We have used a graphene field-effect transistor (G-FET) device whose geometrical capacitance is enhanced by incorporating an aluminum back-gate electrode with a naturally oxidized aluminum surface as an insulating layer. When the graphene, the top-side of the device, is exposed to NO2, the quantum capacitance of graphene and, thus, the measured capacitance of the device, changed in accordance with NO2 concentrations ranging from 1–100 parts per million (ppm). The operational principle of the proposed system is also explained with the changes in gate voltage-dependent capacitance of the G-FET exposed to various concentrations of NO2. Further analyses regarding carrier density changes and potential variances under various concentrations of NO2 are also presented to strengthen the argument. The results demonstrate the feasibility of capacitive NO2 sensing using graphene and the operational principle of capacitive NO2 sensing.A graphene-based capacitive NO However, most of related research has been focused on measuring a change in resistance (or conductance) when chemicals are adsorbed on a graphene surface [Since the discovery of graphene, a two-dimensional carbon atomic layer prepared by mechanical exfoliation of highly oriented pyrolytic graphite, it has garnered the attention of many researchers due to its unique electronic, optical, and mechanical properties ,4,5,6,7.2S, etc. ,9,10,11.2S, etc. ,14,15,162S, etc. , such astrations ,19. In t surface ,21,22,232 sensing has not yet been reported. The total capacitance  of a graphene-based capacitive sensing device consists of the series connection of geometrical capacitance (Cgeo) and quantum capacitance (CQ). The quantum capacitance effect is the main reason behind the changes in Ctotal upon the adsorption of chemical molecules on the graphene surface. However, due to the electrical characteristics of a system of capacitors, the system requires greater Cgeo compared to the minimum CQ , in order for Ctotal to be dominantly determined by CQ. This indicates that the sensitivity of the capacitive sensing device can be improved by enhancing the Cgeo [Although graphene-based capacitive sensing applications have been studied in recent years with various chemicals and gas/vapors ,25,26,27the Cgeo ,28,29. Fthe Cgeo ,31,32,332 sensing performance of an Al back-gated G-FET. Various gas molecules  can be adsorbed on graphene surfaces due to various reasons on graphene surfaces, resulting in changes in electronic response. The issue of functionalizing a graphene surface to increase the selectivity of a target molecule is a completely different issue yet to be resolved. However, we can still test the electrical responses of graphene-based devices by creating isolated environments to test the sensing procedure. Furthermore, the adsorbed gas molecules on graphene can be easily detached by thermal treatment or UV exposure, which allows us to recycle the prepared devices for the test under various conditions. To investigate the capacitive response of graphene for NO2, we restricted the test gases to a NO2/N2 mixture and adopted UV exposure for the recovery process. Water molecules in the air affect the quantum capacitance of graphene by adsorbing it onto graphene surfaces [2 in dry air was detected well using graphene-based devices by measuring its resistance [2 diluted in ambient air as a function of humidity should be further studied.In this work, we report capacitive NOsurfaces . Oxygen sistance . In orde2 sensing performance, and later, the electronic properties of graphene exposed to NO2 molecules will be discussed to strengthen our argument regarding the working principle of the device. Changes in the capacitance of the device were measured before and after the vacuum test chamber was filled with various concentrations of NO2 . Furthermore, the electronic properties, such as residual carrier density and potential variance at different NO2 concentrations, were extracted from the gate voltage-dependent Ctotal. Our results demonstrate the feasibility of capacitive NO2 sensing and, thus, provide preliminary research for capacitive gas sensing using graphene.We focus on capacitive NO2 sensing device was fabricated using CVD-grown graphene on Cu foil (LG electronics). Monolayer graphene was confirmed by Raman spectroscopy, as shown in x) on the surface of the Al gate electrode. Later, this extra thin layer of AlOx would serve as a high-k insulating layer for the graphene-based capacitor with a relatively large geometric capacitance. Next, CVD graphene was transferred onto the sapphire substrate. The Cu foil with top-side graphene was spin-coated with poly(methyl methacrylate) (PMMA) (4% in anisole) and then baked on a hot plate at 120 °C for 10 min. The reverse side of the graphene (lacking PMMA) was etched using O2 reactive-ion etching (RIE). Afterwards, PMMA/graphene on the Cu foil was placed in ammonium persulfate solution (5 wt% in distilled water) for more than 3 h to remove the Cu foil. After finishing the Cu foil etching, the graphene/PMMA film was transferred to distilled water three times to rinse the residual chemicals beneath the graphene. Subsequently, the PMMA-supported graphene film was transferred to the target substrate. Then, the sample was dried in an oven at 60 °C for more than 5 min. The PMMA layer on the substrate was removed by soaking in acetone for more than an hour. The graphene channel was patterned using photolithography and O2 RIE. Finally, 40 nm of Au source and drain electrodes were formed on the graphene channel using photolithography and electron beam evaporation. A cross-sectional view and an optical microscopy image of the fabricated device are shown in A capacitive NO2 99.999%) and 100 ppm NO2  gas, mass flow controllers (MFCs), a UV lamp , a measuring instrument , a mechanical pump, and a quartz window, as shown in 2 sensing performance, various concentrations (1–100 ppm) of NO2 gas are introduced into the test chamber by varying the ratio of the flow rate of 100 ppm NO2 and pure N2 using MFCs at a total flow rate of 10 standard liters per minute (SLM). The gas sensing performance of the device under test (DUT) is monitored by the change in capacitance caused by gas molecule adsorption on the graphene surface. The UV lamp outside the chamber is used during the recovery process; the distance between the device under test and the UV lamp is approximately 7 cm. All electrical measurements were carried out using a Keithley 4200-SCS at room temperature.Gas sensing measurements were carried out in a test chamber equipped with pure nitrogen . Then, all the valves are closed to isolate the chamber. The device is exposed to an NO2 environment for approximately 2 h to check capacitance saturation time (stage ➁). To characterize sensing performance, the response is calculated using the following equation:Cg and C0 are the capacitance before and after the exposure to NO2, respectively [Ctotal is measured by sweeping the back-gate voltage, VBG from +2 V to −2 V, to analyze the electronic properties of graphene exposed to different concentrations of NO2 (stage ➂).Before beginning the sensing measurements, the mechanical pump evacuates the chamber to its initialized state at a pressure of around 10ectively . Then, C2 adsorbates via hole recombination, e.g., hole + NO2− → NO2 (gas), on graphene [2 have been known to exhibit excellent durability and reliability for the UV-assisted recovery process [Thermal treatment and UV illumination can be used to detach adsorbates on graphene ,22,34,36graphene . Though graphene ,38. Furt2 molecules and possible introduction of O2 in the chamber (stage ➃). While pumping, the UV light is turned on to detach the adsorbed gas molecules from the graphene surface (stage ➄) and then tuned off (stage ➅) until the capacitance of the device returns to the initial state (the capacitance before NO2 exposure). Then, the response, gate sweeping, and recovery processes are repeated with different concentrations of NO2.Thus, during the recovery process, the chamber is first evacuated by a mechanical pump to reduce the residual NO2 sensing performance, the real-time capacitive response of the device at different NO2 concentrations was investigated, as shown in 2 concentrations. Note that actual measured capacitance for calculating the response in 2 with a desired concentration from 1 to 100 ppm while measuring the capacitance in real-time. The t = 0 value in 2 after the test chamber is filled with the test gas. The NO2 molecules in the test chamber adhere to the graphene surface and act as electron acceptors; they attracts electrons from graphene, leaving holes [CQ of the graphene exposed to NO2, because CQ of graphene is proportional to the square root of the carrier density of graphene [Cgeo of AlOx and CQ of graphene. This leads to changes in the measured capacitance of the device. The capacitive response immediately increases after the relatively high concentrations of 10–100 ppm NO2 are injected into the test chamber. On the other hand, the response to the relatively low concentrations of 1–5 ppm NO2 begins to change in a few seconds. Relatively high concentrations of NO2 are enough to dope graphene at the moment of the injection, so the response graph shows a dramatic change in value at the very early stages of exposure. The response at low concentrations of NO2 takes some time to dope graphene since, NO2 molecules in diluted gases are not enough to dope graphene at the moment of the injection. However, at the lower concentrations, our device can detect the differences in concentrations with higher precision within a moderately allowed time interval.To evaluate the NOng holes . The holgraphene . The meat = 75 s (roughly when the chamber pressure reaches atmospheric pressure) and 150 s are plotted as a function of NO2 concentration, as shown in 2 concentration in the two regions, namely 1–5 and 10–100 ppm. Although the response time under the higher NO2 concentration mixture is much faster simply due to the introduction of the large number of NO2 molecules in the system, the slope of response graph for lower concentrations (1–5 ppm) at the early stages of exposure is greater compared to that of larger concentration cases (10–100 ppm), which means the change in response provides a more precise distinction between various NO2 concentrations below 10 ppm. We believe that this distinction is drawn simply due to the total number of NO2 molecules our device can hold, and that the numbers can be modulated further by redesigning the size of the active channel. The change in the slope shown in Ctotal as a function of the VG curves. A detailed discussion will be given later. The response at 150 s (solid symbols) is increased with a similar trend to the response at 75 s. The changes between 75 and 150 s at high NO2 concentrations are lower than the low concentrations, which means that the response at high concentrations saturates faster than at low concentrations. To make sure of the effect of the pure N2 as a diluent gas, the response of 0 ppm NO2 was tested. When the test chamber was filled with pure N2, the response was changed by lower than 0.1% for 200 s. Impurities (the rest of 99.999% N2) in pure N2 do not significantly affect the capacitive sensing for 200 s, confirming that the N2 is only used to dilute 100 ppm NO2.The response values at dC/dt) as a function of time. Once the graphene is exposed to 10–100 ppm NO2, dC/dt starts to immediately increase until it reaches the maximum, and then it approaches 0; on the other hand, dC/dt exposed to 1–5 ppm NO2 shows a relatively slowly increase to the maximum and then approaches 0. The dC/dt at 6.6 s  is shown in dC/dt is a nearly linear relation with the NO2 concentration, suggesting that the device can detect a high concentration of NO2 by measuring dC/dt at the early stage. dC/dt at the moment the pressure of the chamber reaches atmospheric pressure (75 s). The dC/dt has an inversely linear relation with NO2 concentrations, except for 0 ppm, which is a different result from 2 concentration saturates faster than at the low NO2 concentration, as expected.2 introductions into the system.Response change for an up to 2 h time span is shown in 2 (99.999% N2) is almost constant for approximately 15 min after the test chamber is filled. The response starts to slowly increase after a quarter hour. We speculate that 99.999% N2 as a diluent containing 0.001% unexpected impurities acting as acceptors slowly dopes graphene. However, when 100 ppm NO2 is diluted by 99.999% N2, responses of 1–50 ppm NO2  do not increase after a quarter hour, indicating that NO2 is binding with graphene far more strongly compared to any other possible impurities that might be present in the balancing gas (N2). This also suggests that N2 is a proper diluent for adjusting NO2 concentration in this test.The response at 0 ppm NOCtotal from 2 V to −2 V after finishing the NO2 sensing measurement (in 2 h), in order to extract carrier density and change in potential fluctuations induced by adsorbed NO2 molecules [2 molecules shift the Dirac point voltage (VDP) to the positive gate voltage side and causes a rounding of the curves near the VDP, which is the voltage with the minimum Ctotal. The shift in VDP can reasonably be explained by charge transfer between the graphene and NO2 molecules [2 molecules act as electron acceptors, which perform a similar function as the applied negative gate voltage [olecules ,33,39. Tolecules . Adsorbe voltage .Ctotal curves is attributed to the enhancement of local geometrical capacitance caused by NO2 molecules near graphene. The NO2 molecules which are polar molecules can be intercalated between graphene and the Al electrode, similar to intercalated H2O molecules between a substrate and graphene [2 molecules enhance the effective dielectric constant of the insulating layer in the graphene/AlOx/Al capacitor. The NO2 molecules on graphene are also one of possible contributions to the enhanced electric field originating from the gate electrode. Due to the low density of the state of graphene, the graphene does not screen all of the electric field, which affects NO2 molecules on graphene when Cgeo is comparable to the minimum CQ. The NO2 molecules on the graphene are aligned by the penetrated electric field, resulting in the enhancement of the effective geometrical capacitance.The upward shift of the graphene . The intCtotal curves in 2 concentration in the chamber increases, the capacitance curves shift up to the right, resulting in changes in Ctotal at VBG = 0 V. This is the mechanism of capacitive NO2 sensing. The dramatic change in the Ctotal curve at 0 ppm near VDP explains the slope in the two regions in R1ppm (the difference between the response with 1 ppm and the response with 0 ppm) after 2 h exposure is 23.8%, suggesting that the device can detect sub-ppm NO2 through the capacitance measurement. The expected response at concentrations between 0 ppm and 1 ppm would be between 11.8% and 35.6%.The 2 adsorbed graphene, the gate voltage-dependent Ctotal in Cgeo, parasitic capacitance, and δV  as fitting parameters [V of graphene exposed to NO2 for 2 h as a function of NO2 concertation is plotted in 2 molecules causes a local potential fluctuation near the adsorbed NO2 molecules. Since the local potential fluctuations are affected by charged impurities near graphene, such as the density of adsorbed NO2 molecules, δV becomes higher as NO2 concentration increases. In addition, the residual carrier densities were calculated from the quantum capacitance minimum extracted from fitting using the equation 2 molecules cause charge transfer from graphene and produce potential fluctuation. Holes produced from the charge transfer shift the VDP to the right. The potential fluctuations round the capacitance curve near VDP. These changes are determined by absorbed NO2 molecules and explain changes in the capacitance at zero gate voltage in To investigate the electronic properties of NOrameters . The ext2 sensing performance and the electronic properties of NO2 adsorbed graphene. The quantum capacitance effect caused by enhanced Cgeo of naturally oxidized Al allowed for capacitive sensing. The capacitance of the device exposed to 1–100 ppm NO2 was changed by 21–51% compared to the initial capacitance at t = 150 s. The ΔR1ppm at t = 150 s is 21%, indicating that the device would detect sub-ppm NO2 by the capacitance measurement of the device. Furthermore, the capacitive NO2 sensing mechanism is explained by the gate voltage-dependent Ctotal. Adsorbed NO2 molecules on the graphene surface shift VDP as a function of concentrations of NO2. Carrier density and potential variations in the device caused by absorbed NO2 molecules were extracted from quantum capacitance by fitting. These results demonstrate the fundamental understanding of the absorbed NO2 effect on graphene from capacitance measurement, as well as that capacitive NO2 sensing is possible by enhancing the Cgeo of graphene-based devices.In this study, Al back-gated G-FET was fabricated to measure capacitive NO


With cellular immunotherapy, the individuals’ medication could ablate (or enhance) any therapeutic benefit of the transferred cells. Murine and human macrophages activated with IL-4 ) improve wound healing and reduce the severity of disease in murine models of colitis. Advancing the position that autologous M(IL4) could be a novel approach to IBD, a critical question arises: will concurrent medication impact the M(IL4)s anti-colitic effect? To address this, we tested if prednisolone, a synthetic, anti-inflammatory glucocorticoid used to induce remission in IBD flares,impacts human M(IL4) phenotype and function.To determine if prednisolone suppresses or enhances a human M(IL4) phenotype as defined by canonical marker molecules and wound healing and anti-colitic activities.in vitro wound healing assay. Retrieved M(IL4) and M were characterized by mRNA expression of CD206 (mannose receptor), RAMP1 (CGRP receptor), and CD14 (LPS co-receptor). One million murine bone marrow-derived M(IL4) or M were injected into BALB/c mice 48h prior to intra-rectal DNBS (3mg), and colitis was assessed 72h-post DNBS.Macrophages were differentiated from the blood monocytes of healthy volunteers using M-CSF (7 days) and treated with GMP-grade IL-4  ± a 24h treatment with prednisolone (1μg/mL). Subsequently, conditioned medium was collected for TGFb measurement by ELISA and for use in a T84 epithelial cell Human M(IL4)s displayed increased mRNA expression of CD206 and RAMP1, and reduced CD14 compared to M(0), with the CD206 and RAMP1 being further increased by prednisolone treatment. M produced more TGF-β than M(IL4) upon LPS stimulation , which would predict an enhanced wound healing capacity. Stimulated M produced more IL-10 than M(IL4). Furthermore, murine M retained an anti-colitic capacity comparable to M(IL4) as determined by disease activity score in the DNBS model.Human M(IL4)s subsequently exposed to the potent immunomodulatory glucocorticoid, prednisolone show increased expression of phenotypic markers and increased output of TGFb and IL-10. Crucially M retained an anti-colic effect in the murine DNBS model of colitis. Interpreting these data, we suggest that the anti-colitic effect of M(IL4) immunotherapy would not be adversely offset by the individuals concomitant use of steroids. Our preliminary findings support pursuing M(IL4) transfers as a novel approach to the management of IBD.OtherHelmsley Charitable TrustNone Declared


Electrochemical exfoliation is an efficient and scalable method to obtain liquid-phase graphene. Graphene in solution, obtained through electrochemical exfoliation or other methods, is typically polydisperse, containing particles of various sizes, which is not optimal for applications. We employed cascade centrifugation to select specific particle sizes in solution and prepared thin films from those graphene particles using the Langmuir–Blodgett assembly. Employing centrifugation speeds of 3, 4, and 5 krpm, further diluting the solutions in different volumes of solvent, we reliably and consistently obtained films of tunable thickness. We show that there is a limit to how thin these films can be, which is imposed by the percolation threshold. The percolation threshold is quantitatively compared to results found in literature that are obtained using other, more complex graphene film fabrication methods, and is found to occur with a percolation exponent and percolative figure of merit that are of the same order as results in literature. A maximum optical transparency of 82.4% at a wavelength of 660 nm is obtained for these films, which is in agreement with earlier works on Langmuir–Blodgett assembled ultrasonic-assisted liquid-phase exfoliated graphene. Our work demonstrates that films that are in all respects on par with films of graphene obtained through other solution-based processes can be produced from inexpensive and widely available centrifugal post-processing of existing commercially available solutions of electrochemically exfoliated graphene. The demonstrated methodology will lower the entry barriers for new research and industrial uses, since it allows researchers with no exfoliation experience to make use of widely available graphene materials. N-methylpyrrolidone (NMP) was first proposed by Coleman et al. [The interest in graphene and other 2D materials keeps growing, especially since the initial delve into fundamental properties was augmented with an outlook towards potential applications . Over thn et al.  as a synn et al. –14.N,N-dimethylacetamide (DMA), γ-butyrolactone (GBL), 1,3-dimethyl-2-imidazolidinone (DMEU), and ortho-dichlorobenzene (o-DCB) [The mechanism of ultrasonic exfoliation involves ultrasonic waves in liquid media creating bubbles or voids in the liquid, which generate shear forces or cavitation bubbles upon collapsing, which then break up the bulk 2D materials into mono- and few-layer nanosheets –16. The  (o-DCB) ,17–18. E (o-DCB) .An alternative to LPE that has subsequently been developed is electrochemical exfoliation, whereby graphene is exfoliated in an electrolyte from an electrode made of graphite . In elecBoth ultrasound-assisted LPE and electrochemical exfoliation result in solutions that contain flakes of different sizes, that is, the solutions are polydisperse. Polydispersity is a significant problem regarding the use of solution-processed graphene, because many applications are size-dependent. On the one hand, for example, for use in composites, flakes with lateral sizes larger than 1 µm are preferred . On the The flake size can either be controlled during exfoliation or selected after exfoliation. Processing parameters that control flake size during exfoliation include the choice and concentration of solvent , a proceAlthough size selection through post-processing with cascade centrifugation has been demonstrated in the context of ultrasonic LPE materials –31, to tg). The obtained centrifugation sediment contained the largest nanosheets of the initial dispersion. The supernatant was subsequently centrifuged at higher rates: 2, 3, 4, and 5 krpm . At each step of centrifugation, the sediments with slightly smaller graphene sheets were gathered and the supernatants were used for the following cascade step. The sediments were collected and redispersed in a specific volume of NMP (Sigma-Aldrich M79603), which ranged between 250 and 1000 µL. This method is schematically described in In order to achieve homogeneous films with defined particle sizes, a dispersion of electrochemically exfoliated graphene from Sixonia Tech GmbH  was processed by cascade centrifugation . The commercially obtained solution contained a dispersion of graphene in NMP. Although many solvents are commercially available, NMP was the solvent of choice because of its favorable properties regarding LB deposition . 1 mL ofIn order to study the optical properties of the produced graphene films, graphene from solution was deposited onto glass substrates with dimensions of ca. 2 cm × 1 cm . AlthougTo study optical properties of the fabricated samples, UV–vis spectroscopy was performed . A xenon lamp was used as a light source. The glass samples were mounted on a holder and inserted into the light path. Optical transmittance was measured by subtracting the signal from a baseline reference signal obtained when a clean glass substrate without graphene was inserted into the light path.Graphene film resistance was measured by inserting the substrates with electrodes into an electrode connector  and the acquiring resistance with a handheld digital multimeter.−4 Pa with electron beam voltages set between 1 and 30 kV, depending on the film. Films that are shown in optical dark-field microscopy and SEM have been made from solutions that have been diluted with 500 µL of NMP.Optical dark-field microscopy of the films was performed with a magnification of 10× (Olympus BX53M). Scanning electron microscopy (SEM) was performed with a FESEM  at a chamber pressure of 1 × 10Films deposited from solutions obtained from different centrifugation protocols are expected to have different thicknesses, due to the different size of the flakes in the solutions. A visual inspection of the images in We quantify the number of scattering centers by analyzing the dark-field images using the software imageJ. First, we set the brightness threshold for what counts as a scattering particle. As these are dark-field images, anything that is not completely dark is counted as a scattering particle. Thus, we set the threshold at a brightness value of 3 . Such an analysis yields results that are consistent with intuitive observation .g) and at 4 krpm (1465g). In these three cases, SEM reveals continuous graphene films. The films contain some wrinkles or flake edges, showing up as bright lines in SEM. Upon closer inspection, the contrast varies slightly across the films, which is likely due to local thickness variations. Overall, the films resemble those made earlier with ultrasonic liquid-phase exfoliation followed by LB deposition [g) is strikingly different from the ones made with lower centrifugation speeds. The 5 krpm (2289g) film has an irregular structure, resembling a fractal coastline. It is also noted that the conductivity of this film is much lower than those of the other three films, as SEM operation quickly leads to surface charging effects.The film structure was further investigated with SEM, shown in t 4 krpm 65g. In tUV–vis spectra of the deposited graphene films at different centrifugation rates, redispersed in specified volumes of NMP, are given in Although atomic force microscopy (AFM) is often employed to characterize graphene films ,12,14,44To analyze the electrical properties of the synthesized graphene films, resistance values of each film were measured. Resistance values, optical transparency values, and the average number of layers in each graphene film are shown in The images in RS obey the following equation:Regardless of how the film thickness is controlled, an analysis of Z0 is the impedance of free space, σop is the optical conductivity, and σdc,B is the bulk dc conductivity of the film. For film thicknesses below the percolation threshold, the transmittance and sheet resistance obey the following equation:where n is the percolation exponent.where Π is the percolative figure of merit (FOM) as described by De and Coleman  and n isn = 3.6, values which are of the same order as values previously observed for graphene [We fit the above two equations to our data in the two observed regimes independently. The solid red line in graphene . The gooThe two regimes exhibit a crossover that likely indicates the percolation threshold. The value of optical transparency at which we observe the threshold is 82.4%, matching the value found in an earlier work that showed particle size selection by control of liquid-phase exfoliation time , possiblSolution-processed graphene holds potential for applications across a diverse range of industries. There exist several production methods for solution-processed graphene, some of which are highly scalable. However, all graphene solutions resulting from those processes are polydisperse, containing a wide distribution of particle sizes, which is unfavorable for applications. It was previously shown that cascade centrifugation can be used as a common method for the separation of graphene particles by size, in the case of ultrasound-assisted liquid-phase exfoliated graphene. In this paper, we demonstrated that cascade centrifugation of electrochemically exfoliated graphene can be used in conjunction with Langmuir–Blodgett assembly to produce thin graphene films. By tuning centrifugation speed and solvent dilution volume, we produced films of different controlled thicknesses. We have shown that the optical transparency and electrical resistance of these films behave similarly to those of other films made of graphene from solution. Namely, the films exhibit a percolation threshold, at which optoelectronic properties experience a critical change. The percolation exponent and percolative FOM are found to be of the same order as in the case of other graphene films found in literature, which indicates that our films are of a similar quality. However, the method that we have used, which relies on cascade centrifugation of a commercially available solution of graphene, is easily accessible to researchers who have no exfoliation experience or equipment. We have shown that ownership of a centrifuge is the only prerequisite for making practical use of widely available graphene in solution.


The objective of this study was to compare the external load of training sessions using as a reference an official competition match in women’s football in order to find if the training sessions replicate the competition demands. Twenty-two semi-professional football players were analyzed during 17 weeks in the first phase of the competitive period of the 2020–2021 season of Spanish women’s football. In addition to the competition , four types of sessions were distinguished: strength or intensity (INT), endurance or extensity (EXT), velocity (VEL), and activation or pre-competitive (PREOM). The external load variables recorded were total distance (TD), high-speed running (HSR), sprint (Sprint), accelerations (ACC2), decelerations (DEC2), player load (PL), distance covered per minute (TDmin), high metabolic load distance (HMLD), and total impacts. The main results were that the external load demanded was different according to the type of session, being, in all cases, much lower than OM. The variables referring to the neuromuscular demands (ACC2 and DEC2) were higher in the INT sessions, the TD variable in the EXT sessions and the velocity variables (HSR and Sprint) in the VEL sessions. We can conclude that there was an alternating horizontal distribution of training loads within the competitive micro-cycle in women’s football, although the order was not the usual one for tactical periodization. In recent decades, there has been exponential global development of women’s football, both in its practice and in the entities and institutions that promote and manage it ,2. This It is essential to know the demands of women’s competition  in orderIn men’s football, the two usual planning strategies are the structured micro-cycle together with tactical periodization ,14. ElitIn order to improve the knowledge of types of load management strategies in women’s football in a novel way, the objective of the present study is to describe the external load in the different types of training and competitive matches in women’s football during a competitive micro-cycle. The starting hypothesis is that there is a horizontal alternation in the conditional demand between the training sessions in the competitive micro-cycle, but none of the sessions replicates the competition demand. In this sense, the results of this study will allow increasing the information on the type of periodization that is proposed in an elite women’s football team, which could help to facilitate load management in women’s football, which is still little known and under research.A total of 22 semi-professional female football players : 65.4 ± 17.9 mm) took part in the study. The data recording was carried out during 17 weeks of the competitive period of the Second Women’s Football Division (Reto Iberdrola) during the 2020–2021 season. Usually, the team completed four days of training and one day of competition per week.In order to obtain position data, the players were monitored with WIMU PRO devices  using the global positioning system (GPS). The GPS device used in this study can operate at 10 Hz, and it is compatible with the Galileo and American satellite constellation, which seems to provide more precision . For the2 ; decelerations of less than −2 m/s2 ; player load , distance traveled per minute ; high metabolic load distance , defined as the distance covered by a player when their metabolic power is above 25.5 W/kg1, ratios per kilogram [The duration of the session was recorded considering only the effective time, that is, the time in which the players were active, excluding times of inactivity . The external load variables were: total distance ; high speed running , established as 60% of the maximum individual velocity of the participants ; sprint kilogram ; and, ton = 12), the types of sessions were differentiated based on the priority conditional objective that was developed: strength or intensity session ; endurance or extensity session ; velocity session ; and activation session . In total, 805 recordings were collected from official matches (n = 49) and 756 training sessions, distributed as follows: VEL = 114, INT = 173, EXT = 239, and PREOM = 230. The number of records per player was 36.8 ± 10.6.Apart from the official matches  and before (OM−) of the OM, following the proposal of previous studies ,14.2). The VEL session was usually the first acquisition session, located 2 days after the OM (OM + 2) and 5 or 6 days before the next official match (OM − 6 or OM − 5), characterized by velocity work, locomotive-oriented and based on tasks of intermittent nature or in waves (with breaks of 1 or 2 min between drills). The tasks were mostly carried out with goalposts and goalkeepers and were made up of a large relative space per player  and 3 or 4 days before the next competition (OM − 3 or OM − 4), characterized by neuromuscular and mechanically-oriented work, based on reduced and positional games, with a relative space per player of less than 100 m2 per player, and with goalposts and goalkeepers.The EXT session was usually the third acquisition session of the micro-cycle, located 4 or 5 days after the OM (OM + 4 or OM + 5) and 2 or 3 days before the next competition (OM − 2 or OM − 3), characterized by cardiovascular work, based on large-format tasks, that is, with a moderate to high number of participants per team (>6), in a relative space equal to or greater than 250 m2 per player) or small  with a polarized orientation and a tactical and strategic approach. Pre-match day (PREOM) was the fine-tuning session. It was always conducted the day before match day (OM − 1). The contents that were developed in it were activation tasks based on activities with a large number of participants (>6) in a medium relative space .The last day of the week was match day (OM). To calculate the load on the day of the competition, the previous warm-up carried out by the players was taken into account in addition to the two parts of the match, which had the following approximate load: duration was 20.1 ± 2.1 min, and the physical demands were as follows: TD: 1280 ± 266 m, HSR: 64.3 ± 37.7 m, Sprint: 5 ± 10.7 m, ACC2: 27.9 ± 9.6 n, DEC2: 28.4 ± 9.6 n, PL 20.6 ± 4.3 au and TDmin 64 ± 12.3  and a customized Microsoft Excel spreadsheet  for Windows.Descriptive statistics data from variables were presented using mean and standard deviation. Tests for normality (Shapiro–Wilk) and equality of variances (Levene) were applied. The null hypothesis was accepted because the distribution of the data met the normality criterion. Furthermore, the variances were homogeneous. Therefore, a one-way ANOVA analysis of variance for independent samples was used to test for differences in the variables between the different sessions . Significant results were then analyzed using post hoc Tukey’s test. Effect size (ES) was also calculated to determine meaningful differences with magnitudes classified as  trivial p < 0.05) between training sessions for all variables. The PREOM session had the lowest (p < 0.05) volume  but with higher density than the VEL and INT sessions. The INT session had the highest volume  but with a lower density than the EXT and PREOM sessions and higher than VEL.As shown in The aim of this study is to describe the distribution of external load in different training sessions and competitions in competitive micro-cycles in women’s football. According to the current state of the art, this is the first study carried out on the description of tactical periodization in women’s football. The results confirm the starting hypotheses because despite describing a distribution of the conditional demands  based on the horizontal alternation during the competitive micro-cycle sessions, none of them replicated the demands of the competition.In line with the results obtained in previous studies carried out in men’s football ,28, the On the other hand, unlike previous studies ,18, the 2 and −1.5–3 m/s2) and high (>3 m/s2 and <−3 m/s2) accelerations and decelerations during training were similar to the competition values in this type of session, in this research work, the competition values were significantly higher than the rest of the types of training sessions. Regarding the INT session, which was usually carried out on the central days of the week (OM − 5 or OM − 4) and prior to the EXT session, a predominance of the force variables (accelerations and decelerations) was described compared to the other sessions, as described in men’s football in previous works ,29. HoweWith regard to the EXT session, which was usually carried out in the middle of the week (OM − 4 or OM − 3), high values were described in TD and PL compared to the other sessions, showing a similar trend as in previous studies ,30. In eDespite the global coincidence of this approach, not all the works ,31,32,33This study describes a variant of tactical periodization, which, while respecting horizontal alternation, proposes a novel distribution in the orientation of content throughout the competitive micro-cycle. The possible practical application of this study is that the content of the training sessions with different conditional orientations could present a different ordering than the one proposed by the tactical periodization. In this way, instead of respecting the usual order of the sessions proposed by the tactical periodization , it could be altered by proposing VEL+INT+EXT when the contextual needs and/or the characteristics of the type of population to which it is directed requires adapting content to optimize their preparation process.It has been verified that none of the training sessions obtained higher values than those of the competition in all the variables . Therefore, coaches should be careful in the return-to-play process and should prepare players adequately because there could be an excessive gap between the demands of training sessions and those of competition, putting players at risk ,12,13.This work has some context limitations. In the first place, since just one team was analyzed, the periodization results should not be interpreted as something generalized in women’s football; more case studies would be required to approach an extrapolation. In addition, the inclusion of internal load variables  could haIn conclusion, this research work provides a description of the profile of training loads throughout a competitive week in women’s football. A horizontal alternation in the stimulation of physical capacities in female football was described, although the order of the training contents varied with respect to the original proposal of tactical periodization. This study could open the possibility of proposing a variant that can better adapt to a particular reality conditioned by the schedules and pitches established by the sports club of the players. In this sense, in ongoing works, a larger sample and other contexts will be included.


Epitaxial lateral overgrowth (ELO) of GaN epilayers on a sapphire substrate was studied by using a laser-patterned graphene interlayer. Monolayer graphene was transferred onto the sapphire substrate using a wet transfer technique, and its quality was confirmed by Raman spectroscopy. The graphene layer was ablated using a femtosecond laser, which produced well-defined patterns without damaging the underlying sapphire substrate. Different types of patterns were produced for ELO of GaN epilayers: stripe patterns were ablated along the  Lack of native substrate has always plagued the development of III-nitride materials and devices. GaN-based structures are usually deposited on various foreign substrates, such as sapphire or SiC; however, the large lattice and thermal mismatch causes high dislocation densities ,2. To imIn this work, we present graphene ablation on sapphire substrates with a femtosecond laser and evaluate if the resulting patterns are suitable for ELO of GaN epilayers. Monolayer graphene is transferred onto the sapphire substrate using a wet method; its quality is assessed by Raman spectroscopy. The laser-ablated patterned structures are analyzed by scanning electron microscopy (SEM). Afterward, the patterned structures are used for the growth of GaN layers, and the structural properties of the epilayers are studied.3) were used as Ga and N precursors, respectively. The epitaxy process was monitored by an in situ laser reflectometry system operating at 650 nm. Epi-ready 2-inch monocrystal (0001) orientation sapphire substrates were used for graphene layer transfer and further deposition of GaN epilayers. Commercially available transfer-ready poly(methyl methacrylate)-coated (PMMA-coated) graphene monolayers from Graphenea Inc.  were used to cover the sapphire substrates. Rapid thermal annealing (RTA) was used for the thermal treatment of the samples. The GaN epilayers were deposited by low-pressure metalorganic vapor phase epitaxy (MOVPE) using a flip-top close-coupled showerhead 3×2” reactor . Trimethylgallium (TMGa) and ammonia . The 532 nm laser excitation source with a power of 1.8 mW was focused on a 0.9 µm diameter spot on the sample surface, and 1800 lines/mm grating was used to record the Raman spectra. The wavenumber axis was calibrated using a polystyrene standard. Femtosecond laser microfabrication setup  based on Carbide laser  and Aerotech xy stages were used to perform laser ablation. The surface of the samples after ablation and epitaxy experiments was studied by SEM . Spatially-resolved cathodoluminescence (CL) measurements were performed using a hybrid CL-SEM system . The electron beam was accelerated to 4 keV; the collected light was dispersed using a spectrometer  and recorded using a CCD camera . All the measurements were performed at room temperature.The key processes involved in the sample preparation are schematically illustrated in 2 were transferred onto the sapphire substrates using the wet transfer procedure. First, the PMMA/graphene cuts were dipped into deionized water and left floating freely on the water surface for 5 min. Afterward, the sapphire substrate was put beneath the floating polymer/graphene piece, which was attached as close as possible to the center of the substrate. The sample was then dried in the air until there were no water droplets visible. Next, the sample was heated inside the RTA oven for 20 min at a temperature of 100 °C under an N2 atmosphere. The PMMA was dissolved by dipping the sample into acetone for 45 min at 40 °C and rinsing in isopropyl alcohol for another 45 min at 40 °C. Finally, the samples were annealed for 8 h using RTA at 300 °C in a vacuum to remove any organic surface residues left [Monolayer graphene pieces of size 1.3 × 1.3 cm−1. However, the intensity of D mode was low compared to G (IG/ID approx. 12), indicating the high quality of the graphene [2D/IG) was 3.8, while the position and full width at half maximum (FWHM) of the 2D peak were 2683 cm−1 and 44 cm−1, respectively, indicating monolayer graphene [The quality of transferred graphene on the sapphire substrate was verified by Raman spectroscopy. The obtained Raman spectrum, shown in graphene ,22. The graphene .v = 6 mm/s. To check the sensitivity of the experimental setup to surface roughness, the initial ablation experiments were performed without using the surface tracking system. 2, which was significantly above the ablation threshold of both graphene  and sapJ/cm2 [2 ). ChangiThe subsequent ablation experiments were performed employing the surface tracking system. To remove 2-µm-wide stripes of graphene without damaging the sapphire substrate, the focal plane was set at the 5 µm distance from the sample surface, and the pulse energy was reduced. The 2 µm width of the graphene window was selected to ensure fast island coalescence during the ELO process: the diameter of the GaN islands reaches a couple of microns before fully merging into a 2D layer . Several patterns with different distances between graphene windows were fabricated, as shown in SEM images in even see a. The evFor the following ELO of GaN epilayers, the graphene layer was divided into four quadrants  were evaluated in each zone using CL. The typical images of the spatial distribution of integrated CL intensity are shown in 9 cm−2 was obtained for GaN grown on square graphene islands (zone B). Somewhat lower values of 8.1 × 108 and 6.6 × 108 cm−2 were estimated for GaN grown on striped graphene pattern . Meanwhile, the lowest DD values of 5 × 108 cm−2 were measured both in GaN deposited on unmodified graphene (zone D) and in reference GaN grown on bare sapphire. The link between graphene pattern and dislocation density can be interpreted by considering the organization of dislocations along the lines parallel to the direction of stripes [The highest average DD value of 1.1 × 10 stripes , as indiTo summarize: femtosecond laser patterning of a graphene monolayer on a sapphire substrate is developed to fabricate a mask for ELO of GaN epilayers. Applicability of patterned graphene for lateral overgrowth is demonstrated: GaN seeds are formed mostly in the graphene windows during the standard MOVPE growth procedure. Growth utilizing masks comprised of graphene stripes confirmed higher GaN growth rate along the


Graphene oxides with different degrees of oxidation are prepared by controlling UV irradiation on graphene, and the charge transport and the evolution of the transport gap are investigated according to the extent of oxidation. With increasing oxygenous defect density  Graphene is a two-dimensional semimetal with high conductivity and mobility ,2, and ioribbons , a bandgoribbons ,5,6 suchC = 50%) ,8,9,10.The most common method to produce GO is based on wet chemistry, consisting of oxidation of graphite in strong acids, followed by a liquid exfoliation ,11. The Transport studies of GO and observation of metal–insulator transition via oxidation or reduction reactions have been reported ,13,14,18T < 400 K. The graphene device with Hall-bar geometry was investigated after UV oxidation by using a quantum design PPMS for lower T down to 2 K and magnetic fields up to 7 tesla.Our experiments were conducted on seven graphene field-effect-transistors (FETs) fabricated using exfoliated graphene on SiOD I(D)D peak phonon, and Here, periment ,27 with A,D=3 nm . As showG peak intensities, G peak intensities can be fitted with the following Equation , consistent with our observation.Regarding or model . Here, Vlows σ∝n a,b, whillat area c–f. Thisgraphene ,29 and σgraphene . The estσh2e2·1πn,32 givese)–hole(h) puddle [The change in the transport mechanism observed in GO FETs happens because the scattering with oxygenous defects becomes more frequent as the extent of the oxidation increases. =1e·dσdn ), decreas method ,33. On t) puddle . The traFπΔnflat ,38,39, o nD=0.29×12 cm−2 aT dependence of T < 400 K. Pristine graphene exhibits metallic behavior in which T. For GO samples with T, and the insulating behavior develops further with increasing l is the mean free path with T-dependence of E) is the density of states of graphene [T < 300 K. T < 145 K, T < 300 K as shown in the inset of T < 15 K, deviating from the linear dependence.In (2D VRH) ,41, follncreases ,41. On tT < 300 K, the T-dependence of the conductivity can be explained by considering 2D VRH and thermal activation conduction (TA) together. The data are fitted well with T-dependence of electrical resistivity In the temperature range of 145 < e regime .(3)ρ=ρcHere, lue of TK. In FiguB increased up to 7 teslas. The negative MR is analyzed with the following equation [equation  includinHere, In summary, we have prepared graphene-oxide FETs with different degrees of oxidation by controlling the UV irradiation time on graphene, and investigated the charge transport and the evolution of the transport gap according to the extent of oxidation. With increasing oxygenous defect density


A major hurdle for single particle cryo-EM in structural determination lies in the specimen preparation impaired by the air-water interface (AWI) and preferential particle-orientation problems. In this work, we develop functionalized graphene grids with various charges via a dediazoniation reaction for cryo-EM specimen preparation. The graphene grids are paraffin-assistant fabricated, which appear with less contaminations compared with those produced by polymer transfer method. By applying onto three different types of macromolecules, we demonstrate that the high-yield charged graphene grids bring macromolecules away from the AWI and enable adjustable particle-orientation distribution for more robust single particle cryo-EM structural determination. Air-water interface and preferential orientation problems are crucial challenges in cryo-EM specimen preparation. Here, the authors utilize graphene-coated EM grids functionalized by salts with various electrostatic properties, successfully overcoming preferred orientation. However, high-quality cryo-specimen preparation still faces many challenges, exemplified by the preferential orientation and air-water interface (AWI) problems2, reducing the success rate and efficiency of high-resolution structure determination by cryo-EM. To alleviate the preferential orientation problem, tilting the cryo-specimen during data collection has been applied3, which, however, may result in large beam-induced motion and inaccurate defocus estimation during data processing, thereby impairing the high-resolution reconstruction. Many efforts have been also made to solve the preferential orientation as well as AWI problem by introducing supporting films9, among which graphene membrane is the most promising one.Cryo-electron microscopy (cryo-EM) technique has been successfully applied to reveal the molecular basis of many essential macromolecules at atomic resolution10, an atomically thin film comprised of sp2-bonded carbon atoms, has a superior electrical and thermal conductivity, mechanical strength and negligible background noise. Graphene membrane of sole nature, however, might induce preferential orientation problem of particles adsorbed to its surface11. The fabrication of clean graphene grids also remains a practical challenge for cryo-EM specimen preparation. Although the graphene functionalization has been used to specifically capture and anchor target particles to avoid the AWI14, the current protocols of the bioactive functionalization process of graphene, based on the electrophilic reaction or conjugation interaction with the π-π bonds, are sub-optimal due to contamination built-up on the graphene surface during the transfer process or storage.GrapheneIn this work, to overcome the graphene functionalization and transfer challenges and solve the preferential orientation problem, we develop a method to produce robust graphene grids functionalized with different electrostatic charges, which provide various electrostatic interaction interface for target macromolecules to bind in and enrich orientations. Applied onto 20S proteasome and ribosome complex, the modified graphene with different charge properties enables particle adsorption on the surface with favorite orientation distribution, thus solving the AWI and preferred orientation problems in cryo-EM specimen. Finally, we successfully determine the cryo-EM structure of Ll.LtrB RNP at 3.2-Å resolution, the highest resolution of the endogenously purified group II intron RNP, whose structural reconstruction is hindered by severe preferential orientation problem on conventional EM grids.15, theoretically making it easier to be cleaned from the graphene surface.As the clean transfer of graphene remains one of the major practical barriers for the application of graphene-based supporting films in cryo-EM, we aimed to develop a robust procedure to transfer graphene on EM grids without contamination. We found that paraffin is an ideal transfer mediator. Paraffin is a chemically inert alkane in solid state with relatively lower melting temperature (50–60 °C) compared to other commonly used transfer mediator such as PMMA, which is normally dissolved and removed by organic solvents. Moreover, the graphene-adsorption energy of paraffin is reported to be much smaller than PMMA16 and then used paraffin to support the graphene during the fabrication procedure  was found on the Raman spectra of the functionalized graphene, suggesting the presence of the modification-induced defects  and SO3--functionalized graphene (SFG) grids have decreased water contact angles of ~67° and ~66°, respectively  to characterize the elemental composition of the functionalized graphene and found the presence of N 1 s Fig. , upper a. p Fig. , down siely Fig. , allowinWe used the charged graphene grids to make cryo-EM specimens of 20S proteasome  The electrostatic interaction is much stronger, compared with other interaction modes such as the van der Waals force, allowing us to effectively manipulate the orientational distribution of target macromolecules. 2) We adopted a dediazoniation reaction to functionalize graphene in moderate conditions with high efficiency. 3) Graphene was modified prior to its transfer onto EM grids to reduce the chance of contamination and then transferred to EM grids with a paraffin-assistant graphene grid fabrication method.The charged graphene grids provide different surfaces for protein molecules or macromolecule complexes to interact with, based on the target molecule’s surface charge properties, therefore allowing multiple orientations on various charged graphene grids and avoiding impairment by the AWI.4, and the mixture was stirred at 0 °C for 5 min. 1 ml 2.2 M sodium nitrite solution was added into the mixture at 0 °C and the solution was stirred vigorously for 1 hr followed by filtration to obtain a crude product. We used acetone to dissolve the solid, used ether to precipitate the product, and finally filtered the product again to get pure diazonium salt. The benzene-sulfonic acid group diazonium salt was dissolved in deionized water and benzene-anilinic group diazonium salt was dissolved in DMSO, both at a final concentration of 20 mM. For better reaction efficiency, 20 µl 1% (by wt) anionic surfactant SDS aqueous solution was added into 100 µl 20 mM diazonium salt solution to form the reactant solution for later graphene functionalization procedures.0.22-gram p-phenylenediamine and 0.35-gram p-aminobenzene sulfonic acid were dissolved in 5 ml deionized water, and then added by 1 ml 50% HBF1H NMR spectra were recorded at room temperature on a Bruker AVANCEIII 400 MHz spectrometer in D2O (for 4-diazoniobenzenesulfonate tetrafluoroborate) or DMSO-d6 (for 4-aminobenzenediazonium tetrafluoroborate), using TMS or solvent peak as a standard. Low-resolution mass spectral analyses were performed with Waters AQUITY UPLCTM/MS.The Graphene was grown on copper foil by the chemical vapor deposition (CVD) method. We firstly cut a piece of graphene-coated copper foil and dropped the diazonium salt solution onto graphene surface, followed by a 30-min incubation at 40 °C. Afterwards, graphene membrane was washed by 1 ml deionized water for 5 times and immersed into deionized water for an additional 0.5–1 h to ensure the complete removal of residual diazonium reagent from graphene surface.2 (11.5 sccm) and Ar (35 sccm) atmosphere for 30 s. We then heated the copper foil to 65 °C, and added a small piece of paraffin (Sigma-Aldrich 18634) onto its functionalized graphene side, which immediately melted and spread to cover the whole foil. After the paraffin spread as a thin film on top of the graphene, the copper foil was cooled down to room temperature and the paraffin solidified. Afterwards, the paraffin-graphene-copper was gently put on the surface of 0.5 M ammonium persulfate (APS) solution with the paraffin side facing up, and incubated for 5 h. After the copper foil was thoroughly etched off, the paraffin-graphene was rinsed with deionized water several times, then transferred into a 40 °C incubator and incubated for at least 1 h, to stretch and diminish wrinkles in the graphene membrane. After that, we used Quantifoil grids  to scoop out the paraffin-coated graphene from the 40 °C deionized water and air-dried the graphene grids in a clean chamber at 40 °C overnight. The paraffin-graphene-grids supported by oil-absorbing paper  were then incubated at 65 °C for 5–10 min to remove most paraffin coating. Afterwards, the grid was soaked into petroleum ether (preheated to 65 °C) and stirred at 60 rpm for 3 h to completely remove the paraffin. Finally, the grids were washed by acetone, isopropyl alcohol (IPA) and water consecutively to clean off residual petroleum ether and air-dried.We firstly removed the unfunctionalized graphene grown on the backside of the copper foil by placing the foil in a Gatan plasma machine (Model 950) with its backside up, and performed the plasma treatment at a O15 as control, to compare and demonstrate the cleanliness of graphene grid fabricated in this work. To better describe the cleanliness of graphene in the holes on EM grid, here we define a hole with more than 5 contamination spots with the area of >400 nm2 as highly contaminated hole, and we found almost all of the holes on the graphene grids fabricated by PMMA-assisted transfer method were highly contaminated. For graphene grid fabricated by paraffin-assisted method, we imaged 141 holes and found 123 holes (~87%) with no or no more than five contamination spots, and 98 holes (~70%) with no or no more than three contamination spots.We used graphene membrane transferred by PMMA-assisted methodX-ray photoelectron spectra was collected on a ESCALAB Xi + (ThermoFisher Scientific), with the monochromatic Al Kα X-ray beam spot of 500 μm. Raman spectra was measured on a LabRAM HR-800 (Horiba) with the 633-nm laser wavelength. For water contact angle measurement, we applied 0.7 μl deionized water onto the grid surface and used OCA15Pro (Dataphysics) to collect the water contact angles. The roughness of the suspended graphene surface was characterized using AFM .Thermoplasma acidophilum 20S proteasome was expressed and purified from Escherichia coli BL21(DE3) strain by following previously reported protocols19. Briefly, we transformed the plasmids into Escherichia coli competent cells, which contained α and β subunits of 20S proteasome, with the N terminal of β subunit tagged by six histidines. After induced expression, Escherichia coli cells were sonicated and the cell lysates were then applied onto affinity nickel column to get purified 20S proteasome sample.Lactococcus lactis strain IL1403 according to the published methods17. We firstly constructed a plasmid containing Ll.LtrB intron with the ORF region in its DIV depleted, its encoded protein LtrA and a chitin binding domain (CBD), which was then transformed into Lactococcus lactis strain IL1403 and allowed for induction. The cell lysates were loaded onto affinity chitin column, followed by dithiothreitol elution and ultracentrifugation to get purified Ll.LtrB RNP sample.Ll.LtrB RNP were purified from E. Coli ribosome was purchased from NEB company (P0763S).For sample vitrification, we firstly pipetted 7 μl sample buffer  onto the functionalized graphene grids, and incubated the grids for 2 min at a chamber of high humidity. Afterwards, the grids were edge-blotted by filter papers to remove the extra buffer, immediately followed by adding 4-μl sample solution, incubated for 2 min and then transferred into Vitrobot Mark IV (ThermoFisher Scientific). The humidity of Vitrobot chamber was set as 100%, and the temperature as 8 °C. The grids were blotted by filter papers (Ted Pella) for 1 s with a force of −2, and then plunge-frozen into liquid ethane. The vitrified specimens were stored in liquid nitrogen. To prepare conventional graphene-supported cryo-specimen, we pipetted 4 μl sample solution onto freshly glow-discharged graphene grid, and blotted the grid for 1~2 s with a force of −2, using Vitrobot Mark IV (ThermoFisher Scientific). After blotting, the grid was frozen into liquid ethane and stored in liquid nitrogen.-/Å2. The movies were motion-corrected by MotionCor220. The CTF values were estimated by CTFFIND421 and the resulted micrographs were imported to Relion3.1 for particle picking and 2D classification22. After discarding junk particles grouped in bad 2D classes, the remaining particles were imported for 3D classification and refinement, to obtain the final 3D reconstructions. The final particle number used for the reconstruction of Ll.LtrB was 399,660, and the resolution was 3.2 Å, estimated by the Fourier Shell Correction (FSC) = 0.143 cutoff criteria. Structural visualization and analysis were performed in UCSF Chimera23.The cryo-EM datasets were collected on a Titan Krios (300 kV) microscope equipped with a Gatan K3 Summit direct electron detector. The pixel size was 0.97 Å and every movie contained 32 frames with a summed dose of 50 e24. We collected 8 frames at each tilt angle, with a summed dose of 3 e-/Å2, which were further motion-corrected by MotionCor220. We used IMOD software25 to align and reconstruct the tomograms.Tilted series were acquired on a Titan Krios (300 kV) microscope equipped with a Gatan K3 Summit direct electron detector, from −60° to +60° with a step of 3° using SerialEMFurther information on research design is available in the Supplementary InformationReporting Summary


Idiopathic pulmonary fibrosis (IPF) is frequently accompanied by comorbidities, with the management of these comorbidities crucial for clinical outcomes. This study investigated the prevalence, incidence, changes over time, and clinical impact of comorbidities in IPF patients, based on nationwide claims data in South Korea.This retrospective cohort study utilised nationwide health claim data in South Korea between 2011 and 2019. Patients with IPF were defined as those with ICD-10 code J84.1 and Rare Intractable Disease code V236 who made at least one claim per year. Patients were classified by sex, age, pirfenidone use and burden of comorbidities, and differences among groups were determined.The yearly prevalence rate of IPF increased from 7.50 to 23.20 per 100,000 people, and the yearly incidence rate increased from 3.56 to 7.91 per 100,000 person-years over time. The most common respiratory comorbidity was chronic obstructive pulmonary disease (37.34%), followed by lung cancer (3.34%), whereas the most common non-respiratory comorbidities were gastro-oesophageal reflux disease (70.83%), dyslipidaemia (62.93%) and hypertension (59.04%). The proportion of some comorbidities differed by sex, age and use of pirfenidone. The proportion of lung cancer was higher in patients treated with pirfenidone, whereas the proportion of anxiety and depression were lower in patients not treated with pirfenidone. Charlson comorbidity index ≥ 4 was associated with increases in hospitalisations and total medical costs.The yearly prevalence and incidence of IPF and comorbidities in Korea increased over time. These comorbidities affected the use of pirfenidone and medical resources.The online version contains supplementary material available at 10.1186/s12890-023-02340-8. Idiopathic pulmonary fibrosis (IPF), defined as a chronic, progressive, fibrotic lung disease of unknown causes, is the most common type of interstitial lung disease . AlthougTreatment options for patients with IPF are often limited, despite the use of antifibrotic agents such as pirfenidone and nintedaninb , 6. BecaThis was a retrospective cohort study based on nationwide health claims data in South Korea. De-identified nationwide health claims data between 2011 and 2019 were collected from the database of the Korean Health Insurance Review and Assessment (HIRA), which includes all health insurance claims from the Korean National Health Insurance System (NHIS) and therefore covers most of the population. The HIRA database presents all information on healthcare utilisation by outpatients and inpatients, including their demographic characteristics, diagnoses, and prescribed medications. Diagnostic codes were determined using the 10th revision of the International Classification of Disease and Related Health Problems (ICD-10). The study was approved by the Institutional Review Board of Asan Medical Center (No. 2020-1000), which waived the requirement for informed consent due to the retrospective nature of the study. The study was performed in accordance with the principles of the Declaration of Helsinki.Patients in the HIRA database were defined as having IPF if they had made at least one claim per year under the ICD-10 code J84.1 of the medical care system. Because patients in South Korea with confirmed IPF were enrolled as the Rare Intractable Diseases (RID), we additionally used RID code V236 to define patients with IPF patients. In South Korea, patients assigned RID code V236 can benefit from reduced medical costs from the NHIS. Hospitals charge the NHIS for the remainder, but if their diagnosis does not meet RID diagnostic criteria, the NHIS refuses to pay these hospital costs. This study therefore assumed that patients were accurately diagnosed at the time of RID registration. Therefore, IPF in the present study was defined as a composite of ICD-10 code J84.1 and RID code V236. The prevalence was estimated by including patients who had made at least one claim per year for IPF. The earliest claim date for IPF was defined as the index date, with incidence calculated based on the year of earliest claim.The demographic characteristics, comorbidities, prescribed pirfenidone, and medical costs of patients with IPF were analysed. Comorbidities were classified as respiratory or non-respiratory. Respiratory comorbidities included COPD, lung cancer, pulmonary embolism, pulmonary hypertension, and obstructive sleep apnoea, whereas non-respiratory comorbidities included GERD, dyslipidaemia, hypertension, diabetes mellitus, ischaemic heart disease, anxiety, depression, and congestive heart failure. The prevalence of comorbidities was assessed at (i) the time of diagnosis, defined as the period from 1 year before to initial diagnosis of IPF; (ii) 1 year after diagnosis, defined as the period from initial diagnosis of IPF to 1 year later; and (iii) 3 years after diagnosis, defined as the period from 1 to 3 years after the initial dignosis of IPF. The occurrence of comorbidities was defined by at least one claim for disease-specific ICD-10 codes , sex, CCI (0–3 vs. ≥ 4) and pirfenidone use, with the latter classified into two groups, those treated with pirfenidone more than 3 months (pirfenidone user group) and those treated for less than 3 months or not treated with pirfenidone (pirfenidone non-user group). In pirfenidone user group, we classified into two groups, those treated with 600 mg oral pirfenidone three times daily for more than 3 months (standard group) and those treated with < 600 mg oral pirfenidone three times daily for more than 3 months (low dose group). Because the cost of pirfenidone has been covered by the NHIS in Korea since October 2015, this classification applied to patients after 2016.p values < 0.05 regarded as statistically significant.Categorical variables are presented as number and percentages, and continuous variables as means and standard deviations. Annual prevalence was calculated as the number of the prevalent cases per 100,000 people. Annual incidence was calculated as the number of newly diagnosed IPF patients per 100,000 person-years. The denominator of prevalence and incidence was based on the annual population census obtained from the Korean Statistical Information Service . PatientThe demographic characteristics of patients with IPF are shown in Table The prevalence of comorbidities was evaluated in the 21,111 patients with IPF by age and sex Tables . The mosClassification by sex showed that, of the respiratory comorbidities, COPD (40.58% vs. 29.32%), lung cancer (4.11% vs. 1.45%), and obstructive sleep apnoea (0.37% vs. 0.16%) were more common in men than in women, whereas pulmonary hypertension (2.43% vs. 1.11%) was more prevalent in women than in men. Evaluation of the non-respiratory comorbidities showed that the rates of diabetes mellitus and ischaemic heart disease were higher in men, whereas GERD, anxiety, depression and congestive heart failure were more frequent in the women group. A higher proportion of patients aged ≥ 70 years had COPD (39.39% vs. 34.20%) and pulmonary embolism (1.92% vs. 1.11%)  and low (CCI 0–3) burdens of comorbidities within 90 and 365 days showed that, within 90 days, patients with high CCI had utilised more resources associated with admission  was the most common respiratory comorbidity, followed by lung cancer (3.34%) and pulmonary embolism (1.60%), whereas the most common non-respiratory comorbidities were GERD 70.83%), dyslipidaemia (62.93%), and hypertension (59.04%). The prevalence rates of these comorbidities were comparable to those in previous studies, although the reported ranges of comorbidity rates were very wide 3%, dysli. When co% was theOf the 12,943 IPF patients since 2016 in this study, 5449 42.10%) had been treated with pirfenidone. This relatively low rate of pirfenidone use may be due to the lack of insurance coverage for this medication prior to October 2015 and/or to our definition of pirfenidone use as treatment with this agent for more than 3 months. A recent real-world cohort study of a Korean population found that only 656 48.77%) of 1345 patients with IPF had been treated with antifibrotic agents, including pirfenidone and nintedanib . This lo.77% of 1.10% had Several studies have evaluated the relationships between comorbidities and clinical outcomes in patients with IPF , 9, 20. The present study had several limitations. First, IPF was defined based on the ICD-10 code at the time of diagnosis. Although actual diagnosis was associated with the operational definition based on ICD-10 code, our use of the latter may overestimate or underestimate the prevalence and incidence rates of IPF and comorbidities . To imprIn conclusion, this study found that the prevalence and incidence rates of IPF in the Korean population increased annually. Many patients with IPF had respiratory and non-respiratory comorbidities, although the prevalence was dependent on sex, age and use of pirfenidone. The prevalence of these comorbidities was higher 3 years after than at the time of IPF diagnosis and affected the use of pirfenidone and medical resources. Additional studies are needed to clarify causal relationships.Additional file 1. International Classification of Diseases, 10th revision codes (ICD-10 codes) for the diseases evaluated in this study.Additional file 2. Prevalence of comorbidities in patients with idiopathic pulmonary fibrosis during the 3 years after initial diagnosis.Additional file 3. Clinical characteristics of patients with idiopathic pulmonary fibrosis based on the pirfenidone dose since 2016.


The burden of cataracts was substantial in the current aging world. However, few epidemiological studies have examined the associations between climate and weather conditions and cataract in older populations. We aimed to investigate the associations of air relative humidity and temperature with cataracts in older adults in China.We used the cohort data from 2002, 2005, 2008, 2011, 2014, and 2018 waves of the Chinese Longitudinal Healthy Longevity Survey (CLHLS). A total of 62,595 Chinese older adults aged between 65 and 105 years were included in the analyses. City-level annual average air humidity and temperature during 2001 and 2017 (before the survey year) was used to measure population exposure. A cataract was self-reports based on the medical record or the doctor's diagnosis and 8,071 older adults had cataract. Covariates included socio-demographic, health status, lifestyles, and chronic conditions. We adopted the Generalized estimation equation (GEE) model to analyze the associations of relative humidity and temperature with cataracts.We found that the average relative humidity  in the past year was inversely associated with cataract likelihoods in older adults and a positive association between temperature  in the past year and cataract likelihoods in older adults. The associations were robust in stratified analyses by sex, urban/rural residence, and education level. Furthermore, we found a nonlinear J-shaped relationship between temperature and cataract prevalence.Our findings provide the evidence that higher temperature and low relative humidity may be associated with cataracts in older adults. Cataract, defined as any opacity of the crystalline lens in the eye that affects clear vision, is a common condition in later life , 2. CataEnvironmental factors-induced adverse health outcomes were well-documented . A largeIn light of the increasing occurrence of extreme climates and extreme temperatures, it is essential to examine their impacts on health, especially that of aging individuals, which will be directly related to future healthy aging and the creation of age-friendly environments. However, epidemiological studies on cataracts in older adults are rare. This study examined the associations of humidity and temperature with cataracts among a nationally representative sample of older adults in China.Data used in this study were derived from the Chinese Longitudinal Healthy Longevity Survey (CLHLS) waves of 2002, 2005, 2008, 2011, 2014, and 2018. The CLHLS applied a multistage, stratified cluster sampling design in 23 out of 31 provinces in China. The goal of CLHLS was to understand better the determinants of healthy longevity in Chinese older adults. Between 1998 and 2018, the CLHLS was conducted in half of the counties and cities (randomly selected) in 23 out of 31 provinces in China. Details of the CLHLS have been described . Hourly data was annually or seasonally averaged to measure residential temperature and relative humidity exposure for each participant in CHLHS, matching by the administrative codes of the cities where the CLHLS samples are resident (non-publicly available information). The temperature and humidity exposures adopted in this study are (1) Average relative humidity over the past year and (2) Average temperature in the past year. To reduce the seasonal bias, we also used (3) Average temperature in the cold months and (4) Average temperature in the warm months to measure the participants' temperature exposure. Similar with the previous publications  and temperature (°C) are derived from the China Meteorological Data Network  for continuous variables and as frequencies and percentages for categorical variables. Associations of relative humidity and temperature with cataracts were analyzed by generalized estimation equation (GEE). We chose GEE because using the observed correlational structure of the data, we can obtain efficient and unbiased regression parameters . We usedA total of 62,595 observations were included in this study. The mean age of the participants was 85.78 ± 11.27 years; the gender composition was generally balanced (44.4% men). The prevalence of cataracts was 12.89%. Nearly half (47.6%) of the older adults lived in urban areas, and nearly half (48.6%) lived in east China. More than half (55.6%) of the participants were illiterate, and about 36.2% of them were married. About 25.4% of the participants have disability in activities of daily living, and 20.9% of them had cognitive impairment .The average relative humidity and temperature are also summarized in The average relative humidity over the past year was negatively correlated with the prevalence of cataracts. In the past year, each 1% rise in average relative humidity was associated with a 1.4% decrease in cataracts . In contrast, the average temperature in the past year was positively related to the risk of cataracts. There was a 4% increase in cataracts with each 1°C increase in average temperature over the past year . Based on the subgroup analyses, we found that the associations were robust across subgroups of cataract risk factors, including sex, urban/rural residence, and level of educational attainment. However, a non-significant effect of average temperature was found for urban population .via following analyses: using the average temperature in the warm months (Model I), the average temperature in the cold months (Model II), the average daily maximum temperature (Model III), and the average daily minimum temperature (Model IV). The results were consistent with our main findings  and 1.04  for annual average relative humidity and temperature, respectively. Furthermore, we found a nonlinear J-shaped relationship between temperature and cataracts prevalence. The associations indicated that extreme heat and low humidity may be linked to a higher incidence of cataracts in older individuals.We found that lower humidity and higher temperatures were associated with an increased risk of cataracts. Although limited epidemiological studies have explored the health effects of humidity, similar findings have been reported in previous studies on other eye diseases. Based on a study of 100,636 participants, relative humidity was negatively related to allergic conjunctivitis , there iRegarding temperature, Miranda et al. reported that senile cataract develops earlier and is more prevalent in warm regions, and that cataract prevalence increases with increasing temperature . AccordiLow humidity may affect cataract prevalence by drying out the airways, resulting in hyperosmolarity, which stimulates nerves to produce reflex responses and may release inflammatory biomarkers . It has 2.5 and NO2) on cataracts. Furthermore, further research from the perspective of molecular biology is needed to investigate the causal relationship between relative humidity and temperature and cataract development.This study contains several limitations. First, relative humidity and temperature are only measured from ambient data at the city level (not from indoors), since personal exposure varies with the ventilation in a house and the movement of people inside the house. Second, except for humidity and temperature, we were unable to assess the possible confounding effects or interactive effects of other environmental exposures  and Natural Science Foundation of Hunan China (2020JJ4087), and the State Laboratory of Sub-tropical Architecture of China (2020ZB10).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


The physical fundamentals for polariton hybridizations depend on the evanescent fields coupling originating from the constituent materials as well as the phase match condition, which can be severely affected by the α-MoO3 thickness and actively tuned by the gate voltages. Hybrid polaritons propagate with in-plane anisotropy that exhibit momentum dispersion characterized by elliptical, hyperboloidal and even flattened iso-frequency contours (IFCs) in the THz range. Our results provide a tunable and flexible anisotropic polariton platform for THz sensing, imaging, and modulation.Electromagnetic field confinement is significant in enhancing light-matter interactions as well as in reducing footprints of photonic devices especially in Terahertz (THz). Polaritons offer a promising platform for the manipulation of light at the deep sub-wavelength scale. However, traditional THz polariton materials lack active tuning and anisotropic propagation simultaneously. In this paper, we design a graphene/α-MoO In THz, the optical response of α-MoOequation : (1)εj=εeference . As showGraphene’s surface conductivity is modeled using the Kubo formula ,22: (2)σe is the electron charge, FE is the Fermi energy, ℏ is the reduced Planck constants, and τ is the momentum relaxation time. The relaxation time of charge carriers in graphene is set to 0.5 ps, which is typical for encapsulated graphene used in optical experiments at ambient conditions. The permittivity of graphene can be written as ε0 is the vacuum permittivity. The real parts of the permittivity tensors of graphene with different Fermi levels are negative in the frequency range from 7 THz to 12 THz  of the real part of the z component of the near-field amplitude Ez. Due to the opposite in-plane permittivity signs of α-MoO3 along [001] and [100] crystal directions, polariton propagation along the [100] direction is forbidden at the low frequency band, which resulted in hyperbolic Ez distribution centered along the crystallographic directions of [001]  and the dispersion of PhPs (without PPs). If there is a crossing point between the dispersions for a given configuration, there will be coupling. Therefore, the graphene Fermi level, α-MoO3 thickness, spectra range, and polariton momenta must be optimized. The permittivity results suggest the potential formation of hybrid plasmon-phonon polariton modes in two excitation bands. We thereby construct a platform for hybridizing isotropic graphene PPs with anisotropic α-MoOlaritons . However3, 3 are not considered in this calculation. The dispersion relation of electromagnetic modes in biaxial α-MoO3 slabs can be described by the following analytical approximations  direction, explaining the canalization of hybrid polaritons along this direction e. When t3 heterostructure can be strongly enhanced, and more importantly, the hybrid polaritons can be actively tuned from elliptic to flattened wavefronts by electrical gating. Moreover, the precise control of light localization is very important in nanophotonics, and the electromagnetic field concentration can be further enhanced by fabricating nanostructures that act as Fabry-Perot resonators. For this purpose, we precisely shaped graphene/α-MoO3 heterostructures into square nanostructures for tailoring electromagnetic field localizations. We adopted a phenomenological cavity model to simulate the near-field amplitude and phase patterns within graphene/α-MoO3 nanostructures [3 nanostructures, the excited polariton wave propagates outward and is reflected by the nanostructure boundaries. The dipole-launched polariton waves interfere with the reflected waves perpendicular to the boundaries. The total field detected can be described as: E0 and p,jE are the dipole-launched polariton wave and that reflected from four boundaries, respectively. Reflected waves are given by R0 and Δφ are the reflectivity and phase shift of the polariton wave, respectively. pγ, jr and pλ represent the polariton damping rate, distance between the nanostructure edge and dipole tip, and the polariton wavelength, respectively. In simulations the amplitude of the dipole-launched polariton wave E0 was set to 1 and the damping rate pγ to 0.4 eV, whereas the polariton wavelength pλ is set based on π was used for the phase shift of the polariton wave [The near-field simulations review found that the hybrid polaritons in graphene/α-MoOructures ,26. A po3 micro-flakes at the cross points in pλ. x and y axes at different wavelengths. Except for the flattened-band dispersion at 10.1 THz, polariton propagation along y axis is forbidden, which results in interference fringes only appearing in the horizonal direction. As the frequency decreases, the number of bright spots decreases and the field localizations are strongest at the regions close to the corner of the nanostructures. The realization of electromagnetic field localization in two dimensions using graphene/α-MoO3 nanostructures can offer several advantages in terms of efficient focusing and manipulating of light at the nanoscale, which can further be integrated with photonic or optoelectronic devices to enhance light-matter interactions. In addition, due to the elliptical-type dispersion, hybrid polaritons propagate along 3 heterostructures. It was found that the polariton hybridization between isotropic graphene PPs and anisotropic α-MoO3 PhPs strongly depends on the α-MoO3 thickness and can be actively tuned by the gate voltages. In addition, the hybrid polaritons propagate with in-plane anisotropy that exhibits momentum dispersion characterized by elliptical, hyperboloidal and even flattened IFCs in the THz range. The near-field results suggest that the localized electromagnetic fields can be enhanced in nanostructures, which provide a flexible anisotropic polariton platform and can further be integrated with photonic crystals or quantum well structures to ulteriorly enhance THz-matter interactions. Our findings demonstrate that active tuning of THz polaritons with certain propagations can be used to steer THz light along specific directions at the nanoscale. More significantly, these results will provide guidelines for THz device fabrications and experiments with dynamically controllable properties.In conclusion, we designed and investigated polariton hybridizations in graphene/α-MoO


In order to maintain the flexibility of PCM without volume shrinkage under the external force, cross-linked graphene aerogel was prepared by the cysteamine vapor method. The cross-linked graphene aerogel had a high stress–strain durability and chemical stability for infiltrating PCM to produce a form-stable PCM composite. The latent heat of PCM is one of the elements to estimate the capacity of PCM thermal energy storage (TES) during the phase transition process. The cross-linked graphene aerogel-supported PCM composite showed a great TES to be utilized in thermal-to-electrical energy harvesting. The cross-linked graphene aerogel also had an excellent mechanical property of preventing damage at a high temperature. The porous materials exhibit internal structures that can infiltrate plenty of solid–liquid working materials into the internal space ,2. That However, one of the recent problems in the graphene aerogel is the volume shrinkage while infiltrating a polymer into the internal space ,16. The It is necessary to utilize graphene aerogel for fabricating form-stable PCM composite due to the leakage problem under the solid–liquid melting process ,30. The In this study, the importance of modified graphene aerogel shows not only the reduction of volume shrinkage but also the prevention of the leakage problem under external force. There are conventional graphene aerogel, PDMS embedded graphene aerogel, and cross-linked graphene aerogel with some typical morphologies, and cross-linked graphene aerogel supported PCM composite shows excellent form-stability under the external force at a high temperature. For the microencapsulation method of fabricating form-stable PCM composite, the replacement of supporting shell material causes a decrease in pure PCM weight fraction, which affects the TES capacity during the phase transition process. The graphene aerogel-supported PCM composite has a high PCM weight fraction and keeps a high TES capacity for utilizing stored energy sufficiently. It indicates that cross-linked graphene aerogel is an optimized supporting material and can be employed to fabricate tremendous form-stable PCM composites.4 oxidized GO had high values in the range of 4–10 pH values, as shown in 0 high-pressure region  that the PEG can be melted at 80 °C. Graphene aerogels need high mechanical properties to sustain the melted PEG in the internal pores. The pure PEG weight fractions of each PEG composite were presented in The fabricated graphene aerogels exhibited high porosities and appropriate mechanical properties. The form-stable test indicates that the graphene aerogels can act as excellent supporting materials to prevent the leakage problem a. In ordIt was demonstrated that the graphene aerogel-supported PEG composites had excellent form stabilities during the melting process. In order to observe the durability of external force at a high temperature, all of the samples were placed on the hot plate under 80 °C. An external force of 2 N was applied, and the resulting deformation of the PEG composites was presented in In this work, we fabricated the graphene aerogel, PDMS embedded graphene aerogel, and cross-linked graphene aerogel (GCA) for 3D porous supporting materials. The modified graphene aerogel, especially the cross-linked GCA sheet, showed a high value of contact angle to infiltrate pure PCM effectively. All the graphene aerogel exhibited nearly 370 of their specific surface area under the BET measurement. High dimensional stability and durability of graphene aerogels are required to infiltrate a large amount of pure PCM, which is over 98%, into the internal space without any volume shrinkage. The modified graphene aerogels exhibited great flexibility to reduce the volume shrinkage effectively. Since the form-stability of PCM composites at high temperatures is essential for applications of PCM. The mechanical properties of the graphene aerogels supported PCM composites were analyzed under the 2N external force during the phase transition process. The cross-linked graphene aerogel could retain the PCM composite without any leakage, even at 80 °C. The cross-linked porous aerogel showed a large potential as a light-weight container for aerospace applications.3), sulfuric acid (H2SO4), potassium permanganate (KMnO4), perchloric acid (HClO4), and citric acid were purchased from Sigma-Aldrich .Polyethylene glycol  was selected as a phase change material (PCM). Graphene nano-platelet , Polydimethylsiloxane (PDMS), nitric acid (HNO3) and potassium permanganate (KMnO4)/perchloric acid (HClO4) solution to oxidize initial GO powders. After removing the excess KMnO4 by adding citric acid, the oxidized GO was purified by centrifugation and freeze-drying process. The oxidized GO aerogel was obtained by mixing with GNP, evaporating solvent, and utilizing the cysteamine vapor method to cause a chemical reaction with GO functional groups [The first step of fabricating porous graphene aerogel was synthesizing graphene oxide (GO). The preparation of GO was followed by the modified Hummers’ method after grinding the graphite ,42. The l groups ,46. As al groups .The graphene oxide (GO) and Graphene nano-platelet (GNP) dispersion abilities were measured by a Zetasizer . The fabricated graphene sheets were put in the contact angle goniometer  to obtain hydrophilic properties. The maximum stress–strain durability of graphene aerogels was observed by a Universal Testing Machine . Field-emission scanning electron microscopy  was used to measure the internal surface structures of graphene aerogels and PCM composites under a 5 kV accelerating voltage. A Brunauer–Emmett–Teller (BET) analyzer  was provided to measure the graphene aerogel internal structures. The intrinsic peaks were confirmed using Fourier-transform infrared spectroscopy  and X-ray diffraction  from 10° to 70° with 3°/min rate (2θ). The phase transition field and latent heat (∆H) of PCM composites were obtained using differential scanning calorimetry  under the range of 15 °C to 90 °C with 10 °C/min. The thermal stability was confirmed by using a thermal gravimetric analyzer .