The sudden turn had only a modest effect on capture and error location, and the analyses suggested that the birds had adjusted their tracking to the novel motion. In Experiment o angle. The occluder produced a disruption in capture success but did not delay capture. Error analyses indicated that the birds did not adjust their tracking to the new motion on turn trials following occlusion. The combined results indicate that pigeons can anticipate the future position of a stimulus, and can adapt to sudden, unpredictable changes in motion but do so better after a disappearance than after an occlusion.Despite being observed throughout the animal kingdom, catching a moving object is a complex task and little is known about the mechanisms that underlie this behavior in non-human animals. Three experiments examined the role of prediction in capture of a moving object by pigeons. In Experiment  Successful capture of a moving object is a complex task. It requires the organism to perceive the object, determine its path of movement, plan the object-directed action and move the capturing device  to the correct location for interception  served as the experimental subjects: Black 50 (B50), Violet 42 (V42), and Green 83 (G83). They had recently participated in experiments where they captured simple linear motions . One wall of the chamber was fitted with an 18 × 25 cm resistive touch screen  that was situated in front of a 15-in TFT monitor that was turned on its side. The monitor was set at a resolution of 640 × 480 pixels for the duration of the experiment. On the opposite wall of the chamber was a magazine pellet dispenser  and clicker . Individual 45-mg pigeon pellets were delivered through a rubber tube into a food cup  that was located 2 cm above the grid floor. A houselight was located on the top-right corner of the wall above the food cup; this delivered diffuse illumination to the pigeon chamber at an intensity of approximately 200 lux . A speaker, which was positioned outside the pigeon chambers, emitted a diffuse 60-dB white noise to mask sounds outside of the room.Responses were recorded from the touch screen via a USB touch screen controller . Control of the feeder and houselight was accomplished by a digital I/O card . A video splitter  allowed simultaneous presentation of images to the control room and operant chamber. Two Viglen Genie P4 computers located in an adjacent room delivered the experimental procedures and recorded data in E-prime v1.1. At the time of each peck, the location of the peck and the position of the stimulus were recorded in the form of XY coordinates with a time tag.Because the pigeons had recent experience capturing linear motions  the stimulus travelled in a linear path for a minimum of 7.5 cm and a maximum of 15.0 cm before making a sudden 90° turn; (2) during the pre-turn motion, any pecks that fell on the stimulus had no consequence; (3) following the turn, the normal training contingencies were in place. Pecks on the stimulus were counted as captures and resulted in termination of the trial and food delivery. Misses resulted in the stimulus continuing towards the screen boundary in a linear path. If the pigeon failed to capture the stimulus, it would move smoothly off screen when it reached the opposite side.There were eight types of turn trials, labeled according to pre- and post-turn motion direction: Right-Down, Left-Down, Right-Up, Left-Up, Down-Right, Up-Right, Down-Left, and Up-Left. There were three presentations of each of the turn trials, which were randomly intermixed among the 60 original training trials, yielding a total of 84 trials. In all other respects, testing was administered in the same manner as original training.Capture responses were assessed with two measures. The proportion of trials ending in a capture was the number of trials ending in a capture divided by the total number of trials in a session. The distance travelled before capture was computed from the point where the stimulus first became eligible for capture to the point of capture. Analyses on turn trials were conducted on pecks that occurred after the turn, once the stimulus was eligible for capture. Because the capture area on turn trials was less than on training trials, the data from training trials were truncated to equate capture opportunity on the two trial types. This was accomplished by only including captures that occurred within the initial portion of the straight trial that was equal in distance to the comparable turn trials ; captures occurring beyond this initial area were excluded from the analysis.For the error analysis, anticipatory and lagging errors were examined by determining the error position relative to the center of the stimulus. Errors in the same dimension as the motion (motion-relevant errors) were expressed as lagging behind the stimulus or leading in front of the stimulus.In addition, to examine within-trial history effects on errors, a regression analysis was conducted. It assessed the extent to which the previous and current motion directions predicted the error positions after the turn or following trial onset on straight trials. The previous motion was coded as the initial motion on both turn and straight trials, and the current motion was coded as the post-turn motion on turn trials and as the initial motion on straight trials. For example, on a rightward motion straight trial, the previous motion was rightward and the current motion was rightward, whereas on a rightward-upward turn trial the initial motion was rightward and the current motion was upward. The errors here were entered as error positions which coded the distance of the peck from the center of the stimulus in both the X and Y planes.For all analyses of turn trials, except the regressions, the data was collapsed across motion direction; this was for two reasons. First, although different motions resulted in somewhat different pecking patterns in the straight portion of the trials, the effect of the turn was highly consistent across motion types. Second, due to the small number of turn tests per session the data on turn trials were more variable than on straight trials. By collapsing across motion types, this resulted in an increase in stability of the test data.F = 1.2. There also was no significant effect of session, F < 1, nor was there any interaction between Turn × Session, F < 1. In examining the individual birds, two of the birds performed slightly worse on turn trials and one bird performed similarly on both trial types.The two measures of capture responses are displayed in Fig. F < 1, but there was an effect of session, F = 3.1, p = .047, but no significant effect of Turn × Session, F = 2.4, p = .091. In examining the individual birds, one bird captured the turn stimulus after a comparable distance to the straight trials, one bird was slightly later in capturing the stimulus on turn trials, and one bird was slightly earlier on turn trials. Overall, there was no consistent pattern to the performance across individuals. Panel c displays the distribution of capture locations. The greatest likelihood of capture occurred at the first point of eligibility and the distribution of capture locations did not appear to differ on straight and turn trials. The capture distributions of the individual birds were highly similar in shape to one another.Panel b displays the mean distance travelled from the start of the motion on straight trials or the turning point on turn trials until the point of capture. There was no indication of any effect of the turn, t(2) = 2.1.Figure Data Analysis) for each motion type following the turn  with the predictors of previous motion direction, current motion direction, and straight versus turn motion type. The dependent variable was the mean error relative to the center of the stimulus  revealed highly similar pecking patterns on the initial component of the trial in comparison to normal straight trials. And, there was no significant interaction of turn and session, indicating that the birds adapted fairly quickly to the turn stimulus. It therefore seems that the constraint did not impinge on the pigeons’ usual tracking performance.The ability to adjust readily to the turn may have been promoted by the constant visual input available on the trial. Experiment Maccaca mulatta) found that they were able to anticipate the reappearance of a moving stimulus when it disappeared behind an occluder, but if it just disappeared (without apparent reason) they were unable to maintain pursuit over the disappearance  at the point of the turn. Thus, the stimulus reappeared on a different trajectory from the initial motion that was displaced by 90°, but at the correct time given a speed of 3.40 cm/s. There was one turn-disappearance trial for each of the eight possible motion combinations in each session.There were a total of 32 test trials that were randomly intermixed with 40 normal training trials for a total of 72 trials. The contingencies of reinforcement were the same during the latter portion of the test trials (after the turn/disappearance) as for the normal training trials. The birds were tested until they met a criterion of at least 70% correct to each of the test types for two consecutive days; this required 40 sessions for Birds B50 and G83 and 50 sessions for Bird V42.Phase 2 was conducted immediately after Phase 1 and was identical in all respects except that the period of disappearance was increased to 1.32 s (or a movement of 4.5 cm). The birds were trained to the same criterion as Phase 1, requiring 40, 22, and 30 sessions for Birds B50, V42, and G83, respectively.Data Analysis). As seen in the figure (panel a), it appears that the disappearance trials, particularly the straight-disappearance trials, resulted in poorer performance than the normal trials, a result that was verified by the statistical analysis, F = 73.3, p < .05. This effect was driven heavily by Bird V42, which displayed a large disruption in performance following stimulus disappearance, particularly on the Straight-Dis trials. The other birds performed more similarly to all four trial types. There was no effect of the turn on capture success, F = 1.1, no effect of session, F < 1, Turn × Disappearance, F < 1, Disappearance × Session, F < 1, Turn × Session, F < 1, or Disappearance × Turn × Session, F = 1.2.Figure F = 12.4, p = .072. There was also an effect of session on performance, F = 6.1, p < .001, due to improvements in performance over the course of testing. Although performance appeared to improve more for the disappearance and turn trials than for the straight trials, there was no interaction of session with the other variables [all Fs < 1.4]. Given that there were changes over sessions, further comparisons were conducted to assess the effect of disappearance on the mean capture location. Disappearance did result in significantly later capture in block 2, 4, 6, and 11 . The effect of the disappearance on capture location can also be seen in the distribution of capture locations, shown in panel c. Stimulus disappearance resulted in a shift in the distribution so that peak in capture did not occur until around 1.5-2.0 cm after the stimulus reappeared; thereafter the capture distribution declined in a manner similar to the normal straight and turn trials. All three birds displayed a deficit in capture location following disappearance, but the magnitude of the effect varied somewhat across birds .The mean distance travelled before capture is shown in panel b. Here, it was disappearance appeared to result in later capture, but this did not achieve statistical significance, F = 14.1, p = .064. The birds also appeared to lag more on the trials in which the stimulus disappeared briefly, however, again there was no significant effect of disappearance, F < 1, nor was there any interaction of turn and disappearance, F < 1.Figure t = 4.79, p < .001. There were no significant effects of the previous motion, β = .04, t = 1.2, straight versus turn, β = .01, t = .16, or disappearance, β= -.06, t = -.72, on the error locations.A regression on the mean error locations with the predictors of previous motion, current motion, straight versus turn, and disappearance versus constant revealed a similar effect to Experiment F = 6.3, p = .127. There was no effect of turn, F = 2.1, nor was there any Disappearance × Turn interaction, F = 1.8. There also was no effect of session, nor any interaction of session with the other variables, all Fs < 1. In examining the individual birds, all three birds displayed a modest deficit in capturing the Straight-Dis and Turn-Dis stimuli, and one bird exhibited an additional deficit in capturing Turn stimuli. The magnitude of the deficits varied across birds from 10%-30%.Panel a of Fig. F = 26.6, p = .036, where the stimulus was caught later on disappearance trials. There was no effect of the turn, F < 1, or Turn × Disappearance interaction, F = 5.1. There also was no change over sessions, F < 1, nor was there any interaction of session with any of the other variables (largest F = 1.2). The distribution of capture locations (panel c) shows that the peak in capture responses occurred approximately 2.5-3 cm later and then declined gradually. All three birds displayed later capture on disappearance trials (both Straight-Dis and Turn-Dis), but the magnitude varied slightly across birds .Panel b reveals an effect of disappearance on mean capture location, F = 43.5, p < .05. There was also a main effect of disappearance, F = 83.7, p < .01, and a significant Turn × Disappearance interaction, F = 82.3, p < .01. Post-hoc analyses revealed that the interaction was due to the birds pecking significantly ahead of the stimulus on the straight normal trials compared to the other trial types.Figure t = -4.70, p < .001. There was no effect of the previous motion, β = .037, t = 1.0, straight versus turn, β = 0.01, t = .02, or disappearance, β = -.05, t = -.62.A regression examining the role of the current and previous motion on errors on normal and disappearance straight and turn trials revealed the same pattern of results as in the previous phase, with a strong effect of the current motion direction on error locations, β = -.17, The disappearance of the stimulus for either 1.5 cm (0.44 s) or 4.5 cm (1.32 s) had only a modest, non-significant effect on capture success, indicating that the disappearance did not disrupt capture. However, both durations of disappearance caused a decrease in capture efficiency and thus the stimulus was caught later on the disappearance trials. In fact, it appeared that the probability of capture at a given point shifted approximately 1.5 cm later for the trials with a 1.5-cm disappearance and 3 cm later for trials with a 4.5-cm disappearance. This suggests that some disruption to the anticipatory mechanism occurred and that longer periods of disappearance were more disruptive.This idea is supported by the error analyses. The short disappearance had a moderate  effect on the mean error position in that the birds lagged behind the stimulus more. When the stimulus disappeared for a greater period (4.5 cm), there was a greater effect on the mean error position with an increase in lagging errors. Regression analyses revealed that the current motion was the only significant predictor of error locations, indicating that the birds were correctly tracking the post-turn motion, even if they were lagging to a greater extent. If there had been a residual effect of the pre-turn motion on performance, then this would have indicated some persistent effect of the early visual information on pecking locations, but this was not the case.Overall, the pattern of results in the present study is consistent with the notion that the disappearance resulted in a period where the pigeons disengaged from tracking the stimulus. When the stimulus reappeared, there was a lag before tracking was re-engaged and this was greater than the normal lag to begin tracking at the start of a trial. This was seen both in the later capture and greater lagging errors, and the effect was larger when the stimulus disappeared for a longer period. But, once tracking was re-engaged, the birds appeared to track the motion appropriately. = 21.9, p = .043. There was no effect of the turn on capture success, F = 4.6, but there was an Occlusion × Turn interaction, F = 230.1, p = .004. Follow-up analyses indicated that indicated that the Straight-Occ trials resulted in poorer performance than the Straight and Turn-Occ trials. There was no effect of session, F = 1.3, nor any interaction of session with the other variables (largest F = 1.4).Capture responses are displayed in Fig. F = 19.2, p = .048. However, there was no effect of turn, session, or any interactions of turn or session with the other variables (largest F = 1.9). In examining the distributions of capture locations (panel c), it appears that occlusion resulted in a modest shift in the peak capture location of around 0.5-1.0 cm. The individual birds all displayed later capture on the occlusion trials, but the magnitude of the effect varied across birds .There also was a significant main effect of occlusion on the mean capture location, shown in panel b, F = 5.7, but did reveal a main effect of occlusion, F = 48.3, p < .05, with birds lagging more on the occluder trials. There was a also a significant Turn × Occlusion interaction, F = 29.7, p <. 05. Post-hoc analyses revealed that the significant interaction was due to the birds pecking significantly further ahead of the stimulus on the straight normal trials compared to the other motions.Figure t = 2.0, p < .05. The current motion direction did not predict errors β = 0.02, t = .5, and there was once again no overall impact of straight versus turn, β = .03, t = .3, but there was a significant effect of occlusion versus non-occlusion on errors, β = -.19, t = -2.0, p < .05.A regression examining the role of the previous and current motion on post-occlusion errors on normal and occluded straight and turn trials revealed that previous motion direction was the primary predictor of error locations, β = 0.09, A 1.5-cm occlusion of the moving stimulus disrupted capture success on straight-occluder trials. This stands in contrast to the general lack of effect that turns and disappearances had on capture success in the previous experiments. A disruption of capture success indicates a loss of stimulus control over tracking behavior, and that there was a general disruption in the tracking process. It is possible that the presence of a novel stimulus on the screen may have caused this disruption by competing with the moving stimulus for attention, as has been observed in infants  increase in lagging errors. The regression analysis conducted on the errors indicated that the birds successfully adjusted their tracking and used the correct motion following unpredictable turns when visual input was available during the turn  over an occlusion is highly adaptive for both predator and prey animals. Despite this clear adaptive value, pigeons are notoriously poor at recognizing partially occluded objects (e.g., Cerella, The use of touchscreen technology allowed us to monitor both capture and error behavior with great accuracy. However, resistive touchscreens do not count every single peck because some lighter intensity pecks fall below the resistance threshold. As such, some information is likely to be missed and this could affect the results, for example, by reducing capture rates. However, given that this issue is present on all trials we do not believe it impacts our overall interpretation of the results. Another issue relates to how pigeons peck when confronted with smaller objects as used here. When smaller grain-like stimuli are presented, pigeons peck in a manner consistent with how they peck at food (Jenkins & Moore, Wilkinson and Kirkpatrick  found thDespite the apparent differences in the mechanisms underlying their tracking behavior, the pigeons’ responses to disappearances and occlusions appear to be similar to those observed in primates (e.g. von Hofsten et al.


Graphene is one of the most favorite materials for materials science research owing to its distinctive chemical and physical properties, such as superior conductivity, extremely larger specific surface area, and good mechanical/chemical stability with the flexible monolayer structure. Graphene is considered as a supreme matrix and electron arbitrator of semiconductor nanoparticles for environmental pollution remediation. The present review looks at the recent progress on the graphene-based metal oxide and ternary composites for photocatalysis application, especially for the application of the environmental remediation. The challenges and perspectives of emerging graphene-based metal oxide nanocomposites for photocatalysis are also discussed. Graphene is a 2-D material composed of layers of carbon atoms crammed into a honeycomb network and has become an escalating star on the prospect of materials science in the past many years ,2,3. GraThe growth of graphene-based composites provides a significant milestone to multiply the application enactment of metal oxide nanomaterials in photocatalysis, as the hybrids have adaptable and suitable properties with superior performances over the individual oxide nanomaterials. With keeping this in mind, considerable efforts have been made on decorating graphene with metal oxides . GraphenIn this context, focusing on the recent developments, an attempt is made in the present review to discuss the advantages and disadvantages of the composite in comparison with pristine graphene.Different metal oxides-GO/rGO composites, their photocatalytic activities and synthesis process are discussed in the following sections as well as in Table 1. However, the photocatalytic performances of metal oxide-GO/rGO composite photocatalysts for the degradation of different pollutants are tabulated in Table 2.3 catalyst dosage has a substantial impression on the photocatalytic activity. The finest Sulfamethoxazole (SMX) removal efficacy was achieved when rGO-WO3-200 loading was 1.0 g L−1, and this study demonstrated that the dosage of the catalyst is crucial for the photocatalytic activity. SMX degradation over rGO-WO3-200 was significantly influenced by neutral pH and displayed the pre-eminent process [3–nanocomposite and WO3 was achieved by 84% and 40% respectively, attributed to the larger specific surface area and lower band gap energy [3 and rGO sheets and suppressing the electron-hole pair recombination in the WO3-rGO nanocomposite [3/GO composites revealed an enhanced photocatalysis under the visible light, which was two-folds of pure WO3 that reduced the recombination of the photogenerated electron-hole pairs and increased the visible light absorption efficiency [rGO-WO process . Photodep energy . Outcomep energy . The enhomposite . WO3/GO 3O4/rGO composite exhibits better photocatalytic activity in a low concentration of Methyl Orange (MO). The higher stocking dose of Co3O4 on Co3O4/rGO contributes more on the activity [α-MoO3 from 2.82 to 2.51 eV [3O4-rGO hybrids were reported to completely oxidize 20 mg/L phenol in 20 min at 25 °C. Origination of sulfate radicals via Co3O4-mediated activation of peroxymonosulfate (PMS) is responsible for this catalytic effect [3O4/N-doped graphene hybrid exhibits similar catalytic activity but superior stability to Pt, in alkaline solutions. This unusual catalytic activity arises from synergetic chemical coupling effects between Co3O4 and graphene [Coactivity . Degrada 2.51 eV . This syc effect . The Co3graphene .2–GO nanocomposites was examined with different ratios of graphene additions and it was found that higher amount of graphene addition decreased the photocatalytic activity [2-GO photocatalyst showed the similar occurrence. This study demonstrated that TiO2-GO cannot offer truly new visions into the assembly of TiO2 carbon composite as high-performance photocatalysts. The TiO2 particles were found to be in anatase phase and a narrow size distribution was dispersed on the surface of graphene sheets uniformly [2 and G-TiO2 was reported with different reaction times, as shown in 2 [2 (P25)-rGO composite was found to be the most proficient photocatalyst for the degradation of Methylene Blue (MB) and the optimum mass ratio was found to be 1/0.2 [A systematic investigation on the photocatalytic properties of TiOactivity . Liquid-niformly . A compaown in 2 . TiO2 , both under UV and visible light. In this report, photoluminescence studies revealed discrete appeasing of the GO photoluminiscence under visible light and near infrared laser excitation. Hence, it was conferred that GO acts as either an electron acceptor or donor of TiOle light .2 (N-P90), nitrogen-doped reduced graphene oxide (N-rGO), as well as their composites were studied for the photocatalytic activity. N-P90/N-rGO showed enhanced photocatalytic activity, and in the presence of this composite, around 80% MB was degraded by visible light irradiation in 160 min. Enhanced photocatalytic performance is observed in N-P90/N-rGO composites for degradation of MB due to photo-induced and electronic interaction between TiO2 and graphene. Comparison of degradation efficiency of MB under visible light irradiation by P90 TiO2, N-P90, and N-P90/rGO is presented in 2 nanofibers (~10 nm diameters) were synthesized from electrospun rice-shaped TiO2 and potassium titanate was achieved from rice-shaped TiO2. The surface area was increased by 2.5 times after the nanofiber formation of TiO2. The results showed that the photodegradation of MO was found to be higher than bare TiO2 (P-25) nanoparticles [2 of different composition ratios were tested and the formulation of catalyst with 1.2 times higher photocatalytic activity than commercial photocatalyst was reported. This catalyst was able to degrade 3 mg/L MB over 10 consecutive cycles with nominal loss in photocatalytic efficacy. Graphene plays a generous part in obstructing the accretion of TiO2 grains upon calcination at high temperature [2 (rGO-TiO2) nanocomposites was synthesized and examined for the photocatalytic activity by decolorization of Rhodamine B dye (RhB) under UV light. In this study, various parameters, such as dye concentration, rGO content, catalytic dose, and pH, were optimized for the decolorization. The catalysts were found to be more active at natural pH of the dye under the UV-illumination for the degradation of RhB dye. The presence of H2O2 and K2S2O8 increased the decolorization. Further, addition of CO32− and Cl− ions decreased the dye degradation rate [Nitrogen-doped P90 TiOarticles . GO/TiO2perature . A set oion rate .2 nanoparticle-attached graphene/carbon composite nanofibers (TiO2-CCNFs) were synthesized and reported as highly active photocatalysts for photocatalytic degradation of MB under the irradiation of visible light. Graphene was suggested to play the role of an electron acceptor and a photosensitizer, resulting in a higher photodegradation rate and reduced electron-hole pair recombination. CNFs having high surface also improved the photocatalytic activity of TiO2 [TiO of TiO2 .An effective scalable method was developed to make nanocomposites of functional graphene sheets (FGS)/ZnO. In this study, poly (vinyl pyrrolidone) (PVP) component was reported to play a crucial role for loading of ZnO nanoparticles onto FGS by connecting Zn ions on the carbon materials and promoting ZnO nucleation and crystal growth in the precursor-prepared route. Further, FGS/ZnO composite was evaluated for photocatalytic activity and was found to be applicable for a number of environmental issues . ZnO/rGO2/graphene sheet (GS) (5%) composites for H2 evolution from water splitting. This study demonstrated that beyond the critical content of GS (5%), photocatalytic activity was decreased by initiating electron-hole recombination centers. Calcination atmosphere was found to be important and better performance was observed for the samples calcined in nitrogen atmosphere [The calcination atmosphere was found to affect the photocatalytic activity of the TiOmosphere .The use of ZnO-graphene composites (Z-GC) was reported to remove dye from water due to the interaction between the graphene sheets and the ZnO nanoparticles . rGO-ZnO2 atmosphere. The superior photocatalytic performance was due to the synergistic effect of the proficient electron inoculation and low charge carriers in the composite, where GO acted as an electron collector and transporter, leading to unceasing generation of reactive oxygen species for the degradation of MB [2S3 bi-component shell anchored on the rGO sheets were synthesized and reported to show a broad and strong photo-absorption in the visible region. These nanorods also manifested better photocatalytic activity for H2 evolution from the glycerol-water mixtures. The superiority in performance is owing to the elevated light absorption and effective charge separation [The ZnO/GO nanocomposite consisting of flower-like ZnO nanoparticles anchored on graphene-oxide. Further, photocatalytic efficiency of ZnO/GO composite progressed by annealing the product in Non of MB . Core-shparation .2O4 in which the graphene acts as a charge carrier to detain the delocalized electrons. Photocatalytic activity was explored with the variation of the dosage and dye concentration [2O4-graphene hybrid materials (CFGHs) showed ferromagnetic behavior and enhanced photodegradation rate and amended adsorbing capacity due to the assimilation of graphene [2O4 and CdS. Escalation in the concentration of MB was observed as the irradiation time increased for CoFe2O4 due to the desorption of MB during irradiation. G-CoFe2O4/CdS easily separated from aqueous solution in an external magnetic field, as seen from the digital photos of Gr-CoFe2O4/CdS after irradiation [2O4-rGO composite unveiled required photocatalytic performance with excellent recycling stability for the degradation of MB, RhB, and MO under visible-light irradiation [Connexion of the graphene suggestively progressed the photocatalytic performance of the CoFentration . CoFe2O4graphene . The phoadiation . The CoFadiation .2O4-rGO (CF–rGO) nanocomposites hold exceptional microwave absorbing properties and high photocatalytic activity for the degradation of various dyes under visible light irradiation [CoFeadiation . 85 CF-1adiation . The phoadiation .2O4-3D TiO2 nanocomposite showed an enhancement in the photodegradation of MB as compared to the commercial rutile-phase TiO2 and the pure urchin-like TiO2 (3D TiO2) microparticles. Results specified that the composite showed relatively consistent photocatalytic activity with slight degradation [2O4 composites [CoFeradation . The phomposites . The pho2O4/Ag under visible light (λ > 400 nm) was investigated by in-situ Fourier transform infrared spectroscopy and the correlated mechanisms were suggested. Superficial degradation ratio of 91.9% over rGO/CoFe2O4/Ag was obtained under visible light illumination of 12 h, while only about 21.7% was obtained with commercial P-25 TiO2 [−1, and then, no noteworthy decrease of time was observed when more catalyst was added. Likewise, use of 2 mL of H2O2 was found to be an optimum amount for the photocatalysis reaction (2O4 nanocomposites was queried for the degradation of 4-Chlorophenol (4-CP) under visible light illumination. Activity of rGO-CoFe2O4 composite was seen in the occurrence of PMS  plane confined in the graphene network exhibited an excellent visible-light-driven photocatalytic activity with an ultrafast degradation rate of 1.924 × 10−7 mol g −1 s −1 for MB [Photocatalytic activity of ZnFeadiation . Graphen1 for MB .2O4-rGO nanocomposite was shown due to the active restraint of the recombination of the photo-excited electron-hole pairs by rGO sheets and the generation of ·OH-free radical [2) [2O4-Ag/rGO nanocomposite was higher by the factor of 14.6 and 5.6 times over its counterparts. Photosensitization effect was prevailed by good interaction ensuing in only 80% removal of EE2 though humic acid [2O4 composite exhibited the remarkable catalytic activity toward MB degradation; in the presence of H2O2, the activity enhanced, and the reaction followed a pseudo-first-order kinetics. The complete MB degradation observed at rGO/ZnFe2O4 composites was attributed to the π–π interaction, hydrogen bonding, and electrostatic interaction exerted between the rGO and ZnFe2O4 [The boosted photocatalytic activity of ZnFe radical . The phoical [2) . The psemic acid . rGO/ZnF ZnFe2O4 .2O4-GO (0.25) hetero-architecture demonstrated a considerable lesser emission intensity. Due to their competent electron-transport property, graphene sheets can deliberately reduce the fluorescence of NiFe2O4 fixed on them. Kinetic results indicated that the rate-determining step is the adsorption course of MB [2O4-GO (0.25) shows the best activity compared to other NiFe2O4-G composites  under visible light due to its combined effects of modulated band gap and covalent bonding between BiFeO3 and graphene [RGO-Bile light . BiFeO3-graphene .3O7 (OH)-RGO composite supported the suggested mechanism of charge separation and transport mechanism. A higher degradation rate was obtained using the nanocomposite prepared with a graphene loading of 3 mgmL−1, and when the rGO loading exceeded 3 mgmL−1, degradation efficacy diminished. This arose as extra rGO sheets gathered and stuck the absorption of incident light [Photoluminescence studies of Nbnt light .2 and ZnO exhibited good photocatalytic activity in UV light due to their wide band gap. These two metal oxides are stable in aqueous conditions during photocatalysis. Further, coupling of graphene with TiO2 and ZnO increases the photocatalytic activity due to increases in the photogenerated charge carriers. Metal oxides with magnetic properties of metal ferrites (MFe2O4) offer an added advantage as photocatalysts since they can be recovered by applying an external magnetic field after catalysis. Metal ferrites  materials are proven to be excellent candidates for visible light photocatalytic H2 generation through water splitting. Recycling ability for metal ferrites are far better compared to nano semiconductors like TiO2 and ZnO. MFe2O4 is a class of semiconductor with narrow band gap, which exhibits characteristic visible light response, possess good photochemical stability, and exhibits excellent optical properties.Pristine TiO2O4 absorbs 42–45% of sunlight, whereas TiO2 and ZnO absorbs 4% of sunlight. MFe2O4 are efficient for the degradation of dye degradation and organic pollutant degradation compared to the other metal oxides , with respect to the catalyst and the light source. In MFe2O4 context, recombination of photogenerated charge carriers is the major limitation in semiconductor photocatalysis as it reduces the overall quantum efficiency. In order to enhance the photocatalytic activity, graphene material is coupled with MFe2O4, where the graphene channels the electrons. Comparison of degradation rate for various photocatalytic reaction systems is incongruous since the nature of catalyst and substrate pollutant molecules are different in each reaction. Ferrite nanoparticles have a strong magnetic property, which can be easily used for magnetic separation after photo-mineralization.MFe2+ ion easily bonds with oxygen by giving an electron and super oxide radical. This super oxide radical can oxidize the organic substrate molecule. The Fe3+ ion and Fe2+ ions can show photo-Fenton reactions in presence of in-situ-generated H2O2. This H2O2 generates hydroxyl-free radicals, which are involved in the degradation of pollutants. Predicted mechanism for the rGO-CoFe2O4 composite is shown in Equations (1)–(4).The photocatalytic efficiency depends on the ratio of the photogenerated charge-carrier transfer rate to the rate of electron-hole recombination. For composite structure, M6 composite shows better photocatalyst compared to other catalysts prepared from hydrothermal method  display that h+, O2−, SO4−, and OH are accountable for decomposition of antibiotics. Overall, irrespective of other parameters, the solvothermal method is best and helps in crystal growing and super saturation is achieved by reducing the temperature in the crystal growth zone.Furthermore, the self-redox properties of iron and manganese atoms in MnFeFurther, noble metal  exhibits surface plasmon resonance (SPR), which is a characteristic feature. The SPR frequency of the metal particles can be tuned into visible light absorption by shifting the size of the deposited metal particles on the catalyst. Deposited metal is involved in multiple crucial roles, such as serving as a passive electron sink with high capacity to store electrons to suppress photogenerated charge carrier recombination, facilitates rapid dioxygen reduction to generate free radicals and direct excitation of metals, especially under visible light, and vectorial electron transfer to the conduction band (CB) of metal oxide. Thereby, showing improvement in the photocatalysis for the removal of various organic pollutants/dyes.Graphene nanosheets act as a substrate to support the metal oxides for photocatalytic activity and graphene-based semiconductor photocatalysts are used for environmental remediation. The morphologies of semiconductors, theoretical electronic-structure calculations, and experimental discovery determinations are necessary on GO to persuade the photocatalytic activity, and composition design is an operative method to enhance the photocatalytic properties. Photocatalytic properties depend on the preparative method, and various parameters like initial concentration, oxidant concentration, pH, particle size, number of GO sheets, and source of light should be explored.The interface regulates the efficacy of the electron-hole separation. Currently, only few methods succeed in unswervingly depicting the interaction of GR and nanoparticles. Atomic force microscopy (AFM), Surface-enhanced Raman scattering (SERS), and scanning transmission electron microscope (STEM) may be the best techniques for determining the interaction of graphene and nanoparticles. Finally, studies on the preparation of a ternary composite as a photocatalyst for both UV and visible-light-driven pollutant photodegradation have been studied and reported. Especially, for the design of ternary composite, magnetic materials such as Fe, Co, Mn, etc., as a dopant, and possessing unique advantages, show a remarkable photocatalytic activity and photostability.Further tasks exist in the application of graphene-based composite for the industrial scale. Some innovative applications of the metal oxide-graphene entail specific understanding between the metal oxides and surface of the graphene, which will have a direct impact on the properties of the composite. Designing a structure for the overall photocatalysis process may require further exploiting of GO by chemically modifying methods. A synthetic approach method of GO-based composite structure by using novel materials has not been achieved to date for photocatalysis, but the solutions to the key challenges appear within reach.In view of this, graphene-based composites possess diverse potential applications, individually having dissimilar desires concerning material properties, and it can be projected that the research on graphene-composite materials will have an optimistic future.


The ability to define an off state in logic electronics is the key ingredient that is impossible to fulfill using a conventional pristine graphene layer, due to the absence of an electronic bandgap. For years, this property has been the missing element for incorporating graphene into next-generation field effect transistors. In this work, we grow high-quality armchair graphene nanoribbons on the sidewalls of 6H-SiC mesa structures. Angle-resolved photoelectron spectroscopy (ARPES) and scanning tunneling spectroscopy measurements reveal the development of a width-dependent semiconducting gap driven by quantum confinement effects. Furthermore, ARPES demonstrates an ideal one-dimensional electronic behavior that is realized in a graphene-based environment, consisting of well-resolved subbands, dispersing and non-dispersing along and across the ribbons respectively. Our experimental findings, coupled with theoretical tight-binding calculations, set the grounds for a deeper exploration of quantum confinement phenomena and may open intriguing avenues for new low-power electronics. Here, the authors investigate armchair graphene nanoribbons by angle-resolved photoelectron spectroscopy, and show the development of a width-dependent semiconducting gap driven by quantum confinement effects, and an ideal one-dimensional electronic behaviour. Namely, GNRs terminated by zigzag edges demonstrate one-dimensional (1D) single-channel ballistic transport at room temperature5, while those with armchair edges promise a width-dependent semiconducting behavior11. In the view of digital applications, the potential of graphene is limited by the lack of a sizeable bandgap in its electronic structure. Hence, the scalable growth of high-quality armchair GNRs (AGNRs) is essential for the development of future graphene-based technologies. Earlier attempts to produce GNRs involved the patterning of pristine graphene layers using standard lithographic means. However, such top-down approaches severely suffer from the lack of atomic-scale precision, resulting in the formation of disordered and rough edges14. In contrast, the surface-assisted bottom-up approach, for instance the self-assembly of molecular precursors on gold surfaces, was demonstrated to create high-quality GNRs17. The drawback of this method lies in its use of a metallic substrate, and thus its impracticality to be incorporated in a technologically relevant application. In this respect, the production of GNRs on a semiconducting substrate such as SiC is an attractive avenue21. Particularly, the selective growth of AGNRs on the sidewalls of SiC mesa structures, is shown to produce well-defined edge morphologies21 and to provide a better control on the overall ribbon widths as compared to the growth of AGNRs on the 19. Therefore, epitaxial sidewall AGNRs stand as promising candidates for the realization of new tunneling-based field effect concepts22, since going beyond conventional metal-oxide-semiconductor field-effect transistor (MOSFET) architectures, almost defect-free semiconductors are mandatory.Graphene nanoribbons (GNRs) are considered to be the fundamental building blocks for future carbon-based nanoelectronics. The functionality of GNRs is governed by the detailed atomic structure of their edges25. A combination of well-resolved dispersing and non-dispersing subbands, parallel and perpendicular to the ribbon direction respectively, is observed. In fact, a characteristic band structure topology, resembling a triangular prism as expected for 1D confined Dirac electrons, was never realized thus far in a graphene-based environment. Tight-binding (TB) simulations reveal that the detailed subbands measured in ARPES originate from a mixture of metallic and semiconducting AGNRs, indicating the development of a width-dependent bandgap. This is further corroborated by scanning tunneling spectroscopy (STS) measurements, which confirm that individual ribbons can display either metallic or semiconducting behavior.In this work, high-quality AGNRs are formed through the periodic modulation of a graphene sheet grown epitaxially on the sidewalls of 6H-SiC mesa structures. Angle-resolved photoelectron spectroscopy (ARPES) reveals peculiar features that differ from previous GNR band structure measurements26. The armchair edge of graphene is naturally oriented parallel to the [1121 with an average inclination of (26 ± 2)°. This periodic array of mini-facets, which is somewhat reminiscent of a “ladder structure”, can be observed in the scanning tunneling microscopy (STM) image of Fig. N-AGNR to denote a ribbon with N dimers across its width, we find ribbons composed of N ~ 18 dimers, corresponding to a width of about 2 nm as shown in the inset of Fig. 5 AGNRs/mm, a density high enough to allow the use of integrating experimental techniques such as low-energy electron diffraction (LEED) and ARPES.Graphene on 6H-SiC develops in a well-defined epitaxial relationship with respect to the substrate26. The periodic array of mini-facets constituting the SiC sidewall  is described in units of the SiC phase S, which is defined as the phase difference between two electron waves scattered from adjacent terraces separated by a step height of a single SiC bilayer. Assuming that the facets consist of mini-steps equivalent to a single SiC bilayer, we would expect the facet rods to intersect the basal rods only at in-phase conditions, i.e., at integer values of S. However, in Fig. Figure see Fig.  while thpz orbitals and preventing the development of a Dirac cone. However, on the vicinal planes, the strong graphene-SiC interaction is lifted due to the different surface registry of the substrate as compared to the basal plane. Consequently, free-standing AGNRs are developed, generating 1D confined Dirac electrons. The mini-facets themselves are unit-cell high (H = 1.5 nm) and have a periodicity of D = (3.1 ± 0.3) nm. The free-standing ribbons covering the mini-facets are approximately 2 nm wide with an average inclination of (29 ± 1)° relative to the basal plane, as estimated from the ARPES measurements discussed below.Based on the properties revealed by AFM, STM, LEED, and SPA-LEED, a structural model is proposed for AGNRs in Fig. 1. Fig. 29, is also made visible near the Fermi level (EF). The Dirac point is situated at EF, which corresponds to electronically neutral AGNRs. The subband energies are determined by locating the positions of the peak maxima in the second derivative of the energy distribution curve (EDC) taken at 6. In particular, ribbons with N = 3p + 2  are predicted to be metallic , whereas all other widths result in semiconducting ribbons whose bandgaps are inversely proportional to their widths6. Accordingly, we consider in Fig. N-AGNRs with N = 16, 18, and 20, whose widths are in the 2 nm range suggested by our STM data , consistent with the band structures from ARPES and TB. A wide-bandgap nanobuffer layer is also identified in Fig. An overview of the electronic band structure of AGNRs is depicted in Fig. Å−1 Fig. . It is ientclass1pt{minimak-space is a result of 1D confinement as discussed above. Such characteristic 1D behaviors have been examined before in other systems, e.g., using self-organized metallic nanowires on semiconductor surfaces31. The intensity modulation observed in the Fermi surface is mainly due to an undesired parasitic graphene layer that, in small patches, has overgrown the SiC sidewalls and has an estimated surface area coverage on the order of a few percent. A 2D graphene sheet that is grown on an inclined facet would correspond to a 2D hexagonal BZ shifted away from normal emission relative to the basal plane as sketched in Fig. 32). At a photon energy of 40 eV and a spectrometer work function of 4.3 eV as used in the experiment, the −1 from their associated Figure mentclass2pt{minimmentclass2pt{minimk-space, we display the 2D reciprocal space map (RSM), which is constructed by drawing the BZs of the parasitic sidewall graphene layers on top of the LEED pattern  eV is introduced in the electronic structure, which is desirable for defining an n-doped 6H-SiC(0001) wafer pieces  were used as substrates for the AGNR growth. They were atomically flattened via hydrogen etching, which was carried out in an inductively heated reactor equipped with a graphite susceptor39. Mesa structures with a lateral dimension of about 150 nm and a height of 20 nm were defined using a combination of e-beam lithography and reactive ion etching . The selective growth of AGNRs was achieved by annealing the samples in an inductively heated furnace at 1800 °C under 850 mbar of argon atmosphere for 1 min21.Nominally on-axis, single crystalline, 2, simultaneously probing around 2000 AGNRs. All ARPES data were acquired in ultra-high vacuum (UHV) at a sample temperature of 80 K. Additional, preliminary ARPES measurements (not shown in the manuscript) were conducted at the 12 endstation of the UE112 beamline at BESSY II, Helmholtz-Zentrum Berlin.ARPES measurements were carried out at the Bloch beamline of the MAX IV synchrotron facility in Lund, Sweden. High-resolution energy-momentum cuts were measured using a high performance deflector-based DA30 hemispherical analyzer from ScientaOmicron. The energy and angular resolutions were set to 15 meV and 0.1° respectively. The spot-size of the beam measured 10 × 24 µmp < 2 × 10−11 mbar) at 80 K using a commercial Omicron LT-STM. The sample was degassed at an elevated temperature of 800 K for several hours before being transferred to the low-temperature chamber. Several electrochemically etched tungsten tips were utilized during the experiments. A lock-in detection technique was used to accumulate and average over 50 STS spectra taken on each individual ribbon.The STM/STS measurements were performed in UHV  tight-binding (TB) Hamiltonianentclass1pt{minimaSupplementary Information


These results demonstrated that FOXC2-AS1 enhances FOXC2 mRNA stability to promote CRC proliferation, migration, and invasion by activation of Ca2+-FAK signaling, which implicates that FOXC2-AS1 may represent a latent effective therapeutic target for CRC progression.Long noncoding RNAs (lncRNAs) have been confirmed, which are involved in tumorigenesis and metastasis in colorectal cancer (CRC). FOXC2 antisense RNA 1 (FOXC2-AS1) was reported, facilitating the proliferation and progression in several cancers. However, the role of FOXC2-AS1 in CRC cell migration and metastasis is not unclear. In this study, we observed that lncRNA FOXC2-AS1 was upregulated in CRC tissues, and its high expression indicated the poor survival in CRC patients. Meanwhile, FOXC2-AS1 was higher in CRC tissues with metastasis than that of nonmetastatic tumor tissues. We found that FOXC2-AS1 was predominately expressed in the nucleus of tissues and cells. FOXC2-AS1 knockdown suppressed CRC cell growth, invasion, and metastasis in vitro and in vivo. Moreover, FOXC2-AS1 could positively regulate the neighboring gene FOXC2 and stabilized FOXC2 mRNA by forming a RNA duplex. Meanwhile, ectopic expression of FOXC2 could obviously alleviate the suppressed effects caused by silencing FOXC2-AS1. For the mechanism, FOXC2-AS1 knockdown could reduce intracellular Ca In recent years, remarkable progress has been made in CRC treatment, but over half of the patients with advanced CRC die from recurrence and metastasis, which led to the unfavorable prognosis3. Thus, a better illustration of the molecular mechanisms regarding CRC progression is urgently needed.Colorectal cancer (CRC) is a common digestive tract malignancy, accounting for about 40% new tumors of the digestive tract. According to the survey, there were nearly 1.3 million new CRC cases and more than 0.6 million deaths worldwide every year4. LncRNAs demonstrated widespread expression in animals and plants, which can exhibit function in cis-regulated expression of neighboring genes or in trans-regulation distal genes5. Numerous literature has confirmed that the dysregulation of lncRNAs is involved in the pathophysiology of numerous human disorders, especially in cancer7. lncRNA H19, MALAT1, HOTAIR, etc., were widely reported to be involved in human tumorigenesis and metastasis10. LncRNA LNMAT1 promotes bladder cancer metastasis through enhancing CCL2- dependent macrophage recruitment11; LncRNA MFI2-AS1 facilitates cell migration and invasion via acting as the ceRNA of miR-574 to regulate MYCBP in CRC12; HOXD-AS1 represses CRC growth and metastasis through suppressing HOXD3-activated integrin β3 transcription and MAPK/AKT pathway13. Thus, lncRNAs may play a key role in cancer metastastis; exploring the molecular mechanisms will be significant for cancer therapy.Long noncoding RNAs (lncRNAs) are a kind of RNA transcripts with lengths >200 nt, and limit or lack protein-coding capacity16. Besides, FOXC2-AS1 was reported to promote doxorubicin resistance in osteosarcoma17. Yet, FOXC2-AS1 is not well-characterized in cancers, including CRC. The expression, function, and mechanism of FOXC2-AS1 has not been reported in CRC.FOXC2-AS1 is an antisense RNA transcript of forkhead box protein C2 (FOXC2). Research has shown that FOXC2-AS1 was highly expressed in breast cancer, prostate cancer, and non-small- cell lung cancer (NSCLC); the upregulated expression of FOXC2-AS1 predicted poor survival, and facilitated proliferation and progression of these carcinomas2+ levels and promote FA formation via activation of Ca2+-FAK signaling pathway, ultimately contributing to CRC proliferation, migration, and invasion. Our investigation revealed the oncogenic role of FOXC2-AS1 through increased intracellular Ca2+ levels and promoted FA formation in CRC, and implied FOXC2-AS1 as a latent prognostic marker and therapeutic target for CRC.In this research, we found that FOXC2-AS1 was obviously upregulated in metastasis CRC tissue, and relates to the clinicopathologic features and poor prognosis in CRC patients. FOXC2-AS1 depletion significantly weakened CRC cell growth, invasion, and metastasis in vitro and in vivo. Moreover, FOXC2-AS1 could promote FOXC2 expression via enhancing its mRNA stability. Further investigation demonstrated that FOXC2-AS1 could increase intracellular Can = 38) and low group (n = 28)  Fig. . The assWe investigate the role of FOXC2-AS1 on CRC cells. First, we detected FOXC2-AS1 endogenous expression in CRC cell lines  and normal colonic cell line NCM460 by qRT-PCR. As shown in Fig. 18, and FOXC2 was the nearest gene, having some overlap sequences with FOXC2-AS1. Thus, we investigated whether FOXC2-AS1 regulates FOXC2 expression. First, FOXC2 expression was detected in CRC tissues and cell lines. It is found that FOXC2 was elevated in CRC specimens, which was consistent with the expression of FOXC2-AS1  in FOXC2-AS1-silenced and control cells. The results displayed that FOXC2-AS1 knockdown does not influence the level of nascent FOXC2 pre-mRNA  Fig. . Therefoown Fig. . Remarkaown Fig. , which i25. Immunofluorescence displayed that the number of FA was obviously reduced in FOXC2-AS1- silenced cells, but increasing FOXC2 expression could alleviate the inhibitory effects . Once activated by cellular signals, the concentrations increase nearly 100 times either through release from inter-Ca2+ stores or through extracellular Ca2+ influx, thus generating Ca2+ signals that activate downstream effectors, such as calmodulin, calmodulin-dependent protein kinase II, and calpain (CAPN)31. Disruption of Ca2+ homeostasis facilitates the formation of malignant phenotypes32. Increasing evidence has demonstrated that Ca2+ plays an important role in tumor proliferation, invasion, and metastasis. For example, PLPP4 depletion suppresses lung carcinoma cell proliferation and tumorigenesis via impeding the influx of intracellular Ca2+33. Gong et al. reported that P2RX6 might function via ATP-induced Ca2+ influx to promote renal carcinoma cell migration and invasion34. ABAT suppressed Basl-like breast cancer by downregulation of intracellular Ca2+ concentration and inactivation of Ca2+-NFAT1 axis35. Here, we found that FOXC2-AS1 knockdown decreases the level of Ca2+ via suppression of FOXC2; moreover, elevating the level of Ca2+ could mitigate the oncogenic effect by FOXC2-AS1 silencing. Consistent with previous reports, our data suggested that FOXC2-AS1/FOXC2 promotes CRC progression by elevating Ca2+ levels.Based on the GESA analysis in an expression-profiling data of FOXC2-overexpressing SW480 cells (GSE58980), we found that FOXC2 expression was positively associated with calcium signaling pathway and FA. As we know, Ca36. FA is recognized dominant in this process, which acts through signal-linkage complexes between the extracellular matrix (ECM) and the cytoskeleton37. The FA assembly and disassembly, named FA turnover, determine cell motility, and the speed of FA dynamic cycle is considered to be the critical step in tumor invasion38. FAK is a well-known nonreceptor protein tyrosine kinase involved in FA dynamics25. FAK contains six tyrosine phosphorylation sites ; FAK regulates cell adhesion and migration, mainly contributing to the autophosphorylation at Tyr397 site, which generates a high-affinity binding site for the SH2 domain of Src39. The interaction of FAK with Src subsequently phosphorylates Tyr407, 576, and 577, which lead to maximizing FAK activity39. Paxillin functions as a scaffolding protein that could recruit regulatory and structural proteins to regulate the dynamics of cell adhesion, modulating the turnover of FA complex and cytoskeletal remodeling, thus promoting cell migration and invasion40. When phosphorylating at Tyr118 by the FAK/Src complex, paxillin was activated; then multiple signaling pathways can coordinate to regulate cytoskeleton formation and promote cell migration41. In breast cancer, lung cancer, neuroblastoma, ovarian cancer, and hepatocellular carcinoma, studies have confirmed that FAK–Src–Paxillin cascade pathway modulates tumor cell proliferation, migration, and metastasis45. Our results proved that FOXC2 overexpression could reverse FOXC2-AS1 silencing-caused FA inhibition and FAK–Src–Paxillin cascade pathway inactivation. Notably, numerous evidence has demonstrated that Ca2+ signaling plays an important role in FA dynamics and FAK phosphorylation48. As shown in our results, elevating the level of Ca2+ mitigated FOXC2-AS1 silencing-induced FA inhibition and FAK–Src–Paxillin cascade pathway inactivation.Cell migration and invasion are a prerequisite for tumor metastasis. The migratory capacity facilitates cancer cells running away from the primary focus and disseminating through nearby circulations. The coordination of cell–substrate adhesion and detachment is necessary for cell migration2+/FAK signaling, and the new signal axis—Ca2+/FAK regulates cell FA and migration. These findings may provide a potentially effective therapeutic target for CRC progression.In summary, our research demonstrated that lncRNA FOXC2-AS1 promotes CRC progression via activation of CaSixty-eight cases of CRC specimens and 35 cases of adjacent tissues were diagnosed and collected in the second Xiangya hospital, Central South University between 2012 and 2016. No patient received chemotherapy and radiotherapy before surgery. The fresh specimens were snap-frozen in liquid nitrogen and then stored at −80 °C. All the patients were followed up at regular intervals after surgery. This research was approved by the Ethics Committee of The Second Xiangya Hospital of Central South University, and informed consent has been obtained by all the enrolled patients. The pathological information was obtained from patients’ medical records.CRC cell lines HT-29, HCT116, SW620, LoVo, and normal colonic epithelial cell line NCM460 were obtained from American Type Culture Collection (ATCC) and cultured following the manufacturer’s instructions.FOXC2-AS1 siRNAs that directly target the sequence were synthesized by Invitrogen ; scramble oligonucleotides were used as a negative control. The sequences of si-FOXC2-AS1 were as follows: si-FOXC2-AS1-1, 5′-GCGUGCCACUUAUUUCCAATA-3′; si-FOXC2-AS1-2, 5′-GCUGCGUAUUCGAUUCUCAGC-3′. FOXC2 cDNA ORF-overexpressed plasmid was purchased from Sino Biological . Plasmids and siRNAs were transfected into cells with Lipofectamine 2000  following the manufacturers’ instructions. Cells were collected 48 h after transfection.TM Kit  according to the product’s protocol. qRT-PCR assay was conducted using SYBR Green I  and performed on the LightCycler480 system . β-actin was applied as an internal control for normalizing. Each reaction was conducted in triplicate. The primer sequences were as follows: FOXC2-AS1 forward: 5′-CCTTCCTGGCTGTTCATCGG-3′, FOXC2-AS1 reverse: 5′-TGGAAATAAGTGGCACGCCC-3′; FOXC2 forward: 5′-CCTACCTGAGCGAGCAGAAT-3′, FOXC2 reverse: 5′-ACCTTGACGAAGCACTCGTT-3′; PDCD4-AS1 forward: 5′-CAGTCTAATGGGCAGAAGGGC-3′, PDCD4-AS1 reverse: 5′- AGGGGCACTGATCACATTCT-3′; β-actin forward: 5′-TTCCTTCCTGGGCATGGAGTC-3′, β-actin reverse: 5′-TCTTCATTGTGCTGGGTGCC-3′.Total RNAs from tissues and cell lines were isolated with Trizol reagent . RNA reverse transcription was performed using GoScript3 cells/well and cultured for 24 h. Next, in each well, we added 20 µl of MTT and incubated at 37 °C for 4 h. After removing the supernatant, each well was replenished with 150 µl of DMSO and incubated at 37 °C for 10 min. Finally, the absorbance was examined at 570 nm with a microplate reader . For the clone-formation experiment, 1 × 103 cells were seeded into a 35-mm dish and cultured for 2 weeks. Then cell colonies were fixed in 4% paraformaldehyde (PFA) and stained with Giemsa dye.For cell-proliferation examination, cells were plated into 96-well plates at the density of 5 × 10Cell migration was detected by wound-healing assay. Simply, cells were seeded in 12-well plates and grown to 100% confluence. Cell wounds were scratched with a 20-μl sterile pipette tip. Afterward, cells were washed with PBS 3 times and cultured with serum-free medium for 24 h. Wound closure was observed and measured the distance between the opposite edges of the wound.5) in serum-free medium were transferred into the upper chamber coated with Matrigel, while the lower chambers were loaded with medium containing 20% FBS. After 48 h, the noninvading cells were wiped using cotton swabs. The invasive cells were fixed by 4% formaldehyde and stained in 0.1% crystal violet for 15 min. Finally, the images were captured under the microscope.Invasion assay was assessed with the BioCoat Matrigel Invasion chamber . Cells  membrane . After blocking by 5% nonfat dry milk, the membrane was incubated with primary antibodies against FOXC2 , p-FAK (Try397) , p-FAK (Try407) , p-FAK (Try576/577) , FAK , p-Src (Try416) , Src , p-Paxillin (Try118) , and Paxillin , overnight at 4 °C. After washing with PBS, the membrane was incubated with HRP-conjugated secondary antibody; the immune signals were examined using enhanced chemiluminescence reagent . GAPDH was used as the loading control.The nuclear and cytosolic fractions of LoVo and SW620 cells were isolated with PARIS Kit  following the manufacturer’s protocol.The probe of FOXC2-AS1-1 was designed and synthesized by BersinBio , and its sequences were 5′-GAGAAUCGAAUACGCAGCCGAUGAACAGCCAG-3′ conjugated with CY3. The slides of cells were fixed by 4% PFA for 20 min, and incubated with protein K for 30 min at 37 °C. After washing with PBS and denaturing at 73 °C for 3 min, the slides were incubated with the hybridization reaction solution that contained 5 ng/μl probes. The hybridization was carried out in a moist chamber at 42 °C overnight. Subsequently, the slides were rinsed with 25% deionized formamide/2 × saline sodium citrate (SSC) at 50 °C and a descending series of SSC at 50 °C. Finally, the slides were stained with DAPI. The signal was observed and photographed under fluorescent microscopy .Fresh CRC samples were immediately fixed with 4% paraformaldehyde (PFA) for 48 h and dehydrated by graded ethanol. After vitrification in dimethylbenzene and embedding in paraffin, the tissues were dewaxed and hydrated. The paraffin-embedded tissues were subjected to FISH referrred to the cell slides.P < 0.05 and false-discovery rate (FDR) < 0.25 were the criterion for identifying statistically enriched genes.The GSEA was launched to analyze gene sets correlated with FOXC2 in CRC. Gene expression data of CRC were downloaded from TCGA database. FOXC2 expression was set into high and low categories based on the median expression value. The FOXC2-correlated gene sets and pathways were explored in the c2.cp.kegg.v7.0.symbols.gmt data set by GSEA v3 soft. 2+ concentration was examined with Fluo-3 AM . Cells were seeded in 12-well plates and incubated with 2 μM Fluo-3 AM diluted in Krebs–Ringer buffer  for 30 min. After removing the culture and washing with Krebs–Ringer buffer, the cells were collected and rinsed by PBS without Ca2+. Finally, the cells were measured by flow cytometry .Intracellular Ca3) were seeded in a 12-well plate paved with sterile slips. After 24 h, the cells were fixed, permeabilized, blocked, and then incubated with anti-paxillin antibody  at a dilution of 1:150 overnight at 4 °C. The next day, the slips were incubated with fluorescently labeled secondary antibody and phalloidin  for 2 h at room temperature. Subsequently, DAPI was used to stain nuclei. Finally, fluorescence was observed and imaged under the confocal laser-scanning microscope .Cells  according to the product’s instructions. Next, qRT-PCR was conducted to detect the level of FOXC2 nascent RNA as mentioned above.Cells were treated with the transcriptional inhibitor actinomycin D  for 0 h, 1 h, 2 h, 4 h, 6 h, and 8 h, respectively. Cells were harvested at different time points, and RNA was isolated. Then, qRT-PCR was used to detect the level of FOXC2 mRNA as mentioned above.Cells were lysed in lysis buffer. Then the lysate was filtrated with a needle and stood on ice for 10 min. Next, the final solution was added with DNase I (Thermo Fisher) 12.5 U/ml and 125 mM NaCl, and divided into two parts. One part was treated with RNase A , while the other was treated with RNase Inhibitor . After incubation at 37 °C for 30 min, the solutions were used for RNA isolation. qRT-PCR was used to detect the level of FOXC2-AS1 and FOXC2 as mentioned above.FOXC2-AS1 and its antisense RNA were in vitro transcribed from the recombinant plasmids pCMV3-FOXC2-AS1 to produce biotin-labeled RNA with the Biotin RNA Labeling Mix . Cell extracts were incubated with RNA probes, followed by streptavidin-mediated RNA pulldown. After RNA isolation, qRT-PCR was applied to investigate the interaction between FOXC2-AS1 and FOXC2.7 LoVo cells transfected with Lv-sh-FOXC2-AS1 , Lv-sh-FOXC2-AS1 + FOXC2, or Lv-NC were injected subcutaneously into the mice right flank. Tumor growth was observed and recorded every 5 days. After 25 days, animals were killed after they were anesthetized and stripped the tumors. Tumor volume was calculated by the formula: (length × width2)/2.The animal experiments were conducted according to the protocols authorized by the Animal Care Committee of the second Xiangya Hospital, Central South University, and were raised with in vivo tumorigenicity. BALB/c nude mice  were randomly divided into three groups (n = 5 for each group). In total, 1 × 106/0.2 ml PBS) were intrasplenically injected into each mice (n = 6 for each group). All mice were euthanized 8 weeks later, and the liver was surgically excised. The liver tissues were fixed in formalin and embedded in paraffin for hematoxylin and eosin (HE) examination. Metastatic nodules were analyzed under microscopy.For in vivo metastasis assays, Lv-sh-FOXC2-AS1, Lv-sh-FOXC2-AS1 + FOXC2, or control Lv-sh-NC lentivirus-transfected LoVo cells . The statistical analysis was conducted by SPSS 18.0 software . Student’s t test was applied to analyze the differential expression between the two groups. A chi-square test was applied to assess the relationship between FOXC2-AS1 expression and clinicopathological characteristics of CRC. Survival analysis was estimated by the Kaplan–Meier method and evaluated using the log-rank test. All the data used and analyzed in this study are included in this article.Figure S1Figure S2Supplementary figure legend


We propose a new method for regulating valley pseudomagnetoresistance in ballistic graphene-based valley field-effect transistors by taking into account the Y-shaped Kekulé lattice distortion and electric barrier. The device involves valley injection and valley detection by ferromagnetic-strain source and drain. The valley manipulation in the channel is achieved via the Y-shaped Kekulé lattice distortion and electric barrier. The central mechanism of these devices lies on Y-shaped Kekulé lattice distortion in graphene can induce a valley precession, thus controlling the valley orientation of channel electrons and hence the current collected at the drain. We found that the tuning external bias voltage makes the valley pseudomagnetoresistance oscillate between positive and negative values and colossal tunneling valley pseudomagnetoresistance of over 30,000% can be achieved. Our results suggest that the synergy of valleytronics and digital logics may provide new paradigms for valleytronic-based information processing and reversible computing. Graphene, being a two-dimensional sheet of carbon atoms, which have excellent carrier mobility and offers the thinnest possible channel for utilizing to design of metal-oxide-semiconductor field-effect transistors . SemenovOn the other hand, Dirac electrons in graphene possess extra valley degree of freedom besides the conventional charge and spin counterparts. Owing to the large momentum difference between the two valleys and the suppression of the intervalley scattering in clean graphene samples –7, the vδt to represent the energy modification to the electron’s hopping corresponding to these three bonds. We assume that the ferromagnetic graphene is made of the same FM metal stripe. The two magnetizations of the source and drain are directed along the current direction (the x axis), which can be in either the parallel (P) or the antiparallel (AP) alignment, with the help of an external in-plane magnetic field. In the Landau gauge, the magnetic vector potential arising from the fringe field has the form [Ay(x)=Ay[Θ(−x)±Θ(x−L)], where the plus(minus) sign corresponds to the P(AP) configuration of magnetizations, Θ(x) is the Heaviside step function. On the other hand, we assume that the same strain are applied on source and drain of the VFETs, which can be induced by a tension on the substrate of the graphene , where AS is the amplitude. In addition, a electic barrier are also applied in the Kek-Y lattice region, which can be tuned by external bias voltage.In this work, we propose a new type of valley field-effect transistors (VFETs) for graphene-based electron. The device design assumes a ferromagnetic-strain (FM-S) source/drain for valley polarized injection/detection, which resembles conventional spin transistor see Fig. a. Valleythe form , 17 \docgraphene . The elae y axis . For defThe low-energy excitation quasiparticles propagation in the VFETs with Kek-Y graphene superlattices can be described by thes following single particle Hamiltonian –121\docσ and τ are the Pauli matrices for the sublattice and the valley, respectively. P= is the momentum of massless Dirac electrons, τz=±1 for K and vF=106m/s is the velocity of Dirac electrons in the pristine graphene, and vτ≃vFδt/3t is the velocity modification term from the bond contraction effect in the Kek-Y lattice . In the graphene channel, the valley degenerate is also lift, but there is an important difference. In contrast to the lead case, where the phases of K and GP and GAP represent the conductance in P and AP configurations, respectively, and Before proceeding with the calculations, we discuss the band structure with lectrons . This in<E<AM+AS , 23. Simmentclasspt{minimaey space . And theey space , which iL=207nm, and restrict the Fermi energy 20 meV<E<140meV, assumed it satisfying |AM−AS|<E<AM+AS. Figure vt with Fermi energy E=80meV and rectangle potential barrier U=−10meV. We can find that GP and GAP have the same oscillation periods but the inverse phases. Therefore, the VPMR oscillates with increase of vt and the negative value VPMR can appear. Those phenomena are similar to the case of the magnetoresistance in ballistic graphene-based quantum tunneling junctions with the spin-orbit interaction [GP and GAP can be explained by the phase difference between the two valley components. When the incident angle ϕ0=0, the phase shift is given by: Δθ determines the orientation of valley polarization before the electron enters the drain, relative to that of the drain state [Δθ=±2nπ,n=1,2,3⋯, the two polarizations are aligned, leading to the conductance GP maximum and VPMR a high positive value . On the other hand, for Δθ=±(2n+1)π,n=0,1,2⋯, they are orthogonal to each other, leading to the conductance GAP minimum and VPMR negative .In the following, we present the numerical results for the FM-S/Kek-Y/FM-S junction in graphene. Throughout the paper, we set channel length Δθ scales are also linear with the Fermi energy and the potential barrier U. Figure E and U value, even when the effective barrier potential U is greater than Fermi energy E. The physical origin for such a phenomenon is related to the Klein tunneling [E and U, some differences can be also found. As E increases, the difference between GP and GAP conductance becomes smaller and smaller, which lead the oscillation amplitude of VPMR to becomes decreased with the increase of Fermi energy. While under the condition Δθ=±nπ is satisfied, the difference between GP and GAP is greater with increasing of U, especially in some location, the GP and GAP conductance presents switching characteristics. The characters are more desirable for the application of VPMR. Remarkably, the observed maximum value of VPMR is over 30,000% at small E. This value greatly exceeds MR of ~ 175% in the ballistic graphene-based quantum tunneling junctions with the spin-orbit interaction [~ 100% in bilayer graphene controlled by external gates [~ 10000% in a merging Dirac cones system [The conductance and VPMR are not only oscillation functions of the hopping-energy modification, they also oscillate with Fermi energy and the effective barrier potential since unneling . Althougeraction  and the al gates , which is system .Fig. 3CIn conclusion, we proposed a type of valley field-effect transistors for graphene-based electron and studied the valley pseudomagnetoresistance through it. We have shown that the oscillation feature of valley pseudomagnetoresistance not only related to the hopping-energy modification and Fermi energy, but also can be tuned largely by the effective barrier potential. The valley pseudomagnetoresistance tuned by external bias voltage benefits the valley field-effect transistor device, and we anticipate that the electric controlled valley quantum devices proposed here can play a role in quantum and quantum-classical hybrid computers.2, WS2, WSe2, etc.) analogs in graphene can also provide an interesting platform for other two-dimensional material-based valley field-effect transistors with Y-shaped Kekulé lattice distortion.Further research could involve the different strain  tunable the valley scattering of electrons and transport in our proposed graphene-based valley field-effect transistors since the stain is useful to control the degree of intervalley scattering in Kekulé patterns . Then, o


In the same sIBM cohort, comorbidity with T2DM was assessed by collecting anthropometric measures and performing an oral glucose tolerance test and insulin determinations. Results were compared to the standard population and other myositis (n = 7 dermatomyositis and n = 7 polymyositis). Mitochondrial contribution into disease was tested by measurement of oxidative/anaerobic and oxidant/antioxidant balances, respiration fluxes, and enzymatic activities in sIBM fibroblasts subjected to different glucose levels. Comorbidity of sIBM with AD was not detected. Clinically, sIBM patients showed signs of misbalanced glucose homeostasis, similar to other myositis. Such misbalance was further confirmed at the molecular level by the metabolic inability of sIBM fibroblasts to adapt to different glucose conditions. Under the standard condition, sIBM fibroblasts showed decreased respiration  and increased anaerobic metabolism . Moreover, when glucose conditions were changed, sIBM fibroblasts presented decreased fold change in mitochondrial enzymatic activities  and increased oxidative stress per mitochondrial activity , underlying scarce metabolic plasticity. These findings do not demonstrate higher prevalence of AD in sIBM patients, but evidences of prediabetogenic conditions were found. Glucose deregulation in myositis suggests the contribution of lifestyle conditions, such as restricted mobility. Additionally, molecular evidences from sIBM fibroblasts confirm that mitochondrial dysfunction may play a role. Monitoring T2DM development and mitochondrial contribution to disease in myositis patients could set a path for novel therapeutic options.Sporadic inclusion body myositis (sIBM) is an inflammatory myopathy associated, among others, with mitochondrial dysfunction. Similar molecular features are found in Alzheimer’s disease (AD) and Type 2 Diabetes Mellitus (T2DM), underlying potential comorbidity. This study aims to evaluate common clinical and molecular hallmarks among sIBM, AD, and T2DM. Comorbidity with AD was assessed in  Inflammatory myopathies are a heterogenic group of diseases characterized by a progressive symmetric weakness and inflammatory infiltrates within the skeletal muscle ,2. RegarThe former, sIBM, is the unique inflammatory myopathy associated with the accumulation of misfolded proteins and ageing . With a In stark contrast with other myositis, sIBM patients show asymmetrical wasting of proximal and distal muscles, in particular quadriceps and finger flexors, leading to difficulty in performing tasks requiring these muscles such as lifting objects or climbing stairs, among others. In addition, neck extensors and pharyngeal involvement may cause dropped head or dysphagia, respectively, affecting 60% of patients over time ,5.At the moment, muscle biopsy remains the definitive diagnostic procedure for sIBM . At the Interestingly, the presence of inflammation, protein depots, and mitochondrial failure seems not to be exclusive for sIBM and, in the last few years, growing evidence has supported the deregulation of these pathways in other chronic and age-related neurodegenerative or metabolic disorders. Among them, the deregulation of mitochondrial function and associated cell consequences has been speculated as the major molecular hallmark underlying a number of these disorders, since mitochondrial dysfunction can trigger, by its own, from bioenergetic failure and oxidative stress production to protein accumulation, inflammation, and cell death , featureIn physiologic conditions, mitochondria are essential for cell survival, playing a central role in calcium homeostasis, bioenergetics, metabolism, and cell respiration. In pathological conditions, they are indeed the main organelle involved in triggering apoptosis and producing reactive oxygen species (ROS).Thus, mitochondrial homeostasis and balance is fundamental for cell function, especially in highly energetic and dependent tissues , where its dysfunction may become evident much earlier.Despite the fact that pathological mitochondrial evidence is frequently observed in muscle biopsies from sIBM patients, mitochondrial abnormalities in this disease have scarcely been assessed at a molecular level ,11. Our Interestingly, mitochondrial abnormalities, energy failure, respiratory chain impairment, and generation of ROS have also been described in brain aging related to neural loss and synaptic alteration . MoreoveAlzheimer’s disease is the commonest cause of dementia, with huge implications for individuals and large sociosanitary and economic associated burdens currently affecting 50 million people. AD is the single biggest cause of dementia, accounting for 50%–75%, and is primarily a condition of later life, roughly doubling in prevalence every 5 years after age 65 . DementiSimilarly to sIBM and AD, the accumulation of proteins and inflammation has also been described as a trigger of type 2 diabetes mellitus (T2DM). In fact, the presence of amyloid deposits within the pancreatic islets is a pathophysiological hallmark of T2DM . MoreoveT2DM is one of the most prevalent and chronic metabolic disorders worldwide. It is characterized by the presence of hyperglycaemia caused by different alterations converging in the development of insulin resistance accompanied by relative to severe insulin deficiency secretion . This diThe link between AD and T2DM is clearly established in literature ,24. HoweIn this study, we aim to evaluate sIBM patients at two levels to assess clinical and molecular comorbidity between sIBM, AD, and T2DM. From a clinical point of view, we will evaluate the neurological and metabolic status in a cohort of sIBM patients to determine the comorbidity between sIBM and AD or T2DM, respectively. In addition, we aim to include a cohort of patients with inflammatory myopathies other than sIBM, such as DM and PM, to evaluate differential or common deregulation with diseases that clinically overlap with sIBM. From a molecular point of view, we aim to evaluate common molecular hallmarks between these degenerative and age-related diseases, focusing our interest on the mitochondrial homeostasis of sIBM fibroblasts that will be subjected to different pathophysiological conditions.The sIBM cases were diagnosed by clinical and pathological tests in the Department of Internal Medicine from the Hospital Clínic of Barcelona . All the patients fulfilled the criteria proposed by the European Neuromuscular Centre for sIBM diagnosis . ExclusiOn inclusion, sIBM patients completed the IBMFRS , a validated disease-specific functional rating scale to assess disease severity. Epidemiological and clinical data  were also recruited.Muscles biopsies were performed for diagnostic purposes indicated for muscle weakness or raised creatine kinase (CK) levels. Surgical muscle biopsies were obtained by trained physicians and the samples were processed routinely in the laboratory as described elsewhere .Fresh samples were frozen in cooled isopentane, sectioned by cryotome at −30 °C, and stained and reacted with different reagents for diagnostic purposes: haematoxylin and eosin, modified Gomori’s trichrome, non-specific esterase, periodic acid-Schiff stain, Oil Red O, acid and alkaline phosphatase, NADH, COX, SDH, and ATPase at pH 4.3, 4.6 and 9.4, as reported elsewhere . In addin = 7) and PM (n = 7). This second cohort was prospectively included in parallel to sIBM patients to evaluate differential or common deregulation with diseases that clinically overlap with sIBM. These patients were also diagnosed at the Department of Internal Medicine from the Hospital Clinic of Barcelona  following the current established clinical and pathological criteria [n = 5), who were prospectively included.According to the standards for diagnosis, results of clinical evaluation for T2DM and AD in sIBM patients were compared to age bracket control data from individuals free of disease or, alternatively, with a cohort of patients with inflammatory myopathies other than sIBM, such as DM , and apolipoprotein E (APOE) genotype.Participants were evaluated by a trained neurologist with a comprehensive neuropsychological battery of tests. Clinical diagnosis was established following the current clinical criteria for mild cognitive impairment (MCI)  and demeThe neuropsychological battery of tests encompassed five cognitive domains. The memory domain included the Free and Cued Selective Reminding Test , the lan3) was acquired at the IDIBAPS’s Imaging core facilities with a 3T whole-body MRI scanner .For each participant, a T1-weighted, high-resolution, MPRAGE structural MRI scan (echo time (TE) 2.98 ms, repetition time (TR) 2300 ms, inversion time 900 ms, flip angle 9°, bandwidth 240 Hz/pixel, matrix 256 × 256, 240 axial slices, isometric voxel size 1⁄4 1.0 mmFor all subjects, global cortical atrophy (GCA), medial temporal lobe atrophy (MTL), and white matter integrity (Fazekas) scores were obtained by a trained radiologist.Subjects underwent a lumbar puncture between 9 a.m. and 12 p.m. In the extraction, 10 mL of CSF was collected. The samples were centrifuged and stored in polypropylene tubes at –80 °C within the first hour after extraction. CSF Aß42 levels, total tau (tau), and phosphorylated tau at threonine-181 (ptau) were measured by enzyme-linked immunosorbent assay kits . The following cut off points were considered: (a) Aß42 ≤ 550 pg/mL, (b) tau ≥ 385 pg/mL, and (c) ptau ≥ 65 pg/mL.Genomic DNA was extracted from the peripheral blood of probands using the QIAamp DNAblood minikit . APOE genotyping was performed by polymerase chain reaction amplification and HhaI restriction enzyme digestion.Procedures for the 2h-OGTT required previous fasting after midnight, to obtain baseline fasting glucose level, and further administration of the oral glucose load (75 g) within a 5 min period. Blood specimens to determine the plasma glucose level were subsequently drawn at 30, 60, 90, and 120 min, timed from the beginning of the glucose load. Patients were established as having impaired fasting glucose  or having impaired glucose tolerance . Criteria for new-onset T2DM was a fasting plasma glucose level greater than 126 mg/dL or 120-min venous plasma glucose level greater than 200 mg/dL on the OGTT.The following data were collected from medical records: circulating baseline levels of insulin (mU/L) and glycated haemoglobin, HbA1c (%). Homeostatic model assessment of insulin resistance (HOMA-IR) was calculated to determine the insulin resistance as: /22.5. Anthropometric and physical examination data concerning weight and height were also recruited to calculate body mass index .2 air incubator until 80% optimal confluence was reached. Fibroblasts were harvested with 2.5% trypsin  and centrifuged at 500 g for 8 min. All functional assays were performed in cells between passages 5 and 10.Fibroblasts were obtained by a skin punch biopsy. Cells were grown in 25 mM glucose Dulbecco’s Modified Eagle’s medium  supplemented with 10% heat-inactivated foetal bovine serum and 1% penicillin-streptomycin at 37 °C in a humidified 5% COTo test fibroblast response to pro-diabetogenic in vitro conditions, cells were cultured under different glucose conditions: high glucose (HG: 25 mM) and low glucose (LG: 5 mM) DMEM media 10 days prior the experiment, mimicking the glucose deregulation observed in T2DM.Expected response to 10 days glucose decrease (from HG to LG) should promote mitochondrial activation to enhance energetic output. To evaluate mitochondria activation in front of glucose reduction, we tested oxidative vs. anaerobic metabolism and oxidant/antioxidant balance, together with mitochondrial respiration and enzymatic activities.® technology  in non-permeabilized fibroblasts, following the manufacturer’s protocols [Mitochondrial respiration is the result of oxygen consumption by the MRC. It was measured in 1 million cells by high-resolution respirometry using Oroboros™ Oxygraph-2Krotocols . Two mea® Blood Analysis System, epoc System Software version 3.32.0 , normalized by cell count and expressed as mmols/L.Lactate measures were performed in 100 µL of fresh supernatant fibroblast culture using the epocTotal Antioxidant Capacity (TAC) was measured in 100 µL of cell supernatant using an OxiSelect™ Total Antioxidant Capacity Assay kit  by spectrophotometry (absorption maximum at 490 nm), normalized by cell count and expressed as µM CRE .Cytochrome c oxidase (COX) function was assessed in 40 µg of total protein from each fibroblast sample according to standardized protocols  and norm™ LPO-586™ kit  through the spectrophotometric measurement of malondialdehyde and 4-hidroxyalkenal, both of which are products of fatty acid peroxide decomposition. Lipid peroxidation was normalized by COX/CS activity as a measure of oxidative stress production per mitochondrial activity.Lipid peroxidation (an indicator of oxidative damage produced by ROS to cellular lipid compounds) was quantified in 150 µg of protein from a fibroblast sample using the OxysResearchp < 0.05.Results were expressed as mean ± standard error of the mean (SEM) or in fold change between conditions (HG vs. LG). Statistical analysis was performed after filtering for outliers to evaluate differential phenotypes depending on group-assignment (patients or controls) by using the non-parametric independent sample Mann-Whitney U test. The signification threshold was set at n = 14 subjects aged between 48 and 90 (mean age 67.70 ± 3.84 years), 9:5 male to female ratio, and all white Caucasian. According to the design of the study, no statistical differences were observed in regards to age, gender, or ethnicity between sIBM patients and the control cohorts. Specifically, PM patients presented a mean age of 69.57 ± 5.39 years and a 1:6 male to female ratio, DM subjects a mean age of 66.67 ± 3.20 years and a 2:5 male to female ratio, and healthy control volunteers aged 57.12 ± 5.38 years and presented a 1:4 male to female ratio, all white Caucasian, according to sIBM demographic characteristics.The sIBM patients included in this study corresponded to At the time of inclusion, the sIBM patient cohort scored 22.00 ± 2.37 out of 40 with the IBMFRS test, indicating moderate-to-advanced sIBM clinical severity.Ten sIBM patients underwent the neurological assessment. Education level ranged between 7 and 18 years and the cognitive reserve questionnaire scored 7 to 18 points. Demographics of the cognitive study are shown in Eight out of the ten subjects that underwent clinical and neuropsychological evaluation were cognitively impaired. From those, 5 were diagnosed with MCI and 3 with dementia (two of them were diagnosed as AD).Three subjects presented symptoms of anxiety and/or depression. MMSE ranged between 6 and 29  and the neuropsychological profile of the study subjects was heterogeneous, although most of them showed an amnestic pattern .Despite reported disturbances in the clinical and neuropsychological assessment, no higher incidence of dementia or AD was detected in sIBM patients when the results were compared to the prevalence of dementia in Europe . See cliEight sIBM subjects underwent the magnetic resonance scan. Medial temporal lobe atrophy (MTL score > 1) was found in 3 subjects  and all individuals presented different levels of white matter deterioration, including two subjects with a Fazekas score ≥ 2  see .However, MTL score > 1 was also found in controls in previous studies . Further42 with normal CSF tau and phosphor-tau levels  showed alterations in 42% (3/7) of the patients, while 14% (1/7) were in the prediabetic range (the plasma glucose level fell between 140 and 200 mg/dL) and the remaining 28% (2/7) felt into the T2DM category (glucose level greater than 200 mg/dL). Moreover, medical records in blood tests from the sIBM population (n = 8) showed increased HbA1c % in 50% (4/8) of these patients (HbA1c greater than 5.7). Finally, when HOMA-IR was assessed in the sIBM cohort (n = 7), 14% (1/7) of the patients were close to insulin resistance values  and PM (n = 7) showing prediabetic state values in 43% (3/7) of cases from both populations. The 2 h venous plasma glucose level in the DM population was greater in 43% (3/7) of patients, which is considered suggestive of a prediabetic state, while the PM cohort showed altered values in 71% (5/7) of patients, 28% (2/7) were considered as in a prediabetic state, and the other 43% (3/7) were consistent with T2DM. Medical data records from blood tests of DM and PM populations also showed altered HbA1c profiles in both cohorts. Particularly, 14% (1/7) of DM and 43% (3/7) of PM had greater values compared to age bracket control data (HbA1c < 5.7%) [Specifically, the basal glucose level was also determined in a cohort of patients with DM ( < 5.7%) . FinallyConsidering the T2DM definition stratified by age from the American College of Endocrinology , the altp = 0.024), increased anaerobic metabolism , and TAC levels , reflecting the mitochondrial dysfunction characteristic of the target tissue of the disease (muscle).Under standard conditions see , sIBM fiCells increase mitochondrial activity in response to the decrease of glucose levels, probably as an attempt to maximize the energetic output in case of glucose restriction. Such activation was observed in control fibroblasts when subjected to glucose changing conditions (HG vs. LG) in terms of respiration. Under high glucose concentration, the mitochondrial respiration rate was low, indicating decreased oxygen consumption and high anaerobic metabolism. When the glucose concentration was diminished, oxygen consumption increased, indicating the activation of the aerobic  metabolism .p = 0.017). Additionally, a lack of mitochondrial activation in sIBM fibroblasts was accompanied by increased fold change proton leak (92.29 ± 73.89 vs. 6.16 ± 83.05) and oxidative stress per mitochondrial activity   and enzymatic activities  .Glucose misbalance manifested at a clinical level in sIBM patients was further confirmed in vitro by the inability of sIBM fibroblasts to adapt to different glucose conditions, thus underlying the clinical and molecular comorbidity between sIBM and T2DM.Despite medical literature postulating potential comorbidity of sIBM with AD and T2DM, scarce data and epidemiological studies (if any) had been conducted to date to evaluate such overlap. Before our investigation, none of the sIBM patients had been diagnosed of T2DM or AD. In the present study, and regardless of the small sample size herein tested, we found null comorbidity between sIBM and AD, but trends towards comorbidity between sIBM and T2DM.Despite sIBM having been described as “Alzheimer in muscle” and both disorders sharing some pathophysiological mechanisms ,15,50,51Interestingly, partial comorbidity between sIBM and T2DM has been demonstrated in this work. Clinically, the metabolic deregulation in sIBM patients is characterized by the initial impairment of glucose metabolism compared with age bracket control data. Despite the fact that only a few of them could be concomitantly diagnosed by sIBM and T2DM, most of the sIBM patients showed clear signs of a prediabetogenic condition that underlie the comorbidity between both diseases.Such coexistence of T2DM in sIBM patients could be due to common etiological links between both disorders. Previous literature describes similar mitochondrial alterations in T2DM and sIBM ,20,52,53At a molecular level, the mitochondrial characterization of fibroblasts from sIBM patients under standard conditions of the present study demonstrate increased anaerobic metabolism and antioxidant capacity at the detriment of mitochondrial oxygen consumption. These results suggest that mitochondrial dysfunction is present in sIBM patients and is not only restricted to the target tissue of the disease (muscle), but may also extend to peripheric alternative tissues, including fibroblasts, as previously demonstrated in the case of blood cells . These fMoreover, we used these fibroblasts to evaluate mitochondrial contribution into sIBM-T2DM comorbidity by exposing sIBM fibroblasts to different glucose conditions. That way, we observed that control fibroblasts showed full mitochondrial plasticity to adapt to different glucose environments, whilst sIBM fibroblasts were mitochondrially unable to adapt its metabolism. Particularly, sIBM fibroblasts were unable to increase their mitochondrial function in terms of respiration and MRC enzymatic activities in front of changes in glucose bioavailability. Mitochondrial dysfunction in sIBM fibroblasts may lead to reduced bioenergetics and consequent cell compromise. In fact, such metabolic stiffness leads sIBM fibroblasts to increased proton leak and oxidative stress, frequently associated with cell damage ,59 and dTo our knowledge, there are no clinical or molecular evidences supporting the preferential affection of muscle tissue in sIBM or the particular targeting of certain muscles (greater in finger flexors and quadriceps), but there is probably a molecular rationale underlying such differential targeting and mitochondria (among others) may stand behind this.Overall, these data demonstrate that mitochondrial dysfunction limit sIBM cell adaption to changing glucose metabolism, being a source of cell damage and the potential pathophysiological link between sIBM and T2DM. These molecular data confirm clinical comorbidity between both disorders, pointing out sIBM as a new population of underdiagnosed T2DM patients. T2DM is a silent infradiagnosed disease and any clue for emerging groups of risks may account for the reduction of further associated consequences. Interestingly, partial deregulated glucose metabolism was also found in other myositis as PM and DM, thus suggesting common lifestyle cues’ contribution in observed comorbidity.Probably, both mitochondrial deregulation and lifestyle habits (such as restricted mobility) may be conditioning the manifestation of metabolic disorders such as T2DM in sIBM patients. Due to its idiosyncrasy, myositis are a group of disorders characterized by weakness and wasting of the muscle and consequently low physical activity or a sedentary lifestyle. This situation is associated with the development of several chronic diseases including T2DM. Additionally, due to skeletal muscle contribution to glucose homeostasis and muscle wasting on account of low patient activity, in a kind of vicious circle, glucose metabolism deregulation may be the final fate of a sedentary lifestyle, in this case, triggered by myositis and mitochondrial dysfunction.Consequently, some general recommendations may be added in the clinical follow-up of patients with myopathy to prevent or manage potential T2DM development in accordance with international standards . They arSome limitations must be acknowledged in the present work. The reduced number of patients included in the study may be the major downfall. Of note, due to the low prevalence of sIBM disorder (considered a rare disease), the study of bigger cohorts is often hampered but recommended. Additionally, further studies are warranted to provide more in-depth understanding of the deregulated pathways underlying glucose impairment in sIBM patients and whether the lipid profile and other features characteristic of the metabolic syndrome are affected in sIBM, even myositis in general, to establish a better correlation between these disorders and the metabolic syndrome. Similarly, the parallel inclusion of fibroblasts from PM or DM patients would have been of interest to deepen the knowledge of common and specific molecular patterns shared among these disorders that clinically overlap.In summary, the conclusions of the present work stand for null comorbidity between sIBM and AD but partial comorbidity between sIBM and T2DM through the impairment of glucose homeostasis, mitochondrial function, and probably lifestyle conditions. Although further investigation is needed to validate this pilot study, these findings provide better knowledge of sIBM complications.On account that effective treatment for sIBM is not yet available, this should be a red flag and the monitorization of the prediabetic state in myositis patients to avoid potential complications in clinical practice associated with the management of both diseases should be considered, thus contributing to improving the quality of life of patients with inflammatory myopathies.