Introns represent almost half of the human genome, yet their vast majority is eliminated from eukaryotic transcripts through RNA splicing. Nevertheless, they feature key elements and functions that deserve further interest. At the level of DNA, introns are genomic segments that can shelter independent transcription units for coding and non-coding RNAs which transcription may interfere with that of the host gene, and regulatory elements that can influence gene expression and splicing itself. From the RNA perspective, some introns can be subjected to alternative splicing. Intron retention appear to provide some plasticity to the nature of the protein produced, its distribution in a given cell type and timing of its translation. Intron retention may also serve as a switch to produce coding or non-coding RNAs from the same transcription unit. Conversely, splicing of introns has been directly implicated in the production of small regulatory RNAs. Hence, splicing of introns also appears to provide plasticity to the type of RNA produced from a genetic locus . We addressed these aspects to add to our understanding of mechanisms that control the fate of introns and could be instrumental in regulating genomic output and hence cell fate. In eukaryotes, the process of making a messenger RNA (mRNA) involves the co-transcriptional excision of introns in the nucleus, whereas joined-exons are exported to the cytoplasm to be translated. As a consequence, introns are inherently non-protein-coding sequences in that they are transcribed but not translated  into protein. Because, and by definition, introns always lie between 2 exons, they were called “Intervening” or INtrons. By opposition, exons are those sequences that are EXpressed and EXported to the cytoplasm.a posteriori, an explanation on how eukaryotes diversify their proteome from just a few genes. We now know that the fate of introns is not simply a matter of being eliminated to allow formation of mRNA coding for proteins.In 1993, the Nobel Prize was awarded to Phillip A. Sharp and Richard J. Roberts for their discovery that genes can be split into segments and, as a consequence, transcripts that originate from them are matured into messenger RNA (mRNA) smaller in size. Hence, introns are segments of a gene between exons, which are transcribed but do not participate in the production of the final protein product as they are removed before translation, allowing joining of exons through a process known as splicing. With the discovery that various combinations of segments may be included in the final RNA molecule, this has changed our view on how genetic information is expressed and provided, Introns are classified in four groups based on splicing mechanisms. 1) Spliceosomal introns are found in coding genes of eukaryotes and utilize spliceosomes (large protein-RNA complexes) for splicing  Spliceos that incSplicing of both Group II and pre-mRNA introns involve the formation of a lariat suggesting that they may be evolutionary related. However, origin and evolution of spliceosomal introns is still a matter of debate, opposing “introns early” to “introns late” theories , the maiBeing eliminated to allow formation of messenger RNA and inherently non-coding, introns have long been kept in the now famous “junk DNA” drawer. However, and maybe against all odds, high sequence conservation among homologous introns of closely related species suggests functional constraints on intronic sequences throughout evolution . Many stWe review here aspects of introns that are perhaps less known but made them see a renewed interest lately.i.e., introns, which cover more than 1.7 billion bases pairs and represent 45% to 55% of the human genome . However, overall, a typical coding gene contains 10.8 exons and 9.8 introns . The absolute record is held by the titin gene, which contains 363 exons . About er genes , but onler genes , were aser genes . The majer genes .BAI3; chr6:69,232,406–69,389,511) gene.Nested genes that produce small regulatory RNAs independently of the transcription of the host gene have also been described ,14. ExamAlthough it has been proposed that a compact genome allows for quicker replication rate in prokaryotes, the functional benefits for mammalian genomes to hold nested genes are still unclear. Still, the presence of nested transcription units in introns raises the questions of their concerted transcription and regulation. It is likely that the host gene and its nested gene transcribed from the opposite strand are inversely regulated and transcribed, owing to promoter competition or steric hindrance of RNA polymerase and transcription factors. In contrast, if the nested and host genes are transcribed from the same strand, it is quite straightforward to predict a concerted model of transcriptional regulation of both genes to allow expression, if not in the same pathways, at least in the same cellular context or in response to the same environmental cues.Through classical mechanisms than we will not detail here, pre-mRNA splicing is precisely controlled in a spatial and temporal manner to generate mature RNAs . If one(1) The recursive splicing consists in the stepwise removal of introns by sequential splicing reactions starting from the most 5' donor site and progressing towards the most 3' acceptor site ,22. To d(2) The intra-splicing or nested splicing first requires internal splicing, using internal canonical donor and acceptor sites, followed by external splicing involving the most outer donor and acceptor sites . This prDMD; chrX:31,137,345–33,357,726) is one of the largest annotated genes, spanning more than 2 Mbp and generating transcripts containing up to 79 coding exons. Strikingly, DMD transcripts are only 14 kb long since more than 99% of the gene sequence is composed of introns, whose lengths vary from 107 nt (intron 14) to 248,401 nt (intron 44) [EPB41) gene in which Exon 1A and exon 2 are joined through two nested splicing events [The human dystrophin gene (tron 44) . Intron g events .In eukaryotes, splicing occurs in the nucleus in a transcription-coupled manner where the spliceosome complex catalyzes intron removal, deposits exon–junction complexes (EJC) to mark sites of intron removal but also participates in nuclear retention of incompletely spliced pre-mRNAs. Quality control of incompletely spliced pre-mRNAs to prevent their translation into aberrant proteins involves at least two mechanisms; the nuclear degradation of pre-mRNAs that failed to be exported to the cytoplasm, and the cytoplasmic degradation through nonsense-mediated decay (NMD) of intron-retaining RNAs with a premature stop codon 5' of an EJC .The major spliceosome comprises 5 small nuclear RNAs (snRNA) and a multitude of associated protein factors to form small nuclear ribonucleoprotein particles (snRNPs), U1, U2, U4, U5, and U6 . SpliceoA minor spliceosome has been reported in plants, insects, and vertebrates, which is specialized in the splicing of a rare class of introns  Transcripts with unspliced minor introns can be found in the cytoplasm, together with components of the minor spliceosome. In addition, the specific inhibition of minor spliceosome components in the cytoplasm, using antisense morpholino conjugated to a nuclear-export sequence, leads to accumulation of unspliced U12-introns in the cytoplasm .(2) The minor spliceosome, unlike its major counterpart, seems to evade mitotic down-regulation, to be able to operate during mitosis and to function independently of transcription ,32,33. I(3) Along the same lines, platelets, which are specialized hemostatic cells that circulate in the blood as anucleate cytoplasts, contain essential spliceosomal proteins and small nuclear ribonucleic acids (snRNAs), as well as a subset of pre-mRNAs that are further spliced into mature message in response to platelet activation . This me(4) Additional support for the functionality of RNA splicing in the cytoplasm was provided in neurons. Neurons have a cell body containing the nucleus and core cytoplasmic components surrounded by highly specialized cytoplasmic extensions called dendrites and axons. Certain not fully spliced intron-retaining mRNAs are directed toward neuronal dendrites where they fully mature ,36. As m(5) Cytoplasmic splicing is one of the major regulatory mechanisms of the unfolded protein response (UPR), although it occurs through unconventional mechanisms . SplicinIn sum, cytoplasmic splicing offers an efficient way to diversify cellular phenotypes without the need for nuclear activation of sets of genes as well as allowing for rapid regulation of local environment and subcellular proteomes and hence, for a quasi instant response to environmental cues.miRNAs play pivotal roles in diverse biological functions, mainly through the repression of target genes. Less than half of intronic miRNAs are transcribed independently of their host gene, as long primary capped and polyadenylated transcripts (pri-miRNAs) by RNA polymerase II (RNA pol II), which undergo a series of maturation steps via the formation of a hairpin-forming precursor (pre-miRNA) until the production of mature miRNAs in the cytoplasm. These intronic miRNAs should be named intragenic or nested miRNAs to distinguish them from independent intergenic miRNAs. In contrast, the other half of so-called intronic miRNAs share promoter and regulatory elements with their host gene ,41,42,43Interestingly, there are cases where small regulatory RNAs are directly produced by way of splicing of the intron from the host transcript and thus, are strictly dependent on both transcription and splicing of the host gene.GRCh38).In the case of snoRNAs, while some have their own promoter and are transcribed by RNA pol II, the majority is produced through splicing of the host intron. Whereas most of the host transcripts are mRNAs, some examples of long non-coding RNA (lncRNA) hosting snoRNAs exist such as the growth arrest-specific transcript 5 (GAS5) that contains 10 different snoRNAs, almost one in each one of its introns  . NoteworAt least two alternative pathways in the biogenesis of miRNAs use splicing to produce pre-miRNA-like transcripts independently of the microprocessor complex: the mirtron and the simtron pathways .i.e., dependency on splicing and debranching machineries, has been experimentally validated for a handful of them [Unlike canonical miRNAs that require the microprocessor complex to produce the hairpin pre-miRNA, the so-called mirtron pathway is thought to require debranching of the lariat-intron produced by splicing, by a debranching enzyme dbr1, as a prerequisite to produce a hairpin pre-miRNA-like structure . Until n of them . More re of them ,52 but nEmbedding of genetic information for small regulatory RNAs within a longer mRNA or ncRNA provides an attractive concerted model for eukaryotic gene regulation, whereby multiple actors in the same pathways or in a given cellular context are produced from interdependent transcripts. The co-linear expression of miRNA and protein-coding genes may also function to ensure negative feedback on the protein-coding gene to prevent its over-expression. Indeed, while some intronic miRNAs are transcribed from their own promoter as independent transcription units, expression of intronic miRNAs has been widely linked to the regulation of expression of their host gene ,53. FurtSplicing of introns may also lead to the production of long non-coding RNAs with a circular structure (ciRNA). In human cells, Zhang and colleagues  have rec. Alternative splicing (AS) is a mechanism that allows the inclusion of non-coding sequences (introns) or excision of coding segments (exons) within mature mRNAs. While AS of introns represents a major event in plants, it accounts for only 2 to 5% of all alternative events in humans ,62. HoweWe already mentioned examples where incompletely spliced intron-retaining transcripts are exported to the cytoplasm where their maturation is completed under certain physiological or cellular contexts. This may seem surprising since, in the majority of cases, intron retention leads to premature stop codons that trigger the cellular surveillance machinery and degradation of such transcripts by the nonsense-mediated decay pathway (NMD) . Why andLY6G5B and LY6G6D genes located in the major histocompatibility complex (MHC) class III region on chromosome 6 can produce leukocyte antigen-6 (Ly6)-producing mRNA as well as intron-retaining non-coding transcripts that seem to be more abundant and stable than the correctly spliced isoform in most cell lines tested, and to escape NMD [Besides the already mentioned example of intron-retention that allows for fine tuning of protein production at the right place and at the right moment in dendritic cells and neurons ,36, intrcape NMD . This meWhen it is tightly coupled to NMD, orchestrated intron retention allows for fine control of expression levels of proteins involved in the immune response . It alsoMore directly, intron-retention participates in the diversification of the proteome, although, as we already mentioned, it is less commonly used in mammals than in plants. et al. [Other examples of intron-retained transcript together with their biological function in mammals can be found in et al. .i.e., RNAs for which both coding capacity and activity as functional RNA have been described [It is now evident that the transcriptional output of mammalian genomes is much more complex than estimates based on the number of protein-coding genes, and that non-coding RNA widely increase the source of regulatory molecules. We recently characterized a new class of ncRNAs, called bifunctional RNAs, escribed ,77,78,79et al. [Historically, the pioneer member of this new class of RNAs is SRA (Steroid Receptor RNA Activator). SRA was first identified as a structural ncRNA molecule in hormone receptor complexes, characterized by discrete stem-loop structures required for its function as a co-activator ,81 as exet al. . A few yet al. ,77,82,83et al. ,77. Wheret al.  which isi.e., in which intron retention disrupts the ORF permitting transcription of a ncRNA [An example of such a genetic locus producing both coding and non-coding RNA, depending primarily on an event of AS of intron , is no l a ncRNA . Specifi a ncRNA . Whereas a ncRNA . More in a ncRNA . Convers a ncRNA ,79. OtheCombined, the recent findings that alternative splicing of introns could produce RNAs with coding capacity or acting as functional ncRNAs and that splicing could directly generate small regulatory RNAs raise the fascinating assumption that a single transcription unit could generate multiple molecules including proteins, long or smaller regulatory ncRNAs such as miRNAs, and bring to light the central role of mammalian introns in the diversification of both proteome and transcriptome depending upon the need of the cell to respond to particular environmental settings.Introns have been regarded for a long time as “junk DNA” and remnants of archaic ancestral genomes. Questions about their origin and function arose immediately after their discovery in 1977, in particular regarding the energy waste that this represents for the cell to transcribe introns if they have to be eliminated from transcripts. Since then, accumulating evidence lent support to their fundamental importance in the regulation of mammalian gene expression programs, from transcriptional initiation, termination and stability, through recruitment of the exon-junction complex to recruitment of chromatin remodelers through the spliceosome.With introns representing more than half of the genome in higher eukaryotes and being absent from prokaryotic genomes, the apparent correlation between the genomic fraction occupied by introns and organism complexity must have some functional significance.In essence, eukaryotic cells use a variety of strategies to control their transcriptional output that employ a large number of regulatory factors that, in turn, must be tightly regulated. We provided examples whereby introns, as genetic entities or RNA segments, facilitate or participate in this amazing regulation feat by sheltering information for small regulatory RNAs allowing for concerted expression of multiple molecules in a given context, influencing where and when a messenger RNA is spliced and translated, preventing or attenuating translation off context or, on the contrary, diversifying the type and function of the molecules produced depending on the internal and external environment.All these intron-linked mechanisms add some levels of sophistication in the way mammalian cells gain their phenotypic variability and opened a whole new field of investigation where introns take central stage and emerge as key elements in shaping cell identity.


Rorippa indica, a wild crucifer, has been previously reported as the first identified plant in the germplasm of Brassicaceae known to be tolerant towards the mustard aphid Lipaphis erysimi Kaltenbach. We herein report the full-length cloning, expression, purification and characterization of a novel R. indica defensin (RiD) and its efficacy against L. erysimi. Structural analysis through homology modeling of RiD showed longer α-helix and 3rd β-sheet as compared to Brassica juncea defensin (BjD). Recombinant RiD and BjD was purified for studying its efficacy against L. erysimi. In the artificial diet based insect bioassay, the LC50 value of RiD against L. erysimi was found to be 9.099 ± 0.621 µg/mL which is far lower than that of BjD (43.51 ± 0.526 µg/mL). This indicates the possibility of RiD having different interacting partner and having better efficacy against L. erysimi over BjD. In the transient localization studies, RiD signal peptide directed the RiD: yellow fluorescent protein (YFP) fusion protein to the apoplastic regions which indicates that it might play a very important role in inhibiting nutrient uptake by aphids which follow mainly extracellular route to pierce through the cells. Hence, the present study has a significant implication for the future pest management program of B. juncea through the development of aphid tolerant transgenic plants.The online version of this article (doi:10.1186/s40064-016-2144-2) contains supplementary material, which is available to authorized users. Brassica juncea (Indian mustard) is one of the most important crops cultivated in India. More than three dozen insect pests are found to be associated with mustard crops in India, of which mustard aphid  is the most dreaded. These pests are able to reduce the yield up to 96 %  Hiern, a wild crucifer and an occasional shade loving weed was noted to be tolerant against mustard aphid. R. indica is found in the Indian subcontinent and Asia  were seen to be differentially regulated  for the present study.Vigna radiata  in transgenic rice  and belong to the γ-thionin family . Homology modelling was done to study the differences between the model structures of RiD and BjD. The RiD and BjD coding sequence were cloned, expressed and purified using prokaryotic expression system. For further characterization, transient localization of RiD was studied and finally the insecticidal potential of expressed RiD as well as that of BjD was checked through artificial diet based insect bioassay against L. erysimi.In the aforementioned background, the present study was conducted to characterize and study the efficacy of RiD against R. indica started to decline sharply with the appearance of the winged  form of the aphids. Among all the sets of infested and non-infested plants as shown in Fig. R. indica was seen to have lower aphid population with increasing days post infestation than B. juncea. The challenged R. indica plants were found to show normal development like the non-infested control plants  Fig. . After 4R. indica as well as in B. juncea was studied using the primers from the EST as previously described  through 9.45 and 9.67-fold at 24 and 48 h respectively  and B. juncea defensin (243 bp). A single intron in the RiD gene and a short region of 393 bp upstream element containing TATA box and other cis-acting elements were also identified , Arabidopsis (GenBank accession—NM106233.3), Sinapis alba (GenBank accession—AY998243) and Brassica rapa (GenBank accession—XM009106448) respectively at nucleotide level . Homology modelling clearly shows that despite having 86 % similarity at the amino acid level, the α-helix and 3rd β-sheet of RiD are little longer as compared with BjD  was used to calculate the electrostatic surface of the proteins. For both RiD and BjD, the outer surfaces of the proteins were seen to have more positive charge Fig. d–g.RiD and BjD were transformed into Rosetta (DE3) pLysS cells individually. The expression constructs produced a 6His-RiD and 6His-BjD fusion protein respectively, which were purified by Ni–NTA column. The maximum expression of RiD and BjD was obtained at 1 mM IPTG at 20° C after 16 h of incubation. As shown in Fig. The recombinant pET28a+ vectors harbouring the coding sequence of R. indica defensin, and other defensin-like proteins. The observed monoisotopic mass of tryptically digested peptides obtained by MALDI-TOF/TOF–MS and their position with respect to matched protein sequence are presented in Table R. indica, gi694199016) with score 47 and 48 % with defensin-like protein 1  with score 50.A MALDI-TOF/TOF–MS analysis was performed to verify the molecular mass of the purified RiD protein. MS analysis yielded a mass of 9067.364 Da for the purified peptide Fig. : Fig. S2R. indica defensin showed that RiD also has a putative N-terminal secretory signal peptide  and thus to the secretory pathway. A SignalP analysis  was calculated from the Abbot’s formula (Abbott P′ = C + P (1 − C), considering a population (P) which would have survived in the absence of the insecticidal agent used in the experiment and where C is natural mortality without the agent. LC50 value of RiD and BjD against L. erysimi were calculated accordingly using the statistical Probit analysis. LC50 value of RiD against L. erysimi was found to be 9.099 ± 0.621 µg/mL, whereas of BjD was found to be 43.51 ± 0.526 µg/mL  genes  has been successfully cloned, expressed, purified and characterized. Analysis of RiD and BjD through homology modelling showed structural difference through the presence of longer α-helix and 3rd β-sheet in the former one, despite having significant similarity in nucleotide level. Plant defensins are reported to have a broad spectrum of biological activities  with LC50 value of 13.47 ± 0.23 µg/mL  with LC50 value 20.7 ± 0.21 µg/mL  with LC50 value of 11.87 ± 0.229 µg/mL , temperature of 25 ± 2 °C and 75 % relative humidity. Mustard aphids (L. erysimi) were collected from the Madhyamgram Experimental Farm, Bose Institute, India. Thirty aphids of third-instar stage were forcedly infested upon each plant (under laboratory conditions previously mentioned), which were used for further experiments. Three plants with 30 aphids each were used as replicates.R. indica and B. juncea at an interval of every 24 h for 7 days, with three plants for each time point. Student’s t test at a significance level of P ≤ 0.05 was carried out to determine the significance of the infestation noted over the time course.The total aphid population was studied in both −∆∆Ct method  of all the infested plants at different time points as well as the non-infested control plants using Trizol reagent . First strand cDNA was synthesized using iScript-cDNA synthesis kit . The obtained cDNA was used for semi-quantitative analysis for the expression of PDF1.2c homolog and real-time PCR analysis using iQ SYBR Green Supermix (Bio-Rad Laboratories) in a iQ5 Multicolor Real-Time PCR Detection System (Bio-Rad Laboratories). Primers were designed on the basis of the previously identified EST (GenBank Accession—JK034054) . Genome walk was carried out according to the protocols of BD genome Walker™ Universal Kit . The primers were designed according to the sequence derived from RACE  to generate 3D coordinates of both RiD and BjD. Both the sequences were searched for a structure using Prime v3.4 module of Schrödinger molecular modelling suite  using RiDexp-F/RiDexp-R (for RiD) and BjDexp-F/BjDexp-R (for BjD) containing the BamHI  and SacI  sites  pLysS cell line . The recombinant cells were induced with 1 mM isopropyl-β-d-thiogalactopyranoside (IPTG) and incubated with constant shaking at 20 °C for 16 h. The recombinant cells were pelleted by centrifugation at 5000g at 4 °C for 30 min, resuspended in lysis buffer  and sonicated 30 times for 30 s each. The cell suspension was centrifuged at 10,000g for 30 min at 4 °C, and the supernatant was incubated for 2 h in 2 mL of nickel–nitrilotriacetic acid (Ni–NTA) column equilibrated with lysis buffer. The column was washed with wash buffer with 20 mM imidazole, pH 7.4 to remove nonspecific proteins, and finally, the target fusion protein was eluted with elution buffer with 250 mM imidazole, pH 7.4. All purification steps were carried out at 4 °C. The expression and purification of the recombinant proteins were analyzed in 12 % SDS-PAGE stained with Coomassie Brilliant Blue. The expressions of the proteins were further confirmed by western blot analysis using anti-His antibody (1: 5000) .The full-length coding sequence of Protein structures may lead to anomalously fast migration in SDS-PAGE  using RiDloc-F and RiDloc-R  was placed on each parafilm and was covered with another parafilm to make a pouch. The entire experiment was done in triplicate. Another experiment was set up with a range of different doses of BjD . Water was used as a no diet control to show that the provided liquid diet (without RiD and BjD) in comparison with water, can support the survival of the aphids. Aphid survivability was then recorded at every 12 h interval. Abbot’s formula was used to calculate the corrected mortality .Second instar nymphs were used for this experiment. Artificial diet was formulated using the original description by Dadd and Mittler . The expy Abbott . Statistthod Chi  was perf


Solvothermal processes are considered efficient approaches for the gram-scale production of graphene. Further modification of graphene by chemical doping is an important approach to tailor its properties. In this work, we successfully synthesized sulfur-doped graphene by using a solvothermal method with dimethyl sulfoxide as a precursor, which is a common laboratory reagent. Nitrogen-doped graphene was produced to demonstrate the generality of this process. These heteroatom-doped graphene materials exhibited high surface areas and high contents of heteroatoms. Furthermore, the lithium-ion storage properties and oxygen reduction reaction catalytic activity of these materials were also investigated. The success of this approach might facilitate the development of other advanced graphene-based materials with relative simplicity, scalability, and cost effectiveness for use in various potential applications. Graphene has attracted great interest in many research areas because of its unique structure and outstanding properties7912132, H2S, CS2, and benzyl disulfide) were studied as sources, and the catalyst activities of S-doped graphene for the oxygen reduction reaction (ORR) were investigated17232415Various elements  were selected in order to introduce into carbon frameworks to produce heteroatom-doped graphene151617181520213CH3SO, DMSO) into S-doped graphene. Furthermore, N-doped graphene was also synthesized using dimethylformamide , which is a N-containing organic molecule. These precursors are commonly used as low-cost solvents. It provides the benefits of a single step reaction with relatively mild synthesis conditions, gram-scale products, and higher contents of the heteroatom. The lithium-ion storage properties and electrocatalytic behaviors of heteroatomic-doped graphene were investigated.In this research, we describe a novel approach to fabricate S-doped graphene via a solvothermal method using S-containing organic molecules as a precursor. This method directly converts dimethyl sulfoxide  and carbon source (CCl4)2 hybridized carbon  mapping was carried out. ectively . The speectively . Graphitectively  correspo2 adsorption and desorption isotherms and pore size distributions of the heteroatom-doped graphene were also analyzed (2 g−1 and 317 m2 g−1 with the pore volumes (PVs) of 0.82 cm3 g−1 and 0.91 cm3 g−1, respectively. After annealing at high temperature (900°C) under an inert atmosphere, the respective BET surface areas of the S-doped graphene and N-doped graphene were increased to 1564 m2 g−1 (PV = 1.53 cm3 g−1) and 2255 m2 g−1 (PV = 1.73 cm3 g−1).The Nanalyzed . The Bru2-hybridization  was observed, and the samples produced a small peak corresponding to C-O bonds, carbonyls (C = O), and carboxylates (O-C = O) 17For elucidating the chemical structure of the S-doped graphene, X-ray photoelectron spectroscopy (XPS) was employed. The XPS spectra of the solvothermal graphene derived from methanol confirmed that carbon and oxygen were the only atoms present. However, sulfur and nitrogen signals could be clearly observed in the XPS spectra of the S-doped graphene and N-doped graphene, respectively . The higO-C = O) 30. The hO-C = O) . Lines a−1 at room temperature (−1). Interestingly, the specific capacities decrease during initial 30 cycles and gradually increase as the number of cycles increases in the S-doped graphene and solvothermal graphene. These activation processes can also be observed in other graphene-based electrodes3111th to 150th cycle was 97.2%, which is a relatively high value, but those of the S-doped and N-doped graphene were much higher . It was demonstrated that half-cell Coulombic efficiency could affect cycle retention in full lithium ion cells3111−1, respectively, at current density of 2500 mA g−1, which correspond to the respective delivered capacities of 44.9%, 40.7%, and 53.1% at a current density of 200 mA g−1. The theoretical capacity of graphene is 744 mAh g−1 through the formation of LiC3 in a condition where all the graphene layers present as monolayers35−1 to more than 1500 mAh g−1  as platinum electrode. The activity of doped carbon for the ORR has been improved by a combination of many factors, which have not yet been clarified4041174344RHE (see RHE) to compare the electron transfer number in 46To investigate the electrocatalytic activity of the S-doped graphene and N-doped graphene for the ORR and to confirm the doping effect, we performed rotating-disk electrode (RDE) voltammetry in 0.1 M KOH electrolyte . The N-doped graphene shows higher ORR activity than pristine solvothermal graphene. Further, N-doped graphene has a similar onset potential -doped graphenes were successfully synthesized by using a simple, single-step solvothermal method with heteroatom-containing organic molecules as precursors. This process provides for a gram-scale production using common laboratory reagents. The solvothermal (S or N)-doped graphenes exhibited high heteroatom content and surface area, and their characterization data indicate that they could be utilized in various energy storage and conversion applications. We believe that heteroatom-doped graphenes can be further extended to applications in sensors, electronics and biological research. We expect that our synthetic approach will be developed to produce doped carbon materials based on other elements  which can then increase the method's potential applications.2 gas flow. The colorless mixture was boiled at around 250°C and was slowly thickened. After boiling for ~24 h, the liquid was converted into a black cake-like substance. Finally, the samples were washed with hydrochloric acid (10 wt%) and deionized water several times, and then dried in an oven at 90°C. The final yield of S-doped graphene is ~35 mg per 1 ml of DMSO  and deionized water several times, and then dried in an oven at 90°C.Solvothermal graphene were prepared according to a literature procedure using methanol as the precursorTEM images and TEM-EDS images were obtained using a JEOL JEM-1010 and JEM-2100F, respectively. AFM images were obtained using a Veeco atomic force microscope. Raman spectroscopy measurements were carried out on a Dongwoo DM500i Raman spectrometer using green (514.5 nm) laser excitation. BET surface area and BJH pore size distributions were measured by using a Micromeritics Tristar 3000. XPS was carried out using a Kratos AXIS-HSi. Elemental analysis was performed using a Thermo Scientific Flash 2000 analyzer at NCIRF .2 for battery applications. For the preparation of working electrodes, a well-mixed slurry of synthesized graphene, Super P and poly (vinylidene fluoride) in N-methyl-2-pyrrolidone (at a weight ratio of 70:10:20) was coated on a copper current collector. After drying, the electrodes were pressed and dried again at 120°C under vacuum. Two electrode 2016-type coin cells were fabricated with prepared working electrodes and lithium foil (as both counter and reference electrodes) in argon filled glove box. 1.0 M LiPF6 dissolved in ethylene carbonate (EC) and diethyl carbonate (DEC) (1:1 in volume ratio) was used as the organic electrolyte. All the voltage ranges for the battery tests in this study were set between 3.0 and 0.01 V (vs. Li+/Li). The specific capacities were calculated only according to the weight of the active material and the weights of additives (binder and conductive agent) were not included.Samples were prepared by heat treatment at 600°C in N2 for electrocatalytic applications. Electrochemical measurements were conducted using an Autolab PGSTAT 101 in a standard three-compartment electrochemical cell. In all experiments, a Ag/AgCl reference electrode was used as the reference electrode, and Pt wire was used as the counter electrode. All potentials referred to in these ORR experiments were converted to the pH-independent reversible hydrogen electrode (RHE) using the hydrogen oxidation reaction. Three kinds of graphene samples (5 mg) and 20 μL of 5 wt% Nafion ionomer were each dispersed 0.4 mL of 2-isopropyl alcohol by sonication for 10 min. For RDE, 7 μL of catalyst ink was loaded onto an RDE with a 0.000196 m2 glassy carbon disk. The electrode rotation speed was 1600 rpm , 0.1 M KOH electrolyte. To calculate the electron transfer number (0.6 VRHE), RDE measurements were conducted using different rotating speeds 400, 900, 1200, and 1600 rpm.Samples were prepared by heat treatment at 600°C in NB.Q. proposed the concept and performed the experiment. S.-H.Y. and A.J. carried out lithium storage properties. D.Y.C. carried out electrocatalytic performances. J.H.P. carried out the AFM data. Y.P., B.Q. and S.-H.Y. co-wrote the manuscript. Y.P. and Y.-E.S. coordinated and supervised the work.Single Source Precursor-based Solvothermal Synthesis of Heteroatom-doped Graphene and Its Energy Storage and Conversion Applications


The superior photoconductive behavior of a simple, cost-effective n-GaN nanorod (NR)-graphene hybrid device structure is demonstrated for the first time. The proposed hybrid structure was synthesized on a Si (111) substrate using the high-quality graphene transfer method and the relatively low-temperature metal-organic chemical vapor deposition (MOCVD) process with a high V/III ratio to protect the graphene layer from thermal damage during the growth of n-GaN nanorods. Defect-free n-GaN NRs were grown on a highly ordered graphene monolayer on Si without forming any metal-catalyst or droplet seeds. The prominent existence of the undamaged monolayer graphene even after the growth of highly dense n-GaN NRs, as determined using Raman spectroscopy and high-resolution transmission electron microscopy (HR-TEM), facilitated the excellent transport of the generated charge carriers through the photoconductive channel. The highly matched n-GaN NR-graphene hybrid structure exhibited enhancement in the photocurrent along with increased sensitivity and photoresponsivity, which were attributed to the extremely low carrier trap density in the photoconductive channel. The vapor-liquid-solid (VLS) technique25In the present study, we sought to form a GaN NR-graphene hybrid structure using the MOCVD technique while maintaining the original properties of the GaN nanorods and graphene. The material and electrical features of this integrated heterostructure were analyzed to assess their potential for developing high-efficiency photoconductive devices. Moreover, the growth of this hybrid structure was achieved on a bare Si substrate without forming silicon dioxide or silicon nitride; furthermore, to avoid contamination, no metal-catalyst was used to grow the GaN NRs. The aim of this study was to demonstrate the highly matched co-existence of well-formed monolayer graphene and GaN NRs in a hybrid structure grown on a Si substrate without using any metal catalyst utilizing the MOCVD process to minimize the production costs. The realization of a high-performance ultraviolet photoconductive device using this hybrid structure was another purpose of this work.−1 and a 2D-band at approximately 2673 cm−1, which are typical peak positions for defect-free graphene282) that predominantly exhibit typical monolayer properties, as identified by the IG/I2D ≤ 0.38 and the FWHM of the 2D band of ≤38 cm−1. Chu et al.2/Si (100) with a G-band/2D-band peak intensity ratio ≤0.4, whereas Kwak et al.G/I2D ≈ 0.38) grown using the CVD process and then transferred to a Si (111) substrate. To our knowledge, this study represents the finest graphene layer transferred directly to a Si substrate reported thus far29S from 4 to 500 μm. A geometry correction was extracted by fitting the measured resistances to the following equation3132The fabrication sequences involved in the process used to synthesize n-GaN NR-graphene hybrid structures are shown in RT is the total resistance between the channel spacing of the contact pads, Rs is the sheet resistance of the graphene films, r1 is the radius of the inner circular contact, S is the channel spacing of the contact pads, and LT is the transfer length. From the linear fit (red line), the estimated sheet resistance and transfer length of the graphene films were Rs ≈ 483 ohm/sq and LT ≈ 13.60 μm, respectively, implying that highly crystalline monolayer graphene films were transferred onto the Si substrate.Here, c-plane. With the integration of a graphene layer, the XRD peaks indicated the existence of GaN NRs grown along the c-plane, i.e., along the (002) and (004) planes (2O3 listed in the Joint Committee on Powder Diffraction Standards (JCPDS) cards No. 50-0792 and 20-0426, respectively.) planes ; the str−1 and 567 cm−1, respectively  images showed that the n-GaN NRs were uniformly grown along the irection . To veriirection ; in addiP is defined as (Imaxdark) at a specific bias voltage22P/Idark) with bias for the bare Si substrate and for n-GaN NRs on the same substrate without and with a graphene layer; the sensitivity was estimated to be approximately 10%, 4%, and 60%, respectively, at a bias voltage of The photocurrent behavior of the bare semi-insulating Si (111) substrate during photoconductivity measurements is shown in  results . The low results , might b results , enhance results  and impr results  of the GP/(Illuminated area × Power density of light)22 and maintained the same dimensions of the illuminated area for all of the channels. At a bias of 2.0 V, the absolute photoresponsivity was calculated to be 20, 21, and 106 mA/W, respectively, for the bare Si substrate, n-GaN NRs grown without graphene and n-GaN NRs with graphene. As et al.214237The same behavior of the photocurrent and sensitivity was repeated when the photoresponsivity of these three channels was compared . The phop ∞ Px235p would increase linearly with increasing optical power when the trap level density is lower. To support this fact, the exponent x should be approximately 1 in the power law. If the photoconductive channel possesses a high density of trap levels, x should be significantly smaller than 1. When the photocurrent values were fitted in 2. The evidence of minimal defects or trap states in the channel reveals why this MOCVD-grown n-GaN NR-graphene hybrid structure on Si is ideal for use in cost-effective, highly efficient photoconductive device applications. The photocurrent values, which are associated with the spectral response, were measured to be almost constant at approximately 36 mA at wavelengths up to an abrupt cut-off wavelength of 380 nm  substrate using a high-quality graphene transfer method and the relatively low-temperature MOCVD process with a high V/III ratio. Raman spectroscopy and HR-TEM images confirmed the presence of monolayer graphene highly matched with n-GaN NRs in the hybrid structure. The excellent electrical conduction properties of the graphene monolayer transferred on Si (111) not only affected the growth of defect-free, highly dense n-GaN NRs but also improved the material quality. As a result, the n-GaN NR-graphene hybrid structure under study represents the superior characteristics of an ultraviolet photoconductive device with extremely low carrier trap density. Moreover, the synthesis of the hybrid structure directly on a Si substrate by the growth of n-GaN NRs without using metal-catalyst or droplet seeds added special significance. To our knowledge, such simple, cost-effective and highly efficient ultraviolet photoconductive devices fabricated using highly matched n-GaN NR-graphene hybrid structures are reported here for the first time.3PO4 (85%) solution to obtain a flat and smooth Cu foil surface with a root-mean-square surface roughness of less than 15 nm (in the scanning area of 100 μm × 100 μm). After the Cu foil was loaded into a low-pressure chemical vapor deposition (LP-CVD) system, it was heated to a process temperature of 1050 °C and maintained for 60 min under 5 sccm of H2 for in situ surface cleaning. A monolayer graphene film was then grown for 15 min under a CH4/H2 mixture  with a reactor pressure of approximately 90 mTorr. After graphene growth, the chamber was rapidly cooled to room temperature with a flow of H2. Following graphene growth, the graphene film was transferred onto a Si substrate for further investigation. In all of our growth experiments, monolayer graphene was observed to grow on both sides of the Cu foil; therefore, the backside of the Cu foil was pre-treated with oxygen plasma to remove the undesired graphene film. To transfer the graphene film to a Si substrate, poly(methyl methacrylate) (PMMA) was spin-coated onto the graphene grown on Cu foil, and the PMMA/graphene/Cu foil was then cured on a hot plate at 180 °C for 1 min. The Cu foil was then etched from the PMMA/graphene/Cu foil assembly using an aqueous solution of (NH4)2S2O8. The PMMA/graphene assembly was subsequently washed with deionized water, transferred onto a semi-insulating Si (111) substrate and dried. The PMMA was then removed using acetone.First, a Cu foil  was electrochemically polished in H3), respectively; silane (SiH4) gas was used for n-type doping, and hydrogen was used as the carrier gas. To grow n-GaN nanorods, TMGa, NH3 and SiH4 were introduced into the reactor chamber for 45 min at flow rates of 0.2 sccm, 3.0 SLM and 10 sccm, respectively . To protect the graphene layer from thermal damage during the growth of the n-GaN nanorods, the growth temperature was fixed at 870 °C, which is relatively low compared to the usual temperatures of greater than 1000 °Cn-GaN nanorods were grown on the graphene-transferred Si (111) substrate using a relatively low-temperature MOCVD process and a high V/III ratio without the formation of any metal-catalyst or droplet seeds. The precursors for gallium and nitrogen were trimethylgallium (TMGa) and ammonia  within the range of 300–550 nm. A monochromator (Oriel Cornerstone 130) was used to provide the monochromatic light incident on the channel.Raman spectroscopy was performed using a WiTec Alpha 300R M-Raman system; the excitation wavelength was 532 nm. The results of the Raman study were used to confirm the existence of highly ordered graphene films. The surface morphology of the n-GaN NRs was analyzed using FE-SEM. A Hitachi S-7400 system was used for the FE-SEM study; it was operated at 15 kV and at a 13° tilt-view. Single-crystal XRD measurements were performed using a Rigaku diffractometer equipped with a Cu-Kα radiation source. The morphology of the n-GaN NR-graphene heterostructures was further revealed by cross-sectional HR-TEM. A JEOL JEM 2010 system operated at 200 kV was used for the HR-TEM study. Samples of the hybrid structure were prepared by being coated with platinum using a dual-beam focused ion beam  technique with a beam current of 65 nA and a resolution of 7 nm at 30 kV. The optical properties of the n-GaN NRs were investigated by photoluminescence (PL) spectroscopy at room temperature using the 325 nm line of a He-Cd laser as an excitation source. For photoconductivity measurements of the fabricated ultraviolet photoconductive device, we utilized a solar simulator (McScience Lab 100) as a light source. This light source generated white light with a maximum power density of 100 mW/cmHow to cite this article: Kang, S. et al. Ultraviolet photoconductive devices with an n-GaN nanorod-graphene hybrid structure synthesized by metal-organic chemical vapor deposition. Sci. Rep.5, 10808; doi: 10.1038/srep10808 (2015).


The enhanced doping effect caused by graphitic nitrogen in the GG produced an asymmetry electron–hole transport characteristic, indicating that the long–range scattering (ionized impurities) plays an important role in determining the carrier transport behavior. Analysis of temperature dependent conductance showed that the carrier transport mechanism in the GG was thermal excitation, whereas that in the PG, was a combination of thermal excitation and variable range hopping.Single–layer and mono–component doped graphene is a crucial platform for a better understanding of the relationship between its intrinsic electronic properties and atomic bonding configurations. Large–scale doped graphene films dominated with graphitic nitrogen (GG) or pyrrolic nitrogen (PG) were synthesized on Cu foils via a free radical reaction at growth temperatures of 230–300 °C and 400–600 °C, respectively. The bonding configurations of N atoms in the graphene lattices were controlled through reaction temperature, and characterized using Raman spectroscopy, X–ray photoelectron spectroscopy and scanning tunneling microscope. The GG exhibited a strong n–type doping behavior, whereas the PG showed a weak n–type doping behavior. Electron mobilities of the GG and PG were in the range of 80.1–340 cm Graphene, a two–dimensional crystal with carbon (C) atoms packed in a hexagonal lattice, has attracted extensive research attention in the past decade, owing to its extraordinary properties123891011121516171991011124/NH3 as precursors, graphitic N dominated graphene was synthesized on Cu foils at 800–1000 °C92H4/NH3 as the precursors23Chemical vapor deposition (CVD) is a powerful method among the synthesis techniques of heteroatom doped graphene films910Herein, we proposed a temperature controlled method for N–doped graphene film growth on Cu foils via a free radical reaction. The graphene was predominantly doped with graphitic and pyrrolic N atoms at different temperatures of 230–300 °C and 400–600 °C, respectively. The bonding configurations of N atoms in the graphene lattice were characterized using Raman spectroscopy, X–ray photoelectron spectroscopy (XPS) and scanning tunneling microscope (STM). The graphitic N doped graphene exhibited a strong n–type transport behavior, whereas the pyrrolic N doped graphene showed a weak n–type transport behavior with a symmetric electron–hole transport. Analysis of temperature dependent conductance showed that the carrier transport mechanism was a thermal excitation process for the graphitic N dominated graphene, whereas it was a combination of thermal excitation and variable range hopping for the pyrrolic N dominated graphene. As–grown mono–component doped graphene film is an important platform for the understanding of the relationship between the bonding configurations and its intrinsic physics of N–doped graphene, as well as development of graphene–based devices.Growth of the N–doped graphene film on Cu foils via the free radical reaction was previously reported by our groupTo reveal the influence of temperature on the growth of N–doped graphene via the free radical reaction, the Cu foil substrates were kept at various temperatures between 230 °C and 600 °C for growth. The morphology of the resultant film was characterized using SEM  and opti126−1 and ~2680 cm−1 are assigned to the G band and 2D band, respectively, which confirm the presence of graphitic carbon−1 activated by the defects via an intervalley double–resonance process is assigned to D bandD/IG) reveals the degree of defects and in–plane crystallite size (LD). Herein, the ratio (ID/IG) decreases as the temperature is increased in the range of 230–400 °C . The downshift of position agrees well with the electron doping feature in the graphene. Otherwise, the change of the fwhm(2D) appeared to be irregular with a larger bandwidth, which can’t be applied alone to identify the doping effectThe position and shape of Raman peaks can also provide doping information in the graphene. Herein, the position of G band was upshifted to the range of 1588.1–1589.0 cmr the GG  comparedsp2 hybridized C–C bond (C1). The peaks at 285.4 ± 0.2 eV and 286.4 ± 0.1 eV correspond to the trigonal phase with a sp2 C (C2) and the tetrahedral phase with a sp3 C (C3), respectively922sp2 C bond (C2). It may be come from pyridinic N or graphitic N atoms in graphene framework as well as the nitro– group connected to graphene framework, considering the spatial and electronic structure after incorporated these N atoms16sp3 C bond (C3). This bonding is originated from new type of incorporation N atoms other than pyridinic N or graphitic N atoms. Meanwhile, this bonding will increase the structural distortion by disrupting the plane framework of graphene. It would be assigned to pyrrolic N bonding configuration. The relative content of each component  is higher than that of C–N (305 kJ mol−1)These bonding configurations of N atoms are determined by bonding energies and their thermal stabilities. The coupling reaction could reduce the energy barrier for dechlorination and leave pyridinic ring radicals intact at low reaction temperatures , although the bonding energy of C–Cl  occurs, the conductance is linearly depended on Vg in both electron– and hole– transport regimes, indicating a long–range scattering (incorporated impurities) in the GG1946μ = (ΔIds·L/W)/(ΔVgs·Vds·Cox), where, Cox is the silica gate capacitance (1.15 × 10−8 F·cm−2 for a gate oxide thickness of 300 nm). As a result, the hole and electron mobilities are in the range of 59.5–429.8 cm2 V−1·s−1 and 80.1–340 cm2 V−1· s−1, respectively. The obtained high mobility of the carriers would be attributed to minimized defect formation upon direct substitution  at Vg = Vg,crossing and Vg,crossing (indicated by arrow). In our transport measurements, the gate voltage at Gcrossing is found to be equal to the Vg,min , implying the potential fluctuations induced by the randomly distributed charged impurities  localize in the graphene system with a homogenously spatial distributions5g,min and Vg,crossing observed in some nitrogen and potassium doped grapheneg regimes for both in the GG and PG  transport5050Incorporation of the pyrrolic nitrogen into graphene introduced not only defects but also a strong disorder resulting in localization of carriers, which is similar to the carriers in the rGO sheetWe have synthesized graphitic and pyrrolic N dominated graphene films by controlling the growth temperature via the free radical reaction. Raman spectra, XPS and STM results confirm the graphitic N and pyrrolic N dominated graphene films are grown at temperatures in the range of 230–300 °C and 400–600 °C, respectively. Furthermore, the graphitic N dominated graphene exhibits a strong n–type doping effect, whereas the pyrrolic N dominated graphene shows a weak n–type doping effect. The enhanced doping effect in graphitic N dominated graphene results in an asymmetry electron–hole transport which suggests that the scattering by ionized incorporated N impurities plays an important role in determining the carrier transport behavior. Temperature dependent conductance shows carriers transport mechanism in the GG was thermal excitation, whereas that in the PG was a combination of thermal excitation and variable range hopping.2 atmosphere (90 sccm) at 15 torr, followed by cooling down to the designated temperature (230–600 °C) and then the pentachloropyridine was heated to 130 °C and kept for various durations (0.5–5 min). After growth, the furnace was cooled down to room temperature. As–grown N–doped graphene was transferred onto the SiO2/Si substrates using a PMMA supported method for the characterization and device fabrications.Cu foils  and pentachloropyridine  were separately placed in a furnace with two–temperature zones. Prior to the growth of graphene, the Cu foils were annealed at 1000 °C for 20 min in H2/Si substrates without further annealing. The SEM images, STM images and XPS data were collected directly from all the doped graphene on Cu foils after growth.As–grown N–doped graphene was characterized using scanning electron microscopy , transmission electron microscopy , optical microscopy (Leica DM4500P), Raman spectroscopy , atomic force microscopy , X–ray photoelectron spectroscopy  radiation as a probe) and scanning tunneling microscope . Raman spectra, optical and AFM images of all the doped graphene were collected from the transferred doped graphene onto the SiO2/Si substrate with a 300 nm thermal oxide layer, and then patterned into micro–ribbonsL) and width (W) were measured to be 14 μm and 24.5 μm, respectively. To obtain a better contact, the devices were thermally annealed at 200 °C in H2/Ar (10/20 sccm) atmosphere for 20 min. Electrical measurements were carried out in a vacuum after pumping for 12 hours at room temperature using a semiconductor analyzer (Keithley 4200–SCS) combined with a Lakeshore probe station. Low temperature measurements were carried out from 5.2 K to 300 K in a vacuum of 1.58 × 10−6 torr.Electrical properties of the N–doped graphene films were evaluated based on a field effect transistor (FET) configuration. The N–doped graphene film was firstly transferred onto a SiOHow to cite this article: Zhang, J. et al. Tunable electronic properties of graphene through controlling bonding configurations of doped nitrogen atoms. Sci. Rep.6, 28330; doi: 10.1038/srep28330 (2016).


Tbx5, a known modifier of CHD, to heart defects on a trisomic backgroun. Mice that were heterozygous for a Tbx5 null allele were crossed with Ts65Dn mice. Thoraxes of progeny were fixed in 10% formalin, embedded in paraffin, and sectioned for analysis of CHD. Gene expression in embryonic hearts was examined by quantitative PCR and in situ hybridization. A TBX5 DNA binding site was verified by luciferase assays.Nearly half of all individuals with Down Syndrome (DS) have some type of congenital heart defect (CHD), suggesting that DS sensitizes to CHD but does not cause it. We used a common mouse model of DS, the Ts65Dn mouse, to study the contribution of Mice that were heterozygous for a Tbx5 null allele were crossed with Ts65Dn mice. Thoraxes of progeny were fixed in 10 % formalin, embedded in paraffin, and sectioned for analysis of CHD. Gene expression in embryonic hearts was examined by quantitative PCR and in situ hybridization. A TBX5 DNA binding site was verified by luciferase assays.Tbx5 null allele with Ts65Dn mice. Mice that were trisomic and carried the Tbx5 mutation  had a significantly increased incidence of overriding aorta compared to their euploid littermates. Ts65Dn;Tbx5+/− mice also showed reduced expression of Pitx2, a molecular marker for the left atrium. Transcript levels of the trisomic Adamts1 gene were decreased in Tbx5+/− mice compared to their euploid littermates. Evidence of a valid binding site for TBX5 upstream of the trisomic Adamts1 locus was also shown.We crossed mice that were heterozygous for a Tbx5 and trisomy affects alignment of the aorta and this effect may stem from deviations from normal left-right patterning in the heart. We have unveiled a previously unknown interaction between the Tbx5 gene and trisomy, suggesting a connection between Tbx5 and trisomic genes important during heart development.Haploinsufficiency of The online version of this article (doi:10.1186/s12861-015-0080-y) contains supplementary material, which is available to authorized users. Congenital heart defects (CHD) comprise the most common congenital anomaly in live births . There iTbx5, a known contributor to heart development, as a modifier and assess its interaction with trisomy. Tbx5, a transcription factor, has well-described effects in cardiac and limb development [Tbx5 has a well-studied role in the morphogenesis of the four heart chambers. It is expressed in the left ventricle (LV) and both atria during chamber maturation and septation [Tbx5 in the left ventricles (LV) of mice suppresses formation of the ventricular septum, resulting in formation of a single ventricle [Tbx5 expression might affect heart development on the “sensitized” trisomic background.We consider the role of elopment , 11. Mutelopment . Tbx5 haeptation . Ectopicentricle . Its impCreld1 and Hey2 genes as disomic modifiers of septal development on this trisomic background [Tbx5 in heart development.Animal models provide important information for understanding the pathogenesis of CHD and the molecular mechanisms that give rise to these conditions. Orthologs of many genes on Hsa21 are found on mouse chromosome 16 (Mmu16), with smaller subsets on Mmu10 and Mmu17 . The mosckground . HaploinTbx5 has been described [Tbx5−/− embryos (Black Swiss/SvJ background) die by embryonic day 10.5 (E10.5) and lack cardiac looping and endocardial cushion formation, among other severe defects [Tbx5+/− mice are greatly influenced by genetic background. Bruneau et al. report a 10 % frequency of the Tbx5 null allele on a 129SvEv/129SvJ background at birth and 28 % on a Black Swiss/129SvJ background, instead of the expected Mendelian ratio of 50 %. Deviation from the expected frequency indicates that prenatal loss has occurred; the different frequencies in different mouse strains suggest that genetic background contributes to the penetrance and expressivity of heart phenotypes in this situation [Tbx5 dosage are susceptible to additional genetic modifiers. The molecular mechanisms by which Tbx5 influences heart development are incompletely described and possible interactions between Tbx5 and genes on Hsa21 are unknown. We hypothesize that Tbx5 acts as a genetic modifier to alter CHD in Ts65Dn mice. Here we provide evidence of an interaction between Tbx5 and trisomy and the effects of that interaction on trisomic gene expression and left-right patterning of the heart.A mouse model with a null allele for escribed . Homozyg defects . The viaituation . TherefoTbx5+/− male mice and B6C3.Ts65Dn females were established to examine the role of Tbx5 as a modifier of CHD. Genetic background of the Tbx5 mice affected viability  background  background. There is some loss of Ts65Dn fetuses during late gestation and the observed frequency of 42 % trisomic pups at P0 . ASD and Gerbode’s defect, an abnormal communication between the right atrium and left ventricle . Three of these animals were affected, with an AVSD, ASD and VSD, respectively  in ~58 % of Ts65Dn;Tbx5ce Table . The pencle Fig. , were alTbx5+/− mice is an interaction between Tbx5 and a trisomic gene(s) that is regulated by this transcription factor. We considered 109 genes that are trisomic in Ts65Dn mice [Adamts1, Dyrk1a, Rcan1, Ripply3, Sh3bgr and Wrb). We used the Transcription Element Search System program  to identify potential TBEs in the first exons and/or promoter regions of those genes [Adamts1 for further investigation.The simplest explanation for the increased incidence of overriding aorta in Ts65Dn;5Dn mice  and gene5Dn mice . Howeverse genes  (see MetAdamts1 encodes an extracellular matrix protease. Transcript levels were increased in Tbx5+/− mice (p = 0.04)  [et al. [Adamts1 transcription start site, was amplified by PCR and cloned into the pGL3 luciferase vector  Fig. , suggestGGTGTVR) . He et a [et al.  found thTbx5  . The putative TBE was mutated to inhibit TBX5 binding and luciferase levels decreased compared to the construct with the intact TBE (p = 1.51 × 10−4).We transfected the construct into cells stably over expressing −4) Fig. . IntroduTbx5 is generally considered to be an inducer of transcription, consistent with the increase in luciferase signal when TBX5 binds the upstream Adamts1 region included in the construct. However, Adamts1 expression is increased when Tbx5 is reduced in Tbx5+/− mice . The expected expression pattern was observed in other organs . Thus, atrial isomerism likely contributes to the significant increase in overriding aorta in the Ts65Dn;Tbx5+/− animals.Overriding aorta and Gerbode’s defect are seen more often in Ts65Dn;somerism . To detents Fig. . These rnts Fig. , where PTbx5 dosage and trisomy resulted in a significant increase in defects of aortic alignment and revealed a potential effect on left-right patterning of the heart. The action of genetic modifiers in heart development is evident from the differences in penetrance and patterns of heart phenotypes in DS, in DS mouse models and when the Tbx5 null allele is bred onto different mouse genetic backgrounds. In addition, Holt-Oram patients who inherit the same TBX5 mutation have variable heart phenotypes, indicating that additional factors affect development [TBX5 displaying OA and a number of additional defects have been described [Tbx5 expression and trisomy with increased OA as a primary outcome.The interaction between elopment . Previouelopment , 9. For escribed . Isolateescribed . Our resPitx2 knockout mice [Pitx2 expression is sufficient to cause septal and valve defects [Pitx2 and Cited2 genes which are involved in left-right patterning exhibit phenotypes similar to the ones reported, including Gerbode’s defect and overriding aorta [Pitx2 is an important determinant of left-right asymmetry of the heart and gut [Pitx2 expression [Pitx2 regulates Tbx5 expression in the abdominal wall during development [Tbx5+/− embryos have reduced Pitx2 expression, which can contribute to atrial isomerism and aortic defects.Disruptions in the left-right patterning pathway frequently lead to the kinds of heart defects seen here. OA is considered to be a milder form of double outlet right ventricle (DORV), the most common defect seen in out mice , arisingout mice . Reducin defects . Mice lang aorta –37. Pitx and gut , 37–40 a and gut , 39, 41.pression , and Pitelopment . Ts65Dn;Tbx5 haploinsufficiency are combined. We suggested previously that a universal response deficit to Sonic hedgehog signaling due to trisomy could explain many of the clinical features of DS [Pitx2 expression [Pitx2 expression seen here.The left-right signaling pathway is affected when trisomy and es of DS ; hedgehopression –47. Exprpression . A respoTbx5 dosage show increased expression of the Adamts1 gene. Adamts1 is important in the development of the extracellular matrix in endocardial cushions [Tbx5 dosage may contribute to the risk of heart defects by a role in transcriptional regulation of the Adamts1 gene, a regulator of the VEGF pathway.Finally, we discovered that mice with reduced cushions , and is cushions . It is icushions . ReducedTbx5+/− mice relative to either genetic defects in isolation. The mechanism likely involves reduced expression of Pitx2, a gene with known roles in laterality. In addition, we demonstrated that a change in Tbx5 dosage affects transcription of Adamts1, suggesting it is a target of TBX5 in normal heart development in vivo.Defects in left-right patterning likely contribute to the significant increase in overriding aorta in Ts65Dn;Tbx5 dosage and trisomy interact to affect the outcome of heart development, potentially altering left-right patterning of the heart. We provided evidence of weak Pitx2 expression, important for left-right patterning, in the Ts65Dn;Tbx5+/− mice. We have also provided evidence that TBX5 plays a role in regulation of the trisomic Adamts1 gene. A valid TBX5 binding site was shown upstream of the Adamts1 locus and mutation of the site significantly affected transcription.We have shown that ad libitum. Ts65Dn mice (B6EiC3H-a/A-Ts(1716)65Dn) were obtained from The Jackson Laboratory and maintained as an advanced intercross on the B6;C3H background. Tbx5 heterozygous null mice (Tbx5+/−) [Tbx5+/− mice onto a C57BL/6J background for at least five generations. All data was taken after mice had been backcrossed for at least five generations. Progeny in Tables Tbx5+/− mice on a B6 background with wild type B6 mice (obtained from Jackson Laboratory) or B6;C3H mice bred in our lab. Progeny in Tables Tbx5+/− males on a B6 background. All procedures were approved by the Institutional Animal Care and Use Committee.Animals were maintained in a virus and antibody-free facility with food and water Tbx5+/−)  were kin3H-a/A-Ts71665Dn) Tbx5 genotyping, three primers designed to amplify either the wild type or null alleles were added together in each reaction [Tbx5 genotyping are as follows: 3 F2: 5′-CCCAGCGGCAGGCGTAGAC -3′; Loxp-F: 5′-GCAGCGCAGTCCTCACCAG -3′; Loxp-R: 5′-AAATTCCAACCCCTTCCACAGAT -3′. The PCR was done under the following cycling conditions: 94 °C 3 min,  for 35 cycles, 72 °C 10 min.We extracted genomic DNA from tail tips of mice by heating at 90 °C for 2 h in 10 mM NaOH, 0.2 mM EDTA and used for genotyping by PCR. Sequences of primers for Ts65Dn genotyping are as follows: C17F: 5′-GTGGCAAGAGACTCAAATTCAAC-3′; C16R: 5′-TGGCTTATTATTATCAGGGCATTT-3′; IMR5: 5′-AAAGTCGCTCTGAGTTGTTAT-3′; IMR6: 5′-GGAGCGGGAGAAATGGATATG-3′. For Ts65Dn genotyping, PCR was done under the following cycling conditions: 95 °C 3 min,  for 31 cycles, 72 °C 5 min. For Tbx5+/− crosses at postnatal day 0 (P0) within hours of birth. We euthanized pups and removed and fixed thoraxes in 10 % formalin for at least 48 h. Tissues were embedded in paraffin, sectioned at 7 μm and stained with hematoxylin/eosin using standard methods. The heart morphology in each animal was scored under a dissecting stereomicroscope  by at least two individuals blinded to genotype. Pictures were taken using the NIS-Elements Br software .We collected the progeny of Ts65Dn x Tbx5+/− mice . Virtual Histology™ is a micro-CT based method that allows for high resolution imaging. Specimens were stained in a proprietary solution before being scanned at 20 μm resolution with a volumetric micro-CT instrument. Using this method tissues are left intact and imaging analysis can be used to create “virtual” histological sections.Virtual Histology™ was performed on six Ts65Dn;We examined 109 genes that are trisomic in Ts65Dn mice for localization of expression in the heart using the following databases: EMAGE gene expression database , VisiGenTbx5 was quantified using a Taqman FAM-labeled pre-designed assay (Mm0195728_m1) from Applied Biosystems and normalized to a Gapdh VIC-labeled assay (Mm99999915_g1) using Taqman Gene Expression master mix. POWER SybrGreen master mix was used for all other expression assays . Primers used are as follows: Adamts1-F: 5′-CACGTGTGACACTCTCGGAA-3′; Adamts1-R: 5′-CGTGCGGCATGTTAAACACA-3′; Gapdh-F: 5′-TGCACCACCAACTGCTTAG-3′; Gapdh-R: 5′-GATGCAGGGATGATGTTC-3′; Pitx2-F: 5′-GCAGCCGTTGAATGTCTCTTC-3′; Pitx2-R: 5′-GTCCGTGAACTCGACCTTTTT-3′. PCR was done under the following cycling conditions: 95 °C 15 min,  for 40 cycles, followed by a melt curve analysis between 95 and 60 °C. Expression of candidate genes was normalized to Gapdh. Fold change expression values were determined using the delta delta Ct method [We extracted total RNA from the hearts of embryos using the RNeasy mini kit . cDNA synthesis was carried out with the First-Strand cDNA synthesis kit  using 1 μg of total RNA as template. PCR was carried out on a 7500 Real-Time PCR System . t method .Tbx5 cDNA clone from the I.M.A.G.E. consortium and subcloned it into the pcDNA3.1+ vector , putting it under the control of the CMV promoter. NIH-3T3 cells were transfected with the pcDNA-Tbx5 construct using Lipofectamine 2000 . Stably transfected cell lines were selected with 1 mg/ml Geneticin . We maintained the cells in Dulbecco’s Modified Eagle Medium with high glucose (4.5 g/L) , 10 % fetal bovine serum, 1 mg/ml Geneticin, 1X glutamine and penicillin/streptomycin at 37 °C and 5 % CO2.We obtained a Adamts1 that was bound by TBX5 in a ChIP study [Adamts1 promoter. The following primers were used to mutate the putative TBE site and insert an EcoRI restriction site, Primer 1: 5′-CACAGCTCGTCACTCTGGGAATTCAAGACGCCGAAACAGCGCTG-3′; Primer 2: 5′-CAGCGCTGTTTCGGCGTCTTGAATTCCCAGAGTGACGAGCTGTG-3′. We transfected the constructs into NIH-3T3 cells constitutively over expressing Tbx5 . Luciferase levels were measured in a 1450 Wallac Jet MicroBeta liquid scintillation and luminescence counter  using the Dual Luciferase Assay Kit .We amplified the portion of the promoter region of IP study , by PCR in situ hybridization using traditional dioxigenin (DIG)-labeled RNA probes detected with alkaline phoshatase (AP)-conjugated anti-dioxigenin antibody using nitro blue tetrazolium chloride / 5-bromo-4-chloro-3-indolyl-phosphate toluidine-salt (NBT/BCIP) substrate. DIG RNA labeling mix, anti-DIG-AP antibody, blocking reagent, NBT/BCIP were all purchased from Roche Applied Science  and used according to the manufacturer’s instructions with minimal modifications. The Pitx2 antisense and sense probes were described earlier [E13.5 embryos were fixed in 4 % paraformaldehyde for 1 h, rinsed in 1X PBS three times for 5 min and then dehydrated in 25, 50 and 70 % ethanol for 1 h each. After dehydration the embryos were stored at −20 °C in 70 % ethanol until embedded in paraffin and sectioned according to standard methods. We performed  earlier . Images t-test using delta Ct values. All tests were 2-tailed and P < 0.05 were considered significant.The incidence of various heart defects for different genotypes was compared by Fisher’s test using GraphPad Prism version 5. A 2×2 contingency table was made comparing the two groups for each separate phenotype. The relative quantification of gene expression from different genotypes was compared by the student’s


P = 0.010), as assessed by low E-cadherin and high vimentin expression. The overall survival of PD-L1+ patients with EMT features was significantly worse than those without EMT features (P = 0.007). In an independent validation cohort (N = 91), as well as in HNSCC cases of The Cancer Genome Atlas (TCGA) and the Cancer Cell Line Encyclopedia, high PD-L1 expression was also associated with the high probability of an EMT signature, referred from the GEO dataset, GSE4824. Survival analysis confirmed PD-L1+/EMT+ patients had a poorer prognosis than PD-L1+/EMT- patients in the TCGA cohort. PD-L1 positivity can thus be divided into two categories according to the absence or presence of EMT. PD-L1 expression is also independently associated with EMT features in HNSCC.Virus-associated malignancies and sarcomatoid cancers correlate with high PD-L1 expression, however, underlying mechanisms remain controversial. We evaluated the correlation between PD-L1 expression and epithelial-mesenchymal transition (EMT) in head and neck squamous cell carcinomas (HNSCC). Tumor tissues from 50 patients with HNSCC were evaluated for PD-L1 by immunohistochemistry, which showed 32 (64.0%) were PD-L1 positive (PD-L1+). Interestingly, PD-L1 expression was significantly associated with EMT ( Discovery of programmed death-ligand 1 (PD-L1) expression in tumors has encouraged research toward more efficient immunologic methods of conquering cancer , 3. The CD274 gene amplification at 9p24.1, which encodes PD-L1 [CDH1) down-regulation and vimentin (encoded by VIM) up-regulation, also correlated with PD-L1 induction [Viruses and epithelial-mesenchymal transition (EMT) are associated with high PD-L1 expression . PD-L1 ues PD-L1 . Anotheres PD-L1 –16. PD-Les PD-L1 . Moreovees PD-L1 . EMT chanduction . HoweverHead and neck squamous cell carcinoma (HNSCC) is a suitable model to investigate the clinicopathologic features associated with PD-L1 up-regulation. Recent advances in genomics have shown that mutational HPV-positive and -negative tumor profiles, which have retained and lost p16 expression, respectively, clearly differ –22. AlthIn the current study, we analyzed 1) PD-L1 expression according to the EMT and p16 statuses of patients with HNSCC, and 2) their clinical implications. A gene expression signature associated with EMT was obtained from the publicly available database, GEO4824 . This siP = 0.013, Figure P = 0.559). The vimentin H-score was significantly higher in PD-L1+/p16- compared with PD-L1-/p16- patients (P = 0.014). However, this trend was not observed in p16+ patients . The number of patients allocated to PD-L1-, PD-L1+/EMT-, and PD-L1+ /EMT+ groups were 32, 32, and 27, respectively , where PD-L1 positivity was significantly higher in patients with EMT-positive tumors  or progression-free survival . However, p16-/EMT+ displayed significantly higher PD-L1 expression compared to p16-/EMT- or p16+/EMT- . However, OS rates were not significantly different, although the 3-year OS rate differed between PD-L1+/EMT+ (40.2%) and PD-L1+/EMT- .OS and PFS rates for PD-L1+/EMT+ patients were significantly worse compared to those for PD-L1- patients from TCGA  and paranasal sinus cancer (N = 2) were included; of these, two patients showed a non-keratinizing type pathology. Therefore, caution is needed in the direct application of the current results to further clinical studies. However, significant statistical results were obtained for the independent validation cohort, as well as for TCGA and the CCLE open databases. The clinical outcomes of anti-PD-1/PD-L1 inhibitor therapy is lacking in this study, although overall prognosis according to PD-L1 expression and its up-stream mechanism should be understood before researching clinical outcomes of the target agent. Although the prognostic impact of PD-L1 expression was not reported, we clearly showed PD-L1 up-regulation according to two distinct up-stream pathways, EMT and HPV/p16, influenced the overall prognosis. The association of EMT with PD-L1 in human cancer is a novel finding in this study.In conclusion, PD-L1 expression is associated with EMT, an independent up-stream pathway distinct from HPV/p16 association. EMT-associated PD-L1 expression confers a significantly poorer prognosis compared to PD-L1 expression not associated with EMT. Clinical investigations using anti-PD-1/PD-L1 inhibitors in patients with EMT-associated PD-L1 up-regulation are warranted.Medical records were retrospectively reviewed for patients diagnosed with locally advanced HNSCC and treated at Seoul National University Hospital between December 2004 and November 2012. Fifty patients with paraffin-embedded tumor samples were included. Firstly, we analysed the association of PD-L1 and EMT in 50 HNSCC patients (training cohort). To confirm preliminary findings, we enrolled a further 91 HNSCC patients , and performed similar analyses.Treatment decisions were determined by a multidisciplinary team , 45. Pat® mAb . IHC was performed using the Ventana Benchmark XT system . When tissue sections showed diffuse and strong nuclear and cytoplasmic staining in ≥ 70% of tumor cells, this was considered positive for p16 [Representative, formalin-fixed, paraffin-embedded tissue blocks from each case were submitted for immunohistochemistry (IHC) using the following antibodies: mouse anti-p16 (E6H4) monoclonal antibody , mouse anti-E-cadherin (36B5) mAb , mouse anti-vimentin (V9) mAb  and rabbit anti-PD-L1 (E1L3N) XP for p16 . For E-chttp://linus.nci.nih.gov/BRB-ArrayTools.html) was used in the analysis of gene expression data [P values were two-sided, and considered significant if P < 0.05. A false discovery rate were applied to control type I errors. All statistical analyses and data generation were carried out using R version 3.1.3 (http://www.r-project.org) and STATA version 12 .The BRB-ArrayTools software program (ion data . A heatmion data . To asseion data  and cellion data , 75 geneion data , and a pion data , 50. Briion data . The robion data . A compa


QLP and LLP) as a function of carrier density are proposed, and the control parameters are defined. Finally, the proposed analytical model is compared with experimental data which indicates good overall agreement.Graphene is an attention-grabbing material in electronics, physics, chemistry, and even biology because of its unique properties such as high surface-area-to-volume ratio. Also, the ability of graphene-based materials to continuously tune charge carriers from holes to electrons makes them promising for biological applications, especially in lipid bilayer-based sensors. Furthermore, changes in charged lipid membrane properties can be electrically detected by a graphene-based electrolyte-gated graphene field effect transistor (GFET). In this paper, a monolayer graphene-based GFET with a focus on the conductance variation caused by membrane electric charges and thickness is studied. Monolayer graphene conductance as an electrical detection platform is suggested for neutral, negative, and positive electric-charged membrane. The electric charge and thickness of the lipid bilayer ( The cAlthough the phospholipid bilayer frequently works well, it can be damaged, and some superfluous materials can penetrate it. Phospholipids have two ends; the first is hydrophilic and attracts water; and the second is hydrophobic and resists water . As the G-Vg characteristic of the graphene-based biosensor. A monolayer graphene-based GFET with a focus on the conductance variation caused by membrane electric charges and thickness is studied. Monolayer graphene conductance as an electrical detection platform is suggested for neutral, negative, and positive electric membrane. The electric charge and thickness of the lipid bilayer (QLP and LLP) as a function of carrier density are proposed and the control parameters are defined.Some smaller molecules, such as carbon dioxide and oxygen, can pass through without difficulty on their own, but larger molecules such as water, sodium, or magnesium cannot pass easily . The intE (k) relation of the GNR is obtained as [The monolayer graphene in an electrolyte-gated biomimetic membrane graphene transistor with a ballistic channel is assumed to monitor the changes in membrane integrity. High-carrier mobility is reported in experiments on the graphene, which is thought to be due to the totally ballistic carrier transportation in the graphene, which leads to a higher transmission probability. By applying the Taylor expansion on graphene band energy near the Fermi point, the ained as .kx is the wave vector along the length of the nanoribbon and β is quantized wave vector given by [M(E) at a given energy which is dependent on the sub bands location can be calculated. By taking the derivatives of wave vector k over the energy E (dk/dE), the number of the mode M(E) is written aswhere given by . Based oL is the length of the nanoribbon. A higher transmission probability causes a higher carrier conductance from source to drain, as provided by the Boltzmann transport equation [where equation ,3:q is the electron charge, Planck's constant is shown by h, E is the energy band structure, M(E) is the number of modes, f is the Fermi-Dirac distribution function and T(E) is the transmission probability. On the other hand, because of the ballistic transport T, the possibility of one inserted electron at one end that can be conveyed to other end is considered equivalent to one (T(E) = 1). The number of modes in accordance with the Landauer formula with respect to the conductance of monolayer graphene can be written aswhere l, k is the wave vector, and where the length of the graphene channel is shown with parameter x = (E - Eg)/kBT and the normalized Fermi energy is η = (EF - Eg)/kBT. Thus, the general conductance model of single-layer graphene obtained is similar to that of silicon reported by Gunlycke [VDS). As shown in Figure where Gunlycke . AccordiVg,min to measure the smallest conductance of the graphene layer, which is identified from the transfer characteristic curve. In total, the Vg,min shift (at the Dirac point) can be considered as a good indicator for lipid bilayer modulation and measurement. Nevertheless, the magnitude of the voltage shift from both positive and negative lipids is comparable when this shift is measured from the position of the minimum conductivity of bare graphene. As shown in Figure Vg,min on the device, and the hole density value can be estimated as decreasing. A negatively exciting membrane demonstrates a very small enhancement in conductivity and a positive change in the Dirac point compared with that of exposed graphene.This is because of an enhancement in the remaining pollution charges caused by the negatively charged membrane. A detection-charged lipid bilayer can be obtained based on a detectable Dirac point shift. In light of this fact, the main objective of the current paper is to present a new model for biomimetic membrane-coated graphene biosensors. In this model, the thickness and the type of coated charge as a function of gate voltage is simulated and control parameters are suggested. Subsequently, to obtain a greater insight into the role of both the thickness and the type of lipid bilayer, GFET modeling is employed to identify the relationship between the conductance and the voltage of the liquid gate, where two electrodes of the sensor, as shown in Figure As depicted in Figure α) and thickness factor (β) are suggested. Subsequently, for better understanding of the role of the lipid bilayer, FET modeling is employed to obtain an equation describing the conductance, electric charge, and thickness, where the suggested structure of the GFET is shown in Figure GLP is considered to be a function of electric charge (QLP) as follows. GLP = GNeutral + αQLP where electric charge factor (α) is assumed, GLP is the channel conductance of graphene with biomimetic membranes of different surface charges, and QLP is the electrical charge of the membrane. Consequently, the supposed conductance model of the graphene-based GFET channel can be written as.Different ions can be adsorbed by changes in the membrane's electric charge and thickness, and subsequently, the sensor will be capable of attracting the ions in the solution which have caused a transformation in the conductance of the graphene-based biosensor. Dependent upon the channel conductance in the biomimetic membrane-coated graphene biosensor, it is concluded that GLP is a function of electric charge and thickness, where GLP is the channel conductance after adding the lipid bilayer. The focus of the present paper is to demonstrate a new model for GFET to measure changes in the membrane's electric charge and thickness. In other words, the conductance of the GFET device as a function of different electric charges and thicknesses is simulated and an electric charge factor  and membrane thickness as follows:Furthermore, the proposed model is strongly supported by the experimental data. According to Figure β) and LLP are the thickness parameter and thickness of the adsorbed lipid bilayer, respectively. In the non-saturation region, the GFET conductance model is involved as a result of gate electrical energy and the perfect conductance-voltage related to the graphene channel of the GFET device, which leads to the modified conductance as:where  indicates the same trends as those reported by [Vg,min with increasing membrane thickness. Comparison of the experimental data depicted with the theoretical data in Figure Vg,min.In Figure GLP and LLP parameters, respectively, in agreement with the reported data which is shown in Table Vg,min did not shift further at greater membrane thicknesses due to the saturation current density of the injected carrier concentration by the charged lipid bilayer.In the suggested model, differently charged lipid bilayers and membrane thicknesses are demonstrated in the form of m,min belongs to the graphene-based biosensor, the control parameter with respect to the iteration method is suggested as:According to the saturation region of the presented conductance model and given that gl1 = 0.4157 and l2 = -0.543. In addition, α for the neutrally, negatively, and positively charged membrane is assumed to be 0, 1, and -1, respectively. Consequently, the justified model for the interaction of charged impurity and the consequence of charged lipid membranes in a biomimetic membrane-coated graphene biosensor is proposed aswhere Vg,min of the ambipolar FET occurs due to the electronic devices on both the n-doping and p-doping materials.The proposed model, coupled with the experimental data, is shown in this work to confirm that the conductivity of the graphene-based biosensor is changed by the electric charge and membrane thickness of the lipid bilayer. In a nutshell, electrolyte-gated graphene field-effect transistor structure was used after chemical vapor deposition (CVD) as the electrical transduction stage because of its high electrical conductivity, optical transparency, and large area, given the likelihood of manufacturing a dual-mode optical and electrical detection system for detecting the changes of membrane properties. Based on what has been discussed, one could firmly claim that, in response to changes of the charged lipid membranes and charges of biomimetic membranes of different thicknesses, a significant shift in QLP) and thickness of the adsorbed lipid bilayer (LLP) is presented. A dramatic decrease in the minimum conductance related to the gate voltage  by both changing the electrical charge from negative to positive and decreasing the lipid thickness has been reported. In the presented model, the Vg, min variation based on the adopted experimental data as an electrical detection platform is considered and the sensor control parameters are defined. The presented model confirms the reported experimental data and in addition facilitates the employment of alpha and beta as biosensor control parameters to predict the behavior of graphene in graphene-based biosensors.The emerging potential of nanostructured graphene-based biosensors in the highly sensitive and effective detection of single-base polymorphism or mutation, which is thought to be the key to diagnosis of genetic diseases and the realization of personalized medicine, has been demonstrated. In a lipid bilayer-based biosensor, the graphene carrier concentration as a function of the lipid bilayer can be modeled. In this research, the total conductance of graphene as a function of the electric charge (The authors declare that they have no competing interests.MJK wrote the manuscript and contributed to the analytical modelling of the presented FET via MATLAB software. Dr. FKCh and Dr. MTA revised the manuscript and coordinated between all the contributors. HKFA, MR, and AH organized the final version of the manuscript. All authors read and approved the final manuscript.