However, these edge states are highly susceptible to edge roughness and interaction with a supporting substrate, complicating the study of their intrinsic electronic and magnetic structure. Here, we focus on atomically precise graphene nanoribbons whose two short zigzag edges host exactly one localized electron each. Using the tip of a scanning tunnelling microscope, the graphene nanoribbons are transferred from the metallic growth substrate onto insulating islands of NaCl in order to decouple their electronic structure from the metal. The absence of charge transfer and hybridization with the substrate is confirmed by scanning tunnelling spectroscopy, which reveals a pair of occupied/unoccupied edge states. Their large energy splitting of 1.9 eV is in accordance with  The zigzag edges of graphene host edge-localized electronic states with aligned electron spins, but these states strongly interact with metallic substrates. Here, the authors measure the electronic structure of graphene nanoribbons after transferring them to an insulating support. Recent advances in the fabrication of precise graphene nanostructures open the door to tailoring their electronic properties to the needs of specific applications. In the case of graphene nanoribbons (GNRs) with armchair edges, a bottom-up approach has been shown to deliver control over width and edge termination down to the atomic level236789151617181918ab initio many-body perturbation theory calculations, which we use to systematically distinguish between edge states localized at the zigzag edges and the energetically and spatially distinct states associated with the armchair edges in the GNRs under study.Here, we focus on the electronic properties of the atomically precise zigzag edges formed at the termini of bottom-up fabricated armchair graphene nanoribbons (AGNRs). To decouple their electronic structure from the metal substrate, on which they are grown, we transfer the AGNRs onto NaCl islands by a scanning tunnelling microscopy (STM)-based multistep manipulation routine. Using scanning tunnelling spectroscopy (STS), we find that electronic decoupling of the edge states establishes a large energy splitting between occupied and unoccupied edge states. This is in accordance with m=7, which are synthesized with atomic-scale precision on a Au(111) single crystal surface using a recently established bottom-up methodn) GNRs, where n specifies their length along the armchair direction in units of carbon zigzag lines. Finite  GNRs host two qualitatively different sets of electronic π-states. One set derives from the Bloch states of the bulk GNR, which are delocalized along the GNR. The short zigzag edges at the termini of the GNRs give rise to another set of states that are localized near the termini6n) GNRs on Au(111) indicated only one, possibly degenerate, edge state near the Fermi level, which may be explained by hole-doping of the GNRWe focus on short AGNRs of width n on a Au11 singleafter the synthesis. Here, we use atomically thin insulating NaCl films that are deposited directly onto the metal surface. In contrast to bulk insulators, these films still allow the electronic properties of adsorbates to be investigated by STM/STS, while considerably reducing their interaction with the metal substraten) GNRs to the Au(111) growth substrate, which enables lateral manipulation and controlled pick-up of individual GNRs by the STM tipI/dV) spectrum exhibits two peaks centred at −0.5 and 1.3 V, as shown in As the synthesis relies on the catalytic activity of the metal surface, the transfer onto an insulating substrate needs to occur GW approximation. The framework provides accurate fundamental gaps, both for bulk insulatorsBeing a theory of the electronic ground state, Kohn–Sham DFT is not designed to describe the charged excitations that take place in STS, which involve the addition/removal of electrons to/from the sample. Although the orbitals of the non-interacting Kohn–Sham system are often found to be accurate approximations of the corresponding quasiparticle wave functionsab initioGW calculations for finite  GNRs, which can be viewed as open-shell molecules with one, singly occupied state localized at each terminus. Their fundamental gap is given by the energy splitting ΔZZ between the occupied and empty edge-localized states. A significant Kohn–Sham gap opens only in the spin-unrestricted formalism, where breaking of spin symmetry gives rise to staggered sublattice potentialsn⩾12. Starting from the PBE orbitals and orbital energies, we compute quasiparticle corrections in the G0W0 approximation. The fundamental gap ΔZZ is found to converge rapidly as a function of length, yielding a value of n⩾12  GNR and the edge states of extended zigzag edges). Direct comparison to experiment would require the inclusion of dynamical screening not only by the electrons of the GNR itself, but also by those of the NaCl monolayer and the underlying Au substrate, which is expected to lead to significant reduction of the fundamental gapZZ=1.9 eV is fully compatible with a fundamental gap of 2.8 eV, reduced by screening from substrate electrons.Here we perform r finite , n GNRs,ZZ is essentially independent of the separation between the zigzag edges, in accordance with the G0W0 predictions (cf. n) GNRs, the additional quantum confinement at the short zigzag edges selects one particular wavelength along the zigzag edge and the corresponding edge state is energetically isolated from the delocalized states of  GNRs.To investigate finite size effects, ribbons of different lengths have been moved onto NaCl islands and inspected. 17ions cf. . Similarions cf. . In the n) GNRs. Their width of 7=3 × 2+1 carbon dimer rows identifies  GNRs as members of the 3m+1-family of AGNRs, which have the largest band gapsI/dV spectra taken at the centre of a  GNR, featuring two sharp peaks at –1.2 and 2.3 V. These peaks indicate the onsets of the highest occupied and the lowest unoccupied bulk states, respectively, yielding a bulk band gap of ΔAC=3.5 eV. In contrast to the energy splitting ΔZZ of the localized edge states, we find that the bulk band gap ΔAC decreases continuously from 3.5 eV for the shortest  GNR to 2.9 eV for the longest  GNR under study, as shown in n) GNRs on Au(111)GW approximation predict a band gap of 3.7±0.1 eV for freestanding  GNRs2AC=2.9 eV is still below this value, it is significantly larger than the 2.4 eV measured for the  GNR on Au(111)We now turn to the delocalized bulk electronic states of decoupled 7, , n GNRs.gest 7, 4 GNR undeme, respectively (me is the free electron mass). We note that the bands appear with different intensity in STS because of the finite tip-sample distance and refer interested readers to the corresponding study on Au(111) for detailsG0W0 approximation. Although the band gap is found to open from 1.6 to 3.6 eV -GNR has been determined via Fourier transformed (FT) STS-GNR has n) graphene nanoribbons from Au(111) to an insulating monolayer of NaCl without introducing any defects. The delocalized electronic states of  GNRs are separated energetically from the states localized at the zigzag termini, making it possible to investigate the effect of electronic decoupling on both classes of states separately. Using STS, the band gap between delocalized states is found to increase from 2.4 to 2.9 eV upon electronic decoupling, whereas no significant modification of the effective masses is observed. Furthermore, we find an energetic splitting of the states localized at the zigzag termini that has long been predicted for freestanding zigzag edges, but is missing completely in  GNRs on Au(111). Both its substantial size of ΔZZ=1.9 eV and its independence on edge separation down to 3 nm are in agreement with ab initio many-body perturbation theory calculations of the GNRs' intrinsic electronic structure, indicating that decoupling by a single layer of NaCl indeed allows to study the intrinsic properties of graphene zigzag edges. We therefore expect that the experimental strategy established here will particularly benefit the eagerly awaited exploration of the low-energy spin physics at graphene zigzag edges using spin-sensitive methods.We have demonstrated the transfer of atomically precise  using an Omicron low-temperature STM. The Au(111) substrate was cleaned by standard argon sputtering and annealing cycles. The GNRs were grown on Au(111) following the recipe by Cai et al.I/dV spectra were recorded using the lock-in technique with Urms=20 mV.Sample preparation and STM measurements were performed in an ultrahigh vacuum system .Supplementary Figures 1-5, Supplementary Table 1, Supplementary Notes 1-3 and Supplementary References


Earlier studies have demonstrated that emotional stimulation modulates attentional processing during goal-directed behavior and related activity of a brain network including the inferior frontal gyrus (IFG) and the caudate nucleus. However, it is not clear how emotional interference modulates behavior and brain physiology during variation in attentional control, a relevant question for everyday life situations in which both emotional stimuli and cognitive load vary. The aim of this study was to investigate the impact of negative emotions on behavior and activity in IFG and caudate nucleus during increasing levels of attentional control. Twenty two healthy subjects underwent event-related functional magnetic resonance imaging while performing a task in which neutral or fearful facial expressions were displayed before stimuli eliciting increasing levels of attentional control processing. Results indicated slower reaction time (RT) and greater right IFG activity when fearful compared with neutral facial expressions preceded the low level of attentional control. On the other hand, fearful facial expressions preceding the intermediate level of attentional control elicited faster behavioral responses and greater activity in the right and left sides of the caudate. Finally, correlation analysis indicated a relationship between behavioral correlates of attentional control after emotional interference and right IFG activity. All together, these results suggest that the impact of negative emotions on attentional processing is differentially elicited at the behavioral and physiological levels as a function of cognitive load. In everyday living, we perceive a complex visual environment and fully analyze many items and events at one time. Attentional control allows the flexible allocation of attentional resources to relevant stimuli while suppressing stimuli that are less relevant. This cognitive process provides a top-down bias for analysis and representation of relevant information in the face of concurrent and non-relevant information  task, which requires increasing levels of attentional control processing associated with a physiological linear increase in prefrontal activity . Inclusion criteria were absence of any psychiatric disorder, as evaluated using the Structural Clinical Interview for Diagnostic and Statistical Manual of Mental Disorders IV, of any significant neurological or medical condition revealed by clinical and magnetic resonance imaging evaluation, of history of head trauma with loss of consciousness, and of pharmacological treatment that could influence cerebral metabolism or blood flow, or drug abuse in the past year. The Wechsler Intelligence Scale—Revised was used to evaluate the intelligent quotient (IQ) . Socio-economic status : All three sizes of arrows were congruent in direction with each other. The cue was the word BIG.Intermediate level of attentional control (INT): The big arrow was incongruent in direction to the small and the medium arrows; the cue was the word SMALL.High level of attentional control (HIGH): The medium sized-arrows were incongruent in direction to the big and the small arrows; the cue was the word SMALL.Subjects performed a task Figure  that wasEmotional interference was obtained using unfamiliar faces with either fearful or neutral facial expressions derived from a validated set of facial pictures  displayed above each attentional stimulus to press a button corresponding to the direction of the large or small arrows (either right or left). They were instructed to respond to the attentional stimuli with the thumb of their right hand using a button box , and to press the response button as fast and accurately as possible. Furthermore, they were asked to move their thumb to a small plastic knob placed between the buttons after each response. All subjects were trained on attentional stimuli of the task prior to the fMRI session to stabilize performance. More specifically, just before fMRI scanning, subjects were instructed on task rules and performed the task outside the fMRI environment until the average behavioral performance did not grossly vary across trials in terms of behavioral accuracy . No instructions related to emotional stimuli were administered.A total of 192 attentional stimuli was preceded by the respective 192 stimuli providing emotional interference, which were 96 fearful and 96 neutral facial expressions. There were 64 stimuli for each level of attentional control. Thirty two stimuli of each attentional level were preceded by facial stimuli with fearful expression, and 32 by facial stimuli with neutral expression. This design resulted in 6 conditions: fearful-low attentional control; fearful-intermediate attentional control; fearful-high attentional control; neutral-low attentional control; neutral-intermediate attentional control; neutral-high attentional control. The order of the stimuli was randomly distributed across the session  while subjects performed the emotional variable attentional control (EVAC) task. The first 4 scans were discarded to allow for signal saturation. post hoc analysis to correct for multiple comparisons. Because of our priori hypothesis, post hoc analysis of the interaction between the level of attentional control and emotional interference was performed for each level of attentional control between fearful and neutral faces.Analysis of variance (ANOVA) was used to compare behavioral data (% correct responses and RT of correct responses). We included only subjects who had an average accuracy above 50% (forced choice between two options). Bonferroni correction was used fort statistic was then used to produce a statistical image for BOLD responses relative to brain processing of stimuli associated with correct responses for each condition. A random effects ANOVA was used at the group level to investigate the main effect of emotional interference, of attentional control stimuli, and their interaction. For all analyses on activity during the EVAC task, we focused our attention on the orbital and triangular parts of IFG and on the caudate nucleus because of their central role in the regulation of cognitive and emotional processes as well as in their integration. Thus, we used a statistical threshold of p < 0.05, k ≥ 5, Family Wise Error (FWE)-corrected for multiple comparisons within brain regions of interest , as defined by the WFU pickatlas software, version 1.04 .Analysis was completed using the event-related module within Statistical Parametric Mapping 5 (SPM5).post hoc analysis was performed on parameter estimates extracted from clusters crossing the statistical threshold of p < 0.05 (FWE corrected) within the predefined ROIs using MarsBar.To further investigate directionality of the interaction between emotional interference and attentional load, F = 31.25; p < 0.000001; average number and % of correct responses: HIGH 26.55, 82.9%; INT 29.52, 92.3%; LOW 31.32, 97.9%), no effect of emotional interference  = 0.98; p = 0.3), and no interaction between level of attentional control and emotional interference  = 1.37; p = 0.2). Post hoc analysis of the main effect of increasing level of attentional control showed a significant difference across all three levels .ANOVA on accuracy data indicated a main effect of increasing level of attentional control  = 103.25; p < 0.000001), no effect of emotional interference  = 0.07; p = 0.7), and an interaction between level of attentional control and emotional interference  = 14.23; p < 0.00002) . Post hoc analysis of the interaction between the level of attentional control and emotional interference indicated a statistically significant difference between RTs at the lower level of attentional control when preceded by presentation of fearful facial expressions . An effect of emotional interference was also present at the intermediate level of attentional control, but in the opposite direction . At the higher level of attentional control, a statistical trend in the same direction of the lower level was found .Furthermore, analysis of RTs revealed a main effect of increasing level of attentional control   .Consistent with previous studies  Figure . Similar Figures . A statip > 0.2). However, there was a significant negative correlation between parameter estimates in right IFG and RTs at the low level of attentional control after neutral facial expressions were shown  . Finally, no significant brain activity-RT correlations were found at the intermediate and high levels of attentional control (p > 0.15).No significant correlations were found between behavioral accuracy data and brain activity in clusters showing an emotional interference by attentional load interaction  Figure . No signOur results indicated differential effects of emotional interference on behavior and activity in prefronto-striatal brain areas during attentional control as a function of cognitive load. In particular, stimuli eliciting the lower level of attentional control were associated with slower RTs as well as greater activity in the right side of the IFG when anticipated by fearful, relative to neutral, facial expressions. However, the interaction between emotional stimuli and the intermediate level of attentional control was associated with the opposite behavioral pattern but with a similar effect on brain activity, this time taking place in both sides of the caudate. Finally, no statistically significant behavioral or imaging effects of emotional interference were found at the higher level of attentional control. Together, these findings suggest that negative emotions modulate goal-directed behavior and related brain activity when low and intermediate levels of attentional control are required, but they do not impact attentional processing when high levels of attentional control are required. More generally, these results suggest that negative emotions may modulate cognitive efficiency during attentional control processing.At the lower level of attentional control, the presentation of fearful, relative to neutral, faces before attentional stimuli was associated with longer RTs. Slower performances of attentional processes by negative emotions has been previously reported  induced by fearful facial expressions modulates detection of other environmental cues. Similar results have been reported recently (Phelps et al., A limitation of this study may be the number of trials used at the high level of attentional control, which could explain the statistical trend we found in behavioral and imaging data between fearful and neutral interference. According to a previous study, 25 trials are sufficient to provide stable activation maps (Murphy and Garavan, In conclusion, this study suggests that emotionally irrelevant stimuli during attentional control processing modulate behavior and brain activity in right IFG and both sides of the caudate as a function of cognitive load. These findings add knowledge to some aspects of the complex functional architecture underlying the integration of emotion and attention in the human brain and could have potential clinical implications. Impairment of emotional and cognitive functions have long been regarded as characterizing several major psychiatric disorders such as schizophrenia, bipolar disorder, depression, and anxiety disorders (Bertolino et al., Dr. Bertolino is a full time employee of Hoffman-La Roche, Ltd. The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Cardiac sarcoidosis is a major cause of death in patients with systemic sarcoidosis. Cardiac manifestations are seen in 2.3% of the patients. Atrioventricular (AV) block is one of the common manifestations of cardiac sarcoidosis. Other presentations of cardiac involvement include congestive heart failure, ventricular arrhythmias, and sudden cardiac death. The presence of AV block in young patients should raise the suspicion of sarcoidosis. AV block may be the only manifestation and patients may not have clinical evidence of pulmonary involvement. Here we describe a young male presented with exercise induced AV block rapidly progressing to complete heart block with recurrent syncope needing urgent pacemaker implantation. Factors that suggested an infiltrative process included his young age, rapidly progressive conduction abnormalities in the ECG in the absence of coronary disease, and previous history of cutaneous sarcoidosis. Sarcoidosis is a multisystem disease of unknown aetiology characterized by the formation of noncaseating granulomas in many tissues. The granulomas mostly affect the lungs and lymph nodes . Cardiac47-year-old male presented to the cardiology out-patient department (OPD) with shortness of breath and chest discomfort on exertion since one week. Chest discomfort was related to exertion and relieved by rest. He has hypertension and hypertriglyceridemia controlled well on oral Amlodipine and Fenofibrate since one year. He does not smoke, drink alcohol, or use any recreational drugs. The patient gives past history of cutaneous sarcoidosis which was diagnosed by biopsy of the lesion one year back. He was evaluated for systemic sarcoidosis and was found to have hilar lymph nodes by computed axial tomography of chest and was advised for a follow-up CT after 6 months. The follow-up CT scan showed improvement and the patient was asymptomatic. Corticosteroids were not started at that time as the patient was concerned about the side effects of steroids on longterm. Hence he was advised for regular follow up. Three months after this evaluation, he presented to our OPD with the present symptoms. Physical examination did not show cutaneous lesions, his pulse was regular at 62 beats per minute and the blood pressure was normal. Cardiovascular examination revealed normal S1 and S2 with no gallop, murmurs, or rub.At the time of presentation his electrocardiography (ECG) revealed sinus rhythm with a rate of 62 per minute with prolonged PR interval and right bundle branch block (RBBB) . CardiacHe has undergone urgent temporary pacemaker implantation. This was followed by permanent pacemaker implantation with Victory XL DDDR pacemaker . He made an uneventful recovery. He remained asymptomatic afterwards. A workup for the autoimmune profile was normal, serum calcium was 8.6 mg/dL, and the serum angiotensin converting enzyme level was elevated. Pacemaker interrogation showed he is fully dependent on pacemaker at 2 weeks after discharge. He remained symptom-free following the procedure. He was evaluated by an internist and a dermatologist after the surgical wound was healed. Based on the clinical presentation and available reports he was started on immunosuppressive therapy with steroids. Presently his ECG shows sinus rhythm with RBBB and pacemaker on demand mode. He is doing well one year after the procedure.Cardiac sarcoidosis is a major cause of death in patients with systemic sarcoidosis. Clinically evident sarcoidosis involving the heart affects 2.3% of patients . Common Cardiac involvement can occur at any point during the course of sarcoidosis. Asymptomatic heart involvement is more common in systemic sarcoidosis. When symptomatic, cardiac involvement can present as CHF, AV block, cardiac arrhythmias, and SCD. The clinical presentation of cardiac sarcoidosis reflects the changes in myocardium caused by granulomatous deposition leading to inflammation and subsequently scar. These become the source of ventricular arrhythmias and conduction block.In a prospective study of 32 patients aged less than 60 years presenting with CHB, 34% had previously undiagnosed cardiac sarcoidosis. This group had higher adverse cardiac events compared with patients without sarcoidosis [The prognosis of symptomatic cardiac sarcoidosis is not clear, but 5-year mortality rates exceed 50% . IncidenReaching a correct diagnosis early in the course of the disease is difficult unless sarcoidosis is also present in other organs. Early administration of immunosuppressant therapy (high dose steroids) may stop or reverse cardiac damage . Our patAn accurate and early diagnosis is essential to enable prompt treatment of arrhythmias. The recovery of his rhythm back to sinus after the institution of steroid also justifies the presumptive diagnosis of cardiac sarcoidosis in this patient. Early institution of steroid therapy is advised in patient with systemic sarcoidosis to prevent fatal cardiac complication even if they are asymptomatic.


A facile production of highly porous graphene foam by using flame treatment of graphene oxide (GO) is proposed. Highly porous architectures with randomly distributed micro-crack and micro-slit were produced due to the high temperature induced ruinous reduction and rapid expansion of GO. Synchronously, abundant oxygen-containing groups (OCGs) on GO sheets could be effectively removed after flame treatment, which renders significantly increased conductivity to the resultant flame reduced GO (FR-GO). The synergistic effect of micro/nanostructuring and the OCGs removal makes FR-GO a promising candidate for electrode materials. Compared with chemically reduced GO (CR-GO), FR-GO delivers much higher specific capacity. It gives us some hints that flame treatment of graphene-based material is a smart strategy for cost-effective production of anode materials for commercial application. Recently, it is also found that optical approach for preparation of porous graphene is also very powerfulet al., achieved a reduced free-standing GO paper as high-rate capable anodes for LIBsAs a new family member of carbon materials, graphene, which is a single layer of two-dimensional (2D) honeycomb-structured carbon lattice, is promising to achieve outstanding electrochemical performance16In this work, we design and fabricate highly porous graphene foam from exfoliated GO paper by using candle flame treatment. The ultra-high temperature of the outer flame was found to be capable of removing the oxygen-containing groups (OCGs) on GO sheets, which not only leads to the formation of highly porous structures but also dramatically improves the conductivity of the flame reduced graphene oxide (FR-GO). The hierarchically porous architecture effectively improves the surface area, which enlarges the contact area between the electrode and the electrolyte. Therefore, the mass transfer has been significantly promoted. It needs to point out that our approach is simple, low-cost, chemical-free and environmental benign for the preparation of porous graphene anodes towards high energy density rechargeable commercial energy storage devices. We also propose that the conductive and porous graphene foam can be widely used in curvilinear electronics, catalytic, biofuel and biomimetic systems.+Notably, the OCGs on the surface of GO sheets would influence the interaction between lithium ions and grapheneTo evaluate the surface wettability change before and after the flame treatments, static water contact angle (CA) of the two samples was measured. As shown in 2, would destroy the GO sheets and generate more defects. Additionally, the emission of carbon species also leads to significant expansion along the lateral direction. It could be clearly observed from the edge of the open pore that the FR-GO sheets curl up, which significantly increases the interlayer spacing. To get further insight into the porous structure along the lateral direction, we further examine the profile structure of the FR-GO sample by SEM. As shown in Taking advantage of the high temperature of the outer flame, the GO paper has been reduced in a drastic manner. As soon as the GO approaches the outer flame, the intense thermal effect would trigger the rapid removal of OCGs, leading to the formation of a highly porous structure due to the drastic expansion. 0 region and in the range of 0.4–1.0, respectively, indicating the presence of mesopores of different pore sizes. Pore size distribution (inset of 2/g) and large pore volumes (1.4 cm3/g). These results show that the FR-GO sample is highly porous, revealing great potential as anode materials for LIBs. For comparison, the GO sample shows a relative low BET surface area of 31 m2/g , Raman spectra, TG curve and X-ray photoelectron spectroscopy (XPS) have been implemented to characterize the FR-GO sample. M images .−1 and 1600 cm−1, which correspond to D and G band, respectively. After flame treatment, the peak intensity changed slightly. The G band is attributed to an E2g mode of graphite associated with the vibration of sp2 bonded carbon atoms, whereas the D band peak is related to the oscillations of carbon atoms with dangling bonds in plane terminations of disordered graphite. After the flame treatment, the intensity ratio of D band and G band, ID/IG, slightly decreased from 1.27 for pristine GO to 0.89 for FR-GO. The removal of oxygen defects would render a much higher content of sp2 carbon, represented by a significantly increased G band peak. The decrease of ID/IG ratio indicates that the flame reduction of GO could effectively remove the oxygen defects and partially reconstruct the sp2 network of graphene.It is well known that Raman spectroscopy has been widely used for characterizing carbon materials. As shown in To further evaluate the residual OCGs on FR-GO sample, thermogravimetric (TG) heating curves of GO and FR-GO have been measured in nitrogen . The weiIn order to analyse the reduction degree of FR-GO sample, we measured the XPS of pristine GO and the resultant FR-GO, respectively. As shown in st, 2nd, 3rd, 50th, 100th, 150th and 200th cycle at 0.33C. In the first cycle, a long voltage region below 0.5 V (vs. Li/Li+) is observed, which is mainly due to the lithium-ions intercalation into the graphene layers33+ on functional groups and its consumption of large amount of Li+ due to the formation of the SEI layer in the first cycle. It was suggested that the CE can be increased dramatically by doping graphene oxide into sucrose to reduce its specific surface area or further enlarge the interlayer lattice distance of FR-GO36The electrochemical performance of FR-GO and chemical reduction graphene oxide (CR-GO) is shown in 2/g) of FR-GO may greatly reduce the transport lengths of Li+, by which Li+ could be electrochemically adsorbed on both sides of graphene sheets383935The excellent performance of the as-synthesized FR-GO could be assigned to the following reasons. First, the highly porous architecture –dimethyl carbonate (DMC)–diethyl carbonate (DEC)  was utilized as the electrolyte and Li foil was used as the counter electrode.Electrochemical measurements of the FR-GO were evaluated with LIR2032 Coin-type half-cells assembled in an argon filled glove box (MB200MOD). The working electrodes were fabricated by a mixture of 10 wt% acetylene black, 70 wt% active materials, and 20 wt% polyvinylidene difluoride (PVDF) binder with N-methyl-2-pyrrolidone (NMP) solution. Following that, the resultant slurry was uniformly coated on a copper foil with a plastic blade, of which it was put in a vacuum oven at 100 °C overnight. A Celgard 2400 microporous polypropylene membrane was applied as the separator, 1.0 mol L-1 from room temperature to 800 °C, Ar has been used as inert gas.The static water contact angle (CA) measurements were made using the Contact Angle System OCA 20  at ambient temperature. The CAs were measured by the sessile-drop method with a water droplet of 5 μL. SEM images were obtained by using a field-emission scanning electron microscope . X-ray photoelectron spectroscopy (XPS) was performed using an ESCALAB 250spectrometer. Powder X-ray diffraction (XRD) patterns were collected on a Rigaku D/MAX 2550 diffractometer with Cu Kα radiation (λ = 1.5418 Å). Raman spectrometer equipped with a liquid-nitrogen-cooled argon ion laser at 514.5 nm (Spectra-Physics Stabilite 2017) as the excitation source; the laser power used was about 10 mW at the samples with an average spot size of 1 μm in diameter. Thermogravimetric (TG) curves were carried out on a NETZSCH STA 449C with a heating rate of 20 °C minHow to cite this article: Jiang, H.-B. et al. Flame treatment of graphene oxides: cost-effective production of nanoporous graphene electrode for Lithium-ion batteries. Sci. Rep.5, 17522; doi: 10.1038/srep17522 (2015).


The ECG on admission shows ectopic atrial tachycardia with multilevel block: variable Mobitz type II exit block to the atrium and Wenckebach phenomena in the atrioventricular node . Chest computed tomography scan also showed mediastinal adenopathy (not shown). This combination of findings narrowed the differential diagnosis to atrial infiltration due to sarcoidosis or lymphoma/tumours. Biopsies collected during minimal thoracotomy established the diagnosis of sarcoidosis (not shown).In cardiac sarcoidosis, conduction disturbances are related to granulomatous infiltration of the conduction system . Their eThis case underlines that when conduction disturbances are seen in younger patients, a search for cardiac structural abnormalities is warranted and sarcoidosis should be suspected .


In vitro models are useful in answering this question by providing information over the spatiotemporal nature of such dynamics. In the present work, we used physical quantities such as growth rate, velocity, and acceleration for the cellular proliferation and identified the fractal structures in tumor cell proliferation dynamics. We provide evidence that the rate of cellular proliferation is of nonlinear nature and exhibits oscillatory behavior. We also calculated the fractal dimensions of our cellular system. Our results show that the temporal transitions from one state to the other also follow nonlinear dynamics. Furthermore, we calculated self-similarity in cellular proliferation, providing the basis for further investigation in this topic. Such systems biology approaches are very useful in understanding the nature of cellular proliferation and growth. From a clinical point of view, our results may be applicable not only to primary tumors but also to tumor metastases.Biological systems are characterized by their potential for dynamic adaptation. One of the challenges for systems biology approaches is their contribution towards the understanding of the dynamics of a growing cell population. Conceptualizing these dynamics in tumor models could help us understand the steps leading to the initiation of the disease and its progression.  Population dynamics and population genetics provide a well-developed mathematical theory of evolution , 2 and mGenes manifest several patterns of differential expression in cancer , 7. Gene in vitro systems provide an excellent opportunity to study effects that are impossible to be measured in vivo. Most importantly, in vitro systems can be studied in the long term. This is required in order to reach conclusions regarding nonlinearity and chaotic behavior of a cellular system. Since primary cell cultures are short-lived when untransformed (15–20 days), the only way to apply such measurements is to use already established cell lines. For this reason, we developed a modeling approach in order to simulate the in vivo conditions, as best as possible. The nature of proliferation dynamics can give insight into the way that not only cells proliferate, but they also differentiate.Data regarding the dynamic nature of a tumor can only be collected after it has been diagnosed. Usually, this is too late for the patient, since all the critical steps for the progression of the tumor have already taken place. Therefore, in vitro cellular systems, using three central nervous system (CNS) tumor and a T-cell acute lymphoblastic leukemia (T-ALL) cell lines. These cells provide an excellent substrate for modeling proliferation dynamics, as previously shown  appeared to manifest the most stable oscillatory pattern, among all measurements that we performed  and the Apollonian Gasket . Our results confirmed two interesting points: (1) cell growth factors follow oscillatory dynamics (of nonlinear nature) and (2) different cell types followed similar dynamics of growth, irrespective of whether they grow as adherent or suspension cells, hinting towards a common mechanism of cellular proliferation.We measured the proliferation of the three CNS tumor cells and the CCRF-CEM cells Figures . In orde Figures . As a re Figures , charact Figures  and acce Figures , respect Figures . Therefo 0.80888 . In ordeIn the present work we identified nonlinear factors of cellular proliferation, in three CNS tumor cell lines and one leukemic cell line. Since our results show that cell growth is of nonlinear nature, we propose an initial theoretical framework for the analysis of such phenomena and for future considerations. This knowledge could be useful in treating tumors, since by understanding the mechanisms of cellular proliferation we could interpret the factors that determine the progression of the disease and/or metastasis. Biological systems are extremely complicated and manifest non-linear/chaotic phenomena. Among others , 32, we  Lyapunov exponents (one of the criteria of chaos existence). Such an example is the understanding of cellular proliferation in which we attempted to contribute with our hints. Here, we show that that different cell types follow similar dynamics with respect to proliferation, however their dynamics follow different trajectories. This difference arises from the probable reason that it is possible that all trajectories can be described by the same function, yet with different constants. Another question arising is that when cellular populations are measured with respect to time, they would give natural numbers corresponding to the exact or approximate number of cells present in the system. At the same time, all other physical variables calculated return oscillations. The question posed, concerns the role of gene expression in controlling cellular proliferation. A possible explanation is that cells follow oscillatory dynamics exactly the same way gene expression does [We have previously described the chaotic patterns of the leukemic cell line that we used . These pion does .Another issue in tumor proliferation dynamics that remains unanswered is the conditions at the time of the disease onset. The only knowledge we have thus far, concerning tumors, originates from the time of clinical presentation, at diagnosis. Practically, we have complete lack of knowledge from the time of tumor initiation to the time of tumor presentation. In that sense, the understanding of the proliferation dynamics of tumor cells is critical since it could provide insight into the understanding of tumor initiation. Supposing we could describe the dynamics of cellular proliferation in a formal, mathematical form, we would be able not only to predict the time zero (the starting point) but also to understand the mechanics of this progression. Additionally, if we suppose that such a formal description could be applied to several tumor types, we could conclude to a general rule of tumor proliferation. To the best of our knowledge, there are no previous works dealing with this subject and posing these questions.The implications of the understanding of proliferation dynamics are immense. To date, we do not have a sufficient theory that could allow us understand and predict cellular growth. This is easy to prove if we ask a simple question: given the population of cells today can we predict the population after 24 hours? The answer is no, since the only way we can do it is by approximation and this can be done only statistically. In other words, if we could predict the future cellular population based on the past population, this could lead us to the starting point of cellular proliferation, information that would be of extreme importance in cancer biology.Concluding, in the present work we aimed to set a framework for the detection of global patterns in cellular proliferation. Considering the fact that our knowledge in tumor biology comes only from the clinical presentation of the disease, the discovery of global models of tumor progression and proliferation could provide more insight in tumor biology and be used for therapeutic or prognostic purposes. Future work should focus on the investigation of the rates of cellular death in the same proliferation models and, most importantly, expand this model to gene expression for the same proliferation models, thus moving from phenotype to the genotype.


A striking puzzle about language use in everyday conversation is that turn-taking latencies are usually very short, whereas planning language production takes much longer. This implies overlap between language comprehension and production processes, but the nature and extent of such overlap has never been studied directly. Combining an interactive quiz paradigm with EEG measurements in an innovative way, we show that production planning processes start as soon as possible, that is, within half a second after the answer to a question can be retrieved . Localization of ERP data shows early activation even of brain areas related to late stages of production planning . Finally, oscillation results suggest an attention switch from comprehension to production around the same time frame. This perspective from interactive language use throws new light on the performance characteristics that language competence involves. There is a striking but largely unexplored puzzle at the heart of the language sciences. The core ecological niche for language use is conversational interaction, where speakers take rapid turns at talking. The puzzle is that the changeover between speakers is very quick, but the latencies involved in planning language production are three to ten times longer (depending on the nature of the turn under construction). This implies that to achieve this timing, participants must start production planning already during the incoming turn. Thus, there must be significant overlap between language comprehension processes and production ones – such dual-tasking forms an interesting challenge for the system since both processes draw on largely the same cognitive and neural resources.Earlier work has established the tight timing of turn-taking in conversational interaction278Given the predictive comprehension and early production implied by turn-taking, a crucial locus of investigation is what cognitive processes are going on in the recipient during an incoming turn to which he or she must respond. Using electroencephalography (EEG) as a non-invasive way of tapping into these processes with an excellent time resolution, we designed an innovative interactive turn-taking paradigm in the form of a quiz game (see Methods). This preserved the essential turn-taking structure, and is a familiar genre of interactive language use. Sitting in a shielded chamber, participants were asked to respond quickly (within 5 seconds) to Dutch quiz questions, and received live feedback from the experimenter. Unbeknownst to the participant the quiz questions themselves were pre-recorded by the same experimenter, providing a high degree of experimental control across participants.007, appears in the famous movies?EARLY: Which character, also called movies, is also called 007?LATE: Which character from the famous To vary when participants could start planning their answers, we created two different conditions.007, in bold) was available early on, whereas in ‘LATE’ questions (see example 2), this information was not available at the equivalent position , but only became available at the very end of the question. The position in the middle of the sentence  is hereafter referred to as the first time-locking point (or TL1) and the onset of the last word of the sentence is referred to as the second time-locking point (or TL2). Answers constituted one word or a small phrase. We were interested in how soon participants would start planning the production of their answer after the critical information became available, especially in the EARLY condition. Crucially, to distinguish production vs. comprehension processes, a control experiment was conducted, in which participants listened to the same quiz questions but instead of answering them, were asked to remember the questions and were tested using probes.In ‘EARLY’ questions (see example 1) the critical information essential to answer the questions  as the main predictor, and random intercepts for participant and item, showed that responses relative to question offset in the EARLY condition  were faster than in the LATE condition  based on the EEG data from 64 electrodes, time-locked to TL1 and TL2. Second, and most importantly, we found a large positive effect, starting around 500 ms after the onset of the critical information in both TL1  and TL2 . In the control experiment, in which participants did not answer the questions, positivities were present as well , but they were reduced in size, as confirmed by statistically comparing the two experiments . Note that the smaller positivities in the control experiments are not caused by the larger N400 effects found there, as shown by two additional analyses  and TL2  were localized at distributed sources in the brain, mostly in the left hemisphere . Local m007). In contrast, in the control experiment where no overt production was required, no frequency effects were found at TL1. At TL2 marginally significant reduced alpha power was found for LATE vs. EARLY questions between about 800 and 1100 ms after onset of the last word (p = .06). An analysis comparing the two experiments confirmed that the reduced alpha effect at TL1 (EARLY vs. LATE) was stronger in the main experiment than in the control experiment , but we found no differences between the experiments at TL2. Since the reduced alpha power was not present or smaller in the control (comprehension) experiment than in the main (production) experiment, this effect also appears to be related to production preparation.In a further analysis, we computed time-frequency representations of the oscillations of the EARLY and LATE questions, using the same time-locking as for the ERPs. We found that power was modulated around the alpha band (about 9–14 Hz) differentially across the two experiments . In the These oscillatory effects might be interpreted in two different ways. First, reduced alpha (and beta) power has been associated with motor preparation , as suggested by source localization of the positivity. The second effect was reduced power in the alpha band that we interpret as a switch of attentional resources from comprehension to answer retrieval and production processes. Importantly, both effects started already around 500 ms after the onset of the information that enables participants to retrieve the answer. This indicates that interlocutors started production planning within half a second of hearing the critical information necessary to start answer preparation – in the EARLY questions on average around 2.4 seconds before the end of the question. Additional analyses time-locked to the response . The 24 remaining participants  had a mean age of 21.3 years old (range 18 to 24). For the control experiment, another 32 participants were recruited from the same pool. Data from 2 participants was excluded from the analysis because of too many artifacts . The 30 remaining participants  had a mean age of 21.1 years old (range 18 to 25). All participants were native speakers of Dutch without hearing impairments. They gave informed consent before participating and received 8 euros per hour for their participation.007, komt voor in de bekende films? “Which character, also called 007, appears in the famous movies?”EARLY: Welk karakter, ook wel films heet ook wel 007? “Which character from the famous movies is also called 007?”LATE: Welk karakter uit de bekende We used 94 experimental Dutch question pairs in two different conditions. In EARLY questions (see example 1), the ‘answer-recognition’ point (the point at which the answer was clear) occurred early in the question, followed by 8.1 syllables on average (range 5–21) or 4.4 words (range 2–10), whereas in LATE questions (see example 2), this point occurred during the last word (or two words). The questions of each pair differed in the order of the constituents, but furthermore contained the same words as much as possible and fitted with exactly the same answer.A pretest (20 participants) ensured that the EARLY questions could be reliably answered correctly immediately after the answer recognition point  and the LATE questions could not reliably be answered correctly before hearing the last word . The same pretest showed that the average cloze probability of the last word was .07 for the LATE condition and .59 for the EARLY condition (t(93) = 13.89, p < .001). Thirty pairs (EARLY and LATE) of relatively difficult quiz questions served as filler questions, to create a more realistic quiz and to create more of a competition for the prizes (see Procedure). A pretest (10 participants) showed that they were answered correctly 31% of the time on average (range: 10–80%).All experimental and filler questions were recorded twice by a research assistant (RA) in the EEG room under the same circumstances as during the EEG experiment, and the best token of each question was chosen for the experiment. The experimental quiz situation was simulated by having a participant present who answered the questions. Some of the filler questions  were recorded with a hesitation or error to create the illusion of live speech. Recorded items were cut around 500 ms before question onset and exactly at question offset using PraatThe experimental questions could differ on the factor Answer . Two lists were created, which were both administered to half of the participants. List 1 contained half of the experimental and filler questions in the EARLY condition and the other half in the LATE condition, which was reversed for list 2. All items were presented in the same order for all participants. The items were divided into 4 blocks of 31 questions each with pauses in between. Question topics were spread over the 4 blocks and each block contained 7 or 8 difficult filler questions, but never twice in a row. The exact same design was used for the main and the control experiment.After EEG preparation, participants sat down in a sound proof booth in front of a computer screen. The same RA that recorded the materials instructed them from outside of the soundproof booth via a sound system. The RA used a microphone and participants heard her via loudspeakers. Participants were first informed that the two participants who answered most questions correctly would win a prize. Then, the RA instructed participants to answer quiz questions that varied from easy to difficult. They were urged to focus on giving the right answer, within a deadline of 5 seconds. They should look at a fixation cross on the screen (without moving or blinking) that was presented from between 1500 and 2000 ms before the question started until a button-press by the RA after the answer was given. Afterwards, the RA gave feedback while the word ‘knipperen’ (blink) was visible on the screen, indicating that participants could blink their eyes. They were told that the complete interaction was live, while in reality the questions were played by a button-press from the RA, while the feedback was given live. Participants started with a practice block of 16 questions, followed by the 4 experimental blocks. After the experiment, participants filled out a short questionnaire and received a debriefing sheet, informing them that the questions had been recorded.In the questionnaire after the experiment, in a first open question, none of the participants reported to have noticed anything strange during the experiment. When asked directly whether they thought (in retrospect) that any of the stimuli they heard could have been recorded, 6 participants (25%) reported that some of the questions could have been recorded. Also 3 other participants (12.5%) reported that the feedback after the questions could have been recorded . To keep enough power, we kept all these participants in the analyses. However, we performed additional ERP and time-frequency analyses for the main experiment without the 6 participants that reported that some of the questions might be recorded, to exclude the possibility that these participants could explain some of the differences we found see .The procedure was the same as in the main experiment except for the following. Participants were instructed that this was a control experiment for an earlier quiz experiment in which participants answered the questions. They were told that the RA read the questions to them live to keep as close to the quiz experiment as possible. Participants were asked not to answer but try to remember the questions. After each block of questions, they saw 10 probe questions on their screen, one by one, and had to indicate for each question whether they had heard it in the previous block or not. Participants received two practice blocks of 20 questions each, to give them an indication of the type of foils they could expect. The foils were very similar to the questions they had heard, differing either in answer or in content. For example, for the quiz question “Which element in coffee is the effective element?”, the foil was: “Which element in coffee wakes you up?”On average 92% of probe questions were remembered or discarded correctly, showing that participants paid attention to the questions. The questions from both conditions were remembered equally well (T < 1). In the questionnaire after the experiment, in a first open question, none of the participants reported to have noticed anything strange during the experiment. When asked directly, only 3 participants reported that some of the questions might have been recorded.EEG was recorded from 61 active Ag/AgCI electrodes using an actiCapFor behavioral data analysis of the quiz questions, we looked at the numbers of errors in the two conditions and we measured the response time from the end of the question to the start of the answer. Incorrect responses  and responses starting with hesitations (5%) were discarded before the analysis. We ran mixed-effects models on these two variables to assess the effect of Condition  in R.007 in example 1). As an equivalent point in the LATE condition, we took the word onset that was closest in time to the same point in the EARLY condition (as measured from sentence onset) for each item . The second time-locking point (TL2) was the onset of the last word in the LATE condition  and in the EARLY condition . See Preprocessing and statistical analysis of EEG data was conducted using FieldtripEpochs for analysis consisted of 200 ms before until maximally 2000 ms after TL1 and 200 before until maximally 1400 ms after TL2 since speech started earlier for TL2 . These preprocessed data entered the event-related potential (ERP) and time-frequency  analyses. For ERPs, epochs were filtered with a low-pass filter of 35 Hz and baselined with a baseline window of 200 ms immediately before the time-locking point. Then, trials of the same condition were averaged per participant. For time-frequency representations, no filtering or baselining was performed, but a linear trend was removed from the data before the analysis. The power of each frequency between 4 and 30 Hz (with steps of 1 Hz) was calculated on the extracted epochs of individual trials using a Hanning taperTo test for statistically significant differences between conditions and reduce the multiple-comparison problem, we used the cluster-based approachTo identify sources underlying the electrode-level effects, a BEM (boundary element headmodel)How to cite this article: Bögels, S. et al. Neural signatures of response planning occur midway through an incoming question in conversation. Sci. Rep.5, 12881; doi: 10.1038/srep12881 (2015).


Graphene exhibits unusual electronic properties, caused by a linear band structure near the Dirac point. This band structure is determined by the stacking sequence in graphene multilayers. Here we present a novel method of microscopically controlling the band structure. This is achieved by epitaxy of graphene on 3C-SiC(111) and 3C-SiC(100) thin films grown on a 3D microfabricated Si(100) substrate (3D-GOS (graphene on silicon)) by anisotropic etching, which produces Si(111) microfacets as well as major Si(100) microterraces. We show that tuning of the interface between the graphene and the 3C-SiC microfacets enables microscopic control of stacking and ultimately of the band structure of 3D-GOS, which is typified by the selective emergence of semiconducting and metallic behaviours on the (111) and (100) portions, respectively. The use of 3D-GOS is thus effective in microscopically unlocking various potentials of graphene depending on the application target, such as electronic or photonic devices. Carrier dynamics in monolayer graphene, a honeycomb lattice consisting of carbon atoms, are described in terms of quantum electrodynamics12368One of the major challenges to such device applications of graphene is that of establishing epitaxial growth of graphene on practical substrates using methods compatible with existing device technologies11131516171516171718191922In addition to these manufacturing advantages, GOS has another more profound advantage: the tunability of the band structure by altering the Si substrate. By controlling the crystallographic orientation of the Si substrate, we can select the surface orientation of the 3C-SiC thin film, which eventually makes it possible to modulate the stacking of graphene and its interface structure with SiC. The latter controls the presence or absence of the interfacial (6√3 × 6√3)-reconstructed buffer layer, which is a precursor for graphene in EG and GOS. On the Si(111) substrate, the buffer layer is present, and GOS is Bernal-stacked17171821In this work, we demonstrate the microscopic tuning of the band structure of epitaxial graphene on a 3D microfabricated Si substrate (3D-GOS) through stacking with the aid of microelectromechanical system (MEMS) technology, specifically anisotropic wet etching which produces (111) and (100) microfacets on a Si(100) substrate.2 film (90 nm) is grown on a Si(100) substrate  by the dry oxidation method. Electron-beam lithography is performed on the sacrificial SiO2 thin film, followed by fast atomic beam etching using SF6 as an etchant272 mask on the surface. Using the SiO2 mask, the surface is anisotropically etched in a 25% tetramethyl ammonium hydroxide (TMAH) aqueous solution at 353 K for 6 min. The etching by the TMAH aqueous solution is used to expose Si(111) microfacets on Si(100) substrates, which are frequently used as the standard fabrication procedure for MEMS devices2 mask with a treatment in a 5% HF aqueous solution. The 3C-SiC thin films (~100 nm in thickness) are grown on microfabricated Si(100) substrate by using gas-source molecular beam epitaxy with monomethylsilane as a gas source at a substrate temperature of 1353 Kin vacuo at 1523 K for 30 min to grow graphene epitaxially on the surface of the 3C-SiC(111) microfacet15161718The formation of 3D-GOS is schematically shown in The interface between graphene and the 3C-SiC microfacets is examined here because the graphene growth mode is determined by the presence or absence of the buffer layer because it works as the template for the Bernal stacking of graphene17183017171333The atomic structure of the interfaces between graphene and the 3C-SiC microfacets has been investigated by using cross-sectional transmission electron microscopy (X-TEM), as shown in To clarify the relation of the observed interface variation caused by microfacets with the graphene stacking, microscopic LEED (μ-LEED) analysis is performed for 3D-GOS, as shown in 1718As seen from the μ-LEED images of the two bevels, a hexagonal LEED pattern from graphene is clearly identified on the 3C-SiC(111) microfacets. The spots due to the specular reflection, labelled as (00) in the images, are located away from the center of the μ-LEED pattern because of the inclination of the 3C-SiC(111) microfacets. The μ-LEED pattern is almost the same as that of the graphene on the flat Si(111) substrate and graphene on 6H-SiC(0001)171836In contrast to epitaxial graphene on the 3C-SiC(111) microfacet, the LEED spots of graphene on both the top and bottom 3C-SiC(100) microterraces are rotated, as observed on the graphene on 3C-SiC(100) thin films on flat Si(100) substrates. The rotation angle between the adjacent spots indicated by the yellow arrows is about 15 degrees. One of the possible mechanisms for this rotated growth of graphene on the 3C-SiC(100) face is the {111} facet-induced mechanismechanism . There a−1), D (~1360 cm−1), and G′ (~2700 cm−1) bands, are visible in the spectra on the graphene on both 3C-SiC(100) microterraces and 3C-SiC(111) microfacets. The G band is the band originating from a single-resonant Raman process at the Γ point39The band structure of 3D-GOS is anticipated to vary with the microfacet and the microterrace orientation owing to a high susceptibility of the band structure to the stacking sequence, by analogy with the susceptibility of the band structure of GOS using flat Si substrates for stacking depending on the crystallographic orientation of the Si substrate17−1. This value is larger than that of epitaxial graphene on 6H-SiC(0001) (37 cm−1)−1) for graphene on a 3C-SiC(100) thin film on a Si(100) substrate172044Among these bands, the G′ band is analysed in detail to clarify the band dispersion41424344394244−1) than that on the 3C-SiC(100) microterrace (70 cm−1). These two features can only be accounted for by considering a set of multiple Lorentzian components, as shown in D/IG of the D band to the G band, which is a good measure of the defect density that broadens the G′ band, is substantially smaller in the graphene on 3C-SiC(111) microfacet (ID/IG = 0.64 ± 0.04) than that on the 3C-SiC(100) microterrace (0.75 ± 0.04). Thus, the G′ band on 3C-SiC(111) cannot be represented by a single Lorentzian. The broadening of the G′ band of the graphene on the 3C-SiC(111) microfacet is thus concluded to be due to presence of multiple components. The presence of multiple Lorentzian components in the G′ band suggests presence of multiple routes for this double-resonant Raman scattering process in the graphene on the 3C-SiC(111) microfacets. Several origins can be considered, which include the band splitting of graphene424347−1 is attributed to the so-called D′ band, induced by defects4742On the other hand, the G′ band of the graphene on the 3C-SiC(111) microfacet is asymmetrical and is much broader  substrate by anisotropic wet etching, which produces Si(111) microfacets as well as Si(100) microterraces. The realization of 3D-GOS is the first step to exploiting graphene-based multifunctional integrated circuits (combining electronics and photonics) compatible with existing Si-based electronics in the next generation of devices.E/ΔE > 104). The photon energy of the SR beam for the measurement is 1000 eV. Using 3D nano-ESCA enables us to obtain a high lateral resolution (70 nm) by focusing X-rays using a Fresnel zone plate. The energy resolution of the spectrometer was set to 300 meV, and the accuracy of the angle resolution is 0.9°. The binding energy scale was calibrated by the photoelectron peaks of a gold foil (Au 4f 7/2). Details of the experimental setup are described in the previous reports3017The spatially-resolved and angle-resolved C1s core-level photoelectron spectra are taken by using the 3D nano-ESCA system installed at the BL07LSU at SPring-83031H.F. performed the 3D-GOS fabrication and all of the microscopic characterizations, designed the study, and participated in the writing of the manuscript. T.I. performed the 3D-GOS fabrication and all of the microscopic characterizations. Y.K. participated in fabricating the 3D-GOS. T.S. participated in the microscopic characterization using 3D nano-ESCA. N.N., K.H. and H.K. performed the microscopic characterization using 3D nano-ESCA, and participated in discussions about the content of the manuscript. M.K., T.O. and T.K. participated in the microscopic characterization using μ-LEED, and participated in discussions about the content of the manuscript. M.O. participated in the microscopic characterization using 3D nano-ESCA, and participated in discussions about the content of the manuscript. M.S. participated in discussions about the content of the manuscript.Supplememtal Information


Our results show that this herbal medicine possesses strong activities including stimulation of angiogenesis, cell proliferation, and cell migration, which provide the scientific basis for its clinically observed curative effects on nonhealing diabetic wounds.“Jinchuang ointment” is a traditional Chinese herbal medicine complex for treatment of incised wounds. For more than ten years, it has been used at China Medical University Hospital  for the treatment of diabetic foot infections and decubitus ulcers. Three different cases are presented in this study. “Jinchuang” ointment is a mixture of natural product complexes from nine different components, making it difficult to analyze its exact chemical compositions. To further characterize the herbal ingredients used in this study, the contents of reference standards present in a subset of the ointment ingredients  were determined by HPLC. Two Medicine Comprehended, published in 1732. Clinical applications of this herbal medicine for diabetic foot infections and decubitus ulcers have been a successful course of treatment in the Division of Chinese Medicine, China Medical University Hospital, Taichung, Taiwan, for more than ten years. Despite its track record of curative effects, there is no literature published in the English language describing the clinical efficacy of “Jinchuang ointment” [ It is well-known that diabetic foot ulcers are extremely difficult to be treated and are the dominant complication leading to amputations . “Jinchu“Jinchuang ointment” is composed of lard, wax, starch, synthetic borneol, camphor, frankincense, dragon's blood, myrrh, and catechu. To further characterize the chemical content of each component in this complex, the ratio of stereoisomers in chemically synthesized borneol used in this study was analyzed by chiral gas chromatography (GC). Meanwhile, the content of reference standards in the herbal components, like frankincense, myrrh, dragon's blood, and catechin, was determined by high performance liquid chromatography (HPLC). Lard is the major component in this complex, and its weight percentage is as high as 67%. It is of great interest to determine the role of lard in this complex. Lard was therefore substituted for synthetic triacylglycerol, coconut oil, Vaseline®, and sesame oil. The activity of these reconstituted complexes was examined in this study. in vitro tube formation assay, a wound healing assay, and a cell proliferation test were carried out to examine the activity of “Jinchuang ointment.” Here, we report the outcomes of treating patients with “Jinchuang ointment,” the results of cell based activity assays, and characterization of herbal components by HPLC.“Jinchuang ointment” is directly applied to the wound surface during treatment. The components in this complex are neither digested nor absorbed in the gastrointestinal tract. It is therefore reasonable to evaluate its bioactivity by direct addition of this complex into media containing cultured human skin or endothelial cells. Wound healing is a very complicated process. In this study, anβ-boswellic acid, catechin, and epicatechin were purchased from the National Institute for the Control of Pharmaceutical and Biological Products . (E)-Guggulsterone and (+)-borneol were obtained from Sigma-Aldrich . (−)-Borneol and (±)-isoborneol were purchased from Alfa Assar . DMEM high glucose with sodium pyruvate media, Catalog number: I-26F55-I, was bought from Amimed® BioConcept . Chemically synthesized borneol and camphor were bought from Cheng Yi Chemical Co., Ltd. . Herbal medicine, dragon's blood, catechu, frankincense, and myrrh were bought from Healthy Beautiful Biotech. Co. Ltd. . Vascular endothelial growth factor (VEGF) was bought from B&D Systems . Antibodies for western blot analysis were bought from Santa Cruz Biotechnology, Inc. . Food-grade lard was obtained from I-Mei Food Company . Synthetic triacylglycerol  was obtained from InterMed Manufacturing Sdn. Bhd. . Food-grade sesame oil was from Fwusow Industry Co. Ltd. . Coconut oil was bought from First Chemical Co., Ltd. . Vaseline was bought from Unilever .The reference standards of dracorhodin perchlorate, acetyl-11-keto-The composition of “Jinchuang ointment” (100 g) is as follows: lard 67.3 g, dragon's blood 2.1 g, catechu 2.1 g, frankincense 2.1 g, myrrh 2.1 g, camphor 6.3 g, borneol 0.1 g, corn starch 8.4 g, and wax 9.5 g. For wound healing and tube formation assays, the DMSO stock solution of “Jinchuang ointment” is prepared as follows: two grams of “Jinchuang ointment” is dissolved in 10 mL DMSO and homogenized by ultrasonication just before use.μm, 250 × 4.6 mm, analytic column  was used for analysis.All experiments were carried out on a Hitachi L-7000 HPLC system, equipped with L-7100 quaternary gradient pump and a L-7450 photo diode array detector. Hitachi HSM software was used for machine controlling, data collecting, and processing. A Mightysil, RP-18, 5 μm syringe filter. For myrrh, 95% ethanol was used for extraction rather than methanol. Other preparation steps were identical to those of dragon's blood, catechu, frankincense, and myrrh.For samples of dragon's blood, catechu, and frankincense, 0.1 g grounded solids were weighed and dissolved in 10 mL of methanol. After ultrasonicating for 30 minutes at room temperature, methanol extracts were transferred to a new glass vial by using disposable glass Pasteur pipettes. 4 mL of methanol was then added and ultrasonicated for another 30 minutes at room temperature. The final volume of the extract was adjusted to 25 mL by adding methanol. Undissolved particles were removed by centrifugation at 2500 ×g for 10 minutes at room temperature and filtrated through a 0.22 3CN containing 0.1% formic acid) and solvent B (90% CH3CN containing 0.1% formic acid) with a flow rate of 1 mL/min [The methanol extract of dragon's blood was separated using a gradient elution of solvent A (10% CH1 mL/min . The elu2O containing 0.1% formic acid), B (10% CH3CN containing 0.1% formic acid), and C (90% CH3CN containing 0.1% formic acid) with a flow rate of 1 mL/min [The catechu methanol extract was separated using a gradient elution of A  and solvent B  with a flow rate of 1 mL/min [The frankincense methanol extract was separated using a gradient elution of solvent A  with a flow rate of 1 mL/min for 25 minutes [To determine the content of (E)-guggulsterones in myrrh, an ethanol extract was separated using an isocratic elution of 0.1% H minutes . The UV μm) was obtained from SGE Analytical Science . Chromatographic conditions were as follows: helium used as a gas carrier; a constant flow of 1.0 mL/min; 2 μL injection volume (splitless model), and a 280°C injector temperature. The GC temperature program was as follows: 90°C (1 min), 90°C to 130°C (41 min), 130°C (10 min), and 200°C (3 min). An Agilent 5973N quadrupole mass spectrometer was operated in selective ion monitoring (SIM) mode, with ionization source by electron impact at 70 eV, transfer line at 280°C, ion source at 280°C, and quadrupole at 150°C.Chiral GC was used to determine the ratio of stereoisomers in synthetic borneol. Analysis was performed using an Agilent GC system model HP 6890N  equipped with a split/splitless injector, liner of silanized quartz with a 4 mm i.d. (effective volume 0.49 mL), and an Agilent 6890 autosampler for 100 vials. A Cydex-B chiral GC column .Confluent HaCaT cells in 12-well plates were starved overnight in DMEM medium. The surface of the plate was scraped with a 200  in vitro wound healing assay by “Jinchuang ointment” were also examined using human microvascular endothelial cells (HMEC-1). Ibidi Culture-Inserts  were placed on the chamber of 24-well cell culture plates. About 70 μL of HMEC-1 (5 × 105 cells/mL) was seeded per well and plates were incubated at 37°C and 5% CO2. After 24-hour incubation, Ibidi Culture-Inserts were removed, and 1 mL of MCDB131 media containing 1 μL DMSO solution of “Jinchuang ointment” was then added into individual wells. The migration of cells was observed by microscopy over a period of 6–24 hours. The gap size was measured by using software Image J.Stimulation effects on4/well) were seeded in 96-well plates. The medium containing “Jinchuang ointment” at various concentrations was then added after cell adhesion. Cells were incubated in DMEM medium for the indicated time. The proliferation of HaCaT cells was subsequently determined by using Cell Proliferation Reagent WST-1 . The statistical method used is Student's t-test.HaCaT cells . Membranes were blocked for 1 h with 5% low-fat milk powder solubilized in phosphate-buffered saline (PBS) containing 0.05% Tween 20. Levels of Cdc25b, Cdc25c, CDK2, CDC D2, Cyclin B, Cyclin D3, and 5 cells/well) was placed into the wells of a 24-well flat bottomed plates precoated with 200 μL Matrigel . Cells were then mixed with 1 mL medium containing “Jinchuang ointment” . Cells were incubated at 37°C for a 24 h exposure [Human umbilical vein endothelial cells (HUVEC) were bought from Bioresource Collection and Research Center . 1 mL of HUVEC . A To the best of our knowledge, there are no English language case reports describing “Jinchuang ointment” treatment for nonhealing diabetic wounds. Three different cases are presented in this study. The first subject, Mrs. Wu, is a 75-year-old female patient with type II insulin-dependent diabetes accompanied by peripheral arterial occlusion disease (PAOD) which led to left lateral leg and ankle necrotizing fasciitis. She was treated with percutaneous transluminal angiography (PTA) to improve lower limb circulation on Feb 6, 2013. As a result of reperfusion injury, the ulcer had enlarged with erythema. After examination, a below-knee amputation was immediately scheduled two weeks later at the Surgery Division, the China Medical University Beigang Hospital. As suggested by a doctor from the Division of Chinese Medicine, she decided to use traditional Chinese medicine to treat her wound. She was referred to the Division of Chinese Medicine for wound management. Normal saline was first used to clean the wound. About 2-3 g “Jinchuang ointment” was applied directly to the wound once daily. Pictures depicting wound healing under treatment with “Jinchuang ointment” are shown in Mr. Tsai, a 71-year-old male, with past history of type II diabetes was diagnosed on September 8, 2013. He had a 3 × 0.8 cm and a 4 × 1.5 cm grade 3 pressure sore in the sacral region. 1 g of “Jinchuang ointment” was applied topically to the wound area once per day. Pictures documenting wound healing under treatment of “Jinchuang ointment” are shown in Mr. Wang, a 64-year-old male with a past history of hypertension, had a chronic wound measuring 6.2 × 5.3 cm which had not healed for more than six months. He received the topical application of “Jinchuang ointment” once per day beginning November 26, 2014. A great improvement was observed after two months of treatment as shown in One of the main problems associated with herbal medicine is the high level of batch to batch variation in the amounts of active components. The content of pure chemical reference standards in herbal products is therefore used as an indicator for the purposes of quality control and standardization. Accordingly, the content of reference standards in dragon's blood, catechu, frankincense, and myrrh used was measured by HPLC in this study. All the calibration curves of reference compounds were linear over the concentration range studied . A linea Daemonorops draco. It possesses antimicrobial, anticancer, and cytotoxic activity [Dracorhodin is a red anthocyanin pigment that is a major component in “dragon's blood” resin of the plantactivity , 10.  FiBoth catechin and epicatechin are phenol-type antioxidants in catechu, an extract of acacia trees. Catechu is a common component of herbal medicine. In addition to their ability to scavenge free radicals in plasma, the health benefits of catechin and epicatechin also include stimulation of fat oxidation, expansion of the brachial artery, and resistance of LDLs to oxidation . At the  Boswellia. In Africa and Asia, it is widely used in incense, perfume, and traditional medicine. Boswellic acids, a series of pentacyclic triterpene molecules, are one of the major components of frankincense. Theiranti-inflammatory properties and ability to induce cancer cell apoptosis have been reported in vitro [β-boswellic acid derivatives and acetyl-boswellic acid [β-boswellic acid in frankincense. The mass percentage of acetyl-11-keto-β-boswellic acid in the frankincense used in this study is 1.62%.Frankincense is a resin from plants in the genusin vitro –17. The lic acid , 18.  Fi Commiphora. The usage of myrrh is similar to that of frankincense. In fact, both myrrh and frankincense are frequently used in concert in many traditional Chinese medicine recipes. In western medicine, myrrh is also used in liniment and healing salves for minor skin ailments. The chemical composition of myrrh is rather complicated [Myrrh is also a resin from plants in the genusplicated . Notablyplicated . Both isplicated . The mas Blumea balsamifera have enjoyed more widespread use. The price of synthetic borneol is the lowest among the sources discussed above. Previous results show that borneol stereoisomers can interact with GABA receptors [Borneol is a bicyclic monoterpene plant secondary metabolite. (+)-Borneol, mainly isolated from the plant family of Dipterocarpaceae, is the major form used for analgesia and anesthesia in traditional Chinese medicine. More recently, chemically synthesized borneol and (−)-borneol isolated from the plant specieseceptors , 23 and eceptors . Chiral μg/mL, 20 μg/mL, and 2 μg/mL “Jinchuang ointment” treatment was 78.2 ± 5.3%, 78.2 ± 4.3%, and 63.8 ± 1.7%, respectively. In contrast, the negative control HaCaT cells without “Jinchuang ointment” treatment only showed 64.4 ± 4.0% wound closure. The positive control treated with 100 ng/mL EGF treatment showed 71.8 ± 1.0% wound closure. It is obvious that application of 200 μg/mL or 20 μg/mL “Jinchuang ointment” is more potent than 100 ng/mL EGF in promoting in vitro wound closure, showing statistically significant differences when compared to both positive and negative controls /initial wound area × 100%. The wound closure percentage with 200 controls .μg/mL “Jinchuang ointment” 24 hours after treatment was 85.0 ± 12.3%, whereas the wound closure percentage observed with the DMSO control was only 43.3 ± 8.2%  by WST-1 assay. The increased percentages of cell proliferation by 200 ectively . These rTo further investigate the underlying mechanisms involved in “Jinchuang ointment”-induced effects on cell proliferation in HaCaT cells, the expression of several key cellular proteins involved in cell cycle progression was investigated by western blot analysis. As shown in  in vitro angiogenic effect of “Jinchuang ointment” on HUVEC cells. As shown in μg/mL “Jinchuang ointment” for 24 hours can efficiently induce endothelial cell capillary tubes and network formation.The process of wound healing has been divided into three different stages, inflammatory, proliferative, and remodeling phases . Angioge in vitro wound healing and tube formation, were used to examine activity. Our results show that this herbal medicine possesses potent activities stimulating cell proliferation, migration, and angiogenesis. This provides a scientific rationale to account for the observed clinical curative effects on wound healing by “Jinchuang ointment.”“Jinchuang ointment” is a Chinese herbal medicine complex. It has been clinically used in the treatment of diabetic foot infection and decubitus ulcers in China Medical University Hospital for more than ten years. Because of its complicated composition, its biological activities have never been investigated. To further characterize its herb ingredients, the content of reference standards present in dragon's blood, catechu, frankincense, and myrrh was determined by HPLC. Two cell based assay platforms,According to current pharmaceutical regulations in Taiwan, only traditional Chinese herbal medicine manufactured from cGMP pharmaceutical factories can be sold in drug stores or by hospital marketing channels. However, for homemade traditional Chinese herbal medicine, they can only be administered to patients by the doctors who made the respective medicine. For this reason, “Jinchuang ointment” cannot be widely used in the Taiwan area since many clinicians are not able or unwilling to prepare this remedy. To facilitate the manufacturing process of “Jinchuang ointment” by cGMP factories, it will be very important to find out the activity indicator markers for components, such as dragon's blood, catechu, frankincense, and myrrh.