Efficient thermal management of modern electronics requires the use of thin films with highly anisotropic thermal conductivity. Such films enable the effective dissipation of excess heat along one direction while simultaneously providing thermal insulation along the perpendicular direction. This study employs non-equilibrium molecular dynamics to investigate the thermal conductivity of bilayer graphene (BLG) sheets, examining both in-plane and cross-plane thermal conductivities. The in-plane thermal conductivity of 10 nm × 10 nm BLG with zigzag and armchair edges at room temperature is found to be around 204 W/m·K and 124 W/m·K, respectively. The in-plane thermal conductivity of BLG increases with sheet length. BLG with zigzag edges consistently exhibits 30–40% higher thermal conductivity than BLG with armchair edges. In addition, increasing temperature from 300 K to 600 K decreases the in-plane thermal conductivity of a 10 nm × 10 nm zigzag BLG by about 34%. Similarly, the application of a 12.5% tensile strain induces a 51% reduction in its thermal conductivity compared to the strain-free values. Armchair configurations exhibit similar responses to variations in temperature and strain, but with less sensitivity. Furthermore, the cross-plane thermal conductivity of BLG at 300 K is estimated to be 0.05 W/m·K, significantly lower than the in-plane results. The cross-plane thermal conductance of BLG decreases with increasing temperatures, specifically, at 600 K, its value is almost 16% of that observed at 300 K. Thermal management at the nanoscale devices is a significant challenge due to the rapid increase in power densities ,2,3. For2 hybridization. The strong sp2 bonding, combined with the lightweight nature of the carbon atoms in the microscopic structure, imparts to graphene exceptional physiochemical characteristics. These include a high Young’s modulus (~1.0 TPa), a large specific surface area (2630 m2/g), and an optical transmittance of 97.7% [2 bonds in nature. Experimental research has shown that the thermal transport properties of SLG depend on various factors, such as the geometry size [In recent years, novel materials, especially low-dimensional materials like quasi-one-dimensional nanowires ,7, nanotof 97.7% ,14,15. Cof 97.7% . These vtry size ,17,18,19try size , and funtry size .ip) and cross-plane thermal conductivity (kcp) of BLG and multilayer graphene. While there have been numerous experimental studies on in-plane thermal transport, research on the cross-plane heat transfer of graphene is relatively limited. In a recent study, Sood et al. [In addition to SLG, bilayer graphene (BLG) and multilayer graphene sheets also exhibit unique behaviors and possess properties that differ from those of SLG. For example, the room temperature thermal conductivity of few-layer graphene varies from 1300 W/m·K to 2800 W/m·K . Multilad et al.  investigThe novel thermal properties of SLG, BLG, and multilayer graphene have been extensively studied by both experimental methods and numerical simulations. While the mentioned experimental research has yielded direct measurements of these properties of graphene, the substantial costs associated with experimentation and the limited ability of nanoscale temperature probing have presented considerable challenges for conducting more extensive investigations. Numerical methods are promising tools to complement experiments and enhance our comprehension of the distinctive thermal properties exhibited by graphene. Molecular dynamic (MD) simulation is one of the main numerical methods for studying the properties of 2D materials, such as graphene. In recent years, there has been a dramatic increase in MD simulations to explore the unique thermal properties of SLG ,27,28,29ip of single- and few-layered graphene nanoribbons. Their findings indicated a reduction in the kip of graphene as the number of layers increased. Cao et al. [ip of multilayer graphene using NEMD simulations. Their results showed that as the size of multilayer graphene decreased, its kip also decreased. Guo et al. [ip of interlayer-bonded BLG, tri-layer graphene, and pyrolytic graphite using NEMD. They discovered that interlayer sp3 bonding reduces thermal conductivity, with reductions of up to 80%. Additionally, they observed that tensile strain further decreases the thermal conductivity of interlayer-bonded graphene by 50%. Zhan et al. [ip of a BLG sheet was approximately 64.8 W/m·K, by employing the reverse non-equilibrium molecular dynamics (RNEMD) method. They also observed an 80% reduction in thermal conductivity with a 1.39% increase in bridge defect. Si et al. [ip of both SLG and multilayer graphene sheets, utilizing Tersoff, REBO, and AIREBO potentials. They concluded that for SLG, the optimized Tersoff potential produced the highest thermal conductivity, closely aligning with experimental results. Regarding multilayer graphene, they concluded that the Tersoff potential provides the best simulation of its thermal conductivity. By using the MD method with REBO and Leonard Jones potentials, Li et al. [ip of misoriented BLG. They explored the effect of interlayer misorientation angle and temperature using NEMD. The study’s conclusion highlighted that thermal conductivities exhibit an approximately linear decrease with the increasing lattice constant of the commensurate misoriented BLG unit cell. Nie et al. [ip and kcp of twisted multilayer graphene sheets utilizing NEMD. The results revealed that the highest thermal conductivity could be achieved when no twist angles were present, for both in-plane and cross-plane conduction. Zhang et al. [ip of BLG by the NEMD, employing different in-layer and interlayer potentials. The Leonard Jones and Kolmogorov–Crespi potentials described the interlayer interactions, while the in-layer interaction potentials, namely Tersoff, REBO, and AIREBO potentials, were compared. Their results showed significant differences in both the magnitude and temperature-related trends of thermal conductivity when different potentials were applied in the simulations.While there have been numerous studies dedicated to MD simulations of SLG, the attention given to MD simulations of BLG or multilayer graphene is relatively limited. Wei et al.  employedo et al.  investigo et al.  determinn et al.  calculati et al.  used NEMi et al.  calculate et al.  examinedip and kcp. Moreover, the effects of several parameters, such as sample dimensions, edge shapes, temperature, and strain, on thermal conductivity are investigated. This study is expected to yield valuable insights for the design and application of BLG in nanoscale devices, particularly where the manipulation of thermal conductivity, whether to suppress it or enhance it, in different directions is essential.Reviewing the aforementioned research reveals that, in comparison to the research on in-plane thermal transport, there has been limited focus on investigating the cross-plane conduction at the interface of graphene–graphene. This signifies an existing gap in our understanding that requires resolution. To the best of the authors’ knowledge, a comprehensive investigation into the in-plane and cross-plane conductivity of BLG has been rarely documented in published studies. Therefore, this work aims to explore the thermal conductivity of BLG through NEMD, considering both kIn this research, we utilized NEMD simulations to calculate the thermal conductivity of BLG. These simulations were executed using the large-scale atomic/molecular massively parallel simulator (LAMMPS) package . Atom viip. The simulation domain comprises three regions: the main simulation region, reservoir regions, and fixed walls. Two fixed walls made up of fixed atoms are positioned at the model’s ends in order to prevent atom escape from the system and limit relative sliding motion between graphene layers. Next to these fixed walls, cold and hot reservoirs (heat sink and heat source) are established to create a temperature gradient. Once the simulation structure reaches an equilibrium state, the atom velocities in the reservoir regions are artificially modified to provide a constant heat flux into the main simulation region, situated between the heat source and heat sink. During each time step, the particle velocities within the heat source and heat sink are uniformly scaled, ensuring that a uniform quantity of kinetic energy is consistently added to or deducted from both reservoirs. This process provides a constant heat flux within the simulation domain, establishing a temperature gradient aligned with the direction of heat flow. From the calculated temperature gradient and heat flow, the thermal conductivity of the graphene can be then determined from the Fourier law. To assess the localized temperatures, the model is segmented into 25 equal divisions along the X-axis direction.The schematic diagram of BLG is shown in cp of the BLG with the NEMD method, the initial configuration of a 6-layer structure needs to be established. The top and bottom layers are set as the fixed layers, and then the heat sink and heat source are positioned as shown in To simulate the kThe optimized Tersoff potential was employed to describe in-layer interactions . In addiip and kcp, the simulation box was first relaxed in an isothermal-isobaric (NPT) ensemble for 100 ps, which proved sufficient to achieve equilibrium. The heat source and heat sink were controlled by Langevin thermostats to maintain constant temperatures. Then, an additional 1 ns was conducted within a microcanonical (NVE) ensemble to ensure the attainment of a desirable linear temperature profile for kip calculations. For simulating kcp, a longer simulation time of 2 ns in the NVE ensemble was necessary to reach a good equilibrium of the temperature profile. This extended duration was required due to the lower thermal conductivity in the cross-plane direction compared to the in-plane direction. Periodic boundary conditions were applied along the X, Y, and Z axis directions.While simulating kA typical temperature profile of the BLG for the in-plane conduction calculations is depicted in ip was computed using Fourier law as specified in Equation (2):ip is the in-plane thermal conductivity coefficient.The kip of BLG, we conducted simulations following the dimensions and operating conditions outlined in the study by Nie et al. [2 with a 0.35 nm interlayer distance. The comparison of results is presented in ip calculated in this study exhibits only slight deviations from the findings of Nie et al. [In order to check the accuracy of the NEMD simulation results for the ke et al. . The inve et al. , with a ip obtained for other sizes of BLG, as reported in the upcoming sections, with the values from prior studies, indicates that our NEMD simulation results are consistent with the findings of previous research [In addition, comparing the range of kresearch ,39.2 and an interlayer distance of 0.335 nm for the calculation of kip. To quantify the impacts of various factors on the kip of BLG, we varied several key parameters such as length, width, temperature, strain, and edge shapes.In our study, we investigated BLG sheets with dimensions of 10 × 10 nmip of BLG with armchair and zigzag edges at 300 K. The thermal conductivities of 10 nm × 10 nm zigzag and armchair BLG are estimated to be 203.6 W/m·K and 124.3 W/m·K, respectively. Contrary to bulk materials, where thermal conductivity is typically size-independent, the results in ip of SLG was observed to increase with the length of the graphene sheet [ne sheet . The lenne sheet . HoweverFurthermore, ip for three different BLG lengths. It is evident from this figure that the thermal conductivity of BLG sheets with zigzag edges is higher at very small widths, particularly for the smallest BLG lengths (10 nm). This phenomenon could be linked to the limited number of phonons present in the system at these small sizes. This scarcity results in fewer available phonon–phonon combinations that satisfy the momentum and energy conservation rules for scattering, leading to reduced phonon–phonon scattering. The higher thermal conductivity at very small widths aligns with findings from other research [research , where tIn addition, Moreover, as observed in Prior research has demonstrated the substantial influence of strain on modulating the thermal conductivity of various 2D materials. The application of strain presents a method to precisely tune the thermal conductivity of materials, as evidenced by these investigations ,34,59. FFurthermore, it is noteworthy that the thermal conductivity of zigzag BLG demonstrates a higher sensitivity to tensile strain compared to armchair BLG. For a tensile strain of 12.5%, the thermal conductivity reduction is 51% in 10 nm, 52% in 14 nm, and 45% in 18 nm compared to the strain-free values for the zigzag BLG. On the other hand, the thermal conductivity reduction is 29% in 10 nm, 32% in 14 nm, and 30% in 18 nm armchair BLG sheets. This behavior is in agreement with the findings in reference , where tcp has been shown to be two to three orders of magnitude lower than its kip [The characteristics of cross-plane thermal conduction differ significantly from those of in-plane conduction in certain materials. For example, in the case of graphite, the k its kip . The the its kip ,62.2·K. To the best of our knowledge, there have been no studies conducted on the thermal conductance of the graphene/graphene interface within BLG. Wei et al. [−9 m2·K/W for the cross-plane thermal resistance of 6-layer graphene at 300 K, as shown in 2·K. In another research, Ni et al. [−9 m2·K/W using equilibrium molecular dynamics simulations, corresponding to a cross-plane thermal conductance of 207 MW/m2·K. In addition, Ding et al. [2 heterostructures using NEMD simulations. For 7-layer graphene, the interfacial thermal conductance at the graphene/graphene interface was calculated as 212.6 MW/m2·K, as shown in Table 2 of Ref. [The effects of temperature on the cross-plane thermal conductance of BLG are illustrated in i et al.  studied i et al.  calculatg et al.  explored of Ref. . Conside of Ref. , it is r of Ref. ,62, indicp of BLG at 300 K is 0.054 W/m·K, which closely aligns with both the reported values of 0.05 W/m·K for BLG (cp is significantly lower than the in-plane results by approximately four orders of magnitude. This anisotropic behavior arises from the high kip of graphene, which is ascribed to the strong covalent sp2 bonding between adjacent carbon atoms. This type of bonding is known to be one of the strongest in nature, marginally stronger than the sp3 bonds in diamond. In contrast, the adjacent graphene layers in multilayer graphene are linked by weak van der Waals interactions.Our simulation results reveal that the corresponding kip, which monotonically decreases with increasing temperature. This observation aligns qualitatively with prior research on the temperature effect on cross-plane heat transfer in few-layered graphene [Furthermore, graphene ,66,67. Tgraphene .ip and kcp of BLG sheets exhibited strong agreement with published data. Furthermore, we conducted a parametric study and thoroughly analyzed the resulting outcomes.This study investigated the in-plane and cross-plane thermal conductivity of BLG sheets using NEMD. We employed the optimized Tersoff potential to describe in-layer interactions, while the van der Waals interaction between different layers was modeled using the Lennard–Jones potential. The obtained results for both kip of 10 nm × 10 nm zigzag and armchair BLG sheets at room temperature are around 203.6 W/m·K and 124.3 W/m·K, respectively. The kip of BLG increased with the length of the sheets: for 10 nm × 30 nm sheets, the values for the zigzag and armchair configurations increased to 1214.15 W/m·K and 773.15 W/m·K, respectively. The influence of sheet width, due to the applied periodic boundary conditions, was less evident on BLG’s thermal conductivity. In addition, the kip of BLG decreased with increasing temperature, with a slightly divergent rate observed between armchair and zigzag configurations. A 10 nm × 10 nm zigzag BLG sample, for instance, saw a reduction from 203.6 W/m·K to 170.8 W/m·K as the temperature increased from 300 K to 600 K. Conversely, the armchair configuration’s thermal conductivity decreased from 124.3 W/m·K to 111.1 W/m·K over the same temperature range. Furthermore, applying uniaxial tensile strain on the BLG sheet led to a reduction in thermal conductivity for both armchair and zigzag configurations, with zigzag BLG displaying a higher sensitivity compared to armchair BLG. A tensile strain of 12.5% on a 10 nm × 10 nm sample resulted in a thermal conductivity reduction of 51% for zigzag BLG and 29% for armchair BLG when compared to strain-free values.Our results showed that the k2·K with the corresponding kcp of 0.054 W/m·K. These findings highlighted that the kcp is significantly lower than the in-plane results by about four orders of magnitude. As temperatures increased in the range of 300 to 600 K, the cross-plane thermal conductance exhibited a reduction. However, the temperature-induced reduction was not highly pronounced and the thermal conductance at 600 K was approximately 16% of that at 300 K.Our findings also demonstrated that the cross-plane thermal conductance of BLG at 300 K is 162.3 MW/mThe high anisotropic heat conduction properties observed in BLG sheets hold promise for applications in the thermal management of nanoscale devices that require controlled regulation of thermal conductivity in different directions.


In addition, an increase of one percentage point in OHT/PHC coverage of municipalities increased the likelihood of avoiding patient referral by 1% . Teleconsulting sessions efficiently avoided patient referral to other care levels. Both contextual and individual factors were associated with avoided referrals by teleconsulting sessions.This cross-sectional analytical study assessed the frequency of avoided referrals of primary care to other care levels by dental teleconsulting and its association with individual and contextual variables using a multilevel approach. It appraised asynchronous dental teleconsulting sessions from the secondary database of the Monitoring and Evaluation System of the Telehealth Results during 2020, during the COVID-19 pandemic. The outcome was “whether referral to secondary care was avoided”. Individual variables were related to teleconsulting and professionals that requested it: sex, dental specialty, and dentistry field. Contextual variables were related to each municipality that requested responses: Municipal Human Development Index, oral health teams (OHTs) in primary health care coverage, dental specialty centers coverage, illiteracy rate, Gini index, longevity, and per capita income. A descriptive analysis was made using the Statistical Package for the Social Sciences. Hierarchical Linear and Nonlinear Modeling software was used to perform multilevel analyses to assess the association of individual and contextual variables with avoiding patient referral to other care levels. Most teleconsulting sessions avoided patient referral to other care levels (65.1%). Contextual variables explained 44.23% of the variance in the outcome. Female dentists were more likely to avoid patient referrals than male dentists (OR = 1.74; CI = 0.99–3.44;  The stimulated gatekeeping of primary health care (PHC) and regulation of referrals to specialized care is a global issue . In BrazDespite implementing strategies to overcome these obstacles, the Brazilian Unified Health System (SUS) network in PHC and specialized care services, such as the dental specialty centers (DSCs), is quite diverse in structure, regional distribution, and financing ,3. It reTelehealth and teledentistry use technological resources for communication in health areas  and may The Brazilian Ministry of Health implemented telehealth in 2006 to strengthen PHC and increase access to specialized health in the country, which is currently known as the Telehealth Brazil Networks Program . Unlike Teleconsulting is one of the telehealth program’s strategies and consists of remote communication between the PHC professional and a specialist in a specific area called a teleconsultant. They exchange information about clinical care, health promotion actions, and work processes by messaging or videoconferences synchronously or asynchronous text messages that must be answered within 72 h. Teleconsulting is offered by telehealth centers, mostly linked to federal universities, potentially covering all Brazilian PHC ,13. TeleIt is crucial to evaluate the resoluteness of the telehealth program’s dental teleconsulting sessions to avoid unnecessary referrals and its associated factors to plan policies that tackle inequalities. Thus, this study aimed to assess the prevalence of patient referrals avoided by dental teleconsulting sessions and their association with individual and contextual variables using a multilevel approach. The null hypothesis was that the prevalence of avoiding patient referral to other care levels by teleconsulting sessions does not differ between municipalities and individual factors.Universidade Federal de Minas Gerais) approved the research under protocol CAAE 17400319.9.0000.5149.A cross-sectional analytical study that appraised asynchronous dental teleconsulting sessions from the secondary database of the Telehealth Brazil Networks Program in 2020 was carried out. This was a population study of the entire Brazilian territory covered by this program. Data collection used an integrative platform that contains information on telehealth centers in the telehealth program: the Monitoring and Evaluation System of the Telehealth Results . Codes fThe outcome was avoiding referral to other care levels after each teleconsulting session, dichotomized into yes/no. The covariables were structured into individual (level 1) and contextual levels (level 2). The variables for the individual level were gathered from the database of the Monitoring and Evaluation System of the Telehealth Results . The vart-test (p < 0.05). Because the interest was focused on explaining the independent effects of each covariate, all potential variables were included in the unadjusted model and adjusted model. This analysis was not focused on prediction [p-values, odds ratios (ORs), and 95% confidence intervals (95% CIs) were estimated in each analysis. The reliability estimate was employed to determine the final multilevel model’s adequacy. All variables theoretically associated with the outcome were kept in the final model.IBM Statistical Package for Social Sciences v. 22.0  was used to perform descriptive analysis, and Hierarchical Linear and Nonlinear Modeling software  to perform a multilevel analysis was used to assess the association of individual and contextual variables with avoiding patient referral to other care levels. The analysis included 1,319 teleconsulting sessions (level 1) from 279 municipalities (level 2). Parameters were estimated using the restricted maximum likelihood method and predictive quasi-likelihood estimation. A multilevel logistic regression model was built. In the first stage, a null model estimated the primary partition of data variability between two levels, and then the individual and contextual characteristics were considered. The variance partition coefficient (VPC) was calculated to determine how much variance in the outcome stems from the municipalities’ differences. Each was individually incorporated into the model before being tested for the association between the outcome and the covariables of levels 1 and 2 using Student’s ediction . The p-vTeleconsulting sessions avoided patient referral to other care levels in 65.1% of the cases. Most PHC professionals that requested responses were female (65.3%) and from the Family Health Strategy (FHS) specialty (47.8%), followed by general dental practitioners (43.8%). Semiology/pharmacology/stomatology was the dentistry field with significant demand (45.7%), followed by general dentistry (40.6%) .Contextual variables were descriptively analyzed .p = 0.000)  . In the p = 0.055). An increase of one percentage point in OHT/PHC coverage of municipalities increased the likelihood of avoiding patient referral by 1%   indicated that female dentists were more likely to avoid patient referrals than male dentists  .This study is original since it nationally assessed potentially associated factors with avoiding referrals by Brazilian dental teleconsulting. It might contribute to achieving a more resolute program and PHC, reducing waiting times to other care levels. The data period selected marks the onset of the COVID-19 pandemic, responsible for creating a new obstacle to accessing health services due to social distancing measures, which generated a repressed demand for treatments in the population .The asynchronous teleconsulting sessions have reportedly been one of the most offered and used ,26, despAs observed in previous studies, female dentists demanded teleconsulting sessions the most ,30. MoreThe significant requests for teleconsulting from FHS dental surgeons and general dental practitioners were expected due to the Brazilian PHC profile , and mosThe null hypothesis was rejected since the null model indicated a difference in the prevalence of avoiding referrals of patients to other care levels by the teleconsulting sessions in the municipalities assessed. The contextual variables partly explained the outcome variance. Contextual factors, such as the organization of local health systems and services, the size of municipalities, institutional support, and the program’s implantation, can influence telehealth utilization by PHC professionals. The heterogeneous incorporation of the telehealth program into the Brazilian territory, which presents marked developmental differences, must be considered to improve the program’s effectiveness and efficiency .One percentage point increase in OHT/PHC coverage of municipalities increased the likelihood of avoiding patient referral by 1%. Municipalities with better OHT/PHC coverage are expected to have a more organized PHC network, which could reflect better potential patient access and capacity to solve problems. Including the OHT in the FHS restructured the PHC model. It reordered health action planning in the public sector, focusing on the territorial and epidemiological needs of the population .Noteworthy is that 75% of the municipalities that requested teleconsulting showed rates of DSC/1,000,000 inhabitants of 0.00. In comparison, most of them had reasonable OHT/PHC coverage rates: the median OHT/PHC coverage was 76.52%, and P75% of the municipalities that requested teleconsulting showed an OHT/PHC coverage of 100%. However, 25% of the municipalities (P25%) still showed OHT/PHC coverage under 41.55%. This rate reinforces the need for a resolutive PHC in the country ,26,33,35Continuing professional development is related to better OHT performances, resulting in better patient care . In thisUsing secondary data, which hindered a richer collection of information regarding whether the avoided referral was unnecessary, and the loss of data due to the lack of PHC professionals’ feedback on the platform were some limitations of this study. The period analyzed was atypical because of the COVID-19 pandemic, with new challenges in the daily practice of health care services. Thus, future studies are needed to explore these issues further and direct interventions to improve the telehealth program’s results.Teleconsulting sessions avoided most patient referrals to other care levels. The contextual variables partly explained the differences in the prevalence of avoided referrals. Female dentists and municipalities with better OHT/PHC coverage were more likely to avoid patient referrals to other care levels by teleconsulting sessions. We should focus on strategies to increase the equity in OHT/PHC coverage in Brazilian municipalities, better support both professionals and patients, and enhance the telehealth program’s resoluteness.


However, the transcriptional regulators and their mechanisms are still unclear, especially in fungal pathogens. Here, we identified Sin3, a component of the histone deacetylase complex, as a transcriptional repressor of ATGs and negative regulator of autophagy induction in the rice fungal pathogen Magnaporthe oryzae. A loss of SIN3 resulted in upregulated expression of ATGs and promoted autophagy with an increased number of autophagosomes under normal growth conditions. Furthermore, we found that Sin3 negatively regulated the transcription of ATG1, ATG13, and ATG17 through direct occupancy and changed levels of histone acetylation. Under nutrient-deficient conditions, the transcription of SIN3 was downregulated, and the reduced occupancy of Sin3 from those ATGs resulted in histone hyperacetylation and activated their transcription and in turn promoted autophagy. Thus, our study uncovers a new mechanism of Sin3 in modulating autophagy through transcriptional regulation.Autophagy is a conserved degradation and recycling pathway in eukaryotes and is important for their normal growth and development. An appropriate status of autophagy is crucial for organisms which is tightly regulated both temporally and continuously. Transcriptional regulation of IMPORTANCE Autophagy is an evolutionarily conserved metabolic process and is required for the growth and pathogenicity of phytopathogenic fungi. The transcriptional regulators and precise mechanisms of regulating autophagy, as well as whether the induction or repression of ATGs is associated with autophagy level, are still poorly understood in M. oryzae. In this study, we revealed that Sin3 acts as a transcriptional repressor of ATGs to negatively regulate autophagy level in M. oryzae. Under the nutrient-rich conditions, Sin3 inhibits autophagy with a basal level through directly repressing the transcription of ATG1-ATG13-ATG17. Upon nutrient-deficient treatment, the transcriptional level of SIN3 would decrease and dissociation of Sin3 from those ATGs associates with histone hyperacetylation and activates their transcriptional expression and in turn contributes to autophagy induction. Our findings are important as we uncover a new mechanism of Sin3 for the first time to negatively modulate autophagy at the transcriptional level in M. oryzae. The –32– by Ume6 , 36. In to ATG32 . In Caene stress , 39. HowM. oryzae  occupancy and transcriptional silencing in . oryzae . In thisM. oryzae, autophagy is required for the degradation of the conidial components during appressorium formation (M. oryzae (autophagy-related genes (ATGs) were significantly upregulated in the Δsin3 strain compared with those in the wild-type (WT) strain, which included ATG1, ATG3, ATG4, ATG8, ATG9, ATG11, ATG12, ATG13, ATG14, ATG15, ATG16, ATG17, ATG23, ATG24, ATG27, ATG26, and ATG29 .Autophagy plays an important role in fungal development and pathogenicity . In M. oormation , 29. In . oryzae . By analATGs in the WT and Δsin3 strains grown in the liquid nutrient-rich complete medium (CM) were determined by reverse transcriptase quantitative PCR (RT-qPCR). Consistent with results of RNA-seq, expression levels of extensive ATGs were significantly upregulated in the Δsin3 strain compared with those of the WT strain under the CM conditions. In M. oryzae, Atg1, Atg13, and Atg17 form an initiation complex during autophagy induction  were detected. The results showed that the upregulated pattern of ATG1, ATG13, and ATG17 was extended in the Δsin3 strain after a 4-h exposure in SD-N medium -Atg8, which is a widely used marker of autophagy activity, was introduced into the WT and Δsin3 strains  staining was performed before microscopic observation with a confocal microscope. In the CM, the fluorescence signal in the GFP-ATG8 strain was located mainly in the punctate structures in cytosol and rarely in the vacuoles, while the fluorescence signal in the Δsin3/GFP-ATG8 strain was located predominantly in vacuoles  staining assay further showed that more autophagosomes were observed in the Δsin3 strains than that in the WT on CM . Furthermore, the number of autophagosomes in the Δsin3/GFP-ATG8 and GFP-ATG8 strains were counted by fluorescence microscopy under normal and autophagy-inductive conditions. Compared with the GFP-ATG8 strain, the number of autophagosomes increased significantly in the Δsin3/GFP-ATG8 strain  has two layers of regulation . One lay3 strain .ATGs, transient luciferase assays were conducted. The expression of luciferase driven by the promoter of ATG1, ATG13, and ATG17 individually was transiently repressed by the overexpression of SIN3  showed that the recombinant Sin3 protein can directly bind to these loci . Furthermore, a chromatin immunoprecipitation followed by quantitative PCR (ChIP-qPCR) assay revealed that compared with the WT strain, the examined regions at the promoters of ATG1, ATG13, and ATG17 had 1.5- to 2-fold enrichment in the SIN3-FLAG/Δsin3 strain . Next, we asked whether direct occupancy on the ATGs by Sin3 would associate with the changed levels of histone acetylation. To answer this question, a ChIP-qPCR assay with an anti-H4K16ac antibody was performed with the Δsin3 and WT strains. Compared with the WT strain, the enrichments of H4K16ac at the promoter regions of ATG1, ATG13, and ATG17 in the Δsin3 strain were 2- to 6-fold higher than that in the WT strain  was created, in which SIN3 was driven by its native promoter and fused with GFP. Immunoblot analysis showed that the relative intensity of Sin3-GFP was also decreased under SD-N conditions compared with that grown in the CM , respectively, were created in the GFP-ATG8 background . In the CM, the fluorescence signal of GFP-Atg8 was located mostly in the punctate structures in the cytosol and rarely in the vacuoles in the GFP-ATG8 strain. In contrast to the distribution of GFP in the WT strain, the GFP-Atg8 was localized partly in the vacuole in the ATG1-OE, ATG13-OE, and ATG17-OE strains    to D. ThSIN3 affects the pathogenicity of M. oryzae, infection assays were performed with susceptible barley and rice seedlings. For the barley leaf infection assay, mycelial plugs from the WT, Δsin3, and complementary strains were inoculated on the detached barley leaves. Four days after inoculation, no visible lesion was formed on the Δsin3-inoculated leaves, while large lesions were formed on the leaves inoculated with the WT and complementary strains , while the typical spindle-shaped blast lesions were formed on the leaves inoculated with the WT and complementary strains  strain for obtaining transformants . Strains and primers used in the experiments were listed in Table S1 and S2 in the supplemental material.formants . The Δsieviously , 48, 49.Hordeum vulgare) was used. For the observation of invasive hypha, 21-day-old healthy rice seedlings  were used for sheath preparation. Conidial suspensions were inoculated into the rice sheath and incubated in the growth chamber with a photoperiod of 16-h light and 8-h dark at 25°C. For the observation of glycogen, conidial suspensions were inoculated on the hydrophobic surface for 0, 4, 8, and 24 h and stained with I2/KI solutions before being photographed  at 25°C for 7 days. The appressorium formation and rice seedling infection assay were conducted as described previously . For theographed . The inoFresh mycelia (0.1 g) cultured in the liquid CM for 2 days were harvested for detecting FLAG or GFP-tagged proteins. Total proteins were extracted with lysis buffer . The resulting proteins were separated by an SDS-PAGE gel, transferred to a polyvinylidene difluoride (PVDF) membrane, and subsequently detected by immunoblotting with an anti-FLAG  or anti-GFP  antibody. The relative intensity of the immunoblots was quantified with Image J software. To detect the autophagy level, the construct of expressing GFP-Atg8 was introduced into the corresponding strains by ATMT . The ressin3 strains were cultured in the CM and SD-N medium. At least 25 hyphal segments were calculated. Conidial autophagy was monitored by incubating conidia of strains expressing GFP-Atg8 on the hydrophobic surface for 0, 8, and 16 h as described previously  for 10 min before being photographed. The fluorescent signals of GFP-Atg8, Sin3-GFP, and H2B-mCherry were captured by a confocal fluorescence microscope (Zeiss LSM700) with 488-nm and 555-nm laser excitation. Vacuoles were observed with 405-nm laser excitation following 7-amino-4-chloromethylcoumarin (CMAC) staining for 30 min. To monitor the translocation of lipid droplets during appressoria formation, conidial suspensions were inoculated on the hydrophobic surfaces and stained with boron-dipyrromethene (BODIPY) solution  for 0, 4, 8, and 24 h before being photographed with a fluorescence microscope  was used as an internal control. Primers used in the experiments were listed and described in Table S2.Mycelia cultured in the liquid CM for 2 days were collected, and total RNA was extracted with TRIzol reagent . Total RNAs were reverse transcribed into cDNA with a commercial kit  according to the manufacturer’s instruction. RT-qPCR was performed with SYBR green qPCR master mix  in the LightCycler480 system (Roche). The constitutively expressed ATG1, ATG13, and ATG17 were cloned into the pGWB435 vector individually by the Gateway technology , incubated at 28°C for 4 h, and then purified with an amylose resin column . Probes were synthesized and labeled with biotin. EMSA was performed with the LightShift chemiluminescent EMSA kit  according to the manufacturer’s instructions. Binding reactions contained 1 μL 10× binding buffer, 2 μg recombinant MBP-Sin3 or MBP alone (negative control), and 1 μL biotin-labeled probes or unlabeled probes. In the competition assays, unlabeled probes were provided in 25-fold excess. The binding reaction mixtures were incubated at 28°C for 20 min, separated on 6% native PAGE gels, and transferred onto IMMOBILON-NY+ transfer membranes (Millipore’ INYC00010). Signals were detected using the chemiluminescent biotin-labeled nucleic acid detection kit . Primers for probes were listed and described in Table S3 in the supplemental material.The full-length coding sequence of Sin3 was fused with MBP and cloned into pMAL-c5X. Subsequently, the fused plasmid was introduced into TUB5 using qPCR. Primers were listed and described in Table S2.ChIP experiments with mycelia were conducted as described previously . Brieflyt test was calculated with Excel software.One-way analysis of variance (ANOVA) was conducted with SigmaStat 3. The two-tailed All data generated or analyzed during this study are available from the corresponding author on reasonable request.


However, the interactionbetween the ionizing electron beam and the sample results in beam-inducedmotion and image distortion, which limit the attainable resolutions.Sample charging is one contributing factor of beam-induced motionsand image distortions, which is normally alleviated by including partof the supporting conducting film within the beam-exposed region.However, routine data collection schemes avoid strategies wherebythe beam is not in contact with the supporting film, whose rationaleis not fully understood. Here we characterize electrostatic chargingof vitreous samples, both in imaging and in diffraction mode. We mitigatesample charging by depositing a single layer of conductive grapheneon top of regular EM grids. We obtained high-resolution single-particleanalysis (SPA) reconstructions at 2 Å when the electron beamonly irradiates the middle of the hole on graphene-coated grids, usingdata collection schemes that previously failed to produce sub 3 Åreconstructions without the graphene layer. We also observe that theSPA data obtained with the graphene-coated grids exhibit a higher b factor and reduced particle movement compared to dataobtained without the graphene layer. This mitigation of charging couldhave broad implications for various EM techniques, including SPA andcryotomography, and for the study of radiation damage and the developmentof future sample carriers. Furthermore, it may facilitate the explorationof more dose-efficient, scanning transmission EM based SPA techniques.Cryogenic electronmicroscopy can provide high-resolution reconstructionsof macromolecules embedded in a thin layer of ice from which atomicmodels can be built  SPA has yielded atomic resolution reconstructions,3 as well as reconstructions from particles smaller than the 38 kDatheoretical size limit.5 SPA data collection has becomemuch faster over the last few years with the availability of fasterdetectors, advanced microscope automation, fringe-free imaging, aberration-freeimage shift, and hole clustering.6 Thishigher throughput has been combined with schemes to improve signal-to-noiseratios (SNR), such as DQE improvement of detectors, energy filters,a lower energy spread of the electron source, enhanced phase contrast,and reduced sample movements.Stimulated by the Resolution Revolutionin cryogenic electron microscopy(cryo-EM),7 However, the physicalmechanisms behind the sample movement are not fully understood. Severalhypotheses have been proposed to explain this beam-induced motion.8 It has been argued that the sample is placedunder compressive stress upon rapid cryocooling. Electron radiationcan induce creep in the presence of this stress that results in domingof the sample in the foil openings.9 Anotherhypothesis implicates new mechanical stress from the breakage of chemicalbonds and the generation of hydrogen gas. Alternatively, electrostaticcharging generates an attractive force that causes bending and warpingof the thin flexible sample layer. Furthermore, it has been shownthat biomolecules can also appear to shift without physically movingthemselves.10 Nonideal and dynamicallychanging lens conditions would result in image distortions in whichmolecules appear to move.11 No matter thecause, movement or distortion during imaging leads to a reduced imagequality by dampening high-resolution signals. Several schemes havebeen proposed to reduce beam-induced motions, such as devitrification,12 vitrification at a low cooling rate and elevatedtemperatures,13 or the use of grids with smallholes to have a low ice thickness/hole diameter ratio.14Practically, it is well-knownthat upon irradiation by electrons,biomolecules embedded in a thin hole-spanning vitreous ice layer areobserved to move.20 The ionizing electron beam produces secondary electrons that escapefrom the sample, thus leaving a positive charge on nonconductive specimens.24 Charging can result in unwanted contrast changes, known as the “beeswarm effect”, characterized by fluctuating granularity causedby random surface charging at low magnification and high-defocus conditions.26 Russo and Henderson describedthat sample charging is a dynamic process that results from the poorconductivity of the specimen under low electron flux conditions.27 The positively charged “footprint”from electron irradiation forms a microlens on the sample, which deflectsincoming electron beams, causing a change in phase contrast. Thiseffect, known as the “Berriman effect”, fades when thebeam scans nearby regions.22 The microlenscan already be formed within a fluence range of ∼10–3 to ∼1 e–/Å2 and may contributeto the defocus change observed in the early frames of micrographs.28Charging of biological samples withinthe TEM has been discussedfor decades.22 Curtis andFerrier noticed that the “bee swarm effect” does nothappen when part of the beam hits the metal grid bar even when thecarbon film and the grid bar within the field of view are not connected.21 Berriman and Rosenthal designed a special seven-holeC2 aperture and demonstrated that secondary electrons emitted fromadjacent areas can reduce the charge on the area of interest.10 Objective apertures can also reduce specimencharge: secondary electrons emitted by the objective aperture cancompensate some of the positive charge of the surface of the specimen.16Sample charging is normallyalleviated by including part of thesupporting conducting film within the region exposed to the electronbeam.31 However, this technique would cause charging on a thin hole-spanningvitreous ice layer. This technique was successful only with specimenssupported by a continuous conductive carbon film, which is undesirablefor SPA due to the SNR reduction.One successful technique to reduce beam-inducedmotion is “spot-scanimaging”, which focuses an electron beam to a diameter of ∼1000Å and scans it over the specimen to capture multiple images.33 gives minimum background noise, and can be overlaid on top of asupport layer. Graphene is used in materials science, and it is increasinglyapplied in the life sciences.40 Ultraflat graphene can result in uniform thin ice layers, allowingfor high-resolution structure determination of sub 100 kDa proteins.38 Others described that graphene can be functionalizedto improve particle density and orientation.43A suitable alternative toamorphous carbon is graphene, which hasorders of magnitude higher conductivity than a carbon layer,In thispaper, we investigate charging effects on vitreous biologicalspecimens and demonstrate that charging can be mitigated by depositinga graphene layer on regular EM grids. We present high-resolution,conventional-TEM SPA reconstructions obtained with such grids anddiscuss the importance of understanding charging for future conventionaland nonconventional SPA schemes. Being able to mitigate charging bydeploying graphene could help to further push the boundaries of resolvingthe high-resolution structures of biomolecules via EM.Mycobacteriumtuberculosis ferritin (BfrB)44 applied to glow-discharged R1.2/1.3 Quantifoil and UltrAuFoil grids.The vitreous ice within the holes of this perforated film acts asan insulator, and a beam size smaller than those of these holes wasused. Similar to Brink et al.,22 we useda defocused diffraction image (DIFF image) to observe the effect ofcharging  and decreased in the underfocused condition(Supplementary Movies 3 and 4). The normalizedDIFF beam size in both overfocus , given that the beam is inside the hole. By comparison, thenormalized DIFF beam sizes from a conductive crossline grating replicasample (Supplementary Movies 5 and 6) remainedconstant throughout irradiation.We usedregular SPA samples and grids: charging 1a.A hybverfocus 1d and unverfocus 1g became–/Å2). At the beginning of irradiation, the diffraction lens wasset in overfocus condition, and the beam was confined within a holeon the vitreous sample (Supplementary Movie 7) highlights thevariation in beam size at various beam locations. There is a noticeablebump at around 50 s, reflecting a rapid charge–discharge processas the beam is scanned from one side of the foil to the other. DIFFmovies collected in the same manner at a diffraction lens under underfocuscondition (Supplementary Movie 8)showed a similar yet inverse trend, wherein the DIFF image beam reachedits maximum (instead of minimum) and remained stable near/on the foiland then decreased (instead of increased) in holes. We repeated theseexperiments with UltrAuFoils and found the same trends (Supplementary Movies 9 and 10).To provide a more in-depth observation of how samplesare chargedand discharged, we collected DIFF movies with a moving stage stayingat each location for 5 s  as a function of time under both overfocus with an extra graphene layer applied on top of them to test whetherthe conductivity from graphene could alleviate charging. We verifiedthe presence of graphene by collecting electron diffraction patternsfrom samples with amorphous ice on graphene 3a. These–/Å2 3c, under–/Å2 3d and un–/Å2 3e conditverfocus 3i and unverfocus 3j conditverfocus 3f, partiverfocus 3g, or toverfocus 3h. Notab–/Å2, after which the beam size remained constant,typical SPA data collection schemes conducted in imaging mode usefluences of tens of e–/Å2, whichmay be more prone to aberrations that are not readily apparent indiffraction mode. Thus, we used the imaging mode with parallel illumination(–/(Å2 s) for 1 s witha Falcon III detector and no objective aperture at a nominal magnificationof 78000× and averaged without patch-track motion correction.We first used a 20 μm C2 aperture to ensure a beam diameter(800 nm) smaller than the hole size (1.2 μm): the beam did nottouch the perforated support film. This setup resulted in severelydistorted images, as if the particles were moving outward from a center. Importantly, when weperformed the same experiments using Quantifoil grids with a graphenelayer we consistently obtained sharp images, independent of the sizeof the beam relative to the foil hole size .While the DIFF experimentsshowed clear charging effects up to a fluence of 1.5 emination1a to fura center4a. Aftera center4b, arguiaperture 4c and thaperture 4d. We foSupplementary Figure 1). However, we could obtain a 2.01 Å reconstructionwith graphene grids . When the beam size was larger than the hole size and touched theconductive support, we achieved reconstruction of maps at 2.12 Åresolution with Quantifoil grids without graphene  and 1.90 Å resolution with graphene-coatedgrids using a similar number of particles , where it can be observed as a change in the size of thebeam .These observations confirm the conclusions in ref —thesedistortions were reversible and restored as soon as the beam touchedthe conductive support film (Supplementary Movie 19), which we attribute to sample discharging. Performing control experimentsin reverse order , and selectingparticles was challenging . Efforts to obtain a proper initial map and reconstructionfrom this data set were unsuccessful. Despite this, we could eventuallyobtain a correct BfrB reconstruction utilizing modern motion-correctiontechniques combined with Bayesian polishing, albeit only at 3.5 Åand a low b factor . This result not only highlights the power of modernimage data processing tools but also provides a warning, as the successof these programs might blind the user to the underlying physicalphenomena that prevented them from getting better data.Surprisingly,whereas the beam size change observed in DIFF mode saturated around1.5 ed in SPA 4a. Whilese order 4c,d showSupplementary Figure 2b). Additionally, for theBfrB protein that we used to perform the experiment, the graphenehelped to concentrate the sample within the holes,43 as we could use a 10 times diluted sample compared to the gridswithout graphene and still obtained a similar number of particlesper micrograph (b factor remained relatively unchanged regardlessof the beam size for the sample on graphene-coated grids (−82and −84 Å2); however, it was significantlyhigher compared to that of Quantifoil grids (−178 Å2) . Despitethe b factor of the data set without graphene onQuantifoil grids with a large beam (−83 Å2)being comparable to data sets with graphene, the resolution valueson the y axis at the same x axisvalues (number of particles) were lower than the other two data sets,and the standard deviation was high . While our results demonstrate that graphene mitigatedcharging and indeed resulted in improvements in movement and resolution deposit less energy in the sample.The findings are consistent with the “spot-scan imaging”approach described by Downing,48 wasobserved in Quantifoil grids with and without graphene  techniques such as ptychography and iDPC.Graphene gridswere prepared from a commercially available graphene-Cu foil, producedby chemical vapor deposition (CVD). First, a thin layer of celluloseacetate butyrate (CAB) was applied via spin coating in a CAB-ethylacetate solution. The CAB-graphene-Cu stack was placed on top of anammonium persulfate etching solution, etching away the Cu. The etchingsolvent was gradually diluted using deionized water to neutralizethe solution and remove the Cu residues from the etched foil. Filterpaper was placed at the bottom of the Petri dish, and the grids wereplaced on top using tweezers. The graphene was transferred onto thegrids by removing the water using a micropipet. The filter paper containingthe graphene-covered grids was placed on a heating plate for 30 minat 35 °C to dry. Finally, the graphene grids were heated in activatedcarbon for 15 h at 300 °C, which removed the CAB layer (whosemelting temperature is between 170 and 240 °C).M. tuberculosis BfrB was prepared according to Gijsbers et al.44 and used at a concentration of 50 mg/mL  for Quantifoil and UltrAuFoil experiments. We used 300 mesh gridswith 1.2 μm diameter holes. A volume of 2.5 mL was applied ontogrids that were glow-discharged under vacuum at a current of 7 mAfor 30 s. For grids with a graphene layer, a volume of 2.5 μLof diluted BfrB  was applied onto mildly glow-dischargedgrids (current of 7 mA for 10 s). Excess liquid was removed by blottingfor 3 s using filter paper followed by plunge freezing in liquid ethaneusing an FEI Vitrobot Mark IV instrument operated under 95% humidityat 4 °C.47 on a 200 keV TecnaiArctica instrument (Thermo Fisher Scientific). The beam was blockedwith the prespecimen beam shutter before exposing the sample for recording.The beam was set to be parallel at a flux of 0.38 e–/(Å2 s) passing through amorphous ice films. TheDIFF image was obtained by defocusing the diffraction lens. No objectiveaperture was used for the DIFF image data collection. Supplementary Movies are available at: https://doi.org/10.6084/m9.figshare.23244299.v1Diffractionimages were collectedwith a Timepix3 hybrid pixel detector60 Movie stacks were corrected for drift (7 × 7 patches) and dose-weightedusing MotionCor2.61 The local contrasttransfer function (CTF) parameters were determined for the drift-correctedmicrographs using Gctf.62 A first set of2D references were generated from manually picked particles in RELION,60 and these were then used for subsequent automaticparticle picking. 63 The resolution of the best final map was 2 Å usingthe gold-standard FSC = 0.143 criterion.64 The maps have been deposited in the Electron Microscopy Data Bankas entry EMD-18010, EMD-18028, EMD-18029, EMD-18030.Cryo-EMsingle-particle data were collected on a Titan Krios instrumentat 300 kV with a BioQuantum K3 Imaging Filter with a 20 eV postcolumnenergy filter. The detector was used in normal counting mode at anominal magnification of 105000×.


Pulmonary sarcoidosis is the most prevalent manifestation of sarcoidosis and the commonest diagnosis in clinics for ILD. Due to the lack of a simple and reliable test, making the diagnosis is often challenging. There are three criteria that must always be considered: (1) compatible clinical presentation; (2) evidence of granuloma formation ; and (3) exclusion of alternative causes of granulomatous disease. There are various tools available for diagnosis, amongst which serum biomarkers like sACE and sIL-2R, HRCT, BAL, EBUS/EUS and sometimes bronchoscopic or surgical lung biopsy are most contributive. However, the degree of invasiveness of the applied test and associated risk to the patient must be weighed against management consequences. In specific situations  or when there is high suspicion based on HRCT in the context of supportive clinical findings, it might be justifiable to decide on a “working diagnosis of sarcoidosis” and to refrain from further invasive procedures for the patient. This should, however, preferably be agreed upon after discussion in an experienced multidisciplinary team and requires close follow-up of the patient. In general, it is advisable to always maintain a healthy dose of skepticism when making the diagnosis of sarcoidosis, especially when the clinical course of disease gives rise to this. Pulmonary sarcoidosis is the most common manifestation of sarcoidosis and the most frequent established diagnosis in the group of interstitial lung diseases (ILD). Clinically, suspicion usually arises through finding of intrathoracic lymph node enlargement and/or diffuse nodular lung disease. Although HRCT is the cornerstone for diagnosis, its imaging findings are currently not considered diagnostically sufficient. Sarcoidosis has many lookalikes and currently remains a diagnosis by exclusion. Moreover, so far, no international agreement on the diagnostic approach has been brought about. Furthermore, usability and availability of diagnostic tools vary around the word. All in all, this makes establishing a diagnosis challenging.The aim of the article is to give an overview of the diagnostic criteria, differential diagnosis, clinical presentations, and approach to the diagnosis of sarcoidosis. Secondly, the value of the currently available tools for diagnosis is discussed. Further, the value of multidisciplinary team discussion and the concept of likelihood and working diagnosis are discussed. Finally, the reader is provided with a diagnostic algorithm based on the recent literature and experience of the author, along with future perspectives.Sarcoidosis is defined as multisystem immune-mediated disease of unknown cause, pathologically characterized by non-caseating granuloma formation in various organs or tissues throughout the body. It commonly affects young and middle-aged adults and usually presents with bilateral hilar lymphadenopathy and/or pulmonary infiltration but can also manifest with ocular manifestations or skin lesions, and the liver, spleen, lymph nodes, salivary glands, heart, nervous system, muscles, bones, and other organs may also be involved ,2.Three major criteria must be met to make a diagnosis of sarcoidosis: (1) a compatible clinical presentation; 2) findings of non-caseating granulomatous inflammation in one or more tissue samples; and 3) the exclusion of alternative causes of granulomatous disease or diseases capable of producing a similar clinical picture [ findings the exclA number of challenges in accomplishing a diagnosis of sarcoidosis are to be addressed.First, there is significant heterogeneity in manifestations of the disease, which are often referred to as clinical phenotypes. There are not only various types of pulmonary involvement but also many extrapulmonary manifestations, and on top of that, many combinations of both are possible in sarcoidosis.The GenPhenReSa (Genotype–Phenotype Relationship in Sarcoidosis) project, a European multicenter study, was designed to map in detail multi-organ involvement in over 2000 European sarcoidosis patients. The study found five distinct clusters according to predominant organ involvement: (1) abdominal organ involvement, (2) ocular–cardiac–cutaneous–central nervous system disease involvement, (3) musculoskeletal–cutaneous involvement, 4) pulmonary and intrathoracic lymph node involvement, and (5) extrapulmonary involvement [ pulmonarSecondly, the defined multisystemic nature of the disease is clinically not always evident. It is well recognized from daily practice in specialized centers that isolated single-organ involvement may occur, especially in cardiac sarcoidosis. Also, other single-organ manifestations with strong suggestion of sarcoidosis may sometimes manifest without clinical evidence of a second organ’s involvement, leading to fundamental discussion about whether the diagnosis of sarcoidosis is acceptable ,6.Thirdly, the diagnosis of sarcoidosis needs exclusion of other causes of granulomatous disease such as tuberculosis, fungal infections, and organic and inorganic exposure-related ILDs such as hypersensitivity pneumonitis and berylliosis. Also, there are other rare diseases that might need consideration in proper context. Differential diagnosis of sarcoidosis is further discussed elsewhere. It should be noted that the likelihood of certain differential diagnosis of (pulmonary) sarcoidosis will differ around the world.Finally, the diagnosis of sarcoidosis can never be 100% sure: It is a diagnosis of exclusion, and this cannot be accomplished with complete confidence. The diagnosis requires clinic-radiographic findings compatible with sarcoidosis, histologic or cytological confirmation of granulomatous inflammation, exclusion of known causes of granulomatous disease, and presence of disease in at least two organs or tissues. The end result of the diagnostic evaluation for sarcoidosis is neither a definitive diagnosis nor an exclusion of the diagnosis but rather a confident likelihood of the disease. In this light, a recent BTS clinical statement on pulmonary sarcoidosis raised the issue that decisions made by individual patients to decline bronchoscopy when there is a highly probable but not definite clinical diagnosis should be supported in most cases, with careful subsequent monitoring .Onset and type of symptoms of pulmonary sarcoidosis can vary largely. Most patients will manifest with gradual onset . Symptoms can be respiratory  but also non-respiratory or combinations . Because the presenting symptoms of sarcoidosis are not specific for the disease, the primary care physician is usually the first health care provider to assess the patient. If a patient with respiratory symptoms does not improve on  treatment for more common diseases like bronchitis, asthma, or chronic obstructive disease, it is usually through chest imaging, revealing signs of lymph adenopathy and/or diffuse lung disease, that referral to a medical specialist will takes place ,9. Also,Although most patients will manifest with gradual onset, a small subgroup of patients will present with symptoms of acute/subacute onset. These symptoms can be either directly related to sarcoidosis or indirectly, i.e., due to secondary complications. A well-known acute clinical manifestation of pulmonary sarcoidosis is Löfgren’s syndrome. Besides acute onset of disease, most commonly with fever, this syndrome is characterized by bilateral hilar lymphadenopathy, erythema nodosum, and/or ankle arthritis or marked periarticular inflammation of the ankles . AnotherOccasionally, acute presentations can be caused by complications related to pulmonary sarcoidosis, such as pneumothorax, pneumonia, or pulmonary embolism . These cAfter presentation of a patient with symptoms and/or signs on imaging that could be compatible with pulmonary sarcoidosis, various other diagnoses need to be considered, especially infectious diseases and malignancy like lymphomas. The differential diagnosis will therefore depend on the level of clinical suspicion and other contextual information  and will usually become narrowed during the course of the diagnostic process. Other diseases should also be excluded that may give the impression of sarcoidosis but are non-granulomatous, such as lymphomas, other malignancies, and immune-mediated conditions like IgG4-related disease or auto-inflammatory syndromes like VEXAS syndrome, the latter being increasingly recognized as a novel entity .When evidence of granulomatous inflammation is found, the differential diagnosis can be categorized into granulomatous disorders of either infectious or noninfectious origin. Differential diagnosis of sarcoidosis requires customization, taking into account not only the individual’s clinical history and presentation but also risk factors, and can depend on geographic situation. Of note, due to migration and the increase of human travel activity over the past decades, infectious causes of granulomatous lung disease that used to be tied to certain continents can now also show up elsewhere  ,17.Finally, as the diagnosis of sarcoidosis is never fully secure, it is advisable to always maintain a healthy degree of skepticism that an alternative diagnosis has been overlooked, especially when the clinical course of the disease gives rise to this .In patients suspected of pulmonary sarcoidosis, various tools are available to secure the diagnosis of sarcoidosis. None of the tests can be regarded as diagnostic proof alone. The extension and the nature of the tests will depend on the degree of ambiguity of the clinical presentation. Usually, a combination of tests lead to sufficient confidence that sarcoidosis may be diagnosed.The first step is an assessment of epidemiological factors, notably the incidence of sarcoidosis and of alternative diagnoses in the region/country and exposure to risk factors . Also, exposure to drugs taken for therapeutic or recreational purposes must be addressed. Family history is of importance, as approximately 10% of sarcoidosis patients report familial occurrence .Subsequent investigations usually include (chest-)imaging, serum biomarkers, bronchoscopy with or without bronchoalveolar lavage, endo sonography, and/or pathologic evaluation of biopsy tissue. Each of these tests are discussed below with focus on their diagnostic value. Of note, to support clinicians as to the probability of sarcoidosis, especially in situations where biopsies might not be easy to perform, it might also be helpful to use clinical scores that support the likelihood of sarcoidosis in front of a compatible presentation ,21.The discovery of electromagnetic radiation and subsequently that of chest radiography made the early pioneers of sarcoidosis aware that the disease was much more than a skin, eye, and joint disease and that lungs and/or intrathoracic lymph nodes were the prime manifestation of the disease .Based on chest radiography, thoracic sarcoidosis has classically been staged in four groups ,24. Stagstage, the term radiographic type is more appropriate for use here [Although the staging has some prognostic significance , it has many limitations. First, interobserver variability is poor, especially between stages with parenchymal involvement. Second, the stages suggest a relationship between disease severity and/or the order in which sarcoidosis may evolve. However, this is far from true, as a patient with stage I might seem to have mild disease but instead can suffer from severe cardiac involvement. Furthermore, although stage I on a chest radiograph is associated with high probability of resolution of intrathoracic lymphadenopathy after 1–2 years, the disease may nevertheless still evolve to progressive sarcoidosis in a minority of patients. Thus, instead of Currently, HRCT is regarded as the most valuable tool for the diagnosis of pulmonary sarcoidosis. With this technique, characteristic features can be visualized, such as “beading” along fissures  and a peImaging of pulmonary sarcoidosis is further discussed elsewhere in this issue.67Ga) in the salivary and lacrimal glands (panda sign) and intrathoracic lymph nodes (lambda sign) has shown to represent distinctive nuclear imaging patterns that are highly specific for sarcoidosis. In the 1990s, the combination of both panda and lambda sign or panda sign in combination with bilateral symmetrical hilar lymphadenopathy on chest X-ray has been suggested to obviate the need for invasive diagnostic procedures [Simultaneous uptake of (67)gallium  positron emission tomography (PET) scanning. It has been shown that FDG-PET is more sensitive than 67Ga imaging in the assessment of sarcoidosis activity. Furthermore, FDG-PET has demonstrated a very good inter observer agreement in contrast to 67Ga imaging [67Ga imaging than for FDG-PET. In 67Ga imaging, the radiation dose is 18.5 mSv compared to 5.6–7.6 mSv for FDG-PET, depending on the patient’s weight [Today however, the use of  imaging . Of notes weight .It is not recommended to apply FDG-PET routinely in the diagnostic work-up of sarcoidosis, but in selected cases, it can be useful in identifying sites for biopsy or in differentiating extinguished fibrotic lesions from treatable inflammatory disease.Recent evaluation of the diagnostic value of different serum biomarkers in sarcoidosis has revealed the best performance of serum angiotensin converting enzyme (sACE), soluble IL-2 (sIL-2R) receptor, and chitotriosidase (CTO). These markers stand out as the most useful diagnostic tools, with significant sensitivity and specificity, although none functions alone as a gold-standard biomarker . The samBronchoscopy can reveal endobronchial lesions due to the mucosal involvement of sarcoidosis. These lesions are typically referred to as “cobble stone lesions” and reveal a high likelihood of finding granulomas upon biopsy.Also, in the absence of visual lesions, there is chance of finding granulomas in random biopsies taken from the endobronchial mucosa of patients suspected of pulmonary sarcoidosis . Even whBronchoalveolar lavage (BAL) is a useful and safe procedure that is widely applied in the diagnostic evaluation of pulmonary sarcoidosis. Cytologic evaluation of BAL fluid shows lymphocytic alveolitis in 90% of patients and therefore contributes to the likelihood of diagnosis . Also, aFinally, the last important role of BAL to be mentioned is, of course, narrowing the differential diagnosis, e.g., by excluding (opportunistic) infections.In most centers, endo sonography will by now have replaced mediastinoscopy as the standard procedure for intrathoracic nodal sampling in the diagnosis of pulmonary sarcoidosis. The latest guidelines justify the preference of endobronchial ultrasound (EBUS)-guided lymph node sampling (87% yield) over mediastinoscopy (98% yield) because it is safer for the patient and usually better tolerated . Also, cRecently, EBUS transbronchial needle aspiration (TBNA) has been compared head-to-head with esophageal endoscopic ultrasound (EUS)-B fine-needle aspiration (FNA) for diagnosing sarcoidosis . The resThere are different techniques to collect tissue from the peripheral lung parenchyma for the diagnosis of sarcoidosis.A summary of different lung tissue sampling procedures, including diagnostic yield, is given in Of note, only in very few cases suspected of sarcoidosis will performing surgical lung biopsy for confirmation of the diagnosis be necessary. In such cases, it may be advisable to first weigh the advantages and disadvantages of a surgical lung biopsy in a multidisciplinary discussion, with special attention to the diagnostic added value and therapeutic consequences.As mentioned above, non-caseating granulomas are the pathological hallmark of sarcoidosis. Typical for sarcoidosis is that the granulomas are well formed, without significant surrounding lymphoid infiltrate. The granulomas are discrete and compact . Although this type of granuloma is characteristic for sarcoidosis and sometimes referred to as “sarcoid granuloma”, it may also be found in other conditions such as Blau’s syndrome, foreign material, drugs, secondary syphilis, common variable immune deficiency, and chronic granulomatous disease ,43. AlsoThe other key feature of sarcoid granulomas is their anatomic distribution. Sarcoid granulomas in the lung are characteristically found along lymphatics, around the bronchovascular bundles, in the interlobular septa, and on the pleural surface. The number of granulomas in each of these locations may vary, but generally, they are more abundant around the bronchovascular bundles. Sarcoid granulomas and/or giant cells may also be found around and sometimes in the wall of pulmonary arteries or veins with a weak-to-absent inflammatory infiltrate and without necrosis of the vessel. The latter pathological finding is, however, not usually clinically associated with pulmonary hypertension or veno-occlusive disease .As finding necrosis in relation to granulomas should always raise high suspicion of an infectious granulomatous disease, the presence of some necrosis in the granulomas of patients with confirmed sarcoidosis has been described in up to 20% of biopsies . NecrotiPulmonary function testing is also central to the evaluation of patients suspected of sarcoidosis but in whom results are not contributing to diagnosis, although they reveal important information on the severity and/or progression of disease and can determine decisions on invasive diagnostic procedures, as mentioned above. Typically, in pulmonary sarcoidosis, all kinds of abnormal ventilatory patterns are possible, including mixed ventilatory defects, which have recently been reported to occur in approximately 10% of patients . Also, iAn algorithm for the diagnostic approach in pulmonary sarcoidosis is given in In case sarcoidosis is regarded highly probable on the basis of collected clinical data, including supportive findings such as elevated serum ACE and/or sIL2R and others , it is nThe ultimate goal of the diagnostic process is to rule out all diagnoses other than sarcoidosis that are consistent with the clinical situation. In some patients, a definite diagnosis may require the continuous gathering of information during follow-up. After diagnosis, a healthy degree of skepticism remains indispensable, especially in the case of an unexpected course of disease during follow-up. In that situation, additional investigations might be needed with reconsideration of the diagnosis.As sarcoidosis is defined as multisystem disease, it would be plausible to involve clinicians of other disciplines than pulmonology in the diagnostic process. The value of multidisciplinary team (MDT) discussion has already been scientifically illustrated and evaluated in ILD  and subshttps://health.ec.europa.eu/european-reference-networks/overview/evaluation-european-reference-networks_en, accessed on 17 September 2023).Nevertheless, many clinicians working in the field of sarcoidosis find an MDT discussion to be of added value, providing a momentum for intra- and interdisciplinary-supported diagnosis or generating new diagnostic considerations. Additionally, MDT discussion may next contribute to peer support for complex treatment decisions, which is especially important in the absence of guidelines with high-quality evidence recommendations. The implementation of MDT discussion in care pathways for sarcoidosis is therefore an important criterium for the evaluation of patient-centered care for sarcoidosis in expert centers across Europe  sarcoidosis receive a standard work-up for diagnosis according to the local care pathway for sarcoidosis, including MDT discussion, depending on the type of major organ involvement .Nonetheless, in general, not all patients have the opportunity to consult a center specialized in ILD. It is, however, the author’s experience that offering MDT conferences  for external patients can play a valuable role in the diagnostic decision and care of these patients. In this way, centers of excellence facilitate greater and more rapid access to sarcoidosis expertise.With ongoing advances in biomolecular technology and the development of artificial intelligence, it is likely that novel diagnostic tools will appear. With no doubt, these will change the methods of diagnosing sarcoidosis in the near future.Interesting new developments in the field of chest imaging have recently been published. Photon-counting CT (ultra-HRCT) has been shown to improve image quality for visualization of certain ILD features, such as traction bronchiolectasis and micro-nodules. This technical advance not only results in lower radiation exposure but may also enhance the diagnosis and prognosis of pulmonary sarcoidosis and ILD in the near future .An intriguing example of the potential of omics in establishing a diagnosis of sarcoidosis was recently found in the eNose study (SpiroNose) . Based oFinally, also research of new and especially combinations of biomarkers is regarded as a promising direction in the field of diagnosis and management of sarcoidosis and might lead to an improved standard of care in the future .The diagnosis of pulmonary sarcoidosis requires sufficient knowledge and experience with the disease, a sharp clinical eye, and a healthy dose of suspicion. Unfortunately, until now, no single, simple diagnostic test has been available. A systematic and multidisciplinary approach, preferably implemented in a local care pathway for sarcoidosis and including MDT discussion, currently provides the best guarantee for establishing the right diagnosis. In this context, based on ATS and BTS recommendations, increased specificity of CT features, and/or cases of acute presentation with specific symptoms, it might also be justifiable to refrain from further invasive procedures and follow-up for the patient.


We found a clear splitting in the first focusing peak whose evolution in charge density and magnetic field is well reproduced by calculations using the SOC strength of ~ 13 meV, and no splitting in the second peak that indicates stronger Rashba SOC. Possible suppression of electron-electron scatterings was found in temperature dependence measurement. Further, we found that Shubnikov-de Haas oscillations exhibit a weaker band splitting, suggesting that it probes different electron dynamics, calling for a new theory. Our study demonstrates an interesting possibility to exploit ballistic electron motion pronounced in graphene for emerging spin-orbitronics.Van der Waals interactions with transition metal dichalcogenides were shown to induce strong spin-orbit coupling (SOC) in graphene, offering great promises to combine large experimental flexibility of graphene with unique tuning capabilities of the SOC. Here, we probe SOC-driven band splitting and electron dynamics in graphene on WSe By combining graphene with transition metal dichalcogenides, such as WSe2, it is possible to induce a large spin-orbit interaction in the graphene layer. Here, Rao et al study the spin-orbit coupling in graphene/WSe2 heterostructures using the ballistic transport based technique, known transverse magnetic focusing. For monolayer graphene, it was theoretically predicted to have two distinctive terms, one that couples out-of-plane spin and valley degrees of freedom  have shown to be highly efficient in inducing strong spin-orbit coupling (SOC) in graphene22. It is particularly interesting as graphene has shown pronounced ballistic transport effects with large tunability, such as transverse magnetic focusing (TMF)25, Veselago lensing27, Fabry-Pérot interference29, and ballistic snake states31 among many others. They have also shown unique features originating from the relativistic nature of Dirac electrons31. However, a vast number of previous studies on graphene-TMDC heterostructures have focused on detecting spin relaxation due to electron scattering rather than the ballistic motion37. Moreover, only a few studies have found direct evidence for the SOC-induced band splitting by measuring beatings in Shubnikov-de Hass (SdH) oscillations (for both mono- and bilayer graphene)38 or tracing changes in quantum capacitance (for bilayer graphene only)11. Not only to understand the effect of the SOC on the electronic properties of the system, such as the band topology40 but also to exploit its full potential on ballistic spintronics22, it is therefore essential to demonstrate the ballistic transport in graphene on TMDCs while simultaneously probing their band structures and electron dynamics.Combined with the high electron mobility and large experimental flexibility of graphene, such a strong interface-induced SOC makes the graphene on TMDCs ideal for ballistic spin-orbitronics where ballistic electron motion can be used to control or detect electron spin through the SOC2 as it can not only probe the SOC-induced band splitting but also investigate electron dynamics simultaneously  at a distance  and no splitting in the second peak, which indicates a stronger Rashba SOC in the system. From the temperature dependence measurement, we also find a possible suppression of electron-electron scatterings that may originate from the dielectric scattering of WSe2 layers and/or the induced SOC. Further, we show that Shubnikov-de Haas oscillations exhibit a weaker band splitting, suggesting that it probes different electron dynamics. Interestingly, a similar behavior was found in studies on 2DEG with SOC21, indicating that this is universal and a new theory is needed to explain the phenomenon. Our study, therefore, places graphene-TMDC heterostructures as an interesting new material platform to explore the effect of SOC on ballistic electron transport in low-dimensional systems.Here, we demonstrate that all these studies are possible in graphene-TMDC heterostructures. We first show a clear splitting of the first focusing peak in graphene on WSe2 such that the graphene is protected from the harsh chemical environments in the following nanofabrication process46. Standard electron beam lithography and lift-off were carried out to make Hall bar devices on 285-nm-thick SiO2 substrate with doped silicon underneath used as a gate to control charge density 2V-1s-1. Especially on the hole side, the symmetry broken quantum-Hall states were observed at magnetic fields as low as ~3 T , monolayer graphene, and multilayer WSementclass2pt{minimlly Fig. .23, we found a large TMF signal constantly exceeding 100 The TMF signal  and thus it can occur even when there are no defect sites in TMDC that can sink charge carriers from graphene and suppress the ballistic transport. Our observation also aligns well with other studies on similar graphene-TMDC heterostructures62.Additionally, the observation of both diffusive SHE at low density and ballistic TMF peak splitting at higher density indicates that the SOC in our sample is induced by proximity with TMDC25. Figure e-e) scattering , followed by electron beam evaporation of 5 nm Cr and 50 nm Au films. The devices were finally shaped into Hall bars by another electron-beam lithography and reactive-ion etching process.The WSeDevices were measured in a 1.5 K cryogen-free variable temperature insert (VTI) with a superconducting magnet. The electrical signals were measured by applying a small low-frequency (17.777 Hz) AC current of 0.1–1 μA between the source and drain terminals and measuring the voltage drop between another two probes using a lock-in amplifier (Stanford Research SR830). The low-noise filters and amplifiers were used to detect small TMF signals. The back gate was controlled by Keithley 2400 source-meter.Supplementary InformationPeer Review File


Objectives: The central sterile supply department (CSSD) is responsible for sterilization processes, and instruments are then stored in a clean room until use. Environmental controls, such as temperature, and relative humidity, are important in preventing the spread of infection during the sterilization process and during storage of medical devices. Data regarding temperature and relative humidity readings in the operating room, and whether instruments remain covered for the duration of the operation, are difficult to obtain. For easy access to relative temperature and humidity data covering all operational intervals, a rapid and convenient notification system was designed using a software application to send temperature and humidity information to the ThingSpeak website. We identified abnormal values for temperature and relative humidity and submitted revisions for all operating periods to facilitate monitoring of the refrigeration system. Methods: We implemented 3 programs: (1) DHT11 humidity and temperature sensor; (2) NodeMCU ESP8266; and (3) Program Arduino IDE. Results: The ESP8266 board connected using WIFI SUTH Mobile, and the DHT11 displayed temperature and humidity. The temperature and humidity data were sent to the website every 10 minutes. When an alarm occurred, it triggered immediate notification via the software application. Conclusions: We designed a temperature and humidity alert system using DHT11. Environmental control was possible using ESP8266, and alerts were triggered in the software application when an anomaly occurred. Data were uploaded to ThingSpeak every 10 minutes. The triple system actually sends alerts through the application and records data every 10 minutes. This system can measure environmental conditions in real time.


Nanoribbons of MoS2/graphene heterostructure are laid over a SiO2 substrate with an applied vertical gate voltage. The computational model shows that we can achieve an absorptance of approximately 50% of the incident light. The absorptance frequency can be tuned through varying the structure and the substrate dimensions, where the nanoribbon width can be varied approximately from 90 nm to 300 nm, while still covering the whole THz range. The structure performance is not affected by high temperatures (500 K and above), so it is thermally stable. The proposed structure represents a low-voltage, easily tunable, low-cost, and small-size THz absorber that can be used in imaging and detection. It is an alternative to expensive THz metamaterial-based absorbers.Terahertz frequency has promising applications in communication, security scanning, medical imaging, and industry. THz absorbers are one of the required components for future THz applications. However, nowadays, obtaining a high absorption, simple structure, and ultrathin absorber is a challenge. In this work, we present a thin THz absorber that can be easily tuned through the whole THz range (0.1–10 THz) by applying a low gate voltage (<1 V). The structure is based on cheap and abundant materials (MoS THz frequency is the range of frequencies lying between 300 GHz and 10 THz that are characterized by their low energy and high penetration. The characteristics of these frequencies have enabled them to be preferable in medical applications, imaging, security scanning, the food industry, and communication ,3. THz f2/Vs) and high optical transparency (97.4%) [2/graphene heterostructure has desirable electronic properties and a high on/off ratio (~108) that open the way for different electronic applications [Graphene, the 2D allotrope of carbon, has a monolayer thickness of approximately 0.34 nm. It is a zero-band-gap semiconductor that has high electron mobility  ,9. Graph7.4% ,9.  (97.4%) ,11. It h (97.4%)  and it i (97.4%) ,13,14. Wications ,16.2 is a transition metal dichalcogenide (TMD) with a bulk indirect bandgap of 1.2 eV and a monolayer direct bandgap of 1.89 eV. This specific bandgap endows MoS2 with unique photoluminescence characteristics where it can be used as a biomarker for cancer and DNA detection [2’s bandgap is larger than the energy of the THz spectrum, no photocarriers are generated when it is excited with THz frequencies [2/graphene heterostructure has more desirable properties than the monostructures of graphene or MoS2. The heterostructure has a larger bandgap than graphene and an enhanced carrier mobility compared to MoS2 [2 in the THz region and the Fermi level tunability of graphene enable MoS2/graphene heterostructures to have promising electronic and optoelectronic applications [2’s optical properties at low energy frequencies [2 and graphene.MoSetection ,19,20,21quencies ; howeverquencies . A MoS2/ to MoS2 ,24. The ications ,26,27. Tquencies ,29, but 2, a lot of work has been carried out to investigate the behaviour of this heterostructure. The MoS2/graphene heterostructure has better absorptance than graphene [2/graphene heterostructure affects its bandgap and its optical properties [2 under electromagnetic excitation. These surface plasmons play an important role in wave absorption. Moreover, the charge transfer through the heterojunction MoS2/graphene enhances the generation recombination process, resulting in better absorption. MoS2/graphene heterostructures are favourable in field effect transistors (FETs) in electronics. In [2/graphene heterostructure was proposed where the charge transfer through the heterojunction results in better chemical sensing and the FET has better stability in dry air. It also has higher sensitivity and better signal-to-noise ratio (SNR). A new transistor based on MoS2/graphene in [2 FETs. A graphene/MoS2 heterojunction FET with a 30 nm channel length has no short channel effects and a has drain-induced barrier lowering of 0.92 V/V. When graphene electrodes were doped with nitrogen, an increase in the device current by 214% was detected and the field effect mobility was four times that of undoped graphene electrodes [2 was previously studied, where low THz absorption peaks were detected in graphene–MoS2 microribbon grating [2 with other materials such as black phosphorous was investigated in [2/BP structure had six absorption bands in the THz region.Due to the favourable characteristics of graphene and MoSgraphene . Applyinoperties ,31. Vertoperties . Local snics. In , a FET sphene in  showed aectrodes . The absgated in , where a2/graphene nanoribbons under vertical gate bias. The heterostructure has enhanced absorptance in the whole THz region. The frequency of absorptance is easily tuned by varying the applied voltage. It is a cheap alternative for metamaterial THz absorbers. It can be utilized in many applications such as the sensing and detection of THz radiations [None of the previous studies discussed the absorptance of the heterostructure in the THz region under the effect of direct voltage application. Tunable THz absorbers can also be developed using metamaterials ; howeverdiations .2 substrate with 5 nm thickness. A monolayer of MoS2 with 0.65 nm thickness stripped into nanoribbons was laid over the graphene nanoribbons to have an infinite array of MoS2/graphene nanoribbons in the horizontal direction. The nanoribbon width, g, was applied to the MoS2 nanoribbons through Au metal contacts which have good compatibility with graphene and MoS2 [. The metal contacts only served as a means of voltage application. They did not have any effect on the characteristics of the design. Since they were very thin (less than 1 nm), they transmitted all the incident light and did not reflect any. A thin layer of h-BN was then laid over the structure. The calculations were based on the finite element method (FEM) used by COMSOL Multiphysics software. The structure was built and simulated using 2D simulation with applying periodic conditions on the vertical sides to solve for an infinite periodic array structure. The wave optics module, electromagnetic wave frequency domain (EWFD), was used.The proposed nanoribbon structure is based on the schematic in the SPPs . The incand MoS2 ,38. The 2 and graphene thickness, taken as 0.65 nm and 0.34 nm, respectively; 2 and graphene, taken as 3.7 and 5.6, respectively. Since monolayer MoS2 has a large bandgap with respect to THz waves, its optical conductivity is represented by the Drude model in Equation (8), where only intraband transitions dominate.2 and 2  and (8) were used to plot the absorption and transmittance of graphene-only nanoribbons and MoS2-only nanoribbons over the SiO2 substrate, as illustrated in 2 in 2, local surface plasmons are excited and a very small absorption peak exists at a low THz frequency, which is due to the low carrier concentration of MoS2. In graphene, the strong excitation of surface plasmons occurs, resulting in higher peaks attributed to its higher carrier concentration. The absorption peak is tuned with voltage due to graphene’s optical conductivity tunability through varying its chemical potential.MoS6.77 THz . The tra2 is slightly affected. The gate bias had no effect on MoS2 bilayer absorption, and in the monolayer, an increase in absorption was reported at 1.85 eV which was far from the THz region (gate bias applied from 50 to 50 V) [2 in the THz range does not affect its performance. Monolayer MoS2 can have absorption peaks at the THz region when doped as n-type or p-type (high carrier concentration) and applying a spin–orbit coupling effect [2 and the simple application of a gate voltage. The refractive index of MoS2 was found to be independent on the vertical gate bias, which is attributed to its large bandgap compared to the THz range and its low carrier concentration [Graphene absorption can be tuned using vertical bias, but MoSto 50 V) . That isg effect ; howeverntration .2/graphene heterostructure, the bandgap increased, forming mini bandgaps [2 and graphene. When applying a positive vertical gate bias on MoS2, holes were encouraged to migrate from the MoS2 to the graphene, which means a wider bandgap for the MoS2/graphene heterostructure and a higher absorptance frequency (the absorptance frequency was shifted to the right) as in 2/graphene nanoribbons, there was a remarkable increase in the absorption peak that can be finely tuned through the whole THz region by applying voltage. The absorption peak at Vg = 0 is nearly the same for graphene-only and the MoS2/graphene heterostructure but it obviously increased with higher gate voltages, indicating that the charge transfer between MoS2 and graphene can be neglected at zero bias, as suggested by [2. Graphene can have TE and TM modes, while MoS2 has only TM modes. The unity between the TM surface plasmon modes of both graphene and MoS2 (the imaginary terms of conductivity of graphene and MoS2 are positive) results in frequency resonance. When this resonance frequency matches the frequency of the incident light, an increase in absorption frequency is detected.In the case of the MoSbandgaps ,50 with 2 due to its higher carrier concentration, which increases when increasing the applied voltage. It is to be noted that the electric field will be high at the corners regardless of the graphene orientation because the conductivity tensor for graphene is the same for both zigzag and armchair [armchair , and alsarmchair ,51.2  frequency increases or shifts towards higher frequencies when the capacitance decreases. In order to have a good voltage tunable absorber for small nanoribbon widths (90 nm) in the THz region, we have to use a small substrate thickness in the range of 5 nm. This can be attributed to the fact that the carrier concentration of graphene is related to the substrate thickness according to Equation (6). We need a small When sectioning graphene into nanoribbons, its bandgap opens up, and when decreasing the nanoribbon width, we obtain larger bandgaps . Here, w2 sheets slightly decreased with the increase in temperature [, curves of MoS2 are not affected when varying the temperature from 4.5 K to 500 K as we go towards the THz region. The proposed nanoribbon structure is stable against temperature variations. Temperature can greatly affect the carrier concentrations in graphene nanoribbons; however, this effect will not be obvious for nanoribbon widths greater than 100 nm . When inperature ; that isperature  showed tWe have presented an ultrathin simple design for a monochromatic, narrow-bandwidth THz absorber with a thickness of approximately 7 nm. The absorptance frequency can be adjusted through the device dimensions (the nanoribbon width) or the applied voltage. When applying 0.4 V, we could achieve ~50% absorptivity at an absorptance frequency of 7.83 THz, a bandwidth of 0.8 THz, and a fractional bandwidth of andwidth b. We couandwidth c. On the2 has high transmittance in the THz range and very low absorption governed by intraband transitions. Graphene-only or MoS2-only nanoribbons cannot achieve good absorptance and tunability in the THz range. Since a MoS2/graphene heterostructure has a wider bandgap than that of graphene and more carrier concentration than MoS2, it can afford better absorption. In this work, we studied the absorption of a large array of infinite MoS2/graphene nanoribbons on a SiO2 substrate in the THz range. The structure showed better absorption than that of graphene-only nanoribbons. The absorption frequency can be easily tuned through the whole THz range by varying the vertical gate voltage based on the optical conductivity tunability of graphene with its chemical potential. The absorption frequency is affected by the nanoribbon width and substrate thickness. The temperature has little effect on shifting the absorption frequency, so the structure performance is stable at different temperatures. The proposed structure represents a cheap, easily fabricated, and tunable THz frequency absorber that can be used in many applications such as sensing, cancer diagnosis [Graphene nanoribbons have low absorption peaks in the THz range, where the absorption peaks can be slightly tuned through applying a gate voltage. MoSiagnosis , and com