Components smaller than the wavelength of electromagnetic waves are called meta-atoms. Thermal emission can be controlled by an artificial structure in which these meta-atoms are arranged on the surface. This artificial structure is called a metasurface, and its optical properties are determined by the materials and shapes of the meta-atoms. However, optical devices may require active control of thermal emission. In the present study, we theoretically and numerically analyze a wavelength-selective emitter using a graphene ribbon metasurface. The graphene ribbon metasurface consists of a graphene ribbon array, potassium bromide thin film, and silver substrate. The geometric parameters of the graphene metasurface are determined based on an equivalent circuit model that agrees well with the results of the electromagnetic field analysis . The proposed emitter causes impedance matching depending on the conductivity of the graphene ribbon in a very narrow wavelength range. The conductivity of graphene can be actively controlled by the gate voltage. Therefore, the proposed emitters may realize near-perfect emission with a high quality factor and active controllable switching for various wavelengths. In addition, the quality factor can be changed by adjusting the electron mobility of graphene. The proposed emitter can be used for optical devices such as thermophotovoltaic systems and biosensing. Thermal emission is a spontaneous and continuous photon emission from the thermal reservoir . TherefoA meta-atom is a component smaller than the wavelength of an electromagnetic wave. An artificial material that realizes functions that are difficult with natural materials by arranging meta-atoms on the surface is called a metasurface. In recent years, nano/micro-scale artificial materials called metasurfaces were used to control thermal emission. Controlling light with metasurfaces has the possibility to realize active switching of narrow-band thermal emission. Active switching of narrow-band thermal emission is a key technology with applications in thermophotovoltaic systems ,4, infraNormally, the optical properties of metasurfaces may be determined by their geometrical shape ,14,15. TIn the present study, we propose a graphene ribbon metasurface to design a device for realizing narrow-band emission peak at the target wavelength and active wavelength control. This structure can be designed based on an equivalent circuit model. We computationally demonstrate that the designed graphene ribbon metasurface can exhibit near-perfect narrow-band thermal emission under active switching using electromagnetic wave analysis. In addition, the peak wavelength and intensity of thermal emission can be controlled by adjusting the Fermi energy of graphene.x-direction and extended infinitely in the y-direction. The metallic substrate was sufficiently thick. This structure has period Λ [µm], graphene ribbon width W [µm], and dielectric thickness d [µm]. The optical conductivity of graphene σg [S] is given as [EF [eV] is the Fermi energy of graphene, ω [rad/s] is the angular frequency in vacuum, τ [ps] is the relaxation time of graphene, ħ [m2kg/s] is the reduced Planck constant, e [C] is the electron charge, kB [m2kg/s2K] is the Boltzmann constant, and T [K] is the temperature of graphene. The Fermi energy of graphene can be tuned by applying a gate voltage.given as (1)σg=lnεg is given as [ε0 [F/m] is the permittivity of vacuum and ∆ [nm] is the thickness of the graphene.The permittivity of graphene given as (2)εg=iσY0 [S] is the admittance of free space, Yin [S] is the admittance of graphene ribbon metasurface, Yd [S] is the admittance of the dielectric layer, Ym [S] is the admittance of the metallic substrate, and R1 [Ω], L1 [H], and C1 [F] are the resistance, inductance, and capacitance, respectively, corresponding to the first-order mode of graphene plasmon. In this equivalent circuit model, the influence of higher-order modes of graphene plasmon is negligible compared to its first-order mode [The equivalent circuit model is useful for designing graphene ribbon metasurfaces and elucidating the underlying mechanism of emissivity enhancement . Figure der mode . The mecYin and Y0 are given as follows:YG [S] is the admittance of graphene, n is the refractive index of the dielectric layer, c0 [m/s] is the speed of light in vacuum, and η0 [Ω] is the impedance of the vacuum (=120π). Each coefficient representing the first-order mode of graphene plasmon is given by the following equations in the wavelength range, where the first term on the right side of Equation (1) may be negligible.effε is the average relative permittivity of the upper and lower media of graphene, and in this structure r1 is a function of the fill factor and takes a value corresponding to W/Λ  is given asThus, the conditional expression for admittance matching for a target wavelength Here, λε) can be calculated from the Kirchhoff’s law, i.e., Since the silver (Ag) substrate is opaque, spectral emissivity  gives the form:If it is desirable that the slope of the structural admittance imaginary part is 0,The admittance imaginary part below graphene and the admittance imaginary part of graphene are expressed from Equations (5), (12)–(14) and (17) in the following.Differentiating Equations (19) and (20) and substituting them into Equations (3) and (18), lead toβ as follows:Define Substituting Equations (8) and (16) into the Equation (22), lead toµ [m2/Vs] is the electron mobility of graphene. According to Equations (7) and (10),According to the relation between relaxation time and Fermi energy in graphene,According to Equations (8), (9) and (12),s1 and s2 are safety factor parameters, and Λ is determined by Equations (25) and (26). Moreover, there are two restrictions in determining the structural parameters: (1) the graphene ribbon width must not exceed the period; (2) because of the effectiveness of the equivalent circuit model, the period must not exceed the wavelength. Due to the above restrictions,λt = 6, 8, and 10 µm were designed. A potassium bromide (KBr) layer was used as the dielectric layer, and an Ag substrate was used as the metallic substrate. Here, the phenomenon can be simplified if the refractive index of the dielectric layer is constant. In addition, unintended emission appears when the dielectric layer has the extinction coefficient. Therefore, KBr was used for the dielectric layer, which has a substantially constant refractive index and almost no extinction coefficient in the near-infrared region. The refractive index and dielectric function of KBr was obtained from the tabulated data from Palik’s databook  and f2 [Hz] are the frequencies at which the emissivity is half the peak emission value on the low-frequency side and high-frequency side of the peak frequency, respectively.Based on the equivalent circuit model, narrow-band emitters at target wavelengths databook . The die) method , which ibsorbers ,27. AlthEF = 0.01 eV is nearly zero in each test case, which implies that active thermal emission switching was also successfully achieved. Therefore, the equivalent circuit theory is significantly effective for designing the graphene ribbon metasurface.β. β. The white region indicates absence of data because the constraint conditions in Equations (27) and (28) are not satisfied. The quality factor depends only on β and is independent of the target wavelengths. Therefore, the design is performed using the smallest β among the β satisfying Equations (27) and (28). As the target wavelength increases, the minimum value of β increases due to the constraint of Equation (28). Therefore, the minimum value of β increases as the target wavelength increases, and the quality factor decreases.Next, we investigated the relationship between the quality factor and the parameter x component of the electric field from the x-z plane around the graphene ribbon at the peak wavelength of 10 µm. EF = 0.01 eV, the normal emissivity is almost zero at any wavelength. This is because graphene plasmon cannot be excited in this situation. Hence, the active switching of emissivity becomes possible by adjusting the Fermi energy in the proposed structure.In addition, the electric field distribution is shown to investigate the mechanism of the emissivity peak. Moreover, the dispersion relation is shown in λt = 10 µm, and the Fermi energy of the graphene was 0.5–1.5 times EF = 1.37 eV. As the Fermi energy was reduced, the peak wavelength of the emissivity shifted to a longer wavelength. This is because the resonance wavelength of the graphene plasmon is red-shifted as the Fermi energy decreases. Moreover, as the Fermi energy decreases, the intensity of the emissivity peak decreases. This can be explained using the equivalent circuit model. As the Fermi energy decreases, the conductivity of graphene decreases. Thereby, the resistance and reactance of graphene, and the admittance of graphene, increase. As a result, emissivity decreases because impedance matching is less likely to occur. Since the proposed structure can change the peak wavelength of emissivity by adjusting the Fermi energy of graphene, it can also be used for wavelength control of the thermal emitter without changing the structure. It is important to note that if the Fermi energy is too high for the experiment, the substrate might be destroyed. In this study, we have determined the Fermi energy of graphene based on previous studies with numerical simulations [To investigate the effect of Fermi energy, normal emissivity spectra at several Fermi energies were calculated, as shown in ulations ,33,34, tβ. The region without data is the region that does not satisfy Equations (27) and (28). The quality factor depends only on β and hardly on the electron mobility of graphene (β increases owing to the constraint of Equation (28) as the electron mobility of graphene increases. Therefore, β increases as the electron mobility of graphene increases, and the quality factor decreases. Since the electron mobility of graphene varies depending on the fabrication method, it is important to select the fabrication method according to the desired bandwidth.To investigate the effect of changes in the electron mobility of graphene on the structure, the emission spectra at several electron mobilities were calculated, as shown in graphene b. Similaλt = 10 µm. In W = 0.355 µm, and in d = 1.64 µm. In the proposed structure, there are many parameters that can affect the emissivity. Therefore, In summary, we proposed a graphene ribbon metasurface for active thermal emission switching. Near perfect thermal emission at the targeted wavelengths could be achieved by adjusting the structural parameters, and emissivity switching was achieved by adjusting the Fermi energy of graphene in those structures. In addition, the proposed structure can be easily designed using an equivalent circuit model based on the impedance matching theory. FP cavities are formed in the proposed structure, and graphene plasmons cause an asymmetric FP resonance. The wavelength and intensity of resonance can be changed by adjusting the Fermi energy of graphene. Furthermore, the quality factor may also be affected by the electron mobility of graphene. This study may be applied to the initial design of a structure for active control, and facilitate the development of graphene-based thermal devices.


RF switches, which use a combination of graphene and two-dimensional high-density electron gas (2DEG) in the AlGaN/GaN system, were proposed and studied in the frequency band from 10 MHz to 114.5 GHz. The switches were integrated into the coplanar waveguide, which allows them to be used in any system without the use of, e.g., bonding, flip-chip and other technologies and avoiding the matching problems. The on-state insertion losses for the designed switches were measured to range from 7.4 to 19.4 dB, depending on the frequency and switch design. Although, at frequencies above 70 GHz, the switches were less effective, the switching effect was still evident with an approximately 4 dB on–off ratio. The best switches exhibited rise and fall switching times of ~25 ns and ~17 ns, respectively. The use of such a switch can provide up to 20 MHz of bandwidth in time-modulated systems, which is an outstanding result for such systems. The proposed equivalent circuit describes well the switching characteristics and can be used to design switches with required parameters. Radio frequency (RF) and terahertz dynamically reconfigurable multi-element devices often require electronic switches. Efficient switches are important components for the development of new communication, sensing, imaging, testing, and instrumentation systems. Applications of such switches include switched-beam reconfigurable antennas, polarization switching, multi-band receivers, transceivers, time division duplexing systems, and test circuits with multiple signal paths.There are a number of technologies available for millimeter-wave switches. Most of them employ transistors based on typical semiconductor manufacturing technologies using silicon or III–V semiconductors ,9,10,11.2 RF switch with a 0–50 GHz operating frequency range was reported in [Graphene and transition metal dichalcogenides also attract attention for designing RF and terahertz switches and other devices ,17. A moorted in . Severalorted in ,20,21,22Graphene can be also used to construct nanoelectro-mechanical systems (NEMS). This concept was investigated in ,24,25. TConsidering variable attenuators as a specific kind of switch, one can notice more examples of graphene-based devices. In , a protoGraphene-based modulators should be also mentioned, since they can be based on switches ,31,32,33What is common in the abovementioned publications on graphene-based switches is that graphene is used as an active switching element: a change in the graphene properties under bias allows the RF power to be switched.In this paper, we propose and study an approach wherein the combination of graphene and two-dimensional high-density electron gas (2DEG) in the AlGaN/GaN system allows us to switch effectively at RF and mm-wave frequencies. The switch was integrated into the planar transmission line. A CPW was chosen because it has smaller losses in comparison with the microstrip line. This approach allowed us to avoid the problems with packaging and interconnects, which can cause high losses, especially at mm waves.The proposed hybrid structure includes a CPW with an embedded graphene/AlGaN/GaN switch. A typical CPW consists of two ground conductors and a signal conductor line in the middle. In the proposed structure, the signal conductor line is made of Ti/Al/Ni/Au on the top of the AlGaN/GaN structure. The signal line is interrupted in the middle and the graphene gate device is placed in the gap between two parts of the signal line see . In the As opposed to a conventional transistor-based design, where a metal gate is used, the properties of a graphene gate also change when the voltage is applied between the graphene gate and 2DEG. Therefore, the properties of both the 2DEG and graphene are controlled by the bias. In other words, the 2DEG also can be considered as a “gate” relative to graphene. Since CVD graphene is of p-type conductivity, a positive voltage on the 2DEG relative to graphene reduces the concentration of holes and increases the graphene resistivity . TherefoxGax1−N (x = 66%), 5 nm AlxGax1−N UID (x = 28%), 10 nm AlGaN:Si (n~1.5 × 1018 cm−3), and a 2 nm UID AlGaN layer. The whole heterostructure was covered by a 2 nm GaN cap layer. A schematic diagram of the fabricated heterostructures is shown in We used AlGaN/GaN epitaxial heterostructures grown by Metalorganic Vapor Phase Epitaxy (MOVPE) on a silicon carbide substrate. MOVPE growth was started from a 38-nm-thick AlN nucleation layer on a commercially available 500-µm-thick semi-insulating SiC substrate. The next layer was 2.3 µm high-resistivity (HR) GaN buffer followed by a 0.7 µm unintentionally doped (UID) GaN layer. The AlGaN barrier consisted of: 1.2 nm Al2 atmosphere. Metallization for the ohmic contacts was deposited on the central line and interrupted in the middle. The ground lines were fabricated simultaneously. In order to be able to tune the width of the gap in the central line, the same Ti/Al/Ni/Au metal stack may be deposited at a later time in order to reduce the gap width. This metal stack was not annealed.The CPW processing was performed using a commercial laser writer system for lithography based on a 405 nm wavelength GaN laser source with a minimum 1 µm linewidth. The first step in the processing was 150 nm mesa etching provided by an Inductively Coupled Plasma–Reactive Ion Etching system. As a result of the etching, the 2DEG remained only under the signal line. Then, ohmic contacts were formed by thermal evaporation of Ti(15 nm)/Al(100 nm)/Ni(40 nm)/Au(50 nm) and rapid thermal annealing at 780 °C for 1 min under a NThe last step was graphene transferring and its patterning. A chemical vapor deposition (CVD) graphene layer was deposited on the whole GaN-based wafer by the high-speed electrochemical delamination technique . The det−1 and 2D mode at 2685 cm−1, which are characteristic of the sp2 hybridization of carbon. The full width at half maximum (FWHM) of the 2D band and the intensity ratio of the 2D/G peaks were used to determine the number of graphene layers and their quality. The FWHM of the 2D peak in −1, which is typical for monolayer graphene. The ratio I2D/IG was over 2, which is characteristic of a monolayer of graphene as well. The obtained results confirmed that the graphene sample was of high quality and defect-free. The spectrum in the range from 1050 cm−1 to 1950 cm−1 showed also features stemming from the SiC substrate.During the CPW fabrication, the quality of the graphene layer was controlled with Raman spectroscopy. The optical microscope images of three of the fabricated devices are shown in n = 1.3–1.4. The subthreshold current, which is determined by the gate leakage current, was very small, even for the transistor with the highest gate area, as shown in t = −3.0 to −3.2 V. This means that at Vg < −3.5 V, the channel is fully depleted and the transmission line is interrupted. It is connected only due to the highly resistive graphene layer and fringing capacitances. At zero gate voltage, the central line is connected by the highly conductive 2DEG.The structures shown in On-chip S-parameter measurements of the graphene/AlGaN/GaN switches in the 70.5–114.5 GHz frequency range were carried out using a measurement setup configured as shown in For the calibration and setting of the measurement plane to the contact pads, the SOLR Cascade 101-190C and LRM Cascade 138-357 standard impedance substrates were used at low and high frequencies, respectively. The S-parameter measurements were performed with bias voltages applied between the central lines and ground. The DC connections were provided through the bias ports of the VNA and through the bias ports of the waveguide probes in the low- and high-frequency configurations, respectively. As the graphene layer was connected to the GND conductors a, a posig = −5 V.L = 2Lgap + Lg = 65 μm exhibits insertion loss between 12.5 and 14.9 dB in the on state across a very wide frequency range. On the other hand, the off-state characteristics show the strong frequency dependence of isolation, which is typical for parasitic capacitive coupling. Isolation values decrease from over 77 dB below 100 MHz to 17.5–19.4 dB in the 70.5–114.5 GHz frequency range. Despite poor isolation at frequencies above 70 GHz, the switching effect is still evident, with an on–off ratio of approximately 4 dB.The structure E4 with the large gap in the central line L = 2Lgap + Lg = 20 μm in the E1 and G1 structures allowed us to achieve lower on-state insertion loss (7.4–14.1 dB) at frequencies above 200 MHz. A smaller gap between extended pads has an adverse impact on the isolation due to increased capacitive coupling between the input and output. As a result, at frequencies above ~100 GHz, switching is not effective. E1 and G1 structures have different dimensions of extension pads, which causes their slightly different behavior at low frequencies.Narrowing the gap in the central line to In order to study the switching properties in the time domain, the DC power supplier was replaced with the function generator.The measured switching times are listed in The measured transmission characteristics shown in The graphene gate is represented by resistors R5-R4-R5 arranged according to the gate shape b. The grSimilarly, pad extension with 2DEG underneath constitutes a capacitor with a single resistive plate R1-C1. Values of circuit elements are given in The comparison of measurements and simulations of the S11 and S21 parameters for the G1 structure is presented in The results of simulations shown in Although the switches in more conventional technologies demonstrate better high-frequency performance, this research is in the early stage and there is room for improvement. The advantage of the application of graphene as a gate in a millimeter-wave switch is that its parameters change with bias at the same time as the channel parameters. Therefore, in the off state, with positive bias on the 2DEG relative to the graphene gate, the conductive layers can be eliminated from the gap in the CPW, making the impact of parasitic connection through fringing capacitances and the gate less significant. The main directions of the future development of this type of structure are the optimization of graphene characteristics and switch geometry as well as the evaluation of shunt architecture to achieve lower insertion losses and a higher operating frequency band.We have proposed and studied the design of a switch that uses a combination of graphene and two-dimensional high-density electron gas (2DEG) in an AlGaN/GaN system in order to provide effective switching at RF frequencies. The switch was integrated into the coplanar waveguide, which was chosen due to lower losses compared to the microstrip line, especially at high frequencies. The presented design is an on-chip solution fabricated in one technological process. The proposed equivalent circuit describes well the switching characteristics and can be used to design switches with required parameters. The switching times are sufficiently low to use this kind of switch in time-modulated systems.


Background: Acupuncture shows benefits for patients with melasma, although no optimal number of sessions have been determined.Methods: The prospective observational study was conducted in melasma patients who were treated with acupuncture procedures two times a week and were evaluated after the 5th and the 10th sessions of acupuncture, with a 1-week follow-up after the last session. Participants Groups A and B received five and 10 acupuncture sessions, respectively. Melasma was assessed by using the melanin index (MI), melasma area and severity index (MASI), patient-reported improvement scores, and acupuncture-related adverse events.Results: Out of 113 participants, 67 received five sessions of acupuncture treatment while 39 received 10 sessions. At 1 week after five sessions of acupuncture in Group A, the mean MI decreased by 28.7 , whereas the median MASI decreased by 3.4  points. At 1 week after ten sessions of acupuncture in Group B, the mean MI decreased by 31.3 , whereas the median MASI decreased by 5.4  points. The first five sessions of acupuncture had a higher incremental effect than the last five sessions, although there was no statistically significant difference. Twenty-nine participants reported minor side effects. Group B had a risk ratio (RR) of having adverse events 1.8 times  compared with Group A.Conclusion: Short acupuncture regimens of 5–10 sessions in melasma seem to be effective and practical with minor side effects. What is already known about this subject?- Acupuncture has effectively improved melasma conditions at low cost with minimal side effects.- Traditionally, one acupuncture course comprises 10 sessions for a chronic condition but dose-response evidence of acupuncture for melasma has been lacking.- The number of acupuncture sessions performed in various randomized controlled trials varied from 21 to 60 over 4–16 weeks.What are the new findings?- Five sessions of acupuncture improved the melanin index (MI) by 28.7 and melasma area and severity index (MASI) by 3.4 points.- About 10 sessions of acupuncture improved MI by 31.3 and MASI by 5.4 points, of which the improvement was mainly from the first five sessions.- Short acupuncture regimens of 5–10 sessions in melasma seem to be effective and practical with minor side effects.How might it impact clinical practice in the foreseeable future?- The dose-response study has not been common but could have clinical and financial implications.Melasma is acquired hyperpigmentation characterized by ill-defined light to dark brownish macules and patches, typically affecting photo-exposed areas of the face –3. The mIn the last decade, there was a paradigm shift in the pathophysiology of melasma which is now considered to be a more complex interplay between epidermal and dermal components including the proliferation of dermal vessels and oxidative damage by reactive oxygen species (ROS) , 5. AparAcupuncture has shown benefits for patients with melasma. The effective rate of acupuncture for melasma has been reported to be around 87–96% –15. In CThis prospective observational cohort was conducted at the Acupuncture Clinic, King Chulalongkorn Memorial Hospital, Bangkok, Thailand, from September–December 2018. We recruited all the melasma patients who came to the clinic for acupuncture treatment and were willing to participate in this observational study. We excluded participants who were pregnant, had severe underlying disease or psychosis, had a bleeding tendency, or had skin infections in the acupoints area. All participants were directed to maintain their usual behavior, such as using the same frequency and amount of sunscreen protection or continuing to not use sunscreen regularly.tm, Maanshan Bond Medical Instruments Co., Ltd., China). Acupuncture was done two times a week by well-trained acupuncturists, licensed with acupuncture Master degrees certified from China. The “De Qi” sensation in every acupoint was mandatory, and the needles were retained 30 min each time. Only manual needle-stimulation was used. The participants were evaluated at every fifth session of acupuncture. Since this was an observational study, we used the usual protocol used in our acupuncture clinic. If the first five sessions of acupuncture did not yield satisfactory results and the patients were willing to continue the treatment, we continued the acupuncture treatment for five more sessions. Therefore, participants were divided into two groups: Group A received five acupuncture sessions and Group B received ten acupuncture sessions.All the patients were treated with fixed acupoints: encircling the melasma area , 2, FengThe participants were evaluated at the first visit, every five sessions of acupuncture, and 1 week after their last session.The primary outcome was the change in the melanin index (MI) measured by Mexameter® MX 18  on the melasma affected part of the face. The MI was calculated as the average of a triplicate measurement in the area where the melasma was darkest.The secondary outcomes consisted of the change in the melasma area and severity index (MASI), patient-reported improvement scores, and related adverse events from acupuncture. The MASI was developed by Kimbrough-Green et al.  and assef(Df + Hf) + 0.3Arm(Drm + Hrm) + 0.3Alm(Dlm + Hlm) + 0.1Ac(Dc + Hc)MASI score = 0.3AThe area of involvement is rated from 0 to 6 while the darkness and homogeneity of pigmentation are rated from 0 to 4; thus, the MASI score ranges from 0 (none) to 48 (severe melasma).The subjective improvement scores were measured by the 6-point Likert Scale, from−1 to +4 . The adverse events due to acupuncture were recorded by interviewing the participants and physical examinations by physicians each visit.All participants were also photographed at every evaluation visit. The MI, MASI, and satisfactory scores were evaluated by the same evaluator who has been trained by a dermatologist.t-test or Wilcoxon signed-rank tests as appropriate. Linear regression was used to compare the results between Group A and Group B after adjusting for unequal baselines. Linear regression was also used to see if any baseline factors are affecting the outcome. For Group B, a mixed model with the subject as a random effect was used to compare the result of the first five sessions (1st−5th) and the last five sessions (6th−10th). Pairwise comparison between the first and last five sessions was made using Scheffe's Test. If the data did not meet the assumptions to use the mixed model or linear regression, the nonparametric test was used. The data were analyzed using the Stata/MP 15.1  software package.The baseline characteristics and data at each time point were analyzed by descriptive statistics. The data after treatment compared with the baseline in each group was analyzed by using a Student's paired The study was registered on August 19, 2018, before recruiting the first case, with the Thai Clinical Trials Registry, Identification Number TCTR20180824002.The study was approved by the Institutional Review Board (IRB), Faculty of Medicine, Chulalongkorn University (IRB No.544/2559). Each participant provided written informed consent.One hundred and thirteen melasma patients came for acupuncture treatment during September–December 2018. The baseline characteristics are shown in The average baseline MI for all the participants in the darkest area was 298.9 (SD 79.7). p < 0.001): from 287.4 (SD 64.8) to 262.3 (SD 62.5). At 1 week after completing five sessions, the mean MI decreased by 28.7  to 258.7 (SD 61.4) from the baseline. In Group B, after the fifth session of acupuncture, the MI decreased by 22.5 : from 315.9 (SD 98.5) to 293.4 (SD 90.6). After the 10th session of acupuncture, the MI decreased by 30.3  to 285.6 (SD 94.7) from the baseline. At 1 week after completing 10 sessions of acupuncture, the mean MI decreased by 31.3  to 284.6 (SD 89.2) from the baseline.In Group A, after the 5th session of acupuncture, the MI decreased by 25 .Comparing the efficacy of the first five sessions (1st to 5th) to the last five sessions (6th to 10th) in Group B, the MI decreased by 22.5 (95%CI −35.5 to −9.5) and 7.8 (95% CI −20.5 to 4.8) respectively. However, there was no statistically significant difference between these two periods  . Group B had a higher baseline MASI score than Group A, as shown in In Group A, after completing five sessions of acupuncture, the median change of the MASI score was −2.9  scores. Moreover, at 1 week post-fifth session, the median change of the MASI score compared with the baseline was −3.4  scores. In Group B, after completing five sessions of acupuncture, the median change of the MASI score was −3.3  scores. After completing 10 sessions of acupuncture, the median change of the MASI score compared with the baseline was −5.1  scores. The median change of the MASI score between the 6th−10th sessions was −1.2  scores. At 1 week after completing 10 sessions of acupuncture, the median change of the MASI score was −5.4  scores.p = 0.08).Due to the non-normal distribution of the MASI scores, the Wilcoxon rank-sum test was used to compare the change of the MASI score between the two groups (The frequency distribution of patient-reported improvement scores at each time point is shown in p =0.05) compared with group A. All the participants were willing to continue the treatment despite the adverse events.There were no major adverse events. Of 106 participants, 29 (27.4%) participants reported minor side effects, wherein 14 (20.9%) participants were from group A and 15 (38.5%) participants were from group B. Five patients reported two symptoms: two had hematoma and pain, two had pain and dizziness, and the last had headache and sweating. The other 24 participants had one discomfort symptom. Group B had a risk ratio (RR) of having adverse events 1.8 times  and MI (0.69 points) , 22. SimSince melasma is a chronic recurrent skin disease, the short practical regimen of acupuncture has many positive clinical and financial implications as in other chronic diseases. White et al.  mentioneWe realized that from Chinese medicine theory, one of the important basic theories is syndrome differentiation and treatment according to each syndrome. However, we found that our short fixed regimen of acupoints yielded effective results for the majority of patients, also making the treatment easier to train and perform. The treatment was likely effective because our selected acupoint included encircling and intralesional melasma area, resulting in a local increase of blood flow . This coIn our study, the minor yet frequent adverse events of acupuncture were pain and hematoma in the facial area. Therefore, when conducting acupuncture on the facial area, clinicians should apply pressure after taking off the needles for a longer period. Higher sessions of treatment seemed to have a higher rate of adverse events.This observational study has several limitations. The data was collected pragmatically, based on the clinical practice protocol of our acupuncture clinic. The patients would receive at least five sessions as the standard short regimen and could decide whether they were willing to continue the treatment. Hence, our study could not be generalized in terms of a direct comparison of clinical efficacies between the two groups. However, the findings from our observations could be suggestive of the dose-response relationship of the acupuncture treatment for melasma. Some factors including duration of melasma, prior acupuncture experience, and family history of melasma did not affect the clinical outcome of the treatment according to the regression result. Also, this study had a relatively short follow-up time so further research should be conducted to see how long the effects last.A short acupuncture regimen of 5–10 sessions in melasma seems to be effective and practical with minor side effects.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by the Institutional Review Board (IRB), Faculty of Medicine, Chulalongkorn University. The patients/participants provided their written informed consent to participate in this study.TS, NS, and KP conceived the idea, designed the study, collected and analyzed the data, drafted, and approved the manuscript. All authors contributed to the article and approved the submitted version.This study was funded by the Ratchadapiseksompotch Fund, Faculty of Medicine, Chulalongkorn University, Grant Number RA 060/60 and RA-MF-12/62.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.


Interstitial lung diseases represent a heterogeneous and wide group of diseases in which factors leading to disease initiation and progression are not fully understood. Recent evidence suggests that the lung microbiome might influence the pathogenesis and progression of interstitial lung diseases. In recent years, the utilization of culture-independent methodologies has allowed the identification of complex and dynamic communities of microbes, in patients with interstitial lung diseases. However, the potential mechanisms by which these changes may drive disease pathogenesis and progression are largely unknown. The aim of this review is to discuss the role of the altered lung microbiome in several interstitial lung diseases. Untangling the host–microbiome interaction in the lung and airway of interstitial lung disease patients is a research priority. Thus, lung dysbiosis is a potentially treatable trait across several interstitial lung diseases, and its proper characterization and treatment might be crucial to change the natural history of these diseases and improve outcomes. Interstitial lung diseases (ILDs) represent a heterogeneous group of diseases encompassing more than 200 entities that affect the lung parenchyma with inflammation and/or fibrosis . FactorsUntil recently, the lung has been considered “sterile” because traditional, culture-based techniques are not sufficiently sensitive to isolate and identify a large number of microbes . Culture3–105 bacteria per gram of tissue [Streptococcus, Prevotella, and Veillonella [Over the last two decades, the microbiome has gained popularity in the scientific community. Joshua Lederberg was among the first to introduce the term “microbiome” to highlight the importance of “the ecological community of commensal, symbiotic, and pathogenic microorganisms that share our body space and have been all but ignored as determinants of health and disease” . It soonf tissue . Common llonella ,19. The llonella ,19. Firsllonella . Thirdlyllonella . Finallyllonella ,19. The llonella .A sampling of the lung microbiome should address key procedural and analytical challenges related to potential contamination from the URT when using bronchoalveolar lavage (BAL) or sputum samples . MoreoveIPF, the most common idiopathic fibrotic ILD, is associated with a mean survival of 4 years from diagnosis, if untreated . DespiteThe first ancillary study that analyzed lung microbiota almost 10 years ago employed the 16S rRNA gene sequencing in a heterogeneous group of ILDs patients, including five with IPF . AlthougPrevotella, Veillonella, and Escherichia spp. The presence of Streptococcus spp. or Staphylococcus spp. at baseline was strongly associated with disease progression, defined by a composite outcome including death, acute exacerbation, lung transplant, or relative decline in forced vital capacity (FVC) >10% or diffusing capacity of the lung for carbon monoxide (DLCO) >15%. However, these microorganisms were found in less than half of the study population and, despite their association with a faster progression, a causal relationship with the development of the disease was not established [In the COMET study , 55 BAL ablished . Further−1 of BAL fluid) of IPF patients, compared with healthy controls and COPD patients. Additionally, an increased bacterial load at the time of diagnosis was correlated with a more rapid progression of IPF and a higher risk of mortality (HR 4.59). A reduced microbial diversity was also observed in the IPF cohort with an abundance for Veillonella, Neisseria, Streptococcus, and Haemophilus spp. Finally, a higher bacterial load was also associated with the carriage of the MUC5B s35705950 T allele, the strongest genetic factor for the development of IPF [A prospective study carried out by Molyneaux et al. aimed at elucidating the role of lung microbiota in the pathogenesis and progression of IPF . This stt of IPF .A single-center retrospective study conducted by Takahashi et al. confirmed that an impaired diversity of lung microbiota was implicated in the progression of IPF . An abunPrevotella and Staphylococcus negatively correlated with increased expression of host immune-response-related signaling pathways. These data provided additional evidence that innate immune responses are aberrant in IPF patients and may be modulated by alterations in the microbial community. Moreover, downregulation of the host immune response by inhibition of signaling pathways was associated with worse survival. This was the first experience showing that host–microbiome interactions enhance fibroblast responsiveness and reduce patients’ survival. Further analysis of the host transcriptome demonstrated an apparent host response to the presence of an altered or more abundant microbiome, suggesting that it may act as persistent stimuli for repetitive alveolar injury in IPF [Further studies that explored the correlation between microbiome and lung inflammation and/or fibrosis suggested a direct or synergistic role of the lower airways microbiome in driving alveolar inflammation and fibrogenesis ,46. In ay in IPF .Recently, O’Dwyer et al. employed digital droplet polymerase chain reaction (PCR), a more sensitive measure optimized for low-biomass sample load, to reanalyze patients enrolled in the COMET study who had BAL-derived DNA available for analysis . The autAll the studies mentioned so far evaluated lung microbiota of IPF patients on BAL collected on a single-time point. In an attempt to quantify the burden and communities in the fibrotic interstitium, Kitsios et al. carried out the microbiome in lung explants (MiLEs-IPF) using lung tissue samples . Lower-lRecently, Yin et al., for the first time, analyzed the virome in patients with stable IPF by using next-generation RNA sequencing . The autThe occurrence of an acute exacerbation (AE) complicates the natural history of a sizeable minority of IPF patients, and it is associated with a median survival “following the event” of approximately 4 months . The risCampylobacter spp. and Stenotrophomonas spp., while a significant decrease was found for Veillonella spp. Based on the significant increase in proteobacteria, which are gastric-associated pathogens, in the BAL fluid during an AE, the authors hypothesized a causative role for aspiration in triggering AE-IPF through continuous alveolar epithelial cell injury.Molyneaux et al. investigated changes in BAL microbiota using a cohort of 15 stable patients with IPF and 20 patients experiencing an AE-IPF . The autEvidence on the role of virome in AE-IPF is lacking, although viruses can play a causative role . RecentlHypersensitivity pneumonitis (HP) results from an immune-mediated reaction in genetically predisposed individuals who are exposed to an inhaled antigen . ChronicStaphylococcus was more abundant in patients with CHP, compared with IPF, although this did not translate to worse clinical outcomes. Notably, no association was found between bacterial burden and survival of CHP subjects. This observation is particularly interesting given the growing evidence that bacterial burden is associated with mortality in IPF [In 2020, Invernizzi et al. evaluated the role of microbiota in patients with HP . Moreovey in IPF ,43,45,46Sarcoidosis is a chronic inflammatory disorder triggered by unknown environmental/infectious agents in genetically predisposed hosts . The curAtopobium spp. and Fusobacterium were detected more frequently in sarcoidosis samples. A subsequent study by Clarke et al. tried to characterize further the microbiome of several tissues in sarcoidosis patients by analyzing BAL, lymph node, and spleen [Cladosporium (a ubiquitous and saprobic fungus) in single types of sarcoidosis samples, mostly BAL, but limited concordance across sample types.The first study exploring lung microbiota in sarcoidosis was carried out by Zimmermann et al. . The autd spleen . The autIn conclusion, whether sarcoidosis pathophysiology drives microbiome changes or dysbiosis drives sarcoidosis progression is still unknown. Further studies are needed, in particular analyzing the interaction among microbiome, host immunity, and genetic predisposition in sarcoidosis.The hypothesis that dysbiosis influences disease progression and outcomes in ILDs has paved the way toward the use of long-term antibiotics in these patients. Retrospective studies showed benefits on clinical outcomes in IPF patients treated with prophylactic azithromycin or doxycycline ,66,67. Mp = 0.03).More recently, three RCTs have explored the role of long-term antibiotics on relevant outcomes in patients with IPF ,71,72 T. In the The EME-TIPAC was a double-blind, placebo-controlled, parallel, randomized trial conducted to evaluate the efficacy of co-trimoxazole in patients with moderate and severe IPF . The priplus antimicrobial therapy with either co-trimoxazole or doxycycline [The CleanUP-IPF study was a multicenter, open-label, randomized trial that compared the standard of care vs. standard of care ycycline . The stuIncreasing evidence suggests that an interaction between the altered microbiome and host immunity might lead to the development and progression of ILDs. Moreover, some comorbidities, such as gastroesophageal reflux (GER), may contribute to altering the microbiome composition and burden . Thus, nRetrospective data have shown a modest benefit of medical and/or surgical treatment of GER in ILD patients, although this has not been a consistent finding ,75,76. EAlthough the gut and lungs are anatomically distinct, they are formed from the same embryonic tissue and their mucosal tissues bear commonalities in embryology, structure, and physiology . PotentiPreliminary evidence suggests that the lung microbiome might influence the natural history of ILDs. The potential mechanisms by which these changes may drive disease pathogenesis and progression are largely unknown. Few longitudinal studies have been performed thus far, and the relationship between microbiome and ILDs is largely associative rather than causative, making translation to clinical applications challenging. Recent evidence showed that also the gut microbiome has a profound influence on lung diseases. Enhancement of the immune response is likely the mechanism by which the gut microbiome is capable of impacting lung homeostasis. However, the absence of an accurate animal model of pulmonary fibrosis hampers our understanding of disease pathogenesis and microbiome interaction. Untangling the host–microbiome interaction in ILD patients is a current research priority. Multicentric, prospective observational studies are needed to characterize the lung microbiome and its interaction with the immune response of the host in order to define pheno-endotypes. Lung dysbiosis is a potentially treatable trait across several ILDs. Although results from RCTs showed that modification of microbiome through antibiotic administration did not improve outcomes in ILDs patients, proper characterization of microbiome and identification of phenotypes might be crucial to modify the trajectory of these dreadful diseases.


Such noticeable properties enable it to realize practical electronic and optoelectronic applications. However, contact engineering for large-area MoS2 films has not yet been established, although contact property is directly associated to the device performance. Herein, we introduce graphene-interlayered Ti contacts (graphene/Ti) into large-area MoS2 device arrays using a wet-transfer method. We achieve MoS2 devices with superior electrical and photoelectrical properties using graphene/Ti contacts, with a field-effect mobility of 18.3 cm2/V∙s, on/off current ratio of 3 × 107, responsivity of 850 A/W, and detectivity of 2 × 1012 Jones. This outstanding performance is attributable to a reduction in the Schottky barrier height of the resultant devices, which arises from the decreased work function of graphene induced by the charge transfer from Ti. Our research offers a direction toward large-scale electronic and optoelectronic applications based on 2D materials.Two-dimensional (2D) molybdenum disulfide (MoS From thf q/(kT) . Using tnditions a, which operties b.2 devices by reducing the SBH [This result correlates with output and transfer characterizations b,c. Wang the SBH .2 (4.45 eV). This means that a higher SBH would be formed between pristine graphene and MoS2. In contrast, the work function of graphene on a Ti film appears to be 4.51 eV, which implies that Ti induces an n-type doping effect on graphene [2 (2/Vs) compared with Ti contacted devices, but the contact effect is weaker than the graphene/Ti contacted device. This suggests that the coupling with graphene and low work function metals (Ti), is significantly beneficial to the MoS2 device performance. To unveil why the SBH is reduced with the use of the graphene/Ti contacts in detail, ultraviolet photoelectron spectroscopy (UPS) measurements were carried out . The worgraphene ,25, and phene [2 a,c. Consphene [2 . Its dev2 devices depending on the metal contacts under light illumination at 470 nm (R), defined as R = Iph/(AP), where Iph is the photocurrent, A is the device area, and P is the light power density  is another figure of merit for the sensitivity of photodetectors, which can be expressed as D* = RA1/2/(2qIdark)1/2, where R is the responsivity, A is the device area, q is the electronic charge, and Idark is the dark current. A detectivity of ~2 × 1012 Jones is achieved for the device with graphene/Ti contacts, which is higher than that of the Ti-contacted devices  and sulfur . Each powder was loaded into two separate alumina crucibles; the MoO3 crucible was positioned at the center of a quartz tube, and the sulfur crucible was placed upstream of the MoO3 crucible. A Si/SiO2 substrate was placed downstream of the MoO3 crucible. The tube furnace was heated to 650 °C and then kept for 40 min under 50 sccm of Ar carrier gas at 300 mTorr. After that, the tube furnace was naturally cooled to room temperature with 200 sccm of Ar gas. A graphene film was synthesized on a Cu catalyst  using CVD by a process similar to the MoS2 synthesis. The detailed synthesis procedure is described in our previous report [The MoS2 devices were fabricated using a standard photolithography method. First, MoS2 films were synthesized on a heavily p-doped Si substrate with a 300 nm-thick SiO2 layer, which acted as the bottom gate and dielectric layer, respectively. To prepare the patterned graphene, we used standard photolithography and an oxygen plasma etching system. Once graphene was patterned, the graphene electrodes were transferred onto the MoS2 film using a wet-transfer method, where a buffered oxide etch solution was used to etch the SiO2 layer. Then, secondary photolithography was performed on the graphene electrodes, followed by metal deposition (50 nm-thick Ti and then 10 nm-thick Au). Channel definition was conducted with a length of 14 μm and width of 40 μm using a third photolithography step, and then an unprotected MoS2 area was etched using Ar plasma treatment. For comparison, a MoS2 device with Ti contacts was prepared by identical processes except for preparation of the graphene electrodes.MoS2 films grown by CVD. To explore the work functions of the MoS2 and graphene films, UPS  measurements were carried out. To characterize the electrical and photoelectrical properties of the resultant devices, we used a semiconducting parameter analyzer  under vacuum (10−5 mbar) at 80–300 K and a visible light lamp (470 nm).We performed optical microscopy , AFM , and Raman and PL spectroscopy  measurements to investigate the physical properties of the MoS2 device arrays with graphene/Ti contacts via a wet-transfer method. Using the graphene/Ti contacts, we achieved remarkable electrical and photoelectrical performances compared with the Ti-contacted devices, including a field-effect mobility of 18.3 cm2/V∙s, on/off current ratio of 3.1 × 107, responsivity of 850 A/W, and detectivity of 2 × 1012 Jones. Such enhancements can be explained by the reduced SBHs coming from a decrease in the work function of the graphene interlayer induced by electron charge transfer from the low-work-function Ti metal. This interaction is more beneficial to boost device performances, compared with only graphene-contacted devices. Our contact engineering enables easy expansion of the doping range of graphene using various metal contacts, and directly combines high-k dielectric layers [In conclusion, we fabricated CVD-grown MoSc layers , achievic layers , thanks


Cryo-electron microscopy (Cryo-EM) has become a routine technology for resolving the structure of biological macromolecules due to the resolution revolution in recent years. The specimens are typically prepared in a very thin layer of vitrified ice suspending in the holes of the perforated amorphous carbon film. However, the samples prepared by directly applying to the conventional support membranes may suffer from partial or complete denaturation caused by sticking to the air–water interface (AWI). With the application in materials, graphene has also been used recently to improve frozen sample preparation instead of a suspended conventional amorphous thin carbon. It has been proven that graphene or graphene oxide and various chemical modifications on its surface can effectively prevent particles from adsorbing to the AWI, which improves the dispersion, adsorbed number, and orientation preference of frozen particles in the ice layer. Their excellent properties and thinner thickness can significantly reduce the background noise, allowing high-resolution three-dimensional reconstructions using a minimum data set. The cryo-electron microscopy (Cryo-EM) technique began in the early 1980s as samples can be embedded in vitrified ice by plunge freezing which was developed by Jacques Dubochet ,3,4. In Although it has been recently reported that the cryo-EM single-particle technique can achieve a resolution as high as 1.22 Å ; this maThe commonly used cryo-EM grid for better automatic data acquisition is a perforated carbon support foil with pre-defined hole size, shape, and regular arrangement, such as Quantifoil grids. One can also use a homemade carbon-coated holey grid with variable hole sizes and bar width. Ordinarily, the coated holey amorphous carbon film is not flat and has a non-uniform thickness . Its surMeanwhile, although amorphous carbon films have the semiconducting property, their electrical conductivity and mechanical strength can be largely decreased due to the manufacturing process . When thUsually, appending an additional ultra-thin carbon layer above the holey carbon film can primarily circumvent the uneven ice thickness caused by the conventional EM grid A and impAs a novel and promising material, graphene emerged for developing nanocomposites around 2010 ,38. The Unlike other sheet crystals, monolayer graphene can still exhibit a relatively high crystalline quality and continuity even in the form of single atomic layers. Graphene is a hexagonally arranged honeycomb-like planar sheet of carbon atoms with adjacent carbon atom spacing of about 0.142 nm, rich in σ-bonds within its planar surface. These σ-bonds contribute to most of the structural stability of graphene. Some unhybridized π-bonds are also prominent outside the graphene carbon planes, which determine graphene’s electrical, thermal, and optical properties . Due to Besides its good mechanical strength and electrical conductivity, monolayer graphene with only 0.34 nm thickness can also significantly minimize electron scattering in transmission electron microscopy. With all these favorable properties, graphene has been applied to cryo-EM as a sample support film to replace the amorphous carbon film, which has already achieved some success and important breakthroughs ,46. HoweThe preparation of graphene-covered grid mainly contains two steps: obtain the high-quality monolayer graphene and transfer it to the grid which is already coated with a holey film made of amorphous carbon or other materials . Since t3 to etch away the copper substrate  on the gold grid to graphene by solvent wetting, and then floats the grid-graphene-copper sandwich in FeClubstrate A 34]. A. A3 to eubstrate B. The enubstrate A. The reubstrate C. The prEven though graphene has many outstanding properties, it should first be made hydrophilic to be usable for biological samples. Besides directly transferring samples to the graphene in the solution , an UV/oGraphene oxide (GO) is a hydrophilic derivative of graphene and is thus considered a near-ideal material for cryo-EM sample preparation. It can be exfoliated from graphite oxide (produced according to the Hummers method ) by soni2 and O2 mixed gas) EM grids with ~1 μm hole size for ~1 min (drop-casting) [2O in a 5:1 ratio helps larger monolayer GO flakes deposit on the EM grids, which also facilitate to reduce the preferred orientation of protein samples in cryo-EM specimen preparation [The next step is to transfer the GO single-layer flakes to EM grid. One of the methods allows directly applying a 4 μL drop of exfoliated GO solution to the glow discharged (using Hcasting) . The GO casting)  and thencasting) . The exccasting) . For concasting) . Therefocasting) . It is fparation ,66.2O mixture enable to enrich particles by adsorption, which is meaningful for samples that are challenging to obtain. More importantly, it may also help to resolve the preferred orientation problem [Depending on the property of the individual protein sample, the behavior in the suspended ice layer varies from protein to protein. Some proteins tend to favorably adsorb on the carbon film rather than distribute in the vitreous ice spanning the holes ,68. In t problem ,65,66.For achieving a high-resolution structure, the vitreous ice in which biological molecules are embedded should be evenly thin to give better contrast and hence a lower defocus range for data acquisition. As a result, the molecules are inevitably exposed to the air at a high surface-to-volume ratio. The air–water interface (AWI) has been considered unfavorable for proteins to maintain their native conformation ,71,72,73To avoid the adverse effects of the AWI on proteins, the following precautions can be considered : (1) staDespite the many excellent properties of graphene and GO support films in frozen sample preparation, some other chemical modifications on their surfaces have been designed to further solve particular problems encountered even with these films applied. Naydenova et al. developed a low-energy plasma surface modification system to allow different covalent functionalizations  across the graphene layer on a single gold grid . The regAlthough the graphene or GO support has many excellent properties, whether to use them depends on the target protein or complex. Many specimens exhibiting stability and that monodisperse in the ice layer have been reconstructed at a high resolution just using regular perforated amorphous carbon film. However, for those problematic specimens that are prone to sticking to the AWI or poor distribution in the holes, the developed graphene or GO film suspended over holes could be the optimal and unique solution with minimal background noise added. In graphene/GO film fabrication, the single-layer, high coverage, easy to operate, and reproducibility should be the most considerable aspects. A higher monolayer coverage is the most crucial factor to the small proteins with molecular weight lower than 100 kDa. Compared with graphene films, GO films may be more widely used because they are naturally hydrophilic and do not require additional hydrophilization, a step that is very important for naturally hydrophobic graphene films. For graphene films, incomplete hydrophilization may affect the uniformity of ice thickness as well as particle dispersion. In addition, whether additional expensive equipment is needed may also be a consideration for the local implementation for both graphene and GO films.For some proteins, adding graphene or GO membrane alone cannot prevent their adsorption to AWI. Thus, it may be required for specific chemical modification on the graphene or GO membrane surface to help the particles completely avoid denaturation or aggregation. In addition, changing the grid material, such as replacing the copper mesh with a gold mesh or replacing the amorphous carbon film-covered gold mesh with a gold foil-covered gold mesh, may be more helpful to reduce the sample movement and further improve the resolution.


This demonstrated that this method is a promising route to enhance the heat dissipation capacity of graphene-based TIMs for practical electronic cooling applications.Given the thermal management problem aroused by increasing power densities of electronic components in the system, graphene-based papers have raised considerable interest for applications as thermal interface materials (TIMs) to solve interfacial heat transfer issues. Significant research efforts have focused on enhancing the through-plane thermal conductivity of graphene paper; however, for practical thermal management applications, reducing the thermal contact resistance between graphene paper and the mating surface is also a challenge to be addressed. Here, a strategy aimed at reducing the thermal contact resistance between graphene paper and the mating surface to realize enhanced heat dissipation was demonstrated. For this, graphene paper was decorated with polydopamine EGaIn nanocapsules using a facile dip-coating process. In practical TIM application, there was a decrease in the thermal contact resistance between the TIMs and mating surface after decoration (from 46 to 15 K mm However, it can be difficult for conventional TIMs to meet the ever-increasing heat dissipation requirement with the continuous improvement of the microelectronics industry.Along with the integration and miniaturization of electronics devices, the thermal management issue caused by the increasing power densities of electronic components is an important area of research ,2,3,4,5.nterface . To addrnterface ,11,12,13ackaging . Thus, ih demand . Current−1 K−1) and advantages for mass production [−1 K−1 [2 W−1). This results in the thermal contact resistance dominating the thermal interface resistance of the TIM (47 K mm2 W−1 of 61 K mm2 W−1). Hence, reducing the thermal contact resistance between graphene paper and the mating surface is an urgent issue for practical thermal management applications [Graphene, a honeycomb two-dimensional material, possesses good thermal and electrical property ,16,17. Toduction ,19,20, goduction ,27,28,29 [−1 K−1 . Nan et  [−1 K−1 . Accordi [−1 K−1 , we founications .2 W−1) under 500 kPa compression. Finally, practical application of the TIM for LED heat dissipation also demonstrated the superior performance of decorated graphene paper for cooling electronic devices.Liquid metal alloys, for instance, EGaIn , have been studied for thermal management because of their high thermal conductivity, low thermal resistance, low viscosity, and low toxicity ,32,33,34Graphene sheets were fabricated from graphite powder  via an intercalation and exfoliation method. Characterization of the graphene sheets was carried out in our previous work . EGaIn (−1). Then, a tip sonicator  was applied to fragment the EGaIn droplet in the solution. The sonication time was 60 min with an applied power of 500 W at a temperature of 298 K. After sonication, we added 1.5 mmol tris base into the suspension to provide a weak alkaline environment (pH = 8.5) for the self-polymerization of dopamine. Step 2: We fabricated the graphene paper via a facile filtration process. First, 500 mg of graphene sheets were uniformly dispersed in ethanol via ultrasonication, and then, the dispersion was filtered to obtain the graphene paper using a Teflon filter membrane with the pore size of 0.22 μm. The graphene paper was further decorated by the suspension from step one via a dip-coating method for 6 h, which was followed by sequentially washing with DI water and ethanol.Step 1: We fabricated PDA-coated EGaIn (PDA-EGaIn) nanocapsules via a facile sonication method, as follows. First, a 1.5 g EGaIn drop was placed in a beaker with 150 mL aqueous dopamine hydrochloride solution  was measured using LFA 467 HyperFlash® system  and specific heat capacity (pC) was evaluated using a differential scanning calorimeter (DSC) . An infrared camera  was used to capture IR images.The morphology of the PDA-EGaIn nanocapsules, graphene sheets, and DGP were investigated via scanning electronic microscopy . The core–shell structure of the PDA-EGaIn nanocapsules was further characterized by Fourier infrared spectroscopy  and transmission electron microscopy . High-angle annular dark-field and energy-dispersive X-Ray spectroscopy (EDS) were carried out using the scanning transmission electron microscopy (STEM) mode of the TEM. The thermal conductivity of the graphene paper can be calculated by the equation −1 (stretching vibration of the aromatic ring and bending vibration of N–H), 1505 cm−1 (shearing vibration of N–H), and 1291 cm−1 (stretching vibration of phenolic C–O) indicate the presence of PDA on the nanodroplet [The morphology of the as-prepared PDA-EGaIn nanocapsules was investigated via scanning electron microscopy (SEM), as shown in odroplet ,45. The 2) and BLT of 250 μm were applied as TIM packaged between the heat source and the heat sink under a packaging pressure of 500 kPa, where heat generated by this measurement system was extracted via the continuous pumping of cooling water through the liquid-cooled Al heat sink to reach an equilibrium state (measure) was recorded using a thermocouple, and the results are shown in −1, respectively, indicating that DGP achieved a 26% enhancement of cooling efficiency compared with GP.The fabricated PDA-EGaIn nanocapsules were used to decorate the surface of graphene paper TIM via a facile dip-coating method to reduce the thermal contact resistance between the graphene paper and mating surface. um state . The reac) between graphene paper before and after decoration, a commercial computational fluid dynamics software (Icepak) was used for in-depth thermal analysis of our test system at an operating power of 20 W, the detailed parameter settings and simulation model of which are shown in efκf) of GP and DGP from the simulation results were 2.8 and 4.3 W m−1 K−1, respectively, as shown in Rc = TIM − RRBulk= BLT/effκ − BLT/bulkκ, where bulkκ is the intrinsic thermal conductivity of graphene paper under a compression of 500 kPa [−1 K−1, which gives a low thermal contact resistance for the DGP of 15 K mm2 W−1. Under the same conditions, the thermal contact resistance of GP was 46 mm2 W−1, which is approximately three times higher than that of DGP, indicating that the decoration process significantly decreased the thermal contact resistance between graphene paper and the mating surface. To reveal the change in the thermal contact resistance (R 500 kPa . The sch2) and BLT (250 μm) were physically fastened between the Al heat sink and LED chip using four screws. Heat generated by the operating LED was extracted by the air-cooling system to reach an equilibrium state. Real-time evolution of the LED temperature was recorded using a calibrated infrared (IR) camera, as shown in Given the practical application of TIMs, the different heat dissipation capacities of GP and DGP were investigated for a high-power light emitting diode . As shown in 2 W−1) between DGP and the mating surface compared with GP (46 K mm2 W−1). The decrease in the thermal contact resistance between the decorated TIM and mating surface was more remarkable at low compression pressures. The clear enhancement in the heat dissipation performance after the TIM is decorated with PDA-EGaIn nanocapsules indicates that this method is promising for improving the heat dissipation capacity of graphene-based TIMs for practical electronic cooling applications.A graphene paper fabricated via a facile filtration method was decorated with PDA-EGaIn nanocapsules, which were prepared via sonication of bulk EGaIn in an aqueous dopamine hydrochloride solution. The dopamine served as a surfactant to stabilize the EGaIn nanodroplets during the sonication process and also served to immobilize the EGaIn nanocapsules on the graphene paper surface. The decorated graphene paper was applied as a TIM and demonstrated a 26% enhancement in cooling efficiency compared with graphene paper. The simulation results indicated that this enhancement was because of the lower thermal contact resistance (15 K mm


Azadirachta indica (neem) is an evergreen tree belonging to the family of Meliaceae. Multiple studies have been confirmed the anti-diabetic and anti-hypertension, anti-hyperlipidemia, and anti-obesity effects of neem. In this review, we reported the protective effects of neem against the complications of metabolic syndrome with a special focus on mechanisms that are involved. It has been shown that neem can control hyperglycemia and hypertension through over-expression of transcription factor nuclear factor erythroid 2–related factor 2 (Nrf2) and anti-oxidant effects. Neem also reduced the glucose uptake through up-regulation of glucose transporter 4 (GLUT4) and inhibition of key intestinal enzymes such as glucosidases. Moreover, neem showed anti-hypertensive effects possibility via the block of calcium channels, up-regulation of endothelial nitric oxide synthase (eNOS), and extracellular signal-regulated kinases 1/2 (ERK1/2) signaling pathway. Anti-oxidant effects play an important role in protective mechanisms of neem against metabolic syndrome and its complications.Metabolic syndrome is a condition associated with obesity, diabetes, dyslipidemia, and high blood pressure. Recently, the use of phytochemicals is suggested in the control and treatment of metabolic syndrome. The  Metabolic syndrome (MetS) is a common metabolic disorder that is described for more than multiple decades. The MetS is also known as insulin resistance syndrome and syndrome X (1). Physical inactivity, smoking, increasing age, obesity, and positive family history are risk factors associated with its development . EpidemiMoreover, several studies have been shown other side effects of MetS such as fatty liver disease , cirrhosCapsicum annuum L.  is an evergreen tree of southeastern Asia that is widely distributed in the Indian subcontinent. The height of this tree is approximately 15-20 m and sometimes even up to 35-40 m. The word A. indica was derived from the Persian language. The Azad means “free,” and the dirakht is meaning “tree,” and “I” refer to “Indian origin.” . Effects of several plants investigated on BP such as loe vera  and Capsnnuum L. . High BP high BP ; (2) the high BP ; (3) Nit high BP ; (4) Nuc high BP . Several high BP , up-regu high BP  and Nrf2 high BP . It has  high BP . The dow high BP . L-NAME  high BP . The met high BP . The cru high BP . Neem restored anti-oxidant enzyme activity, including superoxide dismutase (SOD), glutathione peroxidase (GPx), and glutathione S-transferase (GSTs) in animal models of high BP. Neem also improved glutathione (GSH) and reduced malondialdehyde (MDA) and protein carbonyl (markers of oxidative stress) levels , 32. TheA study has been reported on the effect of neem leaves on high BP of the 90 diabetic patients aged 40-60, which were kept under observation for a month. During the study, patients received 2 g powder of neem daily for three months. A significant reduction was observed in the BP of treated patients .Effects of neem on hyperlipidemiaBerberis vulgaris)    and rosecinalis) . Plasma cinalis) . Hyperlicinalis) . Anti-oxcinalis) . The elecinalis) , 41. Thecinalis) . The meacinalis) . As mentDifferent doses of neem  in streptozotocin (STZ)-diabetic rats decreased serum TC, TG, LDL, VLDL levels and increased serum HDL levels -48. AlsoEffects of neem on obesityObesity and overweight are a serious health problem that is increasing worldwide. Obesity is associated with a life expectancy decrease and a significant increase in mortality . Stress,in vitro system (The aqueous and methanolic extract of stem bark and roots of neem (520 µg/ml) inhibited pancreatic lipase and α- glucosidase in an o system . But, tho system . There aEffects of neem on diabetes Vernonia amygdalina , Nigellasativa L. (Vitis vinifera) ( Allium sativum (garlic)  , and All(garlic)  which ar(garlic) , Pakista(garlic) , Bengal (garlic) , Indones(garlic) , and Nor(garlic) . Glucago(garlic) , 85. Inh(garlic) . In this(garlic) .• Effects of neem in diabetic humanNeem is available as a dietary supplement in an herbal mixture in North America. Treatment with this dietary supplement (2 capsules 3 times per day) for 3 months in type 2 diabetic patients (the ages of 18 and 70) improved glucose control and HbA1c levels . The stu• Effects of neem in alloxan/streptozotocin-induced diabetic animals•• Effects of neem in alloxan-induced diabetic ratsGynura procumbens ethanolic (112.5 mg/kg) extracts (2 times a day for 15 days) increased insulin expression, decreased blood glucose concentration, and improved the morphology of the islets of Langerhans and beta-cells in rats  and inhibits its function. The intraperitoneal injection of alloxan  is a conventional method for the induction of diabetes in rat models . The ora in rats . The aqu in rats , 93. Pol in rats . PHFs us in rats , 95. In •• Effects of neem in alloxan-induced diabetic rabbitsThe hypoglycemic effect of neem  was observed in rabbits that were treated with alloxan  . Also, l•• Effects of neem in streptozotocin-induced diabetic rats.Streptozotocin (STZ) is one of the most diabetogenic agents using in diabetes research. Its mechanisms for the induction of diabetes are inhibition of insulin secretion and the death of the beta-cells . In rat •• Effects of neem in streptozotocin-induced diabetic miceThe decrease of serum glucose levels and increase of glycogen content, plasma insulin, and c-peptide levels with aqueous extract of neem have been shown in mice treated with STZ , 111. Al• Effects of neem in glucose-induced diabetic animalsThe aqueous and ethanolic extracts of neem leaves decreased glucose level and increased insulin secretion in rats were treated with glucose  , 113. AlA. indica (neem) is effective in MetS and anti-oxidant effects appear to play an important role in protective mechanisms of neem against MetS and the complications associated with it. Neem increases the expression of Nrf2-mediated anti-oxidant enzymes and can regulate blood pressure and lipid profile. Also, neem inhibits vascular smooth muscle contraction through the block of calcium channels and decreases high blood pressure. Neem up-regulates eNOS expression as a vasodilator and increases NO level. Moreover, neem reduces vasoconstriction through the regulation of the ERK1/2 signaling pathway. In the diabetic condition, neem up-regulates GLUT4 and reduces the glucose uptake. Neem also inhibits intestinal enzymes such as glucosidases. Understanding the signaling pathways help to expand the use of neem in the treatment of the MetS. However, few studies have been conducted to investigate the anti-diabetic, anti-hypertension, anti-hyperlipidemia, and anti-obesity activities of neem in humans. Therefore, further clinical studies are needed to assess the protective effects of neem.In summary,


Having suitable animal models to study the mechanisms of TDP‐43 aggregation is crucial to develop treatments against disease. We have previously demonstrated that the killifish Nothobranchius furzeri offers the advantage of being the shortest‐lived vertebrate with a clear aging phenotype. Here, we show that the two N. furzeri paralogs of TDP‐43 share high sequence homology with the human protein and recapitulate its cellular and biophysical behavior. During aging, N. furzeri TDP‐43 spontaneously forms insoluble intracellular aggregates with amyloid characteristics and colocalizes with stress granules. Our results propose this organism as a valuable new model of TDP‐43‐related pathologies making it a powerful tool for the study of disease mechanism.Frontotemporal dementia and amyotrophic lateral sclerosis are fatal and incurable neurodegenerative diseases linked to the pathological aggregation of the TDP‐43 protein. This is an essential DNA/RNA N. furzeri associates with stress granules, acquires a doughnut‐like nuclear distribution and forms aggregates exclusively in old animals, mimicking what is observed in amyotrophic lateral schlerosis and other TDP‐43‐related diseases.Our findings demonstrate that TDP‐43 from the fish  ALSamyotrophic lateral sclerosisBSABovine serum albuminCDcircular dichroismFTDfrontotemporal dementiaG3BPGTPase‐activating protein‐binding protein 1IGTPIsopropyl β‐d‐1‐thiogalactopyranosidePBSphosphate buffer solutionRRMRNA recognition motifTDP‐43TAR DNA‐binding protein 43 kDaSDSsodium dodecyl sulphateSGstress granuleTEVTabacco etch virusThTthioflavinTIA1Cytotoxic Granule Associated RNA Binding Protein1TAR DNA‐binding protein 43 kDa (TDP‐43) is an RNA‐binding protein involved in RNA metabolism  and followed by an unstructured C‐terminal tail. Until recently, this region was thought to be the hotspot for protein aggregation because it hosts most of the pathologic mutations . We then demonstrated that the N. furzeri TDP‐43 paralogs are able to aggregate spontaneously in the intact animal producing inclusions that are similar to those observed in humans. The process is aging‐related as it was detected only in old animals. Finally, we showed that the N. furzeri TDP‐43 proteins colocalize in the fish with the stress granule‐contained G3BP, indicating that also N. furzeri TDP‐43 is able to segregate in SGs.In the present study, we explored the suitability of N. furzeri is a unique model system able to recapitulate most of the in vitro and cellular phenotype observed in TDP‐43 human diseases. This evidence opens the possibility to use N. furzeri in ALS studies and provides new insights into the RNA‐to‐TDP‐43 functional relationship.Taken together, our data show that 22.1N. furzeri and human TDP‐43: Nfu_TDP‐43 and Nfu_TDP‐43L share 80% identity with each other, and 75% and 77% identity with Hsa_TDP‐43, respectively  score  resulted soluble in bacteria and could be cleaved and purified from the SUMO tag although with tiny yields. Conversely, Hsa_TDP‐43 could only be expressed in inclusion bodies as previously reported by independent studies  detected at 485 nm. Since, however, all constructs resulted anyway little soluble demonstrating a strong intrinsic tendency to aggregation, we carried out the assays in two different ways. We first used the fusion proteins without cleaving the constructs from the tag. Under these conditions, the fluorescent signal of all three proteins increased steeply within the first 2–3 h indicating fiber formation Figure . After tN. furzeri proteins in full agreement with predictions. However, the possibility to produce the C‐terminus in a soluble recombinant form using the N. furzeri sequences opens new avenues for the study of TDP‐43 aggregation.These data confirm an important role of the C‐terminus in protein aggregation also for the 2.3N. furzeri paralogs did not reach a clear plateau within the time of the experiment (3 days). Apart from these minimal differences, the curves exhibited similar shapes with the intensity of the human protein being noticeably higher than that of the N. furzeri constructs at comparable concentrations. No difference was observed when the assays were repeated in the presence of salt (15 mM KCl) (data not shown).Similar experiments were carried out on constructs containing the two isolated tandem RRM domains (RRM1‐2) to assess the contribution to aggregation of regions of the proteins other than the C‐termini and, specifically, of the RNA‐binding domains. Not unexpectedly, RRM1‐2 proved to be more soluble than the C‐termini and could be efficiently produced in appreciable quantities and high purity Figure . OverallThese results confirm an involvement in aggregation of the RNA‐binding domains as already observed for the human protein  were measured immediately after the final gel filtration step of purification  spectra of purified n Figure . The spen Figure  and withn Figure . When thThermal denaturation was then recorded following the intensity variations of the band at 222 nm as a function of temperature Figure . The curThe three constructs have thus a similar behavior also in terms of a α‐to‐β irreversible transition of the RRM domains triggered by heat destabilization, although with minor differences in their resilience against aggregation.2.5catRAPID signature . We found 550 transcripts predicted to interact with the human protein. Nfu_TDP‐43L was predicted to interact with 936 human transcripts  of which 550 were in common with Hsa_TDP‐43. Nfu_TDP‐43 was predicted to interact with the same 550 human RNAs but, in addition, with 719 additional targets. As a control, N. furzeri methyltransferase was predicted to interact with just 109 targets of which only 21 sequences being in common with Hsa_TDP‐43.s Figure . Two regN. furzeri and H. sapiens TDP‐43 transcriptomes is significant . Analysis of the motifs present in the top 100 RNA targets shared between Nfu_TDP‐43L and Hsa_TDP‐43 indicated an enrichment of GU‐rich sequences   ≈ 0.60 for Nfu_TDP‐43L, 550/(719 + 550) ≈ 0.45 for Nfu_TDP‐43, and 21/(550 + 109) ≈ 0.05 for the control. This tells us that Hsa_TDP‐43 and Nfu_TDP‐43L have more similar propensities than Nfu_TDP‐43 Figure . A FisheN. furzeri RRM1‐2 constructs. We have previously demonstrated that an aptamer (RNA12), known to bind human RRM1‐2 with nanomolar affinities  intercalated between cells with a normal diffuse distribution of the protein (red arrowheads) could be detected only in the old tissue. The progressive plane images taken at a distance of 1 µm (Z1 to 4) underlined the tridimentionality of the perinuclear pathological distribution of the protein around the nuclei (with an average diameter of 5 µm) of the pathological cells Figure . This evN. furzeri brains, by using a Sca/e S immunofluorescence‐optimized methodology (AbSca/e) . As evidenced by the white circles around the doughnut‐like cell cores . It is also an animal that presents clear signs of aging: old fishes appear emaciated, with their spine curved and almost discolored while showing cognitive and locomotor age‐dependent decay at a behavioral level. This makes easy to follow the aging process and relate it to other phenotypes. The possibility to insert human pathogenic mutations in the N. furzeri orthologue gene using CRISPR/Cas9 adds up a further advantage in support to this model , but we did not observe any significant change between young and old subjects. TDP‐43 presence in SGs is not permanent but dynamically regulated in response to cellular metabolism as described in the literature , Nfu_TDP‐43 , and Nfu_TDP‐43L  were cloned in a pET‐SUMO plasmid with an N‐terminal hexa‐His tag, a SUMO tag with a modified polylinker site and a cleavable tobacco etch virus (TEV) protease site. The 4.2Aggregation kinetics of the isolated proteins were determined by the Thioflavin T (ThT)‐binding assay followed by a BMG FLUOstar Omega microplate reader in Greiner Bio‐One CELLSTAR plates. When not carried out immediately after purification, protein aliquots were rapidly thawed from the freezer, spun down, and filtered with a 0.2 µm syringe filter before each assay to ensure removal of any contingent preformed oligomer. The resulting concentration was determined. The solutions were diluted to final concentrations of 10 µM for the RRM1‐2 domains and 6 µM for the C‐termini. The ThT concentration was 20 µM. The plate was sealed with an optic seal to avoid evaporation. The experiments were carried out at 37°C by recording the fluorescence intensity of ThT as a function of time for 48 h using 430 nm as the excitation wavelength and 485 nm for the emission. Experiments in which the fusion protein was cleaved directly in the plate were carried out by adding TEV protease directly to the protein samples using 1:10 enzyme to protein molar ratios. Experiments carried out in the presence of RNA used the fluorescent dye ProteoStat. The aptamer was added to the protein in an RNA to protein 2:1 molar ratio using 10 µM concentrations of RRM1‐2. The excitation wavelength was set at 500 nm and the emission at 600 nm.4.32 flush at 4.0 L/min. All datasets were an average of thirty scans. Far‐UV (190–260 nm) spectra were recorded at 20°C and then gradually heated to 90°C (1°C/min) for the determination of the melting temperature (Tm). The data were plot and the Tm calculated according to literature .CD spectra were recorded on 10 µM samples in phosphate buffer and 15 mM KCl with a JASCO‐1100 spectropolarimeter. The spectra were acquired in 1 mm path length quartz cuvettes under a constant N4.4The protocols of fish maintenance were carried out in accordance with all animal use practices approved by the Italian Ministry of Health (Number 96/2003a) and the local animal welfare committee of the University of Pisa. All experimental procedures were managed, following the prescription of the European (Directive 2010/63/UE) and Italian law (DL 26/04‐03‐2014).Nothobranchius furzeri fishes were hatched and housed locally in a Tecniplast Zebtech system with automatized water flow, pH, and salinity control. All animals were hatched, fed, and maintained as previously described , and 25 tick sections were cut on Superfrost Plus adhesion slides (Thermo Fisher Scientific).Brains from 5‐ and 27‐week‐old animals were microdissected and processed for immunohistochemistry protocols: tissues were first fixed with paradormaldehyde (PFA) 4% in PBS (O/N at 4°C) and cryoprotected with sequential immersions in sucrose 20% and 30% until the tissue precipitated to the tube bottom (a minimum of 6 h/each steps). The tissues were singularly included in Tissue‐Tek4.5N. furzeri. This is a clearing and staining approach for whole‐mount tissues that is faster than the original Scale methods and avoids tissue expansion and preserves lipids  were used.4.7We performed Western Blots both on the purified TDP‐43 domains and on brain extracts to assess recognition of the polyclonal anti‐TDP‐43 antibody and check specificity. We used 35 µg of purified protein samples and ran them in a 12% SDS‐PAGE gel for 2 h at 100V. Afterward, the samples were blotted on a nitrocellulose membrane for 1 h at 100V. The membrane was incubated with the primary antibody overnight at 4°C and then incubated with the secondary antibody for 1 h at room temperature. The membranes were imaged with a Chemidoc XRS scanner using the Quantity one Biorad software.N. furzeri brains, these were extracted, immediately stored at −80° and then homogenized for 30 s using the GelD2 buffer with the addition of protease and phosphatase inhibitors. After centrifugation for 10 min at 28000g, the supernatant was collected, quantified using the BCA kit (Thermofisher), and stored at −80° for further use. To perform the Western Blot, we used 20 µg of protein extract derived from two samples per age and ran them in a 10% SDS‐page gel for 45 min at 150 V. Blotting and incubation procedures were the same as the ones described above.For the Western Blots in 4.8http://icy.bioimageanalysis.org/) for spot analysis. In both cases, we imposed a threshold to isolate the SGs or aggregates and launched the algorithm to identify the granules of TDP‐43 comprised inside the SG/aggregates. We exported the data regarding the number of granules detected (both SG/aggregates and TDP‐43) and the area of the granules. We divided the number of TDP‐43 granules for the number of SG/aggregates to obtain the percentage of SG/aggregates containing TDP‐43 and the area of TDP‐43 for the area of SG/aggregates.To quantify the colocalization of TDP‐43 with G3BP and aggresome staining, we acquired high‐resolution images (2048 × 2048 px) at a Zeiss confocal microscope using a 63× objective. For each animal, we analyzed three images per area . We used the Spot detector algorithm in the open license software Icy  was used to predict the protein tendency to phase separate (Bolognesi et al., catRAPID signature was used to predict the propensity of TDP‐43 to interact with RNA and identify RNA‐binding domains (Livi et al., catRAPID omics was employed to compute the binding targets in the human transcriptome (Agostini et al., The authors declare no conflict of interest.AL, JR, and BE took care of the experimental in vitro experiments, SB carried out all the work on animal samples, GGT is responsible for the bioinformatic analysis, AC is the promoted of the Furzeri project, and ETT and AP supervised the work. All authors contributed to early versions of the manuscript, AP finalized the manuscript, which was then scrutinized by all co‐authors.Supplementary MaterialClick here for additional data file.Video S1Click here for additional data file.Video S2Click here for additional data file.