Idiopathic pulmonary fibrosis (IPF) is a progressive chronic disease characterized by excessing scarring of the lungs leading to irreversible decline in lung function. The aetiology and pathogenesis of the disease are still unclear, although lung fibroblast and epithelial cell activation, as well as the secretion of fibrotic and inflammatory mediators, have been strongly associated with the development and progression of IPF. Significantly, long non-coding RNAs (lncRNAs) are emerging as modulators of multiple biological processes, although their function and mechanism of action in IPF is poorly understood. LncRNAs have been shown to be important regulators of several diseases and their aberrant expression has been linked to the pathophysiology of fibrosis including IPF. This review will provide an overview of this emerging role of lncRNAs in the development of IPF. Fibrosis is a pathophysiological condition that can affect nearly every organ in the human body where irregular and excessive accumulation of scar tissue leads to organ failure and potentially death as seen in the final stages of fibrotic diseases such as pulmonary , cardiacIdiopathic pulmonary fibrosis (IPF) is a progressive chronic interstitial lung disease (ILD) which is characterized by scar tissue accumulation, and therefore thickening of the normal lung walls, leading to impaired gas exchange and restricted ventilation. IPF is a disease of unknown aetiology, which makes the development of effective drug treatments particularly challenging . NonetheAs is the case with most ILDs, inflammation was initially thought to be the major player in IPF until unresponsiveness to anti-inflammatory medications prompted the re-evaluation of this idiom ,9. HowevIPF induces exertional dyspnoea, a feeling of breathlessness, which is one of the most common symptoms experienced. Dyspnoea is often accompanied with a non-productive dry cough which progressively worsens over time until it becomes clear to patients that their daily functionality is compromised. Pulmonary malfunction due to fibrosis can induce other symptoms such as inspiratory crackles, chest discomfort, finger clubbing, weakness, and loss of appetite ,16,17.Under physiological conditions, fibrogenesis is initiated in response to tissue injury and forms part of the wound repair process involved in the restoration of homeostasis. Wound repair is commonly initiated by epithelial injury, leading to activation of the coagulation and inflammation cascades. This in turn results in the activation, recruitment, and proliferation of fibroblasts that are responsible for the release of ECM components. In the final remodeling stage, the wound area is resolved, and normal tissue structure and structural integrity is restored .During fibrosis associated with IPF, any stage of the wound repair process can be dysregulated , resultiAt the present time, the drivers of the aberrant wound healing process leading to fibrosis are unknown. However, certain cells types such as alveolar epithelial cells and fibroblasts, as well as a dysregulated coagulation and inflammatory response have been implicated in disease initiation and progression ,26,27,28Prior to the activation of fibroblasts, it is believed that type I AECs could have been subjected to repetitive injury, which causes damage to the delicate epithelium structure. Type I AECs line more than 90% of the alveolar surface and damage promotes the activation and proliferation of the surfactant-producing type II AECs . This reDuring the normal healing process, the lung tissue eventually regains its original structure and function as the provisional matrix gradually dissipates. In the case of injury of the type I AECs, the hyperplastic type II AECs are thought to undergo regulated apoptosis and trans-differentiation into type I AECs in order to re-establish a fully functional alveolar epithelium .However, if the epithelial basement membrane remains disturbed following extensive damage, the alveoli can collapse and type II AECs fail to undergo re-epithelisation. An aberrant wound repair response can then be initiated during which the epithelial cells, predominantly type II AECs, are thought to release several pro-fibrotic cytokines, growth factors, and other chemokines at the site of injury . The epiOne of the initial concepts relating to IPF pathogenesis was that the disease was initiated and driven by chronic inflammation. However, the role of inflammation was questioned when anti-inflammatory and immunosuppressant therapies failed to improve lung function and survival of IPF patients. A study of 330 IPF patients showed no effect on progression-free survival when treated with the immune-regulatory cytokine interferon-gamma-1β (IFN-γ-1β) . AdditioAlthough the role of inflammation in the initiation and progression of the disease remains unclear, there is plenty of evidence to indicate that IPF is associated with inflammation and changes in the innate and adaptive immune response . StudiesInflammatory cells found in the lungs of IPF patients have been shown to produce elevated levels of reactive oxygen species (ROS) which are thought to contribute to tissue damage in IPF ,47. Mito+ T-helper (TH-17) cells, has been linked with enhanced neutrophil recruitment, also known as neutrophilia, as well as TGF-dependent and IL-1β-driven fibrosis [H-1 effector T cells are thought to exert anti-fibrotic activities through the production of IFN-γ, which has been shown to attenuate fibrosis [H-2 effector T cells are thought to promote fibrosis via the production of pro-inflammatory cytokines such as interleukin 13 (IL-13) which stimulates collagen deposition in fibroblasts [The adaptive immune response has also been linked to the development of IPF. Interestingly, the pro-inflammatory cytokine interleukin 17A (IL-17A) which is expressed by CD4fibrosis . Notablyfibrosis . Additiofibrosis  by inhibfibrosis . TH-2 efroblasts .In this environment, activated platelets, damaged epithelial cells, and recruited inflammatory cells release more pro-fibrotic growth factors and such TGF-β1 drive the fibrotic response. TGF-β1 is considered the driving force of fibrosis as it has multiple properties including activation of fibroblasts, epithelial cell differentiation via epithelial-mesenchymal transition (EMT), and stimulation of the expression of pro-inflammatory cytokines such as IL-1β to further enhance the fibrotic response .The aberrant wound healing response in IPF involves several underling mechanisms which are thought to drive the disease. However, the most well-established and studied concept is the role of TGF-β1 in the development of fibrosis and its effect on fibroblasts and the ECM which play a major role in the fibrotic cascade . TGF-β1 Most importantly, in this fibrotic environment, both fibroblasts and myofibroblasts secrete increased amounts of ECM components to synthesize and maintain the ECM. The ECM is a complex and versatile network of cross-linked and fibrous proteins that form a protective structure under healthy conditions; however, the excessive deposition of the matrix in IPF is thought to exert powerful effects on cell functions via ECM-cell interactions . As suchIPF is a heterogeneous disease that is characterized by complex genetic and environmental interactions that contribute to the development of the disease; and therefore it is now thought that genetically susceptible individuals exposed to environmental stressor stimuli have an increased risk of developing the disease. Both rare and common genetic variants have been associated with sporadic and familial pulmonary fibrosis .Genetic studies have linked IPF in adults to rare genetic variants in surfactant protein-related genes. Mutations have been found in the genes such as surfactant protein C (SFTPC) and A2 (SFTPA2)  which caRare variants in several genes regulating telomere biology have also been identified in IPF patients, particularly mutations affecting telomerase and telomerase-associated proteins such as TERT ,73 and TThe single-nucleotide polymorphism rs35705950, located in the putative promoter region of the MUC5B gene, was also found to play a role in predisposing patients to familial and sporadic forms of IPF by causing increased MUC5B expression . The MUCTranscriptional changes in the lungs of IPF patients have also contributed to our understanding of the disease and its underlying molecular mechanisms. In particular, microarray and sequencing-based approaches have identified an association between IPF and ECM formation, smooth muscle markers, growth factors, chemokines, and immunoglobulins .Several gene expression profiling studies have demonstrated widespread changes in the profile of mRNA (messenger RNA) expression in lung biopsies. A study by Nance et al., identified 873 differentially expressed genes in IPF lung biopsies as compared with controls using RNA sequencing (RNA-seq). Interestingly, 675 of these genes displayed alternative splicing events including those coding for periostin (POSTN) and collagen (COL6A3) . MicroarAs well as biopsies, gene expression in isolated human lung fibroblasts has also been employed to examine and identify novel IPF-related genes and pathways. A recent microarray study identified several IPF-associated genes upregulated in both lung control and IPF fibroblasts in response to TGF-β1 . Anotherl-1R2 were found to be upregulated in both mild and severe IPF patients. Elevated levels of systemic MMP3 and CXCL13 were also identified in the blood of IPF patients using microarrays [In an attempt to identify circulating biomarkers of IPF, transcriptome analysis has also been undertaken on blood serum and plasma. A study by Yang et al.  used microarrays .There are currently two drugs that have been approved for the treatment of IPF, nintedanib and pirfenidone, both of which slow the decline of lung function associated with the disease.l-1β production and lymphocyte counts in bronchoalveolar lavage fluid (BALF) obtained from human fibrotic lungs [Nintedanib, also known as BIBF1120, was originally developed as an anticancer agent and is thought to be a nonspecific tyrosine kinase inhibitor. A study by   showed iic lungs . In 2014ic lungs . The IMPPirfenidone is thought to be an antioxidant and anti-inflammatory compound which was first shown to have anti-fibrotic activities in the bleomycin model of lung fibrosis . It was Initial data from the Human Genome Project indicated that there were approximately 30,000 to 40,000 protein coding genes in the human genome . HoweverThe term ncRNA includes all RNA molecules that do not encode for proteins and a very big proportion of these are the well-known “housekeeping” RNAs including transfer RNAs and ribosomal RNAs, which play a critical role in protein biosynthesis. In addition, there are the small nucleolar RNA (snoRNA), which regulate the transcriptional modification of other RNA species and the small nuclear RNAs (snRNA) that form an important component of the spliceosome complex which removes introns from mRNA during transcription. In total, these “housekeeping RNAs” represent around 85% to 90% of the total RNA whilst the mRNAs account for 5% to 8%. The remaining ncRNAs are speculated to have regulatory roles in various cellular functions and are divided in two classes, i.e., short (<200 nucleotides) which are exemplified by the microRNA (miRNA) and long (>200 nucleotides) non-coding RNAs (lncRNAs) . MicroRNDespite the rapid increase in data relating to lncRNAs, little is known regarding their exact functions, mechanism of action, or even how many different types of lncRNAs exist. Therefore, due to the lack of mechanistic studies, lncRNAs are mainly grouped according to their proximity to protein coding genes. The most significant groups are lincRNAs (long intergenic non-coding RNAs), antisense, intronic, enhancers, and pseudogenes . AlthougLong non-coding RNAs have been implicated in the regulation of epigenetic changes through recruiting and guiding chromatin remodeling complexes to specific genomic loci both in cis and in trans to regulate transcription . The chrFor example, one of the first lncRNAs to be characterized was the mammalian cis-acting XIST which mediates the silencing of one X chromosome in females during development. XIST induces the formation of repressive chromatin and the recruitment of PRC proteins (PRC2) to completely inactivate one of the two X chromosomes by dosage compensation during the early embryonic development of females. Interestingly, other lncRNAs have been shown to interact with and regulate the expression of XIST, with the most prominent being its own natural antisense lncRNA (TSIX) ,110,111.Long non-coding RNAs can also migrate from their site of transcription and regulate the expression of genes in trans either distally located on the same chromosomes or on different chromosomes. Such a lncRNA is the well-studied antisense intergenic lncRNA HOTAIR which is transcribed from the HOXC locus . HOTAIR Long non-coding RNAs have also been associated with the regulation of the monoallelic expression (only one of the two copies of a gene is expressed) of genes according to their parents of origin, also known as genomic imprinting. Imprinted control regions may be associated with the expression of lncRNAs, such as the paternally expressed Airn lncRNA, also known as IGF2R-AS1, which was shown to silence the maternally expressed genes Igf2r/Slc22a2/Slc22a3 . The impThe process of transcription and the associated RNA processing and organization of the nuclear architecture, can also be regulated by lncRNA transcripts. The lncRNAs have been shown to regulate transcription factors and the RNAPII transcription machinery, leading to an increase or suppression of transcription, as well as mRNA processing mechanisms including splicing, capping, and editing . For exaSeveral lncRNAs are transported from the nucleus to the cytoplasm where they are thought to be involved in regulating mRNA stability and translation. As an example, the ubiquitin carboxy-terminal hydrolase L1 antisense RNA 1 (Uchl1-as1) exhibits positive regulation of translation of the Uchl1 protein through the embedded inverted SINEB2 element, although its exact mechanism of action is unclear ,122. In Although our knowledge of how miRNAs regulate disease, including IPF, is well established ; our knoA microarray study by Cao et al.  using thA subsequent study using the same model focused on two differentially expressed lncRNAs, MRAK088388 and MRAK081523, reporting that these could regulate expression of protein coding genes, N4bp2 (Nedd4 binding protein 2) and Plxna4 (plexin-A4), by acting as ceRNAs (sponges) for miR-29b-3p and let-7i-5p. In situ hybridization also confirmed their expression in the cytoplasm of interstitial lung cells .Huang et al.  identifiMore recently, sequencing and microarray analysis of mouse fibrotic lungs identified 513 upregulated and 204 downregulated lncRNAs, of which uc.77 and 2700086A05Rik were found to mediate changes in EMT when overexpressed in human epithelial cells .Wu et al., has demonstrated that silica-induced pulmonary fibrosis can be inhibited by miR-489 in mice. In addition, miR-489 was shown to suppress fibroblast differentiation and inflammation by targeting Smad3 and MyD88, respectively. Interestingly, the upregulation of the lncRNA CHRF (cardiac hypertrophy-related factor) was found to reverse the inhibitory effects of miR-489 in mice, as well as in macrophage and fibroblast cell lines. This suggests that CHRF can play a role in the regulation of miR-489 and the activation of the inflammation and fibrotic signaling pathways .The expression of lncRNA H19 has also been implicated in the development of pulmonary fibrosis in a study by Tang et al.  who founMoreover, the elevated expression of lncRNA AP003419.16 and its adjacent protein-coding gene RPS6KB2  in the peripheral blood of IPF patients has been linked with the increased risk of developing age-associated IPF .Interestingly, the expression of lncRNA NONMMUT065582, also known as pulmonary fibrosis-associated RNA (PFAR), was found to be elevated in the fibrotic lungs of mice as well as mouse fibroblasts . The Lncl-1β-activated human control and IPF lung fibroblasts. Knockdown of IL7AS demonstrated a negative regulatory activity of this lncRNA in IL-6 mRNA expression and protein release (l-8. Both lncRNAs were found to be regulated by the NF-κB pathway [More recently, expression of IL7AS and MIR3142GHG was also shown to be significantly upregulated in I release , whereas pathway .LINC01140 and LINC0090 were also found to be upregulated in isolated IPF fibroblasts, and LINC01140 was also upregulated in IPF biopsies and was shown to be a negative regulator of the inflammatory response . KnockdoAlthough IPF research has made impressive progress in the last decade, it is now evident that understanding the underlying mechanisms of IPF is much more challenging than originally thought. IPF is still an almost invariably lethal disease and a deeper understanding of its pathogenesis is essential for the development of novel and more effective therapies. LncRNAs present a relatively new group of molecules that have been found to regulate gene expression; however, little is known regarding their role in IPF. As a consequence of their cell and tissue-specific expression, the identification of lncRNAs that drive cellular activities within the IPF lungs could present a great opportunity for the development of novel treatment strategies. Several oligonucleotide antisense therapeutics have already proven successful in treating human disease and are being administered to patients, while nucleic acid-targeting drugs have demonstrated great potential in targeting lncRNAs in cancer . Therefo


The corresponding functionalization zones carry bromobenzene-, deutero-, and chloro-addends. We employ statistical Raman spectroscopy together with scanning electron microscopy/energy dispersive X-ray spectroscopy for an unambiguous characterization. The exquisitely ordered nanoarchitectures of these covalently multi-patterned graphene sheets are clearly visualized.Structured covalent two-dimensional patterning of graphene with different chemical functionalities constitutes a major challenge in nanotechnology. At the same time, it opens enormous opportunities towards tailoring of physical and chemical properties with limitless combinations of spatially defined surface functionalities. However, such highly integrated carbon-based architectures (graphene embroidery) are so far elusive. Here, we report a practical realization of molecular graphene embroidery by generating regular multiply functionalized patterns consisting of concentric regions of covalent addend binding. These spatially resolved hetero-architectures are generated by repetitive electron-beam lithography/reduction/covalent-binding sequences starting with polymethyl methacrylate covered graphene deposited on a Si/SiO Covalently 2D-patterning graphene with different chemical functionalities is an attractive way to tailor its physical and chemical properties. Here, the authors realize spatially defined 2D-hetereoarchitectures of graphene via a strategy of molecular embroidering. Key to this endeavor is providing facile and flexible synthetic approaches for 2D-patterning of graphene. So far, shape control over graphene nanostructures has been targeted either by top-down etching10 or by bottom-up synthesis of nanoribbons using low-molecular precursors on surfaces15. Covalent graphene chemistry, in principle, provides another opportunity by addend binding to defined lattice regions. This approach also has the advantage that, next to the spatial structuring of graphene, chemical surface functionalities can be introduced which are located in the proximity of the conductive regions of intact graphene. To date, a few first examples of covalently 2D-patterned graphene have been prepared by a combination of wet-chemical and classical patterning techniques25. However, all covalent patterning protocols established so far are limited to one-step chemical patterning of graphene only. For the realization of more complex 2D-architectures involving the regular arrangement of addend regions with different surface functionalities, a detailed understanding of covalent reaction mechanisms and concepts such as surface activations are required. In the macroscopic world, such a 2D-engineering approach finds an analogy in embroidery. This is a long-existing decoration technique, in which a needle is used as a tool for applying patterns onto fabric and other materials. Following a similar manner, once the chemical needle (as referred to graphene chemistry) was manipulated, the analogous chemical embroidery will provide access to structured graphene nanoarchitectures with a multifunctional array of addend zones next to patterns of intact graphene. Owing to its versatility and chemical tailorability, lithography-assisted chemical functionalization20 is assumed to be one important component for the realization of graphene embroidery. Very recently, we demonstrated a one-step patterned graphene functionalization protocol with diazonium salts involving lithography as a pattern defining tool24. We were able to fabricate well-defined nanostructures down to the submicrometer scale. Another pillar for this success was the realization of comparatively high degrees of functionalization in the addend-binding zones by using reductively activated graphene and allowing for substrate-mediated antaratopic additions.Multiply functionalized and hierarchically patterned graphene sheets with a spatially defined two-dimensional (2D)-architecture are still elusive. The realization of this challenge constitutes the next higher level of graphene chemistry. Such tailored nanosurfaces are highly attractive for high-performance applications, for example, in the fields of optoelectronics and sensors2 substrate. We employ statistical Raman spectroscopy (SRS) together with scanning electron microscopy/energy dispersive X-ray spectroscopy (SEM-EDS) for an unambiguous characterization which clearly shows that covalently functionalized, multi-patterned graphene exhibits an exquisitely ordered nanostructure.Here, we realize graphene embroidery by generating regular multiply functionalized patterns, consisting of concentric regions with covalent addend binding. These spatially resolved 2D-hetero-architectures are generated by repetitive electron-beam lithography (EBL)/reduction/covalent-binding sequences starting with polymethyl methacrylate (PMMA)-covered graphene, deposited on a Si/SiO24. The commercially available graphene monolayer was deposited on a Si/SiO2 wafer by a wet-transfer technique. Here, the PMMA-supported graphene, floating on top of a water surface, was fished onto the prepared Si/SiO2 wafer and subsequently, the initial PMMA coating was removed by acetone and a fresh PMMA double-layer mask was applied by spin-coating. The usage of a reactive Si/SiO2 substrate — reactivity provided by the presence of accessible surface atoms  — is a fundamental prerequisite as it allows for a strain-free antaratopic graphene addition scenario, which is in sharp contrast to supratopic addition reactions that would generate enormous strain in the graphene lattice29. By this technique, a comparatively high degree of functionalization can be established.As a prototype for the sequential multiple functionalization of graphene we developed the generation of three concentric zones carrying bromobenzene-, deutero-, and chloro-addends Fig. . These ta) are generated that are available for the first addend-binding process. The graphene in the PMMA–free zone Ia was activated by a treatment with a Na/K alloy. Briefly, a drop of liquid Na/K alloy was dropped onto the graphene monolayer27. As a result, graphene was negatively charged, which increases the reactivity towards electrophiles. As soon as the Na/K alloy was removed by a gentle stream of argon, the efficient addend binding was accomplished by a reaction with 4-bromobenzene diazonium-tetrafluoroborate31. As a consequence, an array of the first dot-shaped addend zone Ib was generated. The second concentric addend zones were generated following the analog concept. For this purpose, a second lithographic procedure was applied expanding the diameter of the concentric dots to 15 µm (zone IIa). After activation and exposure to deuteroxide, an efficient deuteration of the second patterning zone (zone IIb) was accomplished. This represents the first example for the generation of a deuterated monolayer graphene and remarkably, the achieved degrees of functionalization are comparable to that of deuterated bulk graphite and significantly exceed the numbers reported for deuterated multi-layer graphene obtained by alternative functionalization approaches33. A third diameter expansion of the periodic dots to 20 µm (zone IIIa) allows for the final concentric graphene patterning. Activation with Na/K and subsequent quenching with iodine monochloride (ICl) gave rise to a mild chlorination  generating the third addend zone (zone IIIb). Strikingly, ICl was developed as a facile and mild chlorination source for graphene chlorination. The last step of the entire patterning procedure was the removal of the protective PMMA layer by a treatment with acetone. The final outcome was the first demonstration of a sequential multiple patterning of graphene with different addends bound to spatially defined addend zones. The concept can be considered as a molecular embroidery analogy to the traditional embroidery manufacturing, that is, employing spatially resolved graphene chemistry (needle) to embroider regularly patterned addend zones to graphene (fabric). With this pioneering concept various 2D-patterns beyond this concentric pattern can easily be accomplished by a tailor-made modification of the respective e-beam lithographic masks.The circular patterns in the PMMA mask (array of 40 × 40 circles) for the subsequent three reductive activation/functionalization sequences were generated via EBL. After each individual functionalization step, the complete PMMA mask (which becomes partly damaged by the addition of the electron-trapping reagent benzonitrile) was completely removed by acetone vapor and a fresh PMMA double-layer mask has been applied via spin-coating to enable the next EBL patterning cycle (Supplementary section S3. The bandgap depends on the amount of attached hydrogen/deuterium atoms. On the other hand chlorination causes p-doping36. By the attachment of differently substituted aromatic substituents (σ/π-donor or acceptor) a specific geometrically defined region of the graphene lattice can be addressed directly (here is aromatic acceptor) and fine-tuned with respect to its electrical properties. As a consequence, our constructed multiply patterned graphene with concentric patterns, where each patterned graphene nanodomain has distinct semi-conducting behavior (n-type and p-type), bears promising potential to form graphene p–n junctions. Furthermore, the properties of these type of chemically written p–n junctions may further be fine-tuned by the control of the degree of functionalization in each addend zone . Semiconductor p–n junctions are elementary building blocks for many electronic devices such as transistors, solar cells, photodetectors, and integrated circuits and the potential applications of our constructed graphene p–n junctions in these fields can be reasonably expected. Besides, these constructed p–n junctions are always combined with the electric conductivity of the surround intact graphene regions (zone V), whose geometry can be tailored on demand upon the strategy that we introduce here. In view of this, the blueprint for promising all-graphene nanoscale electronic circuits is laid. Moreover, considering the fact that some aromatic systems are sensitive to light, the 2D-architectures we constructed can write phenyl-based substituents to distinct areas on the graphene flake, resulting in harvesting light of different wavelengths (based on the type of attached chromophore) in different areas on the same electrical conducting substrate. Thus, it may also find applications in nanometer scale optoelectronics such as light sensor. Finally, our innovative approach of writing chemical information into spatially defined patterns may also become of great importance in a field of information storage based on distinct chemical molecules where more than only two states  of information can be stored.For our multiply patterned graphene, where the different functionalities including deuterium, chlorine, and bromobenzene were selectively bound to the graphene surface, the properties of the constructed 2D-hetero-architecture can be tailored. Regular patterns of hydrogen/deuteration-covered regions result in a confinement potential for the carriers in the pristine graphene regions, thus leading to a bandgap opening and characteristic for a n-type semiconductora  shows a pronounced G-band at 1582 cm−1 indicative for a sp2 carbon lattice along with a negligible defect-induced D-band at 1350 cm−1. This is the characteristic feature of an intact monolayer graphene. The absence of a D-band demonstrates that during the EBL process no defects in the graphene lattice were generated. After the reductive arylation a very intense D-band emerged, demonstrating the efficient conversion of sp2 into sp3 carbon atoms within the corresponding carbon lattice of zone Ib. The ID/IG (Raman D-band to G-band intensity ratio) ratio raised from <0.1 to 2.6 after arylation , which were covered by PMMA during the chemical treatment remains completely intact as no D-band was observed. Consequently, our graphene functionalization protocol selectively takes place only in the patterned region. To visualize this spatially resolved 2D-chemistry, large-scale Raman mapping was conducted and the corresponding pattern images can be clearly recognized through ID/IG mapping 39. In contrast, relative to the arylated graphene (zone Ib) deuteration causes n-doping of the remaining conjugated π-system within zone IIb as manifested by a downshift of the Raman G-band to 1567 cm−1 35. The large-scale Raman ID/IG mapping clearly reveals a turquoise pattern of the deuterated zone IIb concentrically arranged around zone Ib , zone IIb (C–D bond cleavage), and zone IIIb (C–Cl bond cleavage) correlate very well with the corresponding bond strengths (BE (C–Cl) = 328 kJ mol−1, BE (C–C) = 332 kJ mol−1, and BE (C–D) = 414 kJ mol−1)44.The thermal behavior of the deuterated zone IIity Fig. . Furtherure Fig. . The decb and IIIb. Therefore, for instance, upon heating to 250 °C, we can tune the ID/IG ratio of zone Ib and zone IIIb to around 1.8 and 1, respectively. The large-scale Raman mapping clearly displayed the pattern image with the corresponding decreased ID/IG ratios, respectively, encoded in yellow for zone Ib and green for zone IIIb .As mentioned above, the reversible defunctionalization process mainly occurs in the temperature range between 200 and 350 °C for zone IIIb Fig. . The 400cle Fig. . Taking b, IIb, and IIIb) exhibit different defunctionalization processes  and Cl (zone IIIb). As expected, SEM-EDS investigation of the Br distribution in zone Ib revealed periodic solid circular dot configurations correlating very well with the extension of zone Ib. When it comes to the element Cl, a distribution related to zone IIIb, namely, a periodic ring configuration, is observed. The SEM-EDS results strongly correlate with the Raman data outlined above. In addition, the Br and Cl atoms are quite homogeneously distributed with the corresponding addend zones Ib and IIIb, demonstrating a homogeneous addend coverage. Moreover, no further element signals are found, which indicates the efficiency of all washing and work-up procedures and underlines the covalent nature of the patterned regionsTo track the chemical nature of this multiply patterned graphene, we carried out SEM-EDS Fig. . Our pat2 substrate. The corresponding functionalization zones carry bromobenzene-, deutero-, and chloro-addends. The successful covalent patterning was unequivocally demonstrated by SRS and SEM-EDS. In addition, the constructed concentric hetero-architectures were very clearly visualized by large-scale Raman ID/IG mapping. Moreover, the different thermal stabilities for each addend zone were revealed by temperature-dependent Raman investigations and the reversibility of the covalent binding, as seen for the chloro- and bromobenzene-functionalities, provide a feasible means for property engineering . The construction principle of 2D-patterning that we have introduced here opens enormous opportunities towards tailoring the physical and chemical properties of graphene sheets with apparently limitless combinations of spatially defined surface functionalities. Such tailored nanosurfaces are highly attractive for high-performance applications, for example, in the fields of optoelectronics, sensors, and catalysis.In summary, we have realized the first prototype of multiply functionalized and hierarchically patterned graphene sheets with a spatially defined 2D-hetereo-architecture. The chemical embroidery was accomplished by repetitive EBL/reduction/antaratopic-covalent-binding sequences starting with PMMA-covered graphene deposited on a Si/SiOThe details on the multi-step covalent patterning and activation/functionalization of graphene and the involved chemicals are given in the Supplementary Information 2 wafers with a heating rate of 10 K min−1.The Raman spectroscopic characterization was performed on a Horiba Jobin Yvon LabRAM Aramis. The spectrometer was calibrated by using crystalline graphite. All measurements were conducted using a laser  with an excitation wavelength of 532 nm, with an acquisition time of 2 s. Spectral data were obtained through a motorized x-y table in a continuous line scan mode (SWIFT-module). The step sizes in the Raman mappings were kept in the range of 0.1–0.5 µm depending on the experiments. The corresponding data processing was performed using software of Lab Spec 5. The temperature-dependent Raman measurements were performed in a Linkam stage THMS 600, equipped with a liquid nitrogen pump MS94 for temperature stabilization under a constant flow of nitrogen. Nitrogen gas needs to be filled for 20 min before starting the measurements to remove air. The measurements were carried out on Si/SiOSEM-EDS was performed on a GeminiSEM 500 equipped with Oxford X-max 150. The working conditions were set at an operating at accelerating voltage of 5 kV, working distance of 7.3 mm, the elevation angle of detector is 35°, and the sample is vertical to the secondary-electron emission.Supplementary InformationPeer Review File


Here, we utilize seeding promoter and low pressure CVD process to enhance sequential WSe2 growth with a reduced growth temperature of 800 °C for reduced compositional fluctuations and high hetero-interface quality. Growth behavior of the sequential WSe2 growth at the edge of patterned graphene is discussed. With optimized growth conditions, high-quality interface of the laterally stitched WSe2-graphene is achieved and characterized with transmission electron microscopy (TEM). Device fabrication and electronic performances of the laterally stitched WSe2-graphene are presented.Nanoelectronics of two-dimensional (2D) materials and related applications are hindered with critical contact issues with the semiconducting monolayers. To solve these issues, a fundamental challenge is selective and controllable fabrication of p-type or ambipolar transistors with a low Schottky barrier. Most p-type transistors are demonstrated with tungsten selenides (WSe Monolayer van der Waals materials, such as graphene and transition metal dichalcogenide (TMD), exhibit excellent electronic performances and atomically thick body without dangling bonds on the surface, which offers potential solutions for fundamental limit of channel materials in Moore’s law, such as short channel effects and various challenges in the scaling , 2. In t2-graphene, enabling inverter and negative-AND (NAND) gates for a complete set of logic circuits based on 2D materials [2) but a high temperature is required for the WSe2 growth because of a higher evaporation temperature of the WO3 precursor [Recently, integration of conductive graphene and semiconducting TMD for improved contacts and novel properties is realized by direct growth of TMD using chemical vapor deposition at the edge of artificially patterned graphene –21. Heteaterials , 17. Theaterials . Most p-recursor –31. A lo2 growth with a reduced growth temperature for reduced compositional fluctuations and high hetero-interface quality [2 growth at the edge of patterned graphene is discussed. With optimized growth conditions, high-quality interface of the laterally stitched WSe2-graphene is achieved and studied with TEM. Device fabrication and electronic performances of the laterally stitched WSe2-graphene are presented.Here, we utilize seeding promoter and low pressure CVD process to enhance sequential WSe quality , 33. Gro2 films were synthesized on sapphire and SiO2/Si substrates in the furnace. Before growth process, the substrates were cleaned with acetone, isopropanol, and then water for 10 min, respectively. Perylene-3,4,9,10-tetracarboxylic acid tetrapotassium salt (PTAS) was uniformly coated on the substrate surface as seeding promoters to enhance the activity and growth rate of the monolayers. High purity solid precursors of WO3  and Se  were placed in two ceramic crucibles, and the substrates were placed face up and next to the WO3 powder. The WSe2 samples were synthesized during 800~900 °C for 10 min with a heating rate of 30 °C min−1 and under a mixture of N2/H2 flow at 1.2 Torr. Graphene is synthesized on Cu foil at 1000 °C for 10 min with a heating rate of 30 °C min−1 and under a mixture of CH4/H2 flow at 4 Torr. The pattern graphene is carried out by e-beam lithography and oxygen plasma etching.Large-area WSe2 devices were fabricated without sample transfer. E-beam lithography process was performed to define the electrodes on the patterned graphene layer. A thin metal layer of Pd (40 nm) was deposited using e-beam evaporation and a following lift-off process was carried out in acetone. Encapsulation layer and gate dielectric of the device are fabricated by using atomic layer deposition (ALD) of thin Al2O3 films (50 nm). A thin metal of Pd (40 nm) was deposited on the dielectric layer to use as the gate electrodes. To improve electronic performances, the devices are annealed at ~ 120 °C for ~ 12 h in a vacuum environment of ~ 10−5 Torr.The graphene-WSe2 nanosheets onto the holey-carbon Cu grid. The TEM images were performed at an accelerate voltage of 80 kV . The electrical measurements were measured using an Agilent B1500a Semiconductor Device Analyzer.Raman spectra and photoluminescence (PL) were obtained by commercial confocal Raman spectroscopy . Wavelength and spot size of the laser are 532 nm and 1–2 μm, respectively. Typical gratings were used with 300 g/mm for PL (low resolution) to get broadband spectrum and (high resolution) 1800 g/mm for Raman signals to get detail information of material. The TEM samples were prepared by using standard PMMA transfer technique to place the graphene-WSe2, sequential growth of the monolayer TMD at the graphene edges is demonstrated in Fig. 2 plasma etching processes are conducted to define the region for sequential growth of the monolayer WSe2. Direct synthesis of monolayer WSe2 at the edges of patterned graphene on sapphire substrate is achieved by low pressure CVD with PTAS as seeding promoters. More detailed information on the synthesis is described in the “2 displays a uniform contrast, which confirms a reduced damage of the pre-patterned graphene after the sequential CVD synthesis of the WSe2 growth. In Fig. 2 indicates a smooth surface morphology of the monolayer heterojunction. Figure 2g mode (WSe2—blue) and the G’ band (graphene—green) as the labels in Fig. 2 is shown in Fig. 2 at the edges of the pre-patterned graphene.To control the synthesis of the lateral heterojunction of graphene and WSe2 synthesis at the patterned graphene is carried out with and without promoters. Figure 2 growth at different temperatures without PTAS as seeding promoter. Above 850 °C, the sequential growth of the WSe2 appears at the graphene edges. A high growth temperature for WSe2 growth is required due to reduced gaseous reactants for the solid precursor of the WO3, as elaborated in previous papers [2 implies random distributed and small size of grains. In contrast, the sequential WSe2 growth at different temperatures with PTAS as seeding promoter is presented in Fig. 2 growth at the graphene edges with larger domain sizes, which is similar to the growth behavior in the TMD-TMD heterojunctions [2 growth at 800 °C, observation of a uniform contrast and higher intensity in Raman mapping of the G’ band (graphene) indicates a reduced damage of the graphene because of the low temperature growth. With increased temperature, a continuous WSe2 film fills in the patterned regions with ideal contact to the edges of the patterned graphene  measurement is performed. In Fig. 2 monolayer. Width of the overlapping region is about 500 nm. An amorphous-like TEM image for the graphene lattice is observed as expected because of unavoidable distortions of graphene with the energetic electron beam. Figure 2 (~ 3.3 Å) is consistent with the parameters in bulk lattices of graphene (2.46 Å) and WSe2 (3.28 Å). The TEM characterizations indicate that the sequential WSe2 growth initiates at the edges of the pre-patterned graphene because higher defect density at the graphene edge enhances the vertical island growth with more nucleation sites. A large lattice mismatch more than 20% between the lattice of graphene and TMD might be responsible for a disorder interface with higher defect density and for combined vertical and lateral TMD growth at the heterojunction. Moreover, the insets in Fig. 2 lattices implies that the sequential growth of the WSe2 favors coherent stacking at the graphene edges.To further investigate the heterojunction of the WSegure 2 ~ .3 Å is c2 stitched at the edges of patterned graphene hetero-device, the device is fabricated without sample transfer. Customized fabrication process based on surface functionality for e-beam lithography on an insulator is developed. Electronic transport performance of the stitched graphene-WSe2 device is studied by connecting metal electrodes (Pd 40 nm) with the patterned graphene and depositing Al2O3 (50 nm) as gate dielectric. Figure 4) and high on-current of approximately a few 100 nA  and W (24 μm) are the channel length and channel width, respectively. Moreover, the output curves of the device at various gate voltages are shown in Fig. I-V curves confirm a good contact between graphene layer and WSe2 layer. An enhanced electronic performance of the stitched TMD-graphene monolayer heterojunctions is achieved because of improved contact properties, suggesting the synthesis for sequential TMD growth at the edges of artificially patterned graphene moves a significant step towards 2D nanoelectronics.To demonstrate the field-effect properties of the as-grown WSerodes Pd 0 nm withrodes Pd 0 nm withrodes Pd 0 nm with2 growth at the edges of the patterned graphene is achieved on sapphire by using promoter-assisted LPCVD. The PTAS promoters significantly reduce growth temperature for ideal sequential WSe2 growth at the graphene edges with larger domain sizes.Sequential WSe2 growth initiates at the edges of the pre-patterned graphene. A highly reduced twisting angle between graphene and WSe2 lattices implies that the sequential WSe2 growth favors coherent stacking at the graphene edges. An enhanced electronic performance of the stitched TMD-graphene monolayer heterojunctions is achieved because of improved contact properties.The TEM characterizations indicate that the sequential WSeAdditional file 1: Figure S1. Scalable synthesis of monolayer WSe2 on sapphire: influence on (a) concentration of the seeding promoters and (b) temperature for the WSe2 growth. Figure S2. Synthesis of the laterally stitched graphene-WS2: (a) optical and (b)AFM image of the graphene-WS2 and (c) PL spectrum of the WS2. Raman mapping of (d) the E2g mode of WS2 and (e) the G’ mode of graphene. (f) PL mapping of the Graphene-WS2. .


APP) and presenilins (PSENs) could potentially lead to the production of longer amyloidogenic Aβ peptides. Amongst these, Aβ1–43 is more prone to aggregation and has higher toxic properties than the long-known Aβ1–42. However, a direct effect on Aβ1–43 in biomaterials of individuals carrying genetic mutations in the known AD genes is yet to be determined.Alzheimer’s disease (AD) mutations in amyloid precursor protein (N = 1431 AD patients (n = 280 early-onset (EO) and n = 1151 late-onset (LO) AD) and 809 control individuals were genetically screened for APP and PSENs. For the first time, Aβ1–43 levels were analysed in cerebrospinal fluid (CSF) of 38 individuals carrying pathogenic or unclear rare mutations or the common PSEN1 p.E318G variant and compared with Aβ1–42 and Aβ1–40 CSF levels.The soluble sAPPα and sAPPβ species were also measured for the first time in mutation carriers.PSEN1, 1.07% APP) and in 0.3% of LOAD patients. Furthermore, 12 known variants with unclear pathogenicity and 11 novel were identified. Pathogenic and unclear mutation carriers showed a significant reduction in CSF Aβ1–43 levels compared to controls . CSF Aβ1–43 levels positively correlated with CSF Aβ1–42 in both pathogenic and unclear carriers and controls . The p.E318G carriers showed reduced Aβ1–43 levels (p < 0.001), though genetic association with AD was not detected. sAPPα and sAPPβ CSF levels were significantly reduced in the group of unclear  and p.E318G carriers , suggesting their possible involvement in AD. Finally, using Aβ1–43 and Aβ1–42 levels, we could re-classify as “likely pathogenic” 3 of the unclear mutations.A known pathogenic mutation was identified in 5.7% of EOAD patients (4.6% 1–43 levels were analysed in CSF of AD patients with genetic mutations in the AD causal genes. The observed reduction of Aβ1–43 in APP and PSENs carriers highlights the pathogenic role of longer Aβ peptides in AD pathogenesis. Alterations in Aβ1–43 could prove useful in understanding the pathogenicity of unclear APP and PSENs variants, a critical step towards a more efficient genetic counselling.This is the first time that Aβ APP), presenilin 1 (PSEN1) and presenilin 2 (PSEN2) have been identified as a cause of both EOAD and LOAD  women, average age at onset (AAO) 73.51 ± 9.81 years, range 29–96 years), ascertained in Belgium through the neurology centres of the clinical partners of the Belgian neurology (BELNEU) consortium. All patients received a diagnosis of possible, probable or definite AD according to the criteria described by the National Institute of Neurological and Communication Disorders and Stroke Alzheimer’s Disease and Related Disorders Association (NINCDS-ADRDA) criteria and the National Institute on Aging–Alzheimer’s Association (NIA-AA) [n = 318) of the patients. Based on the onset age (< 65 years), 19.57% (n = 280) of the patients were classified as having early-onset AD . A Belgian control cohort of 809 individuals  70.12 ± 10.71, 31–96 years) was included in the study. The control individuals were primarily community-dwelling volunteers or spouses of patients. Subjective memory complaints, neurologic or psychiatric antecedents, and a familial history of neurodegeneration were ruled out by means of an interview. Cognitive screening was performed using the Mini-Mental State Examination (cut-off score > 26) [ClinicalTrials.gov identifiers: 54861911ALZ1005 (NCT01978548); 54861911ALZ2002 (NCT02260674)) by Janssen Pharmaceutica NV, Beerse, Belgium [The study population consisted of 1431 AD patients (62.03% [(NIA-AA) . A positre > 26) . The spo Belgium . SubjectAPP, PSEN1 and PSEN2 using a targeted re-sequencing gene-panel as we previously described [PSEN1 rs17125721 A > G, p.E318G variant , considered a possible risk modifier  and Apolipoprotein E (APOE) genotypes were analysed. Variants with a read depth below 20X, with Genoqual value below 99 and with an imbalanced wild-type/variant read depth (cut-off > 3) were considered false positives. All the selected variants were validated by Sanger sequencing (BigDye Terminator Cycle sequencing kit v3.1) on the ABI 3730 DNA Analyser (both Applied Biosystems). Sequences were analysed using SeqManII or novoSNP software packages [APP locus duplications, multiplex amplicon quantification (MAQ) was used on 280 EOAD patients, as previously described [www.vibgeneticservicefacility.be). One sample carrying the APP duplication and four different control samples were used as positive and negative controls, respectively.Genomic DNA of patients and control individuals was analysed for mutations in escribed . We seleescribed . In addipackages . To screPSEN1 p.E318G variant  as well as 64 control individuals . Lumbar puncture, CSF sampling and handling have been performed according to standard protocols [1–42 and Aβ1–40 were measured using enzyme-linked immunosorbent assay (ELISA) with commercially available single parameter ELISA kits at the BIODEM laboratory as previously described [1–43 levels were measured using the amyloid beta (1–43) (FL) Aβ kit (IBL) and both sAPPα and sAPPβ using sAPPα/sAPPβ kit . Concerning T-tau and P-tau181 levels, these were previously measured using hTAU Ag and PHOSPHO-TAU(181P) (INNOTEST), respectively, and the BIODEM laboratory performed the ELISA for the 38 mutation carriers and the Sahlgrenska University Hospital (Sweden) for the 64 control individuals. For this reason, we did not perform statistical analysis on either T-tau or P-tau181 amongst the groups, but we only reported the values of the measurements, which are part of the phenotypic characterization of the cohort. All calibration standards and CSF samples were analysed in duplicate. Only mean values with a coefficient of variation (CV) of the replicates less than or equal to 20% were included in the analysis.CSF were available for a subset of 38 mutation carriers: 18 carriers of pathogenic and VUS mutations (MAF < 1%)  and 20 carriers of rotocols . SamplesPSEN1 p.E318G and APOE ε4 genotype. With a similar model, we further tested the prevalence of APOE ε4 between EOAD and LOAD cohorts and of APOE ε4ε4 between EOAD and LOAD cohorts.Statistical analyses were performed in SPSS version 24, and graphs were made using GraphPad Prism8. A logistic regression analysis was performed in the AD and control cohorts, to determine whether patients had higher probability than controls to carry both the common PSEN1 p.E318G. First, a Kolmogorov-Smirnov test was performed to check for normal distribution. Since most variables did not follow a normal distribution, non-parametric tests were used. Differences amongst groups were tested using Kruskal-Wallis. Post hoc analysis with pairwise comparisons was carried out . To identify whether an association between two markers was present, the Spearman’s rho correlation tests were performed. Correlation coefficients were extracted from each group  separately. After correction for multiple testing (Bonferroni’s correction), p values < 0.005 or below were considered to be statistically significant. Receiver operating characteristic (ROC) curves were additionally performed to assess the diagnostic accuracy of the individual biomarkers, and the area under each ROC curve (AUC) was calculated. Finally, explanatory cut-offs were identified as the concentration of specific biomarker that maximizes sensitivity and specificity of the test (Youden’s index).For the CSF biomarker analysis, the mutation carriers were divided in three different groups: carriers of known pathogenic mutations, of VUS and of 6 lymphoblast cells of the double mutation carrier , of the single carrier  and of 4 non-carriers as negative controls were incubated with 150 mg/mL cycloheximide (CHX) (Sigma-Aldrich) at 37 °C for 4 h. To generate cDNA, total RNA was extracted using the RiboPureTM kit (Life Technologies) followed by a DNase treatment . First-strand complementary DNA was synthesized using the SuperScript III First-Strand Synthesis System (Life Technologies) using random hexamer primers. Full-length PSEN1 transcript sequencing was subsequently performed on a MinION sequencing platform (Oxford Nanopore Technologies (ONT)) based on a modernized cDNA sequencing protocol [PSEN1 full transcript was carried out with Platinum® Taq DNA Polymerase (Clontech Laboratories) using exonic primers, designed with Primer3Plus [PSEN1 spanning sequencing reads were retained for further analysis. Relative quantifications of splice junctions were calculated by dividing the number of junction-supporting reads by the total number of reads spanning the PSEN1 transcript in R .To inhibit non-sense-mediated mRNA decay (NMD), 1 × 10protocol . Brieflymer3Plus , with a mer3Plus , with spAPP, 10 in PSEN1)  and were identified in 19 patients  and 1 control individual . Specifically, 1.07% (3/280) of EOAD patients carried known pathogenic mutations in APP and 4.6% (13/280) in PSEN1. Furthermore, in 0.086% (1/1151) and 0.26% (3/1151) of LOAD patients, known pathogenic mutations were identified in APP and PSEN1, respectively.The genetic screening identified a total of 41 mutations in 54 individuals . The identified variants are listed in Table S1 . FourteeAPP, 2 PSEN1, 5 PSEN2) were detected in 12 patients  and 7 controls . Three were known benign variants  found in 7 patients  and 2 controls . Lastly, 11 rare variants were novel  absent from gnomAD and mutation databases, observed in 8 patients  and 3 control individuals . There was no significant difference between the prevalence of APOE ε4 in EOAD (57%) and LOAD (55%) (p = 0.48). The prevalence of APOE ε4 was higher in EOAD (60%) than LOAD (55%) only on individuals without any APP or PSENs mutations, but this difference was still not significant (p = 0.15). We found a significant difference in the prevalence of APOE ε4ε4 between EOAD (20%) and LOAD (9%) when considering all individuals (p < 0.001). This difference in the prevalence of APOE ε4ε4  was similar when running the same model only on individuals without any APP or PSENs mutations (p < 0.001).In addition, 12 previously reported variants with unclear pathogenicity  and 24 controls . No significant association was found between PSEN1 p.E318G and AD, regardless of APOE ε4 genotype . Furthermore, in the screened cohort, APP locus duplications were not detected.Table S1 . AmongstPSEN1 p.E318G to control individuals. A significant reduction of CSF Aβ1–43 levels was detected in all three carrier groups .The CSF biomarkers were analysed comparing three groups of mutation carriers: 1) known pathogenic, 2) VUS and 3)  known pa VUS and   known p1–43/Aβ1–40 and Aβ1–42/Aβ1–40 ratios were significantly reduced in all three carrier groups  . Aβ1–43 and Aβ1–42 were significantly correlated with Aβ1–40 in the controls and PSEN1 p.E318G group , while there was a correlation between Aβ1–42 with Aβ1–40 in the VUS group, but not significant after Bonferroni’s correction . Both CSF Aβ1–43 and Aβ1–42 were able to differentiate the corresponding controls from the group of known pathogenic and PSEN1 p.E318G  and PSEN1 p.E318G carriers (both p < 0.04) . Aβ1–40 correlated with sAPPα only in the control group  and with sAPPβ in the control  and VUS groups . Aβ1–43/sAPPα and Aβ1–43/sAPPβ CSF levels were significantly lower in PSEN1 p.E318G carriers compared to controls (p < 0.001)   are summarised in Table Aβy) Table . CSF levPSEN1 p.G183V and PSEN1 p.P49L double mutation on PSEN1 alternative transcript generation in patient-derived RNA was examined with ONT MinION sequencing, because of a previously reported effect of this mutation on exon skipping in both HEK293 cells [PSEN1 p.G183V, PSEN1 p.P49L) and the single PSEN1 p.G183V carrier show the skipping of exon 6 in 20% of the sequencing reads from the cDNA obtained from the lymphoblast cells treated with CHX and 5% from the untreated .The effect of the 93 cells  and mice93 cells . Both thted Fig. . The anaAPP, PSEN1 and PSEN2 mutation screening in 1431 AD patients and 809 control individuals identified known pathogenic mutations in 5.7% of EOAD (16/280) and 0.3% of LOAD (4/1151) patients and in 0.12% (1/809) of the controls, in line with what it is usually observed [APP p.A713T mutation considered with unclear pathogenicity but actually showing segregation with the disease in another study [APP mutation is known to have a wide onset age range; therefore, it is not surprising to identify asymptomatic carriers [PSEN1 p.C263F, identified in five patients with onset age range of 53–70, and the PSEN1 p.G183V, previously identified in a patient with frontotemporal dementia and Pick-type tauopathy [APP p.G625_S628del and the PSEN1 p.P355S, both VUS, and patient 16 having the PSEN1 p.G183V and the novel p.P49L.The er study  was asymer study . This APcarriers , 44. Amoauopathy   and Pick-type tauopathy [PSEN1 p.G183V mutation. The PSEN1 p.G183V is located in the last nucleotide of the exonic splice donor site of exon 6, and experiments in HEK293 cells and mice showed the formation of alternative transcripts with skipping of exons 6 and exon 6–7, which are likely degraded by the non-sense-mediated mRNA decay control mechanism (NMD) [PSEN1 p.G183V led to the formation of transcripts lacking exons 6 and 6–7, which are indeed degraded by NMD. Only a small amount of these alternative transcripts remains in the cells (≤ 5%) and they unlikely interfere with the wild-type PSEN1. The presence of the second PSEN1 mutation (p.P49L) could drive the disease in patient 16.Patient 16 received a diagnosis of probable AD (age at onset > 65). Clinically, patient 16 showed amnestic presentation without other remarkable signs or symptoms. Single-photon emission computed tomography (SPECT) displayed a moderate hypoperfusion of the bilateral parietal, temporal and frontal lobe, compatible with AD. Magnetic resonance imaging (MRI) showed age-related atrophy and multiple supratentorial lacunary infarcts. Neuropathological examination showed a mild atrophy of the frontotemporal gyri  , 43. OurAPP, PSEN1 and PSEN2 mutation carriers, both known pathogenic and VUS, was available and used to investigate for the first time the CSF Aβ1–43 levels. A significant reduction of CSF Aβ1–43 levels was observed in carriers of pathogenic mutations (13/18), of VUS (9/13) and of carriers of PSEN1 p.E318G (18/20) compared to controls. This reduction was comparable with CSF Aβ1–42 levels. CSF Aβ1–43 levels positively correlated with Aβ1–42 in all mutation carrier groups but correlated with Aβ1–40 only in PSEN1 p.E318G mutation carriers and controls. Aβ1–43/Aβ1–40 ratio was significantly reduced in all three mutation carrier groups, while Aβ1–43/Aβ1–42 ratio only in the VUS and PSEN1 p.E318G groups, as previously shown for AD patients (independently from the mutation status) [CSF of 38 , of VUS /13 and o1–42 and Aβ1–43) promote aggregation and neurotoxicity, Aβ1–40 appears to act protectively [1–40 levels did not differ significantly between AD patients and controls in a meta-analysis [1–43 with Aβ1–42, but not with Aβ1–40 (except in PSEN1 p.E318G carriers), further supports the involvement of Aβ1–43 in AD. It is however still unclear why Aβ1–40 would be unaltered and lacking of correlation with Aβ1–43 in the mutation carrier groups, as it is related to the process of Aβ1–43. It is plausible that a possible decrease of Aβ1–40 in CSF would only be visible or measurable during the earlier stages of the disease when mostly all neurons are still intact, rather than in later stages , when Aβ production is higher but less neurons are preserved. Alternatively, it is possible that the cleavage of the C99 fragment by γ-secretase may be heavily impaired at the third cleavage step when Aβ1–40 is generated from Aβ1–43, as suggested by Kakuda et al. [PSEN1 p.R278I, which would result in negligible levels of Aβ1–40 and unusually high levels of Aβ1–43. Furthermore, a recent study found a correlation between Aβ peptide length and plaque load (Aβ1–43 > Aβ1–42 > Aβ1–40), indicating that longer Aβ peptides have an increased tendency towards accumulation in the brain [1–41, Aβ1–37 (from Aβ1–43) Aβ1–38 (from Aβ1–42) and Aβ1–34 (from either Aβ1–43 or Aβ1–42) are however needed to better clarify the role of longer Aβ peptides in AD and the pathogenic events linked to Aβ process, degradation and clearance in the presence of APP and PSENs mutations [While longer Aβ peptides , including Aβ1–43, in the pathogenesis of AD is receiving increased attention [PSEN1 p.L435F produces primarily Aβ1–43, which was detected in the brain plaques [APP mutations affect the endopeptidase activity of γ-secretase, leading to Aβ1–42 formation, while PSEN1 mutations inhibit its carboxypeptidase activity, releasing multiple longer peptides including Aβ1–42 and Aβ1–43 [The production of longer Aβ peptides  could provide a more comprehensive interpretation of our results. Lastly, in our study, it was not possible to assess the difference between PSEN1 and APP mutations in affecting γ-secretase and Aβ generation [A potential limitation of this study is the small cohort size used to analyse Aβ 2 years , 25, 48.1–43 were investigated for the first time in carriers of known pathogenic and unclear (VUS) APP and PSENs mutations. We observed a significant reduction of CSF Aβ1–43 levels and a positive correlation with Aβ1–42 in all mutation carrier groups. We suggested the re-classification of three VUS into “likely pathogenic”, as their biomarker levels were comparable to the known pathogenic mutations. We added important information on the debatable genetic modifier PSEN1 p.E318G and we were able to clarify the role of PSEN1 p.G183V, using ONT long-read sequencing, considered so far pathogenic, but probably not involved in AD.In this study, CSF levels of the long Aβ peptide Aβ1–43 load in the brain. For these reasons, Aβ1–43 could be considered an added AD biomarker together with the others already in use. Despite the small study cohort, the data presented here corroborate previous findings on Aβ1–43 [From a clinical perspective, our data could prove useful. A recent study showed that Aβ43 was cleared more than Aβ42 in plaques of patients treated with Aβ immunotherapy . These an Aβ1–43 , 29, 49,CSF levels of sAPPα and sAPPβ were also analysed for the first time in AD mutation carriers, where their levels were reduced compared to controls. Based on the recent analysis on sAPP fragments as regulators of (GABA) neurotransmission, further investigation in AD is important to study more in detail this pathway to open new venues for therapy strategies.Finally, functional work using patient biomaterials can prove valuable to better understand the pathogenicity of unclear AD mutations. This will be useful for patient stratification for clinical trials, genetic counselling and therapy development.Additional file 1: Table S1. List of the identified APP, PSEN1 and PSEN2 mutations. Table S2. Diagnostic accuracy of the different markers and ratios to discriminate between controls and mutation carriers, measured by ROC curve analysis. Table S3. Descriptive features of PSEN1 p.E318G mutation carriers with CSF. Figure S1. Neuropathology of Patient 16. Right lateral (A) and right medial (B) hemispheres showing brain atrophy. Atrophy can be observed also at the ventricles and thalamus (C). The 4G8 staining shows amyloid plaques in the hippocampus CA4 (D). Classic neurofibrillary tangles are present in the hippocampal CA4 (AT8 stain) (E). Figure S2. Area under the curve (AUC) calculated for the three mutation carrier groups compared to controls of Aβ1-43, Aβ1-43/Aβ1-40, Aβ1-42, Aβ1-42/Aβ1-40, Aβ1-40 and Aβ1-43/Aβ1-42. AUC are calculated for the know pathogenic (red), VUS (orange) and PSEN1 p.E318G (green) mutation carrier groups compared to the control group. The AUC values and the ones for sensitivity and specificity are listed in Table S2. Figure S3. Area under the curve (AUC) calculated for the three mutation carrier groups compared to controls of sAPPα, sAPPβ, Aβ1-43/sAPPα and Aβ1-43/sAPPβ. AUC are calculated the know pathogenic (red), VUS (orange) and PSEN1 p.E318G (green) mutation carrier groups compared to the control group. The AUC values and the ones for sensitivity and specificity are listed in Table S2. Figure S4. Transcript analysis of PSEN1 in Patient 16. The bar graph shows the relative quantifications of exon 6-7 in the double carrier , the single carrier , and 4 non-carriers lymphoblast cells CHX treated (CHX) and untreated (UNT). Relative quantifications of splice junctions were calculated by dividing the number of junction-supporting reads by the total number of reads spanning the PSEN1 transcript. The quantifications for both CHX and UNT of the non-carriers are reported as averages .


The government power of actors and their influence on changing policies could result in changes in health outcomes as well as health indicators. Health and wellness are more influenced by the places in which people live, learn, work, and play. Building a safe community environment depends on the governance power of actors. Purpose: The aim of this study is to have an overview of the key actors of the safe communities and compare the governance power of them across a sample of designated safe communities.The present study is a combination of a qualitative study with a document review method and a comparative review study. Initially, upstream documents, reports on stakeholders’ participation in the creation of safe communities were reviewed. The data were analyzed through content analysis. Then through a comparative study, the organizational structures, key actors, and government power of them were compared in a sample of designated safe communities.Collaborative and participatory involvement is an essential factor for the creation of safe communities, which has been emphasized by the upstream and international documents. More convergence of the policies and programs across the spectrum of actors  is needed. The governance power of actors will be flourished by creating a systematic and structured network that will lead to the creation of safe communities.All actors must participate in the creation of a safe community and use their governance power for this purpose. But the evidence indicates that the most important actors, people, is being ignored. It is imperative that authorities have adequate planning to engage in systematic and continuous community participation by advocating for establishing the not for profit organizations and charities to build and maintain networks for safe communities.Governance power, Safe community, Key actors, Community participation


Sarcoidosis is a chronic idiopathic granulomatous inflammatory disease that can affect many major organ systems, primarily the lungs, and hence has remarkable clinical heterogeneity. At least 50% of patients with systemic sarcoidosis develop inflammatory eye disease, and in approximately 21% of cases, it may be the first clinical manifestation. Neuro-ocular involvement occurs in <3% of all sarcoidosis cases, and rarely involves the optic nerve. We describe an unusual case of an intraocular sarcoidosis presenting as an unclear optic nerve mass.A 61-year-old male presented with painful gradual visual loss in the right eye. Previous history included Stage II Hodgkin lymphoma (HL) and concurrent mediastinal sarcoidosis, both in remission 5 years later. On examination, the right eye (RE) vision had no light perception, neovascular glaucoma, attenuated retinal vessels and a non-pigmented optic disc mass. The left eye was normal. The RE showed no response to oral steroids, was painful due to neovascular glaucoma and the concerns of recurrent HL with intraocular manifestations lead to RE enucleation. Macroscopic examination revealed a whitish mass at the optic disc, which histomorphologically showed a non-necrotising granulomatous inflammation consuming the optic nerve head and extending into the optic nerve resection margin. Special stains for microorganisms were negative. The uveal tract was free of inflammation. The morphological features were consistent with optic nerve sarcoidosis. A diagnosis of neuro-ocular sarcoidosis was made, and the patient was commenced on infliximab.Neuro-ocular sarcoidosis is known as the ‘great imitator’ because it can cause a variety of non-specific clinical signs and symptoms, mimicking many other conditions, including lymphomas. Intraocular sarcoidosis is not unusual and typically affects the uvea. Isolated optic nerve sarcoidosis is rare. The challenging aspect of intraocular sarcoidosis is the requirement of prompt treatment to reverse any eye damage and prevent permanent visual loss. Here, optic nerve sarcoidosis was very advanced, and was associated with intracerebral manifestations.Neuro-ocular sarcoidosis is a difficult condition to diagnose and treat. Our case was complicated by the previous history of HL and concurrent mediastinal sarcoidosis which were in remission. In patients with a history of sarcoidosis with new loss of vision and neurological weaknesses oculocerebral involvement must be included in the differential diagnosis even in the absence of typical manifestations of ocular sarcoidosis as in uveal tract involvement. •Optic nerve head (ONH) sarcoidosis without uveal tract involvement is rare.•ONH sarcoidosis can be the first manifestation of systemic sarcoidosis.•ONH sarcoidosis may be the first manifestation of neuro-ocular sarcoidosis.•ONH masses present diagnostic challenges due to the impact of tissue biopsy.•‘Sarcoid-like’ reaction or sarcoid-lymphoma syndrome may occur with concurrent lymphoma. Sarcoidosis can affect many major organs although it predominantly affects the pulmonary system.Herein we describe an unusual case of an isolated ONH tumour presenting as the first manifestation of recurrent systemic sarcoidosis and as the initial manifestation of neuro-ocular sarcoidosis in a male patient with a background history of Hodgkin lymphoma (HL).1.1A 61-year old man was referred to the Liverpool Ocular Oncology Centre (LOOC) in 2018 with a right optic nerve head (ONH) mass. He had a one year history of monocular visual loss and increasing pain in the right eye that had not responded to a four-week course of oral corticosteroids. His previous ophthalmic and general medical history included: Stage II HL involving the left cervical nodes diagnosed in 2013, treated with four cycles of chemotherapy  and field radiotherapy; mediastinal sarcoidosis suspected on an interim positron emission tomography scan during chemotherapy, and confirmed by mediastinal lymph node biopsy in 2014; as well as diabetes mellitus and systemic hypertension.The patient's medication at the time of LOOC consultation included latanoprost, iopidine, atropine drops to the right eye; amlodipine; ramipril and aspirin. On ophthalmological examination, vision in the left eye (LE) was 6/6 aided; anterior segment examination was normal with an intraocular pressure (IOP) of 19 mmHg. There was no light perception (NLP) in the right eye (RE), mild conjunctival injection, a clear cornea with a deep and ‘quiet’ anterior chamber, rubeosis iridis and anterior synechiae. His right IOP was 41 mmHg. The right anterior vitreous was unremarkable. Fundoscopy of the RE revealed attenuated retinal vessels and a non-pigmented mass at the optic disc a. B-scanThe differential diagnoses included: circumscribed intraocular lymphoma, amelanotic melanoma or intraocular sarcoidosis. Given the finding of a steroid unresponsive blind and painful RE, the patient agreed to RE enucleation with implantation of an acrylic prosthesis. The specimen was sent in buffered formalin for histopathological examination.trans-illumination of the eye prior to opening. On dissection of the eye, there was a white-yellow mass measuring 3.3 × 4mm at the ONH.Macroscopic examination revealed an intact, right globe with dimensions of 24.5mm  x 23.3mm  x 23.5mm vertical. The cornea was clear and measured 12mm horizontally and 11.5mm vertically. The optic nerve was 4mm in length. There was no shadow noted on Histomorphological findings of the RE included ONH and nerve replacement by an extensive non-caseating granulomatous infiltrate . The latSpecial stains of the optic nerve lesion for acid fast bacilli (Ziehl-Neelsen) and fungi (Periodic Acid Schiff and Grocott-Gomori's methenamine silver) were negative. The CD68 immunostain highlighted the numerous histiocytes and giant cells . The morSimultaneous to the histomorphological investigation of the RE, a recurrence of mediastinal lymphadenopathy was observed on the patient's computed tomography of the chest. The differential diagnoses were recurrent HL or mediastinal sarcoidosis. The multi-disciplinary team concluded that the radiological features of widespread lymphadenopathy involving hilar and mediastinal regions plus axillary-, iliac- and inguinal nodes together with sub-central bilateral lung nodules were all in keeping with sarcoidosis. The ultimate histological diagnosis of the ONH ‘tumour’ in the RE supported the clinical diagnosis of recurrent systemic sarcoidosis.The patient was discharged to a Hospital Trust closer to his home residence, and joint management of the systemic sarcoidosis was continued by Haematological-, Respiratory Medicine and Neurological teams at this site. In 2019, unfortunately, the patient developed increasing right-sided lower limb weakness, imbalance and worsening tremulousness in the arms and head. Oral prednisolone 45mg was started for six weeks with some improvement in symptoms, but a subsequent referral to a tertiary neuro-sarcoidosis clinic for commencement of infliximab was organised.2Here we present a very unusual case of histomorphologically-confirmed ONH sarcoidosis in a patient with a background history of concurrent HL and mediastinal sarcoidosis, considered to be in remission. Neuro-ophthalmic sarcoidosis presenting initially as isolate optic nerve granulomata, without any involvement of the uveal tract, is rare. In addition, our case demonstrates the diagnostic uncertainties faced in determining recurrent sarcoidosis versus a recurrent lymphoma. The case exemplifies coexistent pathologies, termed sarcoid-like reactions or ‘sarcoid-lymphoma syndrome’, which are not well understood.,Sarcoidosis is a chronic idiopathic granulomatous inflammatory disease that can affect any major organ system, primarily the lungs, and hence has remarkable heterogeneity in clinical presentation, findings and natural history.,In approximately 21% of patients, ocular sarcoidosis may be the first clinical manifestation of the systemic disease.,Optic disc granulomata are very rare manifestations of sarcoidosis and occurs in less than 3% of patients with sarcoidosis.,Subsequent to Hickman's review of the literature, there are eight further cases of ONH granuloma secondary to sarcoidosis reported in the English literature.21,In our case, enucleation was performed due to a blind painful eye unresponsive to steroids. There was extensive infiltration of the optic nerve and involving surgical resection margin by non-caseating epithelioid granulomata, whereas the rest of the eye was ‘quiet’. In contrast to the histology reported in previous cases,,,,Neurosarcoidosis occurs in 5–10% of systemic sarcoidosis cases,,,,Since neuro-ophthalmic sarcoidosis is rare; there are no agreed treatment guidelines. Systemic corticosteroid is the first-line therapy, and results from different studies showing variable patient response.,,Finally, our case was of further interest due to the complexity in reaching a diagnosis due to the patient's background of concurrent cervical HL and mediastinal sarcoidosis. Confusing overlap exists between the clinical manifestations of HL and sarcoidosis leading to challenges in distinguishing the two conditions. Terms to describe the occurrence of HL and sarcoid concurrently or one after the other have been proposed. ‘Sarcoid-like’ reaction refers to the development of non-caseating granulomas in patients with a known malignancy .3Isolated optic nerve sarcoidosis is a rare finding, which may mimic other intra-ocular tumours and result in total loss of vision, and led to enucleation in our case.Biopsy is the most accurate way to confirm the diagnosis; however, biopsy of intraocular tissue or neural tissue is usually not performed due to the invasiveness and impact of optic nerve biopsy. In contrast to the few cases reported, we present findings of sarcoid-related granulomatous infiltrate confined to the ONH and absent in other ocular tissues. Our case also demonstrates the poorly understood phenomenon of sarcoid-lymphoma syndrome.Written consent for the anonymous use of clinical information and biosamples was obtained from the patient. This documentation can be provided on request.No funding or grant supportAll authors attest that they meet the current ICMJE criteria for Authorship.Yamini Krishna: Investigation, Writing, review & editing, Visualization. Louise Christou: Investigation, Writing, review & editing, Visualization. Jakub Khzouz: Visualization, Writing. Rumana Hussain: Patient care; Conceptualization, Investigation. Heinrich Heimann: Patient care; Conceptualization, Supervision. Sarah E. Coupland: Conceptualization; Writing, review & editing; Supervision.The following authors have no financial disclosures: YK, LC, JK, RH, HH, SEC.


Electromagnetic coils are a key component in a variety of systems and are widely used in many industries. Because their insulation usually fails suddenly and can have catastrophic effects, degradation monitoring of coil insulation systems plays a vital role in avoiding unexpected machine shutdown. The existing insulation degradation monitoring methods are based on assessing the change of coil high-frequency electrical parameter response, whereas the effects of the insulation failure mechanisms are not considered, which leads to inconsistency between experimental results. Therefore, this paper investigates degradation monitoring of coil insulation systems under thermal loading conditions from a creep point of view. Inter-turn insulation creep deformation is identified as a quantitative index to manifest insulation degradation changes at the micro level. A method is developed to map coil high-frequency electrical monitoring parameters to inter-turn insulation creep deformation in order to bridge the gap between the micro-level and macro-level changes during the incipient insulation degradation process. Thermally accelerated tests are performed to validate the developed method. The mapping method helps to determine the physical meaning of coil electrical monitoring parameters and presents opportunities for predictive maintenance of machines that incorporate electromagnetic coils. Electromagnetic coils are fundamental energy conversion and transformation components of many systems . AlthougTesting and monitoring methods for insulation systems of electromagnetic coils have been surveyed in Grubic et al. . The cla➢Incipient degradation phase: As a current is passed through the wire, Joule heating causes an increase in the wire temperature, which results in expansion of the conductor  and thus compressive stress on turn-to-turn insulation . Under the long-term effects of compressive and thermal stresses on the turn-to-turn insulation, the insulation materials degrade and lead to a turn-to-turn short.➢Late degradation phase: After the turn-to-turn short occurs, the direct-current (DC) resistance of the coil decreases, and thus current increases, which makes the coil temperature increase. A hot spot can form at the location of the short. Turn-to-turn short faults will spread quickly around the hot spot, which causes the coil to burn out, finally resulting in failure of the complete coil.The on-line monitoring methods can be further divided into inter-turn short fault detection methods and inter-turn insulation degradation monitoring methods. In order to illustrate the relationship between these two methods clearly, insulation deterioration process is analyzed, as shown in Accordingly, the inter-turn short fault detection methods are used in late degradation phase to detect if the turn-to-turn short forms or to assess fault severity by identifying the number of inter-turn shorts, whereas the inter-turn insulation degradation monitoring methods are used in the incipient degradation phase to monitor the health state of insulation before the turn-to-turn short formation.(1) Inter-turn short fault detection methodsMotor current signature analysis (MCSA) is commonly used to detect inter-turn short fault by analyzing the stator current spectrum. The basic principle of MCSA is that the frequency domain features of the stator current will change  in the event of an inter-turn short. The classical MCSA methods include Fourier-transform-based methods ,19, and Other commonly used methods are based on the principle that motor symmetry is changed after the turn-to-turn short formation. The classical methods include the negative-sequence-current-component-based methods , Park’s Artificial-intelligence-based methods are proposed in order to eliminate the interference of noise during fault detection, in which winding current signals with different health states are acquired to train classifiers. Thus, these methods can be used to classify the different short circuit levels by identifying the number of inter-turn shorts. The most common methods are based on artificial neural network (ANN)  and suppMotor magnetic field signals can also be used to detect inter-turn shorts, wherein the motor magnetic field changes in the event of an inter-turn short fault. The classical methods include air-gap-flux-based methods ,27 by inAccording to the analysis mentioned above, the application premise of inter-turn short detection methods is that the turn-to-turn short has already formed. However, once the turn-to-turn short forms, the coil insulation degradation process accelerates, and the impending insulation failure usually occurs suddenly and has catastrophic effects. For example, it has been shown that it takes less than 2 s for a single turn fault to develop into a critical fault in a typical 15-kW induction motor . Therefo(2) Inter-turn insulation degradation monitoring methodsConsidering coil insulation systems usually go through a slow degradation process before a turn-to-turn fault occurs, inter-turn insulation degradation monitoring presents opportunities for continuous technical conditions monitoring and predictive maintenance of machines that incorporate electromagnetic coils. The representative inter-turn insulation degradation monitoring methods include Werynski et al. , who perIn summary, the existing inter-turn insulation degradation monitoring methods detect degradation of insulation used in low-voltage coils by assessing the change of high-frequency electrical parameters  response, whereas the effects of insulation failure mechanism on these electrical parameters were not considered. This brings inconsistency between the experimental results of existing studies. Some representative examples are as follows.(1)There are contradictory claims for the evolving trends of high-frequency electrical parameters in the literature. For example, Perisse et al.  and Savi(2)Perisse et al.  set a faThe difference between the results from experiments in the previous studies may be due to differences in material properties and failure mechanisms. It is premature to make a conclusion on electrical parameter evolving trends or setting failure thresholds without considering insulation failure mechanisms. It is thus important to illustrate the insulation degradation mechanism, examine related material-level changes under certain loading conditions, and associate those changes with evolving electrical parameters when performing on-line degradation monitoring in situ. Therefore, this paper has developed a creep degradation mechanism for inter-turn insulation, aiming at thermal loading conditions that have been shown to be the primary reason for insulation failure . The fraThe remainder of this paper is organized as follows. A cross-sectional view of an electromagnetic coil is shown in IHI) can be defined from the creep degradation point of view. The related definitions are as follows.According to the creep degradation mechanism analysis mentioned above, as shown in ILT):(1) Insulation layer thickness (CD):(2) Inter-turn insulation creep deformation :(3) Insulation health indicator  can be used to analyze the electrical behavior of the electromagnetic coils. The ECM of the coil can be modeled as a resistance, C, where The parasitic capacitance network model, as shown in The turn-to-turn capacitance  and (5) :(4)Ctt=εThe capacitance between the iron core and innermost layer turn of the coil can be calculated according to Equation (6):C and k is an empirical constant that is dependent on the number of layers of the coil.Multisim, a circuit simulation software, was used to find the relationship between d Ctt in , and theC, can be analyzed, as shown in According to theoretical derivation mentioned above, the effect of inter-turn creep deformation on coil high-frequency parameters response, such as coil parasitic capacitance Coil high-frequency electrical parameters can be used as on-line monitoring parameters during the incipient degradation phase because they are a manifestation of insulation degradation on the macro-level and are easy to measure in situ. A method is developed in this section to map coil high-frequency electrical parameters to inter-turn insulation creep deformation, which helps to determine the physical meaning of on-line monitoring parameters and thus presents opportunities to promote the accuracy of remaining useful life prediction of electromagnetic coil insulation.Resonant frequency is a commonly used parameter for coil health monitoring, and this section discusses the mapping method using coil resonant frequency as an example. Considering that (1) Step 1: Mapping from coil resonant frequency to turn-to-turn capacitanceL is the coil inductance and C is the coil capacitance.The resonant frequency, Then,According to Equation (7),So, according to Equation (10), turn-to-turn capacitance (2) Step 2: Mapping from turn-to-turn capacitance to inter-turn insulation creep deformationAssume f|, and the constraint condition is The optimization function:Since Therefore, coil resonant frequency is mapped to the inter-turn insulation creep deformation by the two-step mapping.The purpose of the experiment was to validate the developed creep degradation mechanism and mapping method. The electromagnetic coil was hand-wound using the magnet wire, whose information is shown in The experimental process is described in −3 m, which is equal to the value by magnet wire specification as described in The platform for outer radius measurement is shown in The coil electrical parameter measurement platform is shown in Thus, the calculated result for Next, coil resonant frequency is mapped to inter-turn insulation creep deformation by the mapping method developed in First, according to the magnet wire outer radius measurement results, as shown in Second, the measured and calculated results of inter-turn insulation creep deformation, as shown in IHI by the developed mapping method. As shown in IHI under different aging temperatures can be explored, as shown in IHI and aging time can thus be obtained, as shown in IHI.Finally, coil resonant frequency is used to calculate inter-turn insulation creep deformation and Electromagnetic coils are widely used components in many applications and systems, including solenoids, motors, and transformers. However, coil insulation systems are failure-prone, especially under excessive thermal stresses. Further, the coil insulation degradation process accelerates once a turn-to-turn short has occurred, and the impending insulation failure usually occurs suddenly and has catastrophic effects. Thus, insulation degradation monitoring prior to the formation of turn-to-turn shorts presents opportunities for decreasing the safety risk and economic loss due to the unexpected shutdown of machines that incorporate electromagnetic coils.This paper investigated degradation monitoring of coil insulation systems under thermal loading conditions, which is the primary factor that leads to insulation breakdown, from a creep point of view. Polymer creep curves obtained by inter-turn insulation degradation mechanism analysis and outer radius measurement results of magnet wires during accelerated tests show that the insulation degradation process can be quantified by inter-turn insulation creep deformation on a micro-level. Thus, the insulation health indicator was defined based on insulation layer thickness.A method of mapping coil high-frequency electrical parameters to inter-turn insulation creep deformation was explored by circuit theories and optimization methods, which is validated by comparison between mapped and measured creep deformation during the thermally accelerated tests. The developed method bridges the gap between the micro-level and macro-level changes during the incipient insulation degradation process, and a knowledge base for insulation degradation monitoring can thus be established by converting the electrical monitoring parameter to coil insulation health status. The developed mapping method helps to reveal the practical physical meaning of coil high-frequency electrical parameters and enhance the prognostic ability of existing high-frequency electrical parameter-based insulation degradation monitoring methods.


Photoactuators are promising smart materials that can adapt their shapes upon light illumination. Smart materials with recycling, reusable, and reconfigurable properties are crucial for a sustainable society, and it is important to expand their function. Recently, much effort was made to address the issue of reprocessability and recyclability of photoactuators. Based on the development of polymer chemistry, supramolecular chemistry, and dynamic covalent chemistry, it is now possible to prepare reconfigurable and recyclable photoactuators using azobenzene-containing polymers (azopolymers). Herein, the recent advances on reconfigurable and reprocessable photoactuators, including dynamic crosslinked networks systems and non-covalently crosslinked azobenzene-containing polymers, were reviewed. We discuss the challenges in the field as well as the directions for the development of such photoactuators. ThroughCrosslinking provides materials with mechanical robustness but sacrifices the reprocessing and recycling performance. With the development of the introduction of post-crosslinkable moieties  of the azopolymer. Design and synthesis of linear liquid crystal polymers with robust mechanical properties can address the abovementioned problem.Another strategy to produce the recyclable photoactuators is directly using linear photoresponsive materials or supramolecular non-covalent interactions  without chemical crosslinking networks. The hydrogen bonding-induced physically crosslinked networks in photoactuators based on azopolymers or hydrogels were reprogrammable or recyclable  linear azopolymer was synthesized by ring-opening metathesis polymerization (ROMP) and acted as “photonic muscles” of the microactuator. Subsequently, they further synthesized a linear liquid crystal copolymer  combining the photoresponsive azobenzenes and biphenyl moieties by ROMP  values above room temperature, yet cis azopolymers are liquids with Tg values below room temperature (−1. The low-molecular-weight azopolymers (5–53 kg mol−1) were hard and brittle because their polymer chains lacked entanglements, whereas the high-molecular-weight azopolymers (80–100 kg mol−1) entangled  with different molecular weights by atom transfer radical polymerization (ATRP). High-molecular-weight entangled azopolymers were able to prepare photoactuators  processability, mechanical robustness, and actuating stability: Obviously, crosslinking or entanglement affords mechanical toughness, but it hinders movements of polymer chains, and thus declines reprocessability. Dynamic crosslinked networks provide reprocessability and reconfigurability of the materials. However, during the long-term use of the materials, the dynamic reaction may lead to a reduction in its degree of orientation in actuators, and thus cause performance and reprocessability drops. Constructing new dynamic reaction systems and adjusting the material compositions to achieve a balance between reprocessability, mechanical toughness, and driving stability is of great significance for the practical applications of photoactuators.Develop novel non-covalently crosslinked systems and large-scale preparation on demands: The entangled azopolymer photoactuators cannot be large-scale prepared because it is difficult to polymerize high-molecular-weight azopolymers due to the large steric hindrance of azobenzene and other side reactions. In principle, each polymer has a characteristic average molecular weight between entanglements depending on the chemical structure of the repeating unit. This may also be taken into consideration in designing liquid crystal polymers that favor the use of chain entanglements. At present, the synthesis and design of non-covalently crosslinked photoactuators are still in the preliminary stage, and only a few cases are available. Since oriented polymer chains tend to relax in the isotropic state, the actuation degree may decrease after repeated order-disorder phase transition occurs. To exploit new synthetic methods and explore the impact of various chemical structures on their performance will facilitate the large-scale preparation of photoactuators on demand.Develop microstructured photoactuators: The recent studies showed that multi-smart functions or properties were accessible using microstructured photoactuators, while obtaining a specific function often required a specifically designed microstructure (Lahikainen et al., MC collected and read papers and wrote the manuscript. SL, CL, YL, and SW discussed and revised the manuscript. SW outlined the content of the manuscript. All authors contributed to the article and approved the submitted version.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.