDNA3 vectors were analysed using a tetrazolium-based colorimetric assay   or a MultiTox-fluor multiplex cytotoxicity assay kit (Promega) at 48 h after transfection.All values are expressed as the mean ± SEM. Differences among means were analysed using one-way or two-way ANOVA. When ANOVA analysis revealed significant differences, pair-wise comparisons were performed using Tukey post-hoc test.Supplementary information


The clinical-radiographic distinction between idiopathic pulmonary fibrosis (IPF) and non-specific interstitial pneumonia (NSIP) is challenging. We sought to investigate the gene expression profiles of IPF and NSIP vs. normal controls.n = 22), NSIP (n = 10) and from normal controls (n = 11) was assessed. Microarray analysis included Significance Analysis of Microarray (SAM), Ingenuity Pathway, Gene-Set Enrichment and unsupervised hierarchical clustering analyses. Immunohistochemistry and serology of proteins of interest were conducted.Gene expression from explanted lungs of patients with IPF  contains supplementary material, which is available to authorized users. Idiopathic pulmonary fibrosis (IPF) and non-specific interstitial pneumonia (NSIP) are the most common forms of idiopathic interstitial pneumonia (IIP) . Better The clinical-radiographic distinction is challenging , but is Differential diagnosis between IPF and idiopathic NSIP is based on high-resolution chest CT scan (HRCT) and pathology. Diagnostic radiographic criteria for NSIP are not clearly defined , and as Given the difference in outcomes and response to therapy, we hypothesized that IPF and NSIP exhibit distinct transcriptional profiles, and that specific gene markers may be identified for each condition. As Rosas and Kaminski pointed out, gene expression studies have been highly effective in reclassifying clinically relevant disease phenotypes with similar histologic presentation . The autSpecimens were obtained from the peripheral area of the lower lobe of each lung as soon as the first recipient lung was taken out, snap frozen in liquid nitrogen, and stored at − 80 °C. RNA was extracted and hybridized to the Human Gene 1.0 set array (Affymetrix) from explanted lungs (2001–2008) in 22 patients with clinical diagnosis of sporadic IPF, entirely typical UIP HRCT pattern  and defiThe study was approved by the Human Tissue Committees and Research Ethics Boards of the University Health Network (protocol n.11–0932) and Western University (n.105214).RNA was isolated, labeled, and hybridized to the human gene 1.0 set array according to the manufacturer’s protocols (Affymetrix). Data sets for microarray experiments are available at the Gene Expression Omnibus repository, accession n.GSE110147.Partek software  was used for the preliminary analysis. The q-value  was used to identify differentially expressed genes and was computed with significance analysis of microarray (SAM). For pathways/networks discovery analysis, Ingenuity Pathway Analysis  was used. For Gene-set Enrichment Analysis (GSEA), C5 GeneOntology was used as gene-set database , 18. ForAll IHC slides were reviewed by 2 pathologists. IHC for periostin (POSTN), osteopontin (OPN), p53 and p16, selected ad hoc based on microarray results, was performed using standard techniques.The serum levels of 6 proteins, a priori selected based on literature review ligand-18 [CCL-18]) were measured at baseline and at 4-month intervals for 12 months in 34 patients diagnosed with IPF  and follDetails on sample processing, RNA isolation, complementary DNA synthesis, pre-processing on probe-level data, IPA, GSEA, RT-PCR, IHC and serum measurements are provided in the Additional files P-values < 0.05 were regarded as significant. Prism-4 software package  was used.The Kolmogorov-Smirnov test assessed variables’ distribution. For comparison of groups, either unpaired t test or the Mann-Whitney test, where indicated, was used. Cox proportional hazards regression analysis was used to identify serum biomarkers significantly predicting clinical progression. Receiver operating characteristic (ROC) analysis was used to determine the accuracy of serum biomarkers in predicting clinical progression (c-statistics). There were no significant differences in pulmonary function tests, exercise capacity or pulmonary artery pressures. NSIP patients were significantly younger than IPF patients Table .Table 1DRNA integrity numbers were very similar in the 3 groups: 8.5 ± 0.4 in IPF, 8.8 ± 0.5 in NSIP and 8.6 ± 0.4 in normal controls. Sequential steps in gene expression profiling are shown in Fig. With stringent selection criteria , SAM identified 146 differentially expressed genes, of which 60 upregulated in IPF and 86 upregulated in NSIP. The top 25 upregulated genes in each condition are shown in Tables insulin growth factor binding protein-5 [IGFBP-5], Prominin-1), myofibroblast differentiation and proliferation , collagen deposition , matrix remodeling  peripheral blood mononuclear cells (PBMCs) proliferation and infiltration  and senescence . Most genes were also markedly upregulated in IPF when compared to normal controls. MMP-1 and OPN were the top 2 upregulated genes in IPF compared to controls  in IPF vs. NSIP was confirmed by RT-PCR , ctin ACTA-, OPN), cIPA demonstrated the involvement of relevant genes  in the network “connective tissue disease, organismal injury and abnormality” , the humoral arm of innate immunity  and the recruitment of leukocytes into the lung compartment . The top 25 genes upregulated in NSIP vs. controls are shown in Additional file Genes with significantly increased expression in NSIP, compared to both IPF and normal controls, were involved in regulatory mechanisms of immune reaction, including the alloreactive T cell response  growth factors and SASP matrix remodeling markers. Interestingly, the expression of most genes involved was not significantly increased in IPF  with 79% sensitivity and 73% specificity. Details on ROC analysis are shown in Additional file p = 0.0045). The serum levels of the other biomarkers investigated were not significant predictors of clinical progression , on one hand, and of a combination of senescence and matrix remodelling-related genes  on the other. The existence of this “intermediate” cluster of IPF patients, expressing both NSIP-related and SASP-related genes, cannot be attributed to a mixed histologic pattern, as pathologic diagnosis was confirmed on whole lung explants. Furthermore, all IPF cases in this study featured an entirely typical HRCT pattern of UIP.Considering the interesting observations of radiographic “evolution” of biopsy-proven NSIP cases into an UIP/IPF pattern , and theComprehensive gene expression profiling from whole lung explants aligns separate, well-recognizable histopathologic patterns of IIP with distinct transcriptional profiles and differentially expressed genes, and does not support the notion of NSIP as an early manifestation of IPF, but rather point to a standalone, transcriptionally homogenous condition, even in advanced fibrotic cases requiring LTx. LRP-2 and IFI-44 were identified as candidate molecular markers of NSIP, further validated against normal controls. This, however, does not exclude that “senescent” NSIP itself may represent a risk factor for developing IPF, given the remarkable degree of transcriptional overlap observed in a subgroup of IPF cases. Senescence-related genes are indeed prominent in IPF: we identified IGFBP-5 as a candidate molecular marker specific for IPF; p16 as a candidate fibroblast protein marker of IPF; and POSTN serum levels as better predictors of clinical progression, although not specific for IPF. These findings will likely stimulate validation studies in smaller, routinely available biopsy samples, as well as in larger populations.Additional file 1:Figure S1. A. Probe intensity histogram. B. F ratio: signal-to-noise ratio, IPF vs. NSIP analysis across the whole genome. The bar indicates the average signal for all genes. The height of the bar is the mean square. UIP=IPF (JPG 57 kb)Additional file 2:Figure S2. Principal component analysis, a global analysis across the whole genome. IPF and NSIP groups are shown. UIP=IPF. (JPG 114 kb)Additional file 3:Table S1. Top 25 upregulated genes in IPF vs. control . Table S2. IPF vs. control, Ingenuity Pathway Analysis (IPA). In IPA, “functions” are divided in 3 categories: “disease and disorders”, “molecular and cellular functions” and “physiological system development and function”. Table S3. Top 25 upregulated genes in NSIP vs. control . Table S4. NSIP vs. control, Ingenuity Pathway Analysis (IPA). In IPA, “functions” are divided in 3 categories: “disease and disorders”, “molecular and cellular functions” and “physiological system development and function”. Table S5. Gene expression of cellular senescence biomarkers in IPF vs. NSIP and vs. normal controls. Significantly upregulated genes are indicated with *. Pro- and anti-fibrotic properties of metalloproteinases (MMPs) are indicated in brackets under the gene name. Table S6. Summary of immunohistochemistry findings in IPF and NSIP. Table S7. Receiver operating characteristic (ROC) analysis of serum biomarkers vs. 12-month clinical progression. (DOCX 47 kb)Additional file 4:Figure S3. Comparison of gene expression levels determined by oligonucleotide microarray (left) and by quantitative RT-PCR  in the IPF and NSIP groups: A. MMP-7. B: OPN. C. IGFBP-5. (JPG 81 kb)Additional file 5:Figure S4. Ingenuity network analysis, networks with highest scores. Networks were scored based on the number of network eligible molecules they contained. Network eligible molecules with relatively increased expression are shown in red, whereas molecules with relatively reduced expression are shown in green. The intensity of the color is proportional to the fold change. Non-colored nodes represent genes added by Ingenuity pathway analysis based on its network algorithm but not upregulated in the actual microarray data. A. Network “Connective tissue disease, organismal injury and abnormality, cancer” (score = 29). Network eligible molecules with relatively increased expression in the IPF group are shown in red, whereas molecules with relatively increased expression in the NSIP group are shown in green. B. Network “Anti-microbial response, inflammatory response and cancer” (score = 42). Network eligible molecules with relatively increased expression I the NSIP group are shown in red. (JPG 53 kb)


A combination of carbon nanotubes (CNT) and graphene in the form of macroscopic hybrid buckypaper (HBP), exhibits a unique set of properties that can be exploited for many emerging applications. Here, we present a simple, inexpensive and scalable approach for the synthesis of highly conductive auxetic graphene/CNT HBP via wet-filtration-zipping and demonstrate the electrical, electrochemical and mechanical performance  of synthesized HBP. An overall increase in electrical conductivity of 247% is observed for HBP (50 wt.% graphene and 50 wt.% CNT) as compared to BP (100 wt.% CNT) due to effective electronic percolation through the graphene and CNT. As a negative electrode for lithium-ion batteries, HBP shows 50% higher gravimetric specific capacity and 89% lower charge transfer resistance relative to BP. The graphene content in the HBP influences the mechanical performance providing an auxetic structure to HBP with large negative Poisson’s ratio. The facile green-chemistry approach reported here can be readily applied to any other 1D and 2D materials and solves key challenges associated with existing buckypaper manufacturing methods. The potential of the synthesis method to integrate with current cellulose paper manufacturing technology and its scalability demonstrate the novelty of the work for industrial scale production. The origin of exceptionally high Young’s modulus, 1.2 TPa, electrical conductivity, 106 S/m, and thermal conductivity, 3000 W/mK, are due to the bond strength of sp2 hybridized carbon, out of plane π-orbitals with unique band structure, and phonon transport, respectively6. They are highly sought for advanced energy, structural and electronic applications9. Due to their microscopic nature, they are difficult to handle and therefore need a macroscopic assembly for real world applications. Carbon nanotube papers 37, graphene papers, graphene oxide (GO) papers, reduced graphene oxide (rGO) papers, GO/rGO-CNT hybrid BP (HBP), and graphene-CNT HBP are some of the macroscopic assemblies which represent the outstanding nanoscale material properties in macroscale form, leading to the creation of strong, foldable, auxetic and highly conductive lightweight materials. These assemblies, in particular, HBP combines 1 dimensional (1-D) properties of CNT and 2 dimensional (2-D) properties of graphene in a single 3-dimensional (3-D) macroscopic structure. Moreover, these assemblies are important in various applications, such as in water and air purification39, energy storage40, sensors42, aviation44, medical devices46, composites48 etc. Another interesting aspect of BP/HBP is that the realignment of entangled nanotubes during stretching gives a large negative Poisson’s ratio, and such materials are usually termed auxetic materials. Auxetic materials have various applications, such as press-fit fasteners, curved sandwich panels, flexible impact buffers, soundproof materials, and so on13. Recent progress in multi-walled CNT (MWCNT) and graphene production provides these materials at scalable quantity50. Concomitantly, the manufacturing of BP/HBP also needs a low-cost, simple, scalable procedure to convert the CNT and graphene powder into BP/HBP.Carbon nanotubes (CNT) and graphene consist of st method, BP is directly produced during CNT growth by chemical vapor deposition - also known as CVD method21 or post-CNT synthesis using dry techniques of shear pressing, alcohol drenching and pressing, domino pushing and CNT drawing. A disadvantage of this technique is that the resulting BP is typically not thick. Moreover, the production process is not roll-to-roll. In the 2nd method, BP is produced by powder compaction and frit-compression. However, the powder compaction requires a mold, thus the created BP may not be flexible nor foldable. Despite the thicker BP compared to 1st method, frit-compression technique requires a relatively high cost membrane which hinders scalability. The 3rd method involves techniques such as drop casting30, rod coating27, air spraying, and tape-casting26. Yun et al. described the fabrication of BP using tape-casting on a mold followed by vacuum oven heating15. The need of mold as well as vacuum heating process may hamper this method’s scalability. Moreover, it is problematic to detach the CNT layer/film from the supporting substrate to get freestanding BP due to low surface energy of CNT. Recently, to overcome this problem, Susantyoko et al. developed a surface-engineered tape cast method to manufacture flexible, freestanding, and foldable BP on a roll-to-roll system without the usage of mold16. However, a special need for surface-engineered conveyor belt limits its use in the laboratories. The 4th method is the membrane filtration38, which can be effortlessly installed in laboratories. Despite the common usage of this method, challenges of the membrane-filtration method include membranes with a relatively high-cost; requirement of hazardous chemicals; requirement of vacuum; requirement of high pressure; long filtration time due to the high fluid flow resistance (particularly for thick BP); relatively small BP diameter ; and relative non-uniform BP thickness. Because membrane filtration has a low-throughput, it may not be a potential candidate for mass production of BP. We solved these challenges using environment-friendly water based wet-filtration-zipping. In this method water with the surfactant is used to disperse the hydrophobic MWCNT and graphene to obtain their well dispersed solution. We use inexpensive cellulose filter papers with relatively large pore size (11 µm) and thickness (180 µm) to obtain MWCNT and graphene sediment on it by gravity. Further, MWCNT-graphene sediment was zipped due to elasto-capillary densification and thus a freestanding BP/HBP54 is formed. Our process has the potential to integrate with the current industrial paper manufacturing technology. Other advantages are scalability, uniform thickness (at micron level), and potential of the synthesis route to accommodate any other 1D and 2D materials .The HBP production is new and mostly utilizes the existing BP production methods. Four major BP production methods have hitherto been reported in the literature. In the 155. However, there are few limitations of graphitic anode such as low specific capacity and mediocre rate capability, warranting further research to advance the performance of carbon-based anode materials. CNT and graphene are promising candidates for large capacity lithium-ion batteries due to their excellent conductivity as well as stability. In addition to the high specific surface area, the two sides of graphene sheets, and internal as well as external walls of CNT can be fully utilized to store lithium ions to form LiC3, thereby raising anode theoretical specific capacity (744 mA h g−1) to more than twofold of the graphite (has a lower surface area and forms LiC6 during lithiation) specific capacity (372 mA h g−1). Consequently, the right selection of material and appropriate architectural design modification are crucial for excellent battery performance. It is expected that the combination of CNT and graphene in HBP will improve the anode performance. Therefore, the anodic performance of binder free HBP is evaluated using cyclic voltammetry, electrochemical impedance spectroscopy and galvanostatic cycling measurements with lithium metal as the counter electrode.Another aim of this work is to study the electrical, electrochemical and mechanical performance of BP/HBP. We have demonstrated potential application of HBP as an anode for lithium-ion batteries. The anode is one of the most important part of lithium-ion batteries since the anode characteristics influence their electrochemical performance. Incumbent anode utilizes graphitic carbon because of its advantageous features, such as the excellent electrical conductivity, beneficial hierarchical structure for the intercalation of lithium ion, low-cost and abundant availabilityAs such, without any chemical bonding and binder, these free-standing self-assemblies are held together due to entanglement and weak van der Waals interaction. It is important to understand the tensile and fracture behavior of such assemblies. Therefore we also focused on evaluating their tensile properties and fracture resistance in mode I and mode III, and identified the corresponding failure mechanisms.50. MWCNT are synthesized by a continuous atmospheric CVD system with a glass fiber fabric substrate49. The diameter of MWCNT is within 10–30 nm range with 3–10 walls  is much higher than that of the ethanol (23 mN/m) and acetone (25.2 mN/m) and therefore higher densification is expected in the case of water. It is expected that the weak van der Waals forces, entangled network of MWCNT and covalent bonding in MWCNT shared walls are responsible for holding the whole BP/HBP assembly. Moreover, MWCNT and GC are not functionalized and therefore other chemical bonding such as hydrogen and hydroxyl is absent. The lateral dimensions of BP/HBP depend on the size of the metal tank whereas the thickness of BP/HBP depends on the amount of MWCNT and GC thus allowing scalability over lateral size and thickness  produced at industrial scale are used for the fabrication of buckypaper (BP) and hybrid buckypaper (HBP)Surface morphology of as-prepared (uncompressed) and compressed BP observed in SEM is shown in Fig. As prepared (uncompressed) and compressed HBP show a distinct surface morphology as compared to BP Fig. . Apart f2–106 S/cm. The difference in the electrical properties of our HBP and the MWCNT/graphene is due to the discontinuity between the individual MWCNT/GC and the weak electrical conductivity between the contacts of MWCNT–MWCNT, MWCNT–GC, GC-GC. In comparison with the electrical properties of published work of BP/HBP, our BP exhibits properties similar to BP with randomly oriented CNT, and our HBP with HBP comprising self-assembly of CNT-graphene. However, our BP/HBP exhibits marginal properties compared to the BP with oriented CNT and HBP with chemically bonded CNT-graphene. The difference in these properties comes from various parameters such as graphene type , CNT diameter and length, alignment, density, graphene flake size, chemical treatment, processing condition, fabrication method, nature of bonding between CNT and graphene, etc.The GC content and compressed HBP show an ameliorating effect on the electrical conductivity Fig. . The ele−1 discharge rate had 1st, 2nd, 50th and 100th cycle specific capacity of 821.4, 204.8, 149.6 and 149.7 mAhg−1, respectively. BP has specific capacity of 1065.5, 218.8, 88.8 and 101.9 mAhg−1 at 1st, 2nd, 50th and 100th cycle, respectively. The superior specific capacity of HPB after cycles can be attributed to secondary lithium-ion storage mechanism in the disordered stacks of graphene sheets. The specific capacity drop from 1st cycle to 2nd cycle of 616.5 mAhg−1 of HBP is lower than that of BP of 846.653 mAhg−1. Electrochemical impedance spectroscopy (EIS) was performed and the data was fitted using equivalent circuit in Fig. 57. However, being a freestanding structure, the HPB does not have dead weights from metal current collectors, which set it apart from references57. In short, the right selection of material, and appropriate architectural design modification are crucial for excellent performance of battery. As a result, the combination of MWCNT and graphene in HBP improves the anode performance.Figure 58.BP and HBP are highly desirable next generation carbon-based materials which have potential to replace the carbon fibers in multifunctional composites. It incorporates the outstanding properties of 1D CNT and 2D graphene. In this regard, mechanical properties of both BP, and HBP are important. Tensile response of BP and HBP are shown in Fig. 60. The compression further improves the mechanical performance of BP and HBP  test. The details of the tests are provided in the Methods section, and geometric parameters and dimensions of DENT specimen are provided in the Supplementary Information Figure . Figure entclass1pt{minimaet al. observed the stick-slip tearing in graphene oxide papers intercalated with CNT. The stick-slip behavior has also been observed in thermoplastic elastomers and polyvinyl alcohol gel sheets66, and in layered structures with low interfacial properties67. The as-prepared BP shows tearing toughness of 879 J/m2 whereas the as-prepared HBP shows the tearing toughness of 288 J/m2  anode shows 50% higher specific capacity and 89% lower charge transfer resistance relative to BP (100 wt.% MWCNT). In cyclic test, HBP anode shows excellent Columbic efficiency and very low cycle to cycle capacity fading. The GC content in the HBP influences the tensile performance giving a large negative Poisson’s ratio. The presence of GC flakes in the MWCNT network aids higher transverse expansion with load and results in negative Poisson’s ratio. The fracture resistance of BP and HBP is studied in two different modes, including mode I fracture using the DENT test and mode III tearing test using the trouser test. The BP shows the average fracture toughness 1.11 MPa mGraphene CrystalTM KAUST-Saudi Arabia. The details about the GC are provided in the patent. The GC are graphene flakes with less than 10 layers containing defect-free graphene lattice (sp2 hybridized C-C bonding) over the full range of basal plane. The flake size is less than 300 µm. Multi-walled carbon nanotubes/nanostructures (MWCNT) were obtained from Applied Nanostructured Solutions, LLC. MWCNT flakes consist of bundles of aligned MWCNT. Sodium dodecyl sulfate (CH3(CH2)11OSO3Na) surfactant was purchased from Sigma Aldrich (CAS Number 151-21-3).Few-layer graphene flakes in a powder form ) were obtained from spin-off Schematic of wet-filtration assembly is shown in Fig. The fabrication of BP/HBP starts with dispersing MWCNT and GC in water using the surfactant. In a typical experiment, MWCNT and GC were weighted in the range of 100 mg to 1000 mg using microbalance with the accuracy of 1 mg. The weight ratio of MWCNT and GC was varied to obtain the HBP. A BP contains only MWCNT whereas HBP contains MWCNT and GC in a different weight ratio. We also tried to obtain only GC paper but it was difficult to peel it off from the filter paper due to the weak interactions between two GC. In a glass beaker, 1 g SDS was dissolved in a 1 l water. The weighted MWCNT and GC were very easily mixed with water due to the presence of surfactant. The solution was subjected to 24 h ultra-sonication using a tip-sonicator with a 15 s ‘on’ and 10 s ‘off’ pulse of 40% amplitude and 500 W power. It was observed after 24 h that the dispersion clear black solution is obtained, moreover, the solution stays stable for the several days indicating proper dispersion of MWCNT and GC in the water   and transmission electron microscope . Monochromator and image corrector were used to acquire high-resolution TEM (HR-TEM) images at 80 kV. Spectroscopic characterizations were carried out using Raman confocal spectroscopy using WITec confocal Raman spectrometer with an excitation wavelength of 532 nm  was taken using a precision balance  with a readability of 0.01 mg. Coin cells of 2032-type were assembled using a crimping machine (MSK-110 Coin Cell Crimping Machine) inside a glovebox (MBraun MB-Labstar 1450/780). The liquid electrolyte used was 1 M LiPF6 in EC:EMC (1:1 vol%) with 2 wt.% FEC. A half-cell configuration was used in which a lithium metal foil serves as counter and reference electrodes. Coin cells were tested using a battery tester (Maccor Battery Test System Series 4000) inside an environmental chamber (CSZ Model MC-3 Chamber) at a constant temperature of 25 °C. Battery testing was performed using constant current charge discharge (CCCD) at various rates with a potential limit between 0.01 and 3 V vs. Li/Li+. The BP/HBP sheets were also cut as 5 mm disk samples to be assembled in coin cell configuration for rate-capability and cyclic voltammetry tests. Cyclic voltammetry was conducted at scan rate of 0.1 mV s−1 from 0.01–3.0 V vs. Li/Li+, using a multi-channel potentiostat/galvanostat (Princeton Applied Research PMC-1000) without iR compensation. The specific capacities were calculated by considering the mass of the whole electrodes = mass of MWCNT + mass of graphene. Electrochemical impedance spectroscopy (EIS) measurements were performed obtained using an Autolab PGSTAT302N; the acquisition of the impedance spectra was done near open circuit potential (at 1.4 V) with ac frequencies between 106 Hz to 2.3 × 10−3 Hz and ac amplitude of 10 mV.The BP/HBP were cut as 12 mm diameter disks. The mass loading (average of 3.1 mg cm3 were performed in Zwick-Roell Z005 device with a 20 N load cell at ambient temperature (~20°C). The tensile load was applied at a constant crosshead speed of 200 μm/min. To reduce stress localization in the grip zone, tab made from thick adhesive tape was used. DIC technique was used for full-field measurement of strain on the specimens. White speckle patterns of acrylic paint were applied on the black surface of BP/HBP using an airbrush prior to testing. Consecutive speckle images were acquired as a function of load using a monochrome 5.0 MP camera for strain evaluation. The average engineering strain in the axial and lateral directions over the gauge length zone was evaluated using Vic-2D software. These strain values were then used to construct the engineering stress‒engineering strain curve and to evaluate the Poisson’s ratio. Further details on DIC experimental setup can be found in58.Tensile tests of the BP/HBP with sample size of 60 × 6 × (thickness) mm68:The mode I fracture toughness of BP/HBP was evaluated using double edge notched tension (DENT) specimen configuration. The tensile load was applied at a constant crosshead speed of 200 μm/min using the Zwick-Roell machine with a 20 N load cell at ambient temperature (~20°C). The specimen with ligament length mentclass2pt{minimThe strain energy release rate F is the mean force over the entire ligament length; and The trouser tear tests were performed to evaluate the resistance to tear propagation of BP/HBP. The tests were conducted on the Zwick-Roel Z005 machine with 20 N load cell at cross-head speed of 2 mm/min. The critical tearing energy Supplementary Information


In this paper, a series of fluorinated graphene materials are prepared by using N, O-doped graphene aerogel as precursor via a direct fluorination method, and the effect of fluorination temperature on the FG structure is investigated. The prepared FG samples are systematically characterized by scanning and transmission electron microscopy, X-ray photoelectron spectroscopy, X-ray diffraction, Fourier transform infrared spectroscopy, and Raman spectroscopy. It is found that the structure of FG, including features such as layer size, chemical composition, chemical bond state of the component elements, etc., is significantly related to the fluorination temperature. With the change of the fluorination temperature, fluorine atoms enter the graphene framework by a substitution process of the N, O-containing groups, including residual phenol, ether, carbonyl groups, or C–N groups, and the addition to CC bonds, subsequently forming a fluoride with different fluorine contents. The fluorine content increases as the fluorination temperature increases from 200 °C to 300 °C, but decreases at a fluorination temperature of 350 °C due to the decomposition of the fluorinated graphene. The prepared FG samples are used as cathode material for lithium primary batteries. The FG sample prepared at 300 °C gives a high specific capacity of 632 mAh g Graphene-based materials have attracted great attention in nanotechnology because of their amazingly attractive properties, such as high theoretical specific surface area (2630 m2 g−1), superior electron mobility, easy self-assembling into three-dimensional (3D) macroscopic materials with controlled microstructures, high mechanical, chemical, thermal, and electrochemical stabilities, etc. Therefore, graphene-based materials have already been widely applied in many fields, including but not limited to nanoelectronics . T. T2 and–3 hybridization of carbon atoms) to semi-ionic (sp2 hybridization of carbon atoms), as shown in 2 hybridization structure of FG-350 results in a weakness of the intensity of its C–F bonds. From the O1s spectra of RGOA  and C–NH–C (400.3 eV), which are formed via the reaction between EDA and the oxygen-containing groups. The change of the N1s peak in  of RGOA b, we cou−1, 1662 cm−1, 1532 cm−1 and 1114 cm−1, which are attributed to the stretching vibration peak of –OH, the C=O stretching vibrational peak, the bending vibration absorption peak belonging to carbonyl, and C–O–C vibration absorption peak, indicating the existence of –OH, –COOH, C–O–C, –C=O. Moreover, the peaks at about 1386 cm−1 and 1580 cm−1 are assigned to –CH2 and N–H groups. The formation of these groups can be attributed to the EDA-mediated nucleophilic ring-opening reaction of epoxy groups on the surface of GO according to the previous report [−1 corresponding to the characteristic vibrational mode of C–F bond [−1, 1217 cm−1, 1226 cm−1, and 1190 cm−1 for FG-200, FG-250, FG-300, and FG-350, respectively. The C–F peak of FG-350 shows a significant red-shift, and the peak at 1572 cm−1 increased, further indicating the weakness of C–F bond and the increase of O content. The change trend exactly coincided with the XPS results. The chemical structures of RGOA and FG were further studied by infrared spectroscopy a. It cans report . After fC–F bond . The exa2 and –CF3 groups on the surface, thus resulting in the partial recovery of π–π stacking of the graphene sheets for FG-350. Moreover, a weak peak at about 8.6°, corresponding to a large layer spacing of about 1.1 nm, was found for FG-350. This could be explained by the high content of N, O elements (the oxygenated groups and bridge-linked amide groups). The crystal structures of FG and RGOA were also characterized by the Raman spectra, which are shown in −1 and 1590 cm−1, respectively. The presence of D peaks is a typical feature of chemically synthesized graphene. In general, the D peaks are closely related to the disorder in the graphene, and the G peaks are attributed to the first-order scattering of the stretching vibration mode E2g observed for the sp2 carbon domain. A slight upshift of the G band from 1593 to 1605 cm−1 for the fluorinated samples was attributed to the formation of the C–F bond [As shown in C–F bond .+/Li. As shown in −1. No discharge potential platform was observed for FG-200 and FG-350, which may be related to its low fluorine content [−1, 375 mAh g−1, 569 and 30 mAh g−1 for FG-200, FG-250, FG-300, and FG-350, respectively. These values are proportional to the fluorine content, but much lower than the theoretical ones according to the F/C ratio, since the fluorinated samples investigated here possess many –CF2 and –CF3 groups, which are proved to be non-electroactive [−1. As shown in −1 and 430 mAh g−1 at 10 mA g−1 and 500 mAh g−1, corresponding to energy densities of 1485 Wh kg−1 and 731 Wh kg−1, respectively. It should be noted that there no apparent output potential delay was found at a high current density of 500 mA g−1 due to the large compact surface between the Li+ and the 2D fluorinated graphene. Finally, the discharge behavior of FG-350 was investigated, and the results are shown in + [The prepared fluorinated graphene materials were investigated as Li-primary battery cathode.  content . The disroactive . Next, wown in + .2 hybridization structure. Due to the large amount of oxygen groups in RGOA, CF2 and CF3 are produced during the fluorination process, which reduces the electrochemical performance of the fluorinated graphene as cathode materials of lithium primary batteries. The discharge capacity of FG-300 is calculated to be 632 mAh g−1 at 10 mA g−1, corresponding to energy densities of 1485 Wh kg−1, which are the highest ones among the investigated samples.We have developed four fluorinated graphene materials by using a scalable method of direct fluorination. N, O co-doped fluorinated graphene nanosheets exhibit high fluorine content and amorphous microstructures with two-dimensional nanostructures. The structure of the fluorinated graphene is proved to be very sensitive to the fluorination temperature. As the fluorination temperature increases from 200 °C to 350 °C, the fluorine content firstly increases and then significantly decreases. The high fluorination temperature leads to a decomposition of C–F, thus resulting the loss of fluorine and partial restore of sp