Sporadic inclusion body myositis (sIBM) is the commonest idiopathic inflammatory muscle disease in people over 50 years old. It is characterized by slowly progressive muscle weakness and atrophy, with typical pathological changes of inflammation, degeneration and mitochondrial abnormality in affected muscle fibres. The cause(s) of sIBM are still unknown, but are considered complex, with the contribution of multiple factors such as environmental triggers, ageing and genetic susceptibility. This review summarizes the current understanding of the genetic contributions to sIBM and provides some insights for future research in this mysterious disease with the advantage of the rapid development of advanced genetic technology. An international sIBM genetic study is ongoing and whole-exome sequencing will be applied in a large cohort of sIBM patients with the aim of unravelling important genetic risk factors for sIBM. Sporadic inclusion body myositis (sIBM) is the commonest form of idiopathic inflammatory myopathy among individuals aged over 50. It has The clinical diagnosis of sIBM is confirmed by muscle biopsy assisted by the clinical phenotype, electromyography (EMG), determination of serum muscle-enzyme levels and muscle imaging with MRI. CurrentsIBM is a complex disease. The primary pathogenic mechanism is still hotly debated and it is believed that many factors including environmental factors contribute to the disease process. There are many clinically and/or histologically sIBM-like muscle diseases with known genetic defects, which may provide clues to understand the pathogenesis of sIBM. In this review we will comprehensively summarize known and candidate genetic susceptibility risk factors for sIBM.. Human leukocyte antigen (HLA) class II genotypes were reported in a strong association with some families, particularly the DR3 allele (DRB*0301/0302) in four western families[DR15(2)/DR4 (DRB1*1502/0405) in two Japanese sisters[There are several IBM case reports of two or more affected siblings in the same family-26 and ofamilies,25 and tHereditary inclusion body myopathies (hIBM) include several autosomal-recessive and autosomal-dominant muscle disorders with various clinical presentations, but with a number of pathological features similar to those of sIBM, including rimmed vacuoles, protein accumulations and tubulofilaments on electron microscopy. Patients with hIBM can be distinguished from sIBM by the earlier age of onset, rarity of inflammatory features and negative MHC class I expression, hence the term ‘myopathy’ instead of ‘myositis’. The hIBBeyond the above conditions, there are various other neuromuscular disorders showing rimmed vacuoles in the muscle tissue, which make the differential diagnosis of sIBM even wider. Mutated genes have also been found in these called ‘rimmed vacuolar myopathies’, such as oculopharyngeal muscular dystrophy (OMIM#164300), X-linked recessive Emery-Dreifuss muscular dystrophy OMIM#310300), limb girdle muscular dystrophy (LGMD) type 2G (OMIM#601954), LGMD type 1A (OMIM#159000), LGMD type 1G (OMIM#609115), and rigid spine syndrome (OMIM#602771). Some in0300, limMYH2 (gene associated with hIBM3) significantly increased the risk of developing sIBM (RR = 12.2). However, none of the patients with that variant reported joint contractures or external ophthalmoplegia, a family history of IBM or consanguineous marriage[MYH2 gene mutation described in non-familial cases. Immunohistochemical studies of myosin heavy chain isoforms showed atrophy or loss of muscle fibres expressing myosin heavy chain IIa or IIx, which may provide important clues for establishing the pathogenicity of this variant. In the same study, a novel missense mutation c.1719G > A (p.V566M) in the LIM domain binding 3 gene , coding for Z-band alternatively spliced PDZ-motif-containing protein, was reported in one of the patients[LDB3 mutations might cause abnormalities in the Z-band in skeletal muscle resulting in the disease observed in this patient. Further studies with a larger number of cases will be needed to clarify the contribution of these genes to the pathogenesis of sIBM. In addition, investigations should be carried out on more hereditary myopathies candidate genes, including α-B crystalline (CRYAB), dystrophin (DMD) and myotilin (MYOT) in the future.Overall, these common features raise the possibility that sIBM and these diseases share certain etiologic factors. Indeed, a recent study, reportemarriage. This wapatients. This fiCurrently there are two most popular theories for the pathogenesis of sIBM – an autoimmune pathway and a degeneration pathway. In addition, ageing is also considered an important factor contributing to mitochondrial abnormalities Figure .+ T cells and T cell-mediated cytotoxicity, which ultimately result in damage or death of muscle fibres[The theory of an inflammation pathway stands for viral or inflammatory triggering factors leading to the clonal expansion of CD8e fibres. This the fibres. Degenere fibres. The dege fibres.However, sIBM is poorly responsive to even vigorous immunosuppression. Even where there is histopathological evidence that inflammation was reduced, this was not accompanied by clinical improvement. Therefore, some investigators are supporting a degenerative hypothesis over inflammation as the primary pathogenesis of sIBM. The identification of aberrant protein aggregates in sIBM vacuolated muscle fibres has shown the remarkable parallels of those features in brain tissue of Alzheimer’s disease (AD) and Parkinson’s disease with Lewy bodies. This theory suggests that inflammation is secondary to the degeneration-associated processes in sIBM muscle fibres includinDespite sIBM not being an inherited Mendelian disease, multiple genetic risk factors are being shown likely to play important roles in the development and progression of sIBM. A list of possible susceptibility genes that could be important candidate genes for understanding the pathogenesis of sIBM is shown in Table HLA-DR3 and the extended 8.1 ancestral haplotype (AH) with sIBM[HLA-A*01, -B*0801, -DRB1*0301, -DRB1*0101, -DRB1*1301, -DQB1*0201, -DQA1*05[HLA-DRB1*0301 and *1301 also influence on the age at onset and the severity of muscle weakness[HLA-DRB1 locus may contribute to both disease susceptibility and clinical phenotype. Other alleles and haplotypes have also been associated with increased risk of sIBM, including the 35.2AH  in Caucasians[HLA-B*5201, DRB1*1502) in Japanese[HLA-DR53 and HLA-DRB1*0401, *0701, *0901, *1101, *1501, HLA-DRB1*0301/*0401 and HLA-DRB1*0301/*0701 diplotypes, and HLA-DRB1*1501, were found protective for sIBM[Garlepp and colleagues first reported the strong association of ith sIBM. This as -DQA1*05-65. Furtucasians and the Japanese. Interesfor sIBM,65,67.HLA-DR and C4[HLA-DRB1 and pre-B-cell leukemia homeobox2 [BTNL2) to telomeric of HLA-DRB1 including three protein-coding genes – part of BTNL2, HLA-DRA and HLA-DRB3[NOTCH4; a transmembrane receptor which regulates cell fate decisions), advanced glycosylation end product-specific receptor gene , testis-specific basic protein gene , and G-protein signalling modulator 3 gene [A potential region of MHC-associated susceptibility genes was first defined between DR and C4, and thes HOX12). A furth HLA-DRB3. Other g as G18),63.NOTCH4 gene polymorphisms (rs422951 and rs72555375) have shown a strong association (OR > 2) with sIBM in two independent Caucasian cohorts[Two  cohorts. These v cohorts.The MHC contains important genetic contents which have been reported as being associated with sIBM and among them 8.1AH which is predominant in Caucasians, supporting an immune-mediated mechanism underlying sIBM. Identification of the actual genes within the MHC will be crucial to understand the role of MHC in the pathogenesis of sIBM.NT5C1A) was recently identified in sIBM by two independent groups[NT5C1A and NT5C1B genes[An autoantibody against the cytoplasmic 5′-nucleotidase 1A , and AβPP-mRNA in both sIBM and hIBM muscle fibres[APP gene in sIBM have not been fully understood. The fact that mutations in exons 16 and 17 of APP gene have been found to be pathogenic in AD, resulting in early-onset familial AD, suggests that mutations in this gene might also play a role in sIBM. In line with a previous small study[APP mutations were found in our 58 sIBM cases.It has been widely recognized that there are abnormal accumulations of beta-amyloid (Aβ) protein, N-terminal and C-terminal of beta-amyloid precursor protein  and presenilin 2 (PSEN2) genes are also found to be responsible for early onset familial AD, with mutations in PSEN1 gene as the most common cause[PSEN1 and PSEN2 have not been investigated in either sIBM or hIBM cases.Abnormal accumulation of presenilin proteins occurs in the brains of both sporadic and familial AD patients. Autosomal dominant inheritance of mutations in presenilin 1 [DYSF gene might link to the pathogenesis of sIBM though the possible mechanisms is unknown[Dysferlin is a modular type II transmembrane protein newly identified as a binding partner of AβPP that co-aggregates with Aβ42 in sIBM muscle fibres. In norm(LGMD2B). Similar(LGMD2B). In addi unknown.APOE ϵ4 allele is recognized to be an important risk factor for developing late-onset AD. Several studies have been carried out to investigate the roles of APOE alleles in sIBM. Only one study of a cohort of 14 Australian sIBM patients suggested that ϵ4 allele may be a susceptibility factor for the development of sIBM[APOE ϵ4 allele as a susceptibility factor for developing sIBM. However, the authors demonstrated a non-significant trend towards an earlier age at onset in patients with the APOE ϵ2 allele[ApoE protein deposits but not mRNA have been identified in both sIBM and hIBM muscle fibres,83. The  of sIBM. However of sIBM,51,85-87 of sIBM, includi2 allele.APOE ϵ4 allele. Interestingly, we found a statistically significant decreased frequency of APOE genotype ϵ2/ϵ3  in the patient group compared with our British control individuals (unpublished data), and thus we found an association between ϵ2 allele and a reduced risk of developing sIBM  is consistent with the above meta-analysis with respect to the association with MAPT) gene mutations have been identified in many different tauopathies including AD, frontotemporal dementia and Parkinsonism linked to chromosome 17 (FTDP-17), progressive supranuclear palsy (PSP) and corticobasal degeneration (CBD). The majority of them affect exons 9–13[MAPT mutations, a common extended haplotype (H1) in MAPT appears to be a risk factor for sporadic PSP[MAPT intron 9 is inherited as a part of the less common H2 sub-haplotype[del-In9) in 81 sIBM DNA cases and 159 British healthy controls, but no significant differences were found between the two groups.Accumulation of phosphorylated-tau is widely known to be another major protein present in sIBM muscle aggregates, which is strikingly similar with those found in taupathies which share hyperphosphorylated deposited tau protein in the brain. Differeons 9–13. In addiadic PSP. A 238 baplotype. We analSERPINA3, previously known as ACT) is considered a major acute-phase protein and belongs to the serpin superfamily specifically inhibiting serine proteases. Similarly to AD brains, abnormal accumulations of α1ACT, located closely with β-amyloid protein containing ubiquitinated amyloid plaques, have been described in sIBM muscle[SERPINA3/AA (rs4934) was found to alter the AD risk associated with APOE ϵ4/ϵ4 genotype[APOE ϵ4/ϵ4 genotype.Alpha-1-antichymotyrpsin  gene was more prevalent in sIBM , when compared with controls[Prion protein and mRNA  have been described as being abnormally accumulated in vacuolated muscle fibres in both sIBM and hIBM,93. Lampcontrols. But thicontrols and our TARDBP have been detected in both sporadic and familial forms of ALS[TARDBP gene have not yet been examined in a sIBM.A predominantly nuclear heterogeneous nuclear ribonucleoprotein (hnRNP) TAR DNA-binding protein-43 (TDP-43) is found as a major component of ubiquitinated inclusions in amyotrophic lateral sclerosis (ALS) and frontotemporal lobar dementia (FTLD-U). Mutations in s of ALS. Severals of ALS,96-98 has of ALS. Similars of ALS. The simhnRNPA1 and hnRNPA2B1 have been identified in familial multisystem proteinopathy with similar myonuclei depletion and sarcoplasmic aggregates[Abnormalities in the distribution of two other hnRNPs, hnRNPA1 and hnRNPA2B1, were identified in sIBM muscle fibres. Similargregates. These gC9orf72 has been identified as the most common genetic cause of familial ALS, FTLD or a combination of both phenotypes and TDP-43-based pathology (FTLD-TDP)[C9orf72 may involve disrupted protein degradation[C9orf72 might also play a role in the pathogenesis of sIBM. Fifty-eight sIBM cases were analysed for C9orf72 repeat by our group, but no expanded repeats were found.An expanded GGGGCC hexanucleotide repeat in the noncoding region of TLD-TDP). In a reradation. TherefoTYMP, previously known as ECGF1), solute carrier family 25  member 4 gene , chromosome 10 open reading frame 2 (C10orf2) and polymerase gamma 1 (POLG1), which are important for mtDNA maintenance and replication[Mitochondrial abnormalities are another important pathological feature in sIBM muscle biopsies, consisting of ragged-red fibres and mostly showing enzyme histochemical deficiency of COX activity. These changes are more prevalent in sIBM than in polymyositis, dermatomyositis and normal ageing muscle fibres. It is tlication. Putativlication,104. ButHLA-DR3 positive[HLA-DR3, which is in a genomic region strongly associated with sIBM. Further studies are required to investigate whether this variant plays a pathogenic role in sIBM and/or its possible interaction with other genetic factors.In addition to mtDNA deletions, mutations at mtDNA nucleotide positions 3192, 3196, 3397 and 4336, which are associated with late-onset AD, are possible risk factors for sIBM. In sIBM cases only the frequency of the common 16311C variant has been found more frequent than in AD and controls, but these differences were not statistically significant. Interespositive, suggestTOMM40) which is adjacent to and in linkage disequilibrium with the APOE locus on chromosome 19, has been implicated in AD[TOMM40 encodes an outer mitochondrial membrane translocase facilitating the transport of unfolded proteins such as amyloid-β from the cytosol into the mitochondrial intermembrane space[TOMM40 gene together with the APOE genotypes has been shown to influence disease susceptibility of AD[APOE ϵ3 allele with a very long (VL) polyT repeat alleles in TOMM40 had reduced risk of sIBM compared to controls, and this was also associated with a later age at onset of symptoms[TOMM40 and/or APOE to influence disease susceptibility. This also comes to a hypothesis that genetic variants of TOMM40 could be associated with altered mitochondrial pore function and transport of proteins into mitochondria. This could result in energy metabolism changes and increased reactive oxygen species formation, further contributing to the impairment in mitochondria and degeneration in muscle fibres[Recently a gene called ‘Translocase of Outer Mitochondrial Membrane 40’ (ed in AD. TOMM40 ne space. A polyTty of AD. It has symptoms. The rs1e fibres.TREM2 were recently found to be associated with increased risk of Alzheimer’s disease by using whole-genome and whole-exome sequencing (WES)[HLA gene, fragment analysis for repeat variants, and comparative genomic hybridization (CGH) analysis for copy number variants which cannot be detected by WES.Over the past five years, next-generation sequencing technologies have already shown considerable promises in the identification of rare variants associated not only with Mendelian but also with complex diseases. For example, rare variants in ng (WES). Up to rng (WES). Therefong (WES), but no In the future, besides genetic defects identified in sIBM, RNA sequencing in muscle tissue and lymphoblast lines will identify transcriptomic profiles and expression quantitative trait loci (eQTL) in sIBM and other inflammatory myopathies. The application of RNA sequencing will also allow us to study and dissect splicing abnormalities which may not be detected by exon arrays. Investigating the transcriptomic and genomic factors will be important in revealing dysregulated molecular pathways in these disorders, identifying a diagnostic signature in blood and revealing biomarkers for future therapeutic strategies. A searchable database to the research community is also planned to build up to integrate WES, RNA sequencing and eQTL data, further to aligned with the biobank database with disease progression, response to treatment and muscle imaging.In conclusion, sIBM remains an enigmatic muscle disorder even though it has been about half a century since the first case was described. The primary pathogenesis of sIBM is still controversial, but it is more likely to due to multifactorial mechanisms, including immune changes, degenerative changes, mitochondrial abnormalities and the ageing milieu in the muscle fibres, which involve a complex interaction between environmental triggers  and genetic susceptibility. Despite the strikingly phenotypic similarities between sIBM muscle fibres and the brains of AD, research on the candidate genes encoding the deposited proteins have not shown any positive result so far. Genes located within the MHC region are still the most strongly associated candidates with sIBM. Genes linked to hIBM and pathologically sIBM-like myopathies may provide important clues for sIBM genetic research. Effective WES and RNA sequencing analysis of a large sIBM cohort will significantly contribute to our knowledge in this under-explored area.References for this Review were identified by searches in PubMed between 1969 and May 2014. The search terms “inclusion body myositis”, “inclusion body myopathy”, “IBM”, “rimmed vacuolar myopathy”, and “inflammatory myopathy” were used. References were also identified from the bibliography of the identified articles. Only papers published in English or with available English translations of relevant data were reviewed. The final reference list was generated on the basis of relevance to the topics covered in this Review.The authors declare that they have no competing interests.QG contributed to the literature search and to drafting the manuscript. All authors contributed to the critical revision process and approved the final manuscript. MGH, HH, and PM are principal investigators of the International IBM Consortium Genetic Study (IIBMCGS).


With rising energy concerns, efficient energy conversion and storage devices are required to provide a sustainable, green energy supply. Solar cells hold promise as energy conversion devices due to their utilization of readily accessible solar energy; however, the output of solar cells can be non-continuous and unstable. Therefore, it is necessary to combine solar cells with compatible energy storage devices to realize a stable power supply. To this end, supercapacitors, highly efficient energy storage devices, can be integrated with solar cells to mitigate the power fluctuations. Here, we report on the development of a solar cell-supercapacitor hybrid device as a solution to this energy requirement. A high-performance, cotton-textile-enabled asymmetric supercapacitor is integrated with a flexible solar cell via a scalable roll-to-roll manufacturing approach to fabricate a self-sustaining power pack, demonstrating its potential to continuously power future electronic devices. Activated cotton textiles are promising for wearable energy storage due to their flexibility and high conductivity, but scalability is an issue. Here, the authors combine supercapacitors and solar cells on activated cotton textiles via scalable roll-to-roll assembly to make self-sustaining power packs. With ever-increasing global energy consumption and the depletion of fossil fuels, finding a sustainable and clean energy supply has become one of the most important scientific and technological challenges facing humanity todayCompared with lithium-ion batteries, the relatively low energy density of supercapacitors limits their wide application. Developing new electrode materials with well-defined nano-architecture and high active surface area is key to improving the electrochemical performance of supercapacitors. To date, according to the different mechanisms of charge storage, typical supercapacitor electrode materials can be categorized into electric double-layer capacitive materials including various carbon materials3567910IIx1-MIIIx(OH)2]x+[An−]x/n·mH2O, where MII and MIII are divalent and trivalent metal cations and An− are the charge-balancing anions1921et al. reported that nickel-aluminium LDH (Ni-Al LDH) deposited on nickel foam exhibits an ultrahigh specific capacitance of 2,123 F g−1 at 1 A g−1  film had a significantly enhanced specific capacitance of 2,682 F g−1 at 3 A g−1  and a capacitor-type electrode as power source 13g−1 ref. . In our Consumer demand for portable/wearable electronics has triggered a technological race to drive innovation in flexible mobile phones, bendable displays, electronic skin and distributed sensors34353738394344in situ on flexible ACTs to realize ACT/Co-Al LDH hybrid by a facile hydrothermal method. A highly conductive graphene coating is wrapped around ACT fibres to form ACT/graphene composite by a simple dipping, drying and reducing process. A flexible, all-solid-state asymmetric supercapacitor (ACT/Co-Al LDH//ACT/graphene) is assembled using the nanostructured ACT/Co-Al LDH as the positive electrode, ACT/graphene as the negative electrode, and PVA-KOH gel as both the solid-state electrolyte and separator. The asymmetric cell works synergically to achieve excellent electrochemical performance in terms of its working potential (1.6 V), energy density (55.04 Wh kg−1), power density (5.44 kW kg−1) and cycling stability . Moreover, we demonstrate a practical roll-to-roll manufacturing approach to combine the flexible asymmetric supercapacitor with a flexible solar cell to fabricate an integrated self-sustaining power pack, which is scalable for industrial manufacturing. Importantly, the assembled power pack can power a commercial light-emitting-diode (LED) continuously, with or without sunlight, demonstrating its potential for flexible, self-powered energy devices.Cotton textile, a source of flexible, ‘green', renewable, breathable clothing, has been shown to be an excellent wearable platform for constructing flexible energy storage devices as activated cotton textiles (ACTs) exhibit eminent flexibility and excellent conductivity46in situ on ACT fibres using a facile hydrothermal method (step ii), which resulted in a pink coating on the surface of ACT fibres. in situ anchored on ACT fibres not only enhances the contact between the active material and ACT substrate but also serves as a reservoir for electrolyte ions, shortening ion diffusion path and facilitating charge transfer, jointly enhancing the electrochemical performance.2 g−1. A lattice spacing of 0.26 nm (The structure and morphology of the Co-Al LDH nanosheets were further investigated by transmission electron microscopy (TEM) and X-ray diffraction (XRD). TEM inspection  reveals  0.26 nm  can be a 0.26 nm . The cor 0.26 nm  shows he 0.26 nm  shows a −1) were first prepared by direct conversion of a cotton T-shirt. As shown in π–π stacking or hydrogen bonding49d-spacing of ∼0.37 nm in the HRTEM image suggest the existence of defects in the graphene nanosheets. The crystalline nature of the graphene nanosheets was validated by the well-defined diffraction spots in the SAED pattern. More importantly, the ACT fibres showed a porous structure as revealed by the TEM image  compared with the ACT/graphite oxide with the ID/IG ratio of ∼1.03. The increased ID/IG ratio is ascribed to the re-establishment of graphene network during the reduction process, indicating smaller sizes, more defects and disordered structures of the graphene1453The as-prepared ACT/Co-Al LDH was used as the positive electrode, ACT/graphene as the negative electrode and the PVA-KOH gel as both the solid electrolyte and separator to construct a flexible asymmetric supercapacitor. The energy storage performances of the assembled cells and individual electrodes were studied by cyclic voltammetry (CV), galvanostatic (GV) charge/discharge and electrochemical impedance spectroscopy (EIS) in both two-electrode and three-electrode testing systems.−1 with the potential windows ranging from −0.2 to 0.6 V are shown in The three-electrode testing system was used to study the electrochemical reaction mechanisms of the as-prepared ACT, ACT/graphene and ACT/Co-Al LDH electrodes. The CV curves of the aforementioned electrodes at the scan rate of 25 mV s−1. Compared with the triangular-shaped charge/discharge curves of the ACT and ACT/graphene electrodes, the ACT/Co-Al LDH electrode exhibited two charge/discharge voltage plateaus, which are the pseudocapacitive plateaus from the Co-Al LDH. The specific capacitance values of the ACT, ACT/graphene and ACT/Co-Al LDH electrodes were measured to be 90.1, 136 and 977.3 F g−1, respectively. The CV curves of the ACT/Co-Al LDH electrode at the scan rates of 25, 50, 75 and 100 mV s−1 in 6 M KOH electrolyte solution are shown in −1 are 977.3, 863, 727 and 545 F g−1, respectively, with a capacitance retention of ∼55.8% even when the current density increased from 2.5 to 20 A g−1.−1. The rectangular-shaped CV curves of the ACT/graphene symmetric cell suggest that the ACT/graphene mainly worked as an electrochemical double-layered capacitor. The broad peak at 0.25 V of the ACT/Co-Al LDH symmetric cell indicates the Faradic reaction from the battery-type Co-Al LDH. The CV area of the ACT/Co-Al LDH//ACT/graphene asymmetric cell was much larger than that of the ACT/graphene//ACT/graphene and ACT/Co-Al LDH//ACT/Co-Al LDH symmetric cells, suggesting the larger specific capacitance of ACT/Co-Al LDH//ACT/graphene asymmetric cell resulting from the high accessibility of electrolyte ions. −1 for the ACT/Co-Al LDH//ACT/graphene asymmetric cell, ACT/Co-Al LDH symmetric cell and ACT/graphene symmetric cell at the current density of 12.5 mA cm−2, respectively. −1. The working voltage of the ACT/Co-Al LDH//ACT/graphene asymmetric cell can be extended to 1.6 V, which is essential for practical application. −1 at an operation window of 1.6 V. The current density increases with increasing scan rate. All the CV curves at different scan rates in −1 was achieved even when the current density increased from 7.5 to 50 mA cm−2, indicating the excellent rate performance of the ACT/Co-Al LDH//ACT/graphene asymmetric cell. The rate performance and corresponding coulombic efficiency of ACT/graphene//ACT/graphene and ACT/Co-Al LDH//ACT/Co-Al LDH symmetric cells as well as ACT/Co-Al LDH//ACT/graphene asymmetric cell are comprehensively compared in −2) in a wide range of current densities (7.5–50 mA cm−2). Whereas the ACT/Co-Al LDH symmetric cell showed poor rate performance (35% capacitance retention) and low coulombic efficiency (91.8% at 50 mA cm−2). Compared with the ACT/Co-Al LDH symmetric cell, the ACT/Co-Al LDH//ACT/graphene asymmetric cell exhibited not only improved capacitance retention (52.3% capacitance retention) but also enhanced coulombic efficiency (98.5% at 50 mA cm−2). At high charge/discharge rates the ions on electrode decreased rapidly with increasing rate, and the ions in the electrolyte diffused too slowly to satisfy the need of ions near the solid–liquid interface, leading to the decrease of capacitanceRate performance and coulombic efficiency are important factors for the real power application of supercapacitors. −1 at the power density of 387.9 W kg−1 and maintained 28.72 Wh kg−1 at the power density of 5.44 kW kg−1. For an in-depth understanding of the electrochemical behaviour of the assembled symmetric and asymmetric cells, the EIS tests were carried out on the aforementioned cells (Cdl) and the charge transfer resistance (Rct) at the electrode and electrolyte interface, corresponding to the charge transfer-limiting processRct was directly measured from the diameter of the semicircle arc in the Niquist plot. Clearly, compared with the ACT/graphene (1.1 Ω) and ACT/Co-Al LDH (1.89 Ω) symmetric cells, the relatively smaller Rct of the asymmetric cell resulted from the synergistic effect of the ACT/graphene and hierarchical ACT/Co-Al LDH architectures that facilitated the access of electrolyte ions to the active surface and shortened the ion diffusion path.Energy density and power density are important factors for evaluating the practical application of supercapacitors. ed cells . A simil−2 using a GV charge–discharge technique. An obvious capacity decay (∼35%) was observed for the ACT/Co-Al LDH symmetric cell, whereas there was a capacity increase (∼8%) for the ACT/graphene symmetric cell after 2,000 cycles. For the ACT/Co-Al LDH//ACT/graphene asymmetric cell, ∼16% decay of its original capacitance was noted after 2,000 cycles. The asymmetric cell exhibited linear and symmetrical characteristics in its charge/discharge curves, and no obvious ‘IR drop' was observed, indicating its low internal resistance . A flexible, all-solid-state asymmetric supercapacitor (ACT/Co-Al LDH//ACT/graphene) was assembled using the nanostructured ACT/Co-Al LDH as the positive electrode, ACT/graphene as the negative electrode and PVA-KOH gel as both the solid-state electrolyte and separator. The hierarchical flower-like Co-Al LDH nanoarrays with interconnected nano-petals and pompon-like nano-stamens provided a highly open and porous scaffold-like structure, facilitating the transportation of electrolyte ions. In addition, the wrapped graphene on porous ACTs rendered a highly accessible surface area and good electrical conductivity, endowing the flexible asymmetric supercapacitor with higher rate performance. The assembled ACT/Co-Al LDH//ACT/graphene asymmetric supercapacitor exhibited an excellent combination of electrochemical and mechanical performance. Moreover, we demonstrated a low-cost, roll-to-roll manufacturing approach to combine the flexible asymmetric supercapacitor with a flexible solar cell to build an integrated, self-sustaining power pack, which is scalable for industrial manufacturing. Importantly, such hybrid energy storage devices could continuously power a commercial LED, demonstrating a great potential for the future of self-powered nanotechnology.Two-dimensional battery-type Co-Al LDH nanosheets were anchored in situ on ACT fibres via a simple hydrothermal process. Briefly, 0.582 g of Co(NO3)2·6H2O, 0.518 g of Al(NO3)3·6H2O, 0.296 g of NH4F and 0.6 g of urea were dissolved in 36 ml distilled water. The resulting solution was transferred into a 50-ml Teflon-lined stainless autoclave with a piece of vertically suspended ACT (1 × 2 cm2) in the solution. Then, the autoclave was placed in an electric oven at 100 °C for 24 h. Finally, the as-prepared products were washed thoroughly with ethanol and distilled water and dried at 80 °C overnight to produce ACT/Co-Al LDH composite.All of the chemicals were used after purchasing without further purification. A piece of commercial cotton T-shirt was first cleaned using distilled water in an ultrasonic bath before activation. Activation of the cotton T-shirt was performed as described in detail in our previous report−1. A piece of ACT (1 × 2 cm2) was then soaked with the graphene oxide aqueous colloid. After drying at 80 °C for 6 h, the as-prepared ACT with graphene oxide coating was heated at 450 °C for 1 h with argon/hydrogen mixture gas  to reduce flexible ACT/graphene oxide to ACT/graphene-negative electrode.Graphite oxide was synthesized by exfoliating natural graphite flakes using a modified Hummers methodKα radiation . The Raman spectra of the ACT/graphene were recorded by a Renishaw InVia Raman microscope at 785 nm. Surface areas of the active materials were measured by physical adsorption of N2 at 77 K  and calculated by the BET method.The morphology and microstructure of the as-prepared products were characterized by SEM (FEI Quanta 650), TEM (JEOL 2000FX), HRTEM (FEI Titan), and atomic force microscopy (Nanoscope IIIa). The crystallographic structure of the synthesized materials was determined by a PANalytical X'Pert Pro Multi-Purpose Diffractometer equipped with Cu 2) and a saturated calomel electrode were used as the counter and reference electrodes, respectively. The mass of the ACT, ACT/graphene and ACT/Co-Al LDH was measured to be 2, 3.45 and 4.1 mg cm−2, respectively. The asymmetric supercapacitor used in this paper was assembled with two pieces of flexible, face-to-face electrodes (ACT/Co-Al LDH-positive and ACT/graphene-negative) separated by the solid-state, polymer PVA/KOH gel electrolyte (ACT/Co-Al LDH//ACT/graphene). The polymer gel electrolyte was prepared by mixing 3 g KOH and 6 g PVA in 60 ml deionized water by magnetically stirring the solution at 80 °C until the solution became clear. Then, the above solution was transferred into a flat Petri-dish to naturally solidify to PVA/KOH gel film electrolyte. Two pieces of flexible electrodes separated by the PVA/KOH gel film were used to assemble the flexible asymmetric cell. The as-obtained solid-state, polymer PVA/KOH gel film severed as both the separator and electrolyte. The electrochemical properties of the assembled flexible asymmetric cell were measured using a CHI 660E electrochemical workstation. CV, GV charge/discharge curves and EIS in the frequency range from 100 kHz to 0.05 Hz with an AC perturbation of 5 mV were used to evaluate the electrochemical performance of the flexible solid-state asymmetric supercapacitors. The integrated flexible energy conversion and storage unit was assembled by combining a piece of commercial flexible solar panel  with an energy storage unit consisting of two pieces of asymmetric cell connected in series using an electrical rolling machine . Two pieces of assembled asymmetric flexible supercapacitors were connected in series and then coated with double-side adhesive tapes. At the same time, the bottom surface of the flexible solar cell was also coated with double-side adhesive tapes. As illustrated in The electrochemical performances of the prepared ACT, ACT/graphene and ACT/Co-Al LDH electrodes were characterized by both three-electrode and two-electrode systems. The three-electrode tests were carried out in 6 M KOH aqueous electrolyte at room temperature. The prepared ACT, ACT/graphene and ACT/Co-Al LDH were used as the working electrodes, platinum foil , I (A), t (s), m (g) and ΔV (V) are the specific capacitance, the discharge current, the discharge time, the mass of the active material and the total potential window, respectively.where E) and power (P) density derived from GV tests were calculated from the following equations:Energy density (E (Wh/kg), C (F/g), V (V), P (W/kg) and t (s) are the energy density, specific capacitance, potential window, power density and discharge time, respectively.where How to cite this article: Gao, Z. et al. Cotton-textile-enabled flexible self-sustaining power packs via roll-to-roll fabrication. Nat. Commun. 7:11586 doi: 10.1038/ncomms11586 (2016).Supplementary Figures 1-3.


Activation of the HSR clears insoluble TDP-43 and increases cell survival.Insoluble TDP-43 inclusions are the pathological hallmark of ALS and tau-negative frontotemporal lobar degeneration. Chen  TARDBP) neuronal cytoplasmic inclusions are the pathological hallmark in ∼95% of amyotrophic lateral sclerosis and ∼60% of frontotemporal lobar degeneration cases. We sought to explore the role for the heat shock response in the clearance of insoluble TDP-43 in a cellular model of disease and to validate our findings in transgenic mice and human amyotrophic lateral sclerosis tissues. The heat shock response is a stress-responsive protective mechanism regulated by the transcription factor heat shock factor 1 (HSF1), which increases the expression of chaperones that refold damaged misfolded proteins or facilitate their degradation. Here we show that manipulation of the heat shock response by expression of dominant active HSF1 results in a dramatic reduction of insoluble and hyperphosphorylated TDP-43 that enhances cell survival, whereas expression of dominant negative HSF1 leads to enhanced TDP-43 aggregation and hyperphosphorylation. To determine which chaperones were mediating TDP-43 clearance we over-expressed a range of heat shock proteins (HSPs) and identified DNAJB2a  as a potent anti-aggregation chaperone for TDP-43. DNAJB2a has a J domain, allowing it to interact with HSP70, and ubiquitin interacting motifs, which enable it to engage the degradation of its client proteins. Using functionally deleted DNAJB2a constructs we demonstrated that TDP-43 clearance was J domain-dependent and was not affected by ubiquitin interacting motif deletion or proteasome inhibition. This indicates that TDP-43 is maintained in a soluble state by DNAJB2a, leaving the total levels of TDP-43 unchanged. Additionally, we have demonstrated that the levels of HSF1 and heat shock proteins are significantly reduced in affected neuronal tissues from a TDP-43 transgenic mouse model of amyotrophic lateral sclerosis and patients with sporadic amyotrophic lateral sclerosis. This implies that the HSF1-mediated DNAJB2a/HSP70 heat shock response pathway is compromised in amyotrophic lateral sclerosis. Defective refolding of TDP-43 is predicted to aggravate the TDP-43 proteinopathy. The finding that the pathological accumulation of insoluble TDP-43 can be reduced by the activation of HSF1/HSP pathways presents an exciting opportunity for the development of novel therapeutics.Detergent-resistant, ubiquitinated and hyperphosphorylated Tar DNA binding protein 43 (TDP-43, encoded by Amyotrophic lateral sclerosis (ALS) is the most common adult-onset motor neuron degenerative disease, characterized by progressive motor neuron loss from the spinal cord, brainstem and motor cortex, leading to muscle weakness and eventual respiratory failure. The average age of disease onset is 55 years and affected individuals typically die within 3 to 5 years of diagnosis. No effective treatments are available and consequently there is a high unmet need for novel therapeutic development  and this process is facilitated by molecular chaperones. When cells are acutely stressed by insults that affect proteostasis, such as heat shock, they induce a transcriptional programme orchestrated by the transcription factor heat shock factor 1 (HSF1) to upregulate selected chaperones, known as heat shock proteins (HSPs), to protect them from being overwhelmed by the accumulation of aberrant proteins. HSPs are categorized into several families according to their function and size -independent, HSP70-dependent mechanism. This suggests that the HSF1-DNAJB2a pathway refolds TDP-43 and returns it to its natural physiological state rather than facilitating its degradation. We also demonstrated that the levels of HSF1 and HSPs are markedly reduced in both a transgenic TDP-43 mouse model and in spinal cord tissues from sporadic ALS patients. These results highlight the potential of HSF1 and HSP activation as an important therapeutic strategy for the TDP-43 proteinopathies.The green fluorescent protein (GFP)-TDP-43 constructs were generated and used in a previous study A were gPrimary antibodies used in this study included: rabbit anti-ubiquitin, K48-specific ; mouse anti-phospho TDP-43 ; rabbit anti-mouse TDP-43 . Cells were left for 48 h after transfection to be harvested for analysis unless otherwise stated.HEK293T and SH-SY5Y cells were cultured using Dulbecco’s modified Eagle medium (DMEM) and DMEM/F12 (Life Technologies) supplemented with 10% foetal bovine serum (Life Technologies), and maintained at 37°C, 5% COWTand TDP-43Q331Kmice as previously described  Act 1986, and were approved by the Kings College, London ethics review panel. Compound transgenic mice were generated by crossing TDP-43WTxQ331Kanimals and their single and non-transgenic littermates (n = 3–4) or 50 mg of spinal cord post-mortem tissues from controls or patients with sporadic ALS were homogenized in the appropriate amount of RIPA buffer , sonicated and stored at −80 °C. Protein concentration was determined with DC™ Protein Assay Kit (Bio-Rad). Western blotting was run with 10 μg of protein lysate.Whole spinal cords from 8-week-old TDP-43WTxQ331Kmice and age-matched littermates were anaesthetized and transcardially perfused with phosphate-buffered saline (PBS) followed by 4% paraformaldehyde (PFA) in phosphate buffer. Spinal cord was postfixed in 4% PFA in 15% sucrose for 5 h, cryoprotected in 30% sucrose for 24 h and cut into 30 µm sections on a cryostat. Sections were incubated in rabbit anti-HSF1 antibody , washed and incubated with a biotinylated anti-rabbit antibody  and then incubated in an ABC kit (Vector). Sections were imaged using a Zeiss light microscope and Axiovision software.Eight-week-old, end-stage TDP-43Cells were harvested in immunopurification buffer . After centrifuging (14 000 rpm for 30 s at 4°C), the supernatant was collected and incubated with immunopurification antibody and Dynabead protein A (Life Technologies) overnight at 4°C. The Dynabead® protein A-antibody-protein complex was purified using magnetic separation and washed with immunopurification buffer before eluted in loading buffer.gfor 20 min at 4 °C. After centrifugation, the supernatant was collected as the RIPA solubility fraction. The pellet, after being washed once with RIPA buffer, was then suspended in 20% of the original lysis volume with urea buffer  and collected as the insoluble, detergent-resistant fraction.The fractionation for protein solubility was performed using a protocol described byhttp://imagej.nih.gov/ij/). Integrated band intensities were normalized to that of loading control or the RIPA fraction.Protein quantification and western blotting were performed as described before  for 20 min and washed with PBS three times for 5 min. Cells were permeablized by incubation in PBS containing 0.5% Triton™ X-100 (Sigma) for 15 min at room temperature, followed by blocking in PBS containing 1% donkey serum for 1 h at room temperature. Cells were incubated with primary antibody diluted in blocking solution overnight at 4°C. After washing in PBS, cells were subsequently incubated with fluorescent secondary antibodies diluted in blocking solution for 1 h at room temperature. DAPI (Sigma) was then used to stain for nuclei before being mounted on coverslips using FluroSave .For cell survival assay, SH-SY5Y cells were trypsinized and stained with Calcein Violet 450 AM Viability Dye  for 30 min at room temperature. After washing twice with PBS, cells were resuspended in PBS and analysed with the BD FACSCanto II (BD Biosciences).This study used dominant active and negative forms of HSF1 to investigate the effect of the HSR and HSP upregulation on TDP-43 proteinopathy in cellular models. HSF1 is a transcription factor that upregulates HSPs during the stress response. In the unstressed condition, HSF1 is bound to HSP90 via its regulatory domain and is kept in an inactive form. During cellular stress, HSF1 dissociates from HSP90, trimerizes, translocates into the nucleus, and binds to the heat shock element (HSE) through its DNA binding domain (DBD), where it promotes the transcription of downstream genes via its transactive domain (TAD) A. This P < 0.001). Conversely, HSF1(−) significantly increased the level of insoluble TDP-43 by ∼250% (P < 0.001). The mutation status of TDP-43 when co-transfected with HSF1 did not significantly change TDP-43 protein solubility, nor did the presence of a mutation in TDP-43 when transfected alone (P < 0.01 for dNLS-TDP-43), suggesting the expression of HSF1(−) results in an increased tendency for overexpressed cytosolic TDP-43 to shift from soluble monomer to insoluble aggregates.The expression level of endogenous TDP-43 in our cells was not high enough to cause any proteinopathy phenotype similar to that seen in disease tissues, and was not found in the insoluble fraction A and B.d alone C. HSF1 substantially enhances the clearance of insoluble TDP-43, whereas inhibition of the HSR via HSF1(−) expression results in an increase in insoluble TDP-43, likely resulting from an inhibition of the endogenous, basal HSR.P < 0.01) and enhanced by HSF1(−) (P < 0.001). No phospho-TDP-43 staining was identified in GFP-vector transfected cells alone (P = 0.012). The genotype of the overexpressed TDP-43 had no significant impact on TDP-43 phosphorylationper se reduction in cell death. Overexpression of TDP-43 in HEK293 cells did not show cytotoxicity (data not shown), supporting the observation in ALS studies that neuronal cells are particularly sensitive to TDP-43 cytotoxicity.As reported in other cellular and transgenic animal studies, overexpression of TDP-43 can be cytotoxic to some cells, especially neurons , indicaIn summary, we have demonstrated that overexpressing TDP-43 in cells recapitulates pathological features of the TDP-43 proteinopathies and that the presence of these features results in an increase in cell death. Manipulating the heat shock response (HSR) by expression of HSF1(+) reduces this pathological TDP-43, and improves cell survival. In contrast, when cells are deprived of their basal heat shock response activity by the expression of HSF1(−), the degree of TDP-43 proteinopathy is intensified.HSF1 is a transcription factor that is responsible for the induction of a wide range of HSPs and other stress-responsive proteins upon its activation B. To deDNAJB2and also known as HSJ1a, is a target of HSF1  suffering from the most severe motor deficits and TDP-43 pathology, and a drastically reduced life span compared to both single and non-transgenic mice (To study the role of TDP-43 in the development and progression of ALS, lines of transgenic mice expressing either wild-type-TDP-43, Q331K-TDP-43 or both were established (Q331Ksingle transgenic mice that displayed a mild, non-lethal motor defect at later stage (WTxQ331Kmice (WTxQ331Kmice, whereas HSP90 remained unaffected. Immunohistochemical staining also showed reduced levels of HSF1 in the remaining motor neurons in the end-stage diseased mice spinal cord sections, (Interestingly, HSF1 was decreased in the TDP-431Kmice A. As a ctions, B indicaExamination of HSF1 and the HSPs in human tissue identified a significant decrease in HSP70 D and HSTaken together, our results suggest that the HSR is compromised in both the TDP-43 transgenic ALS mouse model and in patient tissues, which could be initiated either directly or indirectly by long term TDP-43 protein overexpression. Additionally, these results demonstrate that a reduction in the HSR leads to decreased TDP-43 refolding efficiency, promoting the accumulation of phosphorylated insoluble TDP-43.In this study, we generated a cellular model of the human TDP-43 proteinopathies that developed detergent-resistant, hyperphosphorylated cytoplasmic insoluble TDP-43 aggregates that are cytotoxic. Co-expression of a dominant positive version of the master regulator of the heat shock response HSF1 induced a dramatic clearance of insoluble TDP-43 aggregates without affecting the levels of soluble TDP-43. Conversely the dominant negative version of HSF1 lead to a massive accumulation of insoluble and hyperphosphorylated TDP-43 aggregates confirming a vital role of the HSF1-HSP axis in physiological TDP-43 proteostasis. We have shown that HSF1-induced TDP-43 clearance is partly mediated by HSP70 and its co-chaperone DNAJB2a in a proteasome- and autophagy-independent manner. We propose that DNAJB2a recognizes and binds to insoluble TDP-43 taking it to the chaperone protein HSP70, which then refolds the protein. Once it has been returned to its soluble, properly folded status, TDP-43 is then released to perform its biological functions . BecausWe have also shown that levels of several HSR proteins are reduced in transgenic mice and sporadic ALS tissues implying that the HSF1–HSP axis may be compromised due to the disease process. Q331K TDP-43 transgenic mice develop a slowly progressive motor phenotype and have moderately reduced levels of HSF1 and HSP-40, whereas the severely-affected double transgenic mice have a dramatic loss of HSF1, HSP40 and HSP70. In human spinal cord tissues the levels of HSP70 and HSP40 were significantly reduced but not HSF1.The data from our cellular, animal and human tissue studies indicate that the HSF1/HSP70/DNAJB2a pathway plays a vital role in the physiological maintenance of TDP-43 proteostasis. In the disease state, however, the HSR appears to be compromised and may contribute to the accumulation of insoluble TDP-43 protein aggregates and exacerbate defective proteostasis .Molecular chaperones are essential for maintaining proteostasis B. As thInteractions between HSPs and TDP-43 have been documented previously.TDP-43 is an aggregation-prone protein that is found to form detergent-resistant aggregates or stress granules during conditions of cellular stress such as heat shock B; howevHSP90 binds to and inactivates HSF1 by preventing it from entering the nucleus and inducing the HSR. HSP90 inhibitors such as 17-AAG have therefore been tested in cellular and animal models of neurodegeneration with some success (Supplementary DataClick here for additional data file.


Neuregulin 1 (NRG1) and NMDARs play important roles in various neuronal functions including neural development. NMDARs also promote many cellular events such as proliferation and survival of neuroblasts before synapse formation. Although many recent studies have indicated that NRG1 regulates NMDAR function in cortical neurons, the effect of NRG1 on NMDAR activation before synapse formation is not well studied.A receptor-mediated TrkB activation. Co-immunoprecipitation and proximity ligase assay showed that TrkB interacts with ErbB4 (NRG1 receptor) and the TrkB-ErbB4 interaction was increased following NRG1 treatment. A significant reduction in TrkB-ErbB4 interaction was observed in the prefrontal cortex of schizophrenia subjects. We found significant increase in released BDNF levels following NRG1 treatment, which was inhibited by ErbB4 inhibitor, AG1478. In addition, pretreatment with BDNF neutralizing antibody, but not control IgG abolished NRG1-induced increases in phospho-TrkB and phospho-NR2B levels. Moreover, studies using TrkB mutants showed that intercellular domain of TrkB is necessary for TrkB-ErbB4 interaction and NR2B activation.NRG1 induces activation of NMDAR subunit NR2B, and tropomyosin-related kinase receptor B (TrkB), the receptor for BDNF via activation of phospholipase C-gamma (PLC-γ) in immature primary cortical neurons. Our data using TrkB inhibitor (K252a), TrkB siRNA and TrkB−/− neurons demonstrated that TrkB inhibition suppresses NRG1-induced NR2B activation in neurons. We found that NRG1 stimulation leads to GABABDNF/TrkB signaling plays an important role in the NRG1-stimulated NR2B regulation. These findings could be of relevance to many neurodevelopmental disorders, as NRG1 and BDNF signaling pathways have been implicated in autism and schizophrenia. Activation of NMDA type glutamate receptors (NMDARs) facilitate a number of signaling pathways involved in neuronal development, learning, and memory . The dev2+ signaling is known to mediate NRG1-induced NMDAR regulation in neurons . We. We24]. AR-mediated release of BDNF from neurons, which then act on TrkB-expressing neurons promoting NR2B activation via PLCγ . We found that NRG1 treatment stimulates TrkB phosphorylation in neurons, which was inhibited by a BDNF neutralizing antibody, and BDNF inhibition abolished NRG1-induced increase in TrkB-ErbB4 interaction. NRG1-induced ErbB4 activation is known to increase GABA release . Our finBoth NRG1 and BDNF play important role in neurodevelopment and synaptic plasticity. Since NRG1 and BDNF are also present in non-neuronal cells such as oligodendrocytes and astrocytes, it is important to identify whether the processes similar to the ones found in the current study are functioning in those cell types. Moreover, it will be interesting to examine whether TrkB-ErbB4 interaction plays any role in GABAergic function in interneurons. The signaling mechanism described in this study, in which the regulation of NR2B activation by NRG1 is mediated by BDNF/TrkB signaling, may function in developing neural networks to enable NRG1 to modulate synaptogenesis, and growth of dendrites and axons prior to the formation of functional synapses. It will also be important to determine the extent to which this mechanism persists in the adult nervous system and contributes to the regulation of synaptic plasticity and cognitive function by NRG1 and BDNF. Given the alterations in NRG1 and BDNF signaling pathways result in neuronal dysfunctions as well as the implication of NMDARs in neuroplasticity, our findings on the role of TrkB in NRG1-stimulated NR2B phosphorylation could be of relevance to many neurodevelopmental disorders, as NRG1 and BDNF signaling pathways have been implicated in autism and schizophrenia.−/−) were provided by Dr. Barbara Rohrer, Medical University of South Carolina, Charleston, SC and the colony was maintained in our animal housing facility at the Georgia Regents University. All experiments were done in compliance with Georgia Regents University animal welfare guidelines.Timed pregnant CD-1 mice were purchased from Charles River Laboratories . TrkB knockout  by PCR of tail biopsy DNA . PCR reaction was performed over 35 cycles using GoTaq® Green Master Mix (Promega). Primers utilized were as follows: trkb-n2: 5′-ATGTCGCCCTGGCTGAAGTG; trkbc8: 5′-ACTGACATCCGTAAGCCAGT; pgk3-1: 5′-GGTTCTAAGTACTGTGGTTTCC. Annealing temperature was set at 60°C. The products of the PCR reaction were visualized using agarose gel electrophoresis.TrkBA receptor antagonist, picrotoxin (Tocris) at 100 μM. Inhibitor, agonist or antagonist was added 20 or 30 min prior to NRG1 treatment.Primary cortical neurons were prepared at E16. Neurons were cultured in Neurobasal medium containing, B27 supplement, 10% Fetal Bovine Serum (FBS), penicillin/streptomycin mixture of antibiotics and Glutamax for 4 days. The following pharmacological treatments were used: Neuregulinβ1  at 5 nM ; the Trk4 , anti-pErbB4 , anti-TrkB , anti-pTrkB  , an, an4  and anti-PV staining was performed overnight at 4°C followed by rinsing with PBS and incubation for 1 hr at room temperature with Cy2 and Cy3 conjugated secondary antibodies (Jackson Immunoresearch). Cells were then washed and mounted using ProLong® Gold Antifade Mountant containing DAPI (Molecular Probes). Images were taken using a LCM confocal microscope (Zeiss).In Situ reagents (Sigma). The cortical neurons (~2 × 104 cells/well) were seeded into 24-well plate on poly-D-lysine coated cover slip. After treatment, the cortical neurons were washed twice with ice-cold 1 × PBS and fixed with 4% paraformaldehyde and 120 mM sucrose in PBS at room temperature for 20 min. After permeabilization the cells were incubated in the blocking buffer (provided with the kit) overnight at 37°C in a humidified chamber. The cells were incubated with the primary antibodies, anti-TrkB  and anti-ErbB4  diluted in the antibody diluents for 2 hours at room temperature followed by washing in Buffer A (supplied with the kit) 3 times for 15 minutes and incubation with the PLA probes for one hour at 37°C in a humid chamber. The antibodies were omitted in the PLA control group. The cells were again subjected to a 10 minute wash and a 5 minute wash in Buffer A. The ligation reaction was carried out at 37°C for one hour in a humid chamber followed by a 10 and 5 minute wash in Buffer A. The cells were then incubated with the amplification-polymerase solution for two hours at 37°C in a darkened humidified chamber. After washing with 1x Buffer B (supplied with the kit) for 10 minutes followed by a 1 minute wash with 0.01X buffer B the cells were mounted using the mounting media supplied with the kit. Images were collected using Zen 2012 lite imaging software from several fields of view per experiment. The number of PLA signals per cell (indentified as red spots) was counted from three Z-plane images using ImageJ (NIH).The PLA was performed using Duolink Brain-derived neurotrophic factor was measured with a conventional sandwich ELISA using the BDNF Emax Immuno-Assay System  according to the protocol of the manufacturer.pBiFC-TrkB-FL and pBiFC-TrkB-∆-ICD constructs were kindly provided by Dr Maruyama, Okinawa Institute of Science and Technology, Japan . These cWe used 19 nt siRNA  named Ntrk2 for mouse TrkB and 19 nt siRNA (GCAAGAAGUUCCUCCAGUA) for mouse PLCγ (M-040978-01-0005) from Dharmacon Research Inc. The mouse control siRNA used was 19 nt quadruplex with two 3′ overhanging nucleotides . Transfection of both siRNAs (50 nM) was performed in cultured cortical neurons using Amaxa 4D-Nucleofector Protocol (Lonza) 48 h before the NRG1 treatment.We obtained postmortem prefrontal cortex samples from schizophrenia (N = 15) and control (N = 15) subjects from the Human Brain and Spinal Fluid Resource Center . Description on the demographic details of samples is published elsewhere . The samp) values of less than 5% were considered significant.Quantified data are presented as mean ± SEM and analyzed by GraphPad PRISM. Statistical comparisons between two groups were made using t tests. Comparisons among multiple groups were made using one-way or two-way ANOVA, with Bonferroni’s post hoc analyses to identify significant differences between groups. The probability (The authors declare that they have no competing interests.AP designed the study. CDP performed the experiments and analyzed the data. AP wrote the manuscript. Both authors approved the final version of the manuscript.


The limitations of our cognitive resources necessitate the selection of relevant information from the incoming visual stream. This selection and prioritizing of stimuli allows the organism to adapt to the current conditions. However, the characteristics of this process vary with time and depend on numerous external and internal factors. The present study was aimed at determining how the emotional state affects effective connectivity between visual, attentional and control brain areas during the perception of affective visual stimuli. The Directed Transfer Function was applied on a 32-electrode EEG recording to quantify the direction and intensity of the information flow during two sessions: positive and negative. These data were correlated with a self-report of the emotional state. We demonstrated that the current mood, as measured by self-report, is a factor which affects the patterns of effective cortical connectivity. An increase in prefrontal top-down control over the visual and attentional areas was revealed in a state of tension. It was accompanied by increased outflow within and from the areas recognized as the ventral attentional network. By contrast, a positive emotional state was associated with heightened flow from the parietal to the occipital area. The functional significance of the revealed effects is discussed. The processing of emotional stimuli is a complex and non-linear multi-stage process. On different levels, it is affected by both characteristics of the stimuli itself and numerous cognitive and affective factors such as expectations, cognitive set, previous experience, current mood, or learned strategies of emotional control , focuses on the information that is relevant to the current task or goal-related requirements. It represents top-down control over the perceptual system and is responsible for both spatial and feature-related attention. Anatomically, it is identified as the dorsal fronto-parietal network and is composed of bilateral frontal eye fields (FEF) and intra-parietal sulci with superior parietal lobules  applied on the EEG recording  was expected as a marker of facilitation of bottom-up processes and simultaneous inhibition of top-down attentional control associated with the increased role of stimuli salience. Finally, it was posited that negative stimuli – biologically relevant as they signal potential danger and thus require immediate responses – would reveal stronger effects than those observed for positive ones.Additional computations were performed using prestimulus alpha power over the visual cortex to integrate the connectivity analysis with classic EEG methods. Its level during an idle period, just before the presentation of the stimulus, is thought to reflect the active top-down influence that modulates the sensitivity of the perceptual system  and approved by the Ethical Committee of the Institute of Psychology, Jagiellonian University. Informed consent was obtained from all subjects. Forty-two undergraduate student volunteers , participated in the study after signing a written consent form. All of them were medication-free with no reported history of neurological and psychiatric diseases – or of substance abuse – and had normal or corrected-to-normal vision. During the procedure, the subjects were seated in an air-conditioned soundproof cabin before a 24″ LCD monitor.There were two randomly sequenced sessions, one positive and one negative, which were determined by the valence of the slides displayed. Forty emotional pictures were chosen from the International Affective Picture System (IAPS) database . Based on EEG montage brain atlases , Tension Arousal  and Hedonic Tone (HT: a pleasant and enjoyable state).Next – after ensuring normal distributions of data by means of a series of Kolmogorov-Smirnov tests – the relationships between the subjective scales and DTF values were estimated separately in positive and negative sessions using Pearson’s r correlations. Hence, flow increases or decreases refer to positive or negative correlations respectively, with the subjective scales given observed for the extracted epochs. Finally, because there were multiple comparisons involved, the False Discovery Rate (FDR) procedure  and lower energetic arousal (EA), as well as with more negative valence  were seen as increased flow from the CP6 to CP2 electrode for both sessions, and from the CP2 to the O1 electrode only in the negative session. Detailed correlation coefficients with corresponding p-values (both raw and FDR-adjusted) are shown in Tables p = 0.005), while the positive one revealed no significant relationships.Significant negative correlation between the alpha power and average DTF from the left prefrontal cortex to the visual area was observed in the negative session , as a scale strongly linked to negative valence, turned out to be the most influential factor affecting the strength of connectivity. Some results for Hedonic Tone (HT), which, on the other hand, is strongly linked to positive valence, were also observed. In contrast, Energetic Arousal (EA) did not yield any significant results. Since the EA scale is associated mostly with the energetic and not the valence aspect of the emotional state  as well as to the visual cortex (O/PO electrodes). This vast increase in signal propagation can be interpreted as a heightened top-down influence enforced on the attentional and perceptual systems. Such a pattern could possibly underlie mood-related sensitization to potential threat signals, which can be referred to as “hyper-vigilance”. Indeed, the strengthening of top-down connections is suggested as a phenomenon accompanying the prioritized processing of affective content  as a control center. It has been suggested that the left DLPFC plays a dominant role in continuously monitoring the demands of the current situation and imposing automatic control over the attentional systems  to certain other PFC regions. This was also accompanied by strengthening of connectivity with the right TPJ (located under the CP6 electrode). Importantly, these effects were visible for both sessions, which indicates that they may appear prior to full stimuli evaluation as a tonic sensitization to potential changes in the environment. Indeed, the role of the right IFG in modulating different features of visual attention by influencing the occipital and parietal areas has been widely shown  to the CP2 electrode (over the right IPS). Thus the hypothesis that the path of communication between both attentional networks involves the right TPJ as an interface  was found to be associated with a heightened flow from the electrodes over the right IPS to the occipital area. This effect possibly relates to the extension of the spatial scope of attention in a positive mood, which is accompanied by a simultaneous decrease in sensitivity to negative signals. Some data suggest the causal role of the right IPS in regulating this trade-off by strengthening the top-down spatial features of attention . This would seem to be a plausible neural mechanism underlying the sensitization of the stimuli-driven attention. On the other hand, positive mood  was related to increased flow from the right parietal area to the visual cortex, which could underlie changes in attentional scope.Most importantly, we managed to demonstrate that the current mood, as measured by self-report, is a factor that affects the patterns of effective cortical connectivity. A multiple-item checklist tool was chosen for use in the study, which was intended for more implicit mood measurement, where the researcher’s intentions are not easily disclosed. The intermixing of emotional stimuli with adjectival items could be less susceptible to the influence of demand characteristics, since the consideration of possible experimental expectations in subjects is limited when they are focused on a pending procedure. These factors suggest that the accuracy of such quasi-simultaneous measurement is possible and better than a retrospective check (Hurlburt and Heavey Some other limitations and future directions should also be mentioned. Firstly, the limited spatial accuracy of EEG should be borne in mind when interpreting the results. In this way the proposed correspondence between electrodes and cortical areas should be treated as putative but not certain. Secondly, the indirect inference about attentional characteristics was based on existing theoretical models that link the emotional state with changes in perception. Further verification of the results is recommended in future studies, where different task demands and attentional manipulation can be applied.


Melanin-concentrating hormone (MCH) regulates food intake through activation of the receptor, MCHR1. We have identified AMG 076 as an orally bioavailable potent and selective small molecule antagonist of MCHR1. In mouse models of obesity, AMG 076 caused a reduction in body weight gain in wild-type (MCHR1+/+) but not in knockout (MCHR1−/−) mice. The body weight reduction was associated with decreases in food intake and increases in energy expenditure. Importantly, we show that these MCHR1-dependent effects of AMG 076 were also reflected in improved metabolic phenotypes, increased glucose tolerance and insulin sensitivity. Preliminary data on effects of AMG 076 in obese cynomolgus monkeys are also presented. Obesity has become a global health epidemic affecting people of virtually all ages and socioeconomic groups. Obesity contributes to the deaths of ∼300,000 adults in the US each year, making it one of the leading causes of preventable mortality , a neuropeptide highly conserved among vertebrates, plays an important role in the regulation of food intake and energy balance in mammals. MCH is synthesized in the hypothalamus and zona incerta Nahon  of the bOnly one MCH receptor, MCHR1, is expressed in rodents. MCHR1 was identified as a G protein-coupled receptor -11-Methyl-9-trifluoromethyl-1,3,4,4a,5,6,11,11a-octahydro-pyridocarbazol-2-yl)-ethyl]-cyclohexane carboxylic acid was synthesized at Amgen  was purchased from PerkinElmer  and MCH peptide from Phoenix Pharmaceuticals . Inhibition of MCH-induced Ca2+ mobilization was assayed in HEK293 cells stably expressing human MCHR1 (HEK293-MCHR1), using a FLIPR assay kit from Molecular Devices  according to the manufacturer's protocol. IC50 was determined by nonlinear regression curve fitting program (GraphPad Prism). Inhibition of MCH-induced Ca2+ mobilization in multiple species was measured by aequorin assay in CHO cells transiently expressing MCHR1 of different species. Concentrations of MCH used for the FLIPR and aequorin assays were 12 nM and 35 nM, respectively. All assays were repeated at least three times for Ki and IC50 determination and representative experiments are presented.Radioligand membrane binding and aequorin assays were carried out as described previously .The selectivity of AMG 076 was evaluated in radioligand displacement assays against a panel of 64 GPCRs, transporters and ion channels by Eurofins Panlabs . The affinity of AMG 076 for 5HT2C was also evaluated by Amgen in radioligand displacement-binding assay using 5HT2C membrane and specific radioligand -MCH with a Ki of 0.6 ± 0.10 nM  was associated with a significant increase in measured VO2 compared with the MCHR1(+/+) mice that received vehicle. In contrast, no effect of AMG 076 (100 mg kg−1 day−1) treatment was observed in the MCHR1(−/−) mice compared with the MCHR1(−/−) mice that received vehicle. In summary, these data show that the prevention of high-fat diet-induced body weight gain by AMG 076 is MCHR1-dependent and is at least partly mediated through an increase in energy expenditure.AMG 076 was evaluated for its ability to prevent the accumulation of body fat in C57BL/6 mice transferred onto a high-fat diet. To confirm that the effects of AMG 076 were mediated specifically through MCHR1, MCHR1(−/−) mice with the same genetic background  or sibutramine (10 mg kg−1 day−1). The effect of AMG 076 and sibutramine on body weight was assessed over the period from day 0 to 144  reached statistical significance by day 2, 7, and 3, respectively, and remained statistically significant through day 144. Dose-related exposures of AMG 076 in plasma were confirmed as summarized in Table The diet-induced obese (DIO) mouse model is a well-established animal model for testing anti-obesity agents . Sibutramine also reduced food intake (P < 0.05), but to a lesser extent than AMG 076.Food intake measurements were initiated 3 weeks into the study and continued through day 115 of treatments. A dose-related reduction was observed in daily food intake in response to AMG 076 treatment relative to vehicle during the course of treatment .P < 0.05), was reached only by the 1 mg/kg group. Five of the six monkeys given the vehicle gained significant weight, and at least three monkeys in each of the AMG 076 dose groups showed a commensurate decrease in body weight and BMI. Intra-abdominal fat (IAF) content was measured on day 88 of the treatment using abdominal CT scan and food intake was measured daily. Although a downtrend in IAF and food intake similar to that of body weight and BMI was observed with the AMG 076 treatment groups, the changes did not reach statistical significance by any of the treatment groups  mice treated with AMG 076 up to 100 mg kgWe did not observe significant effect of AMG 076 treatment on food intake in lean mice on high-fat diet. We, however, detected a significant reduction in food intake in DIO mice treated with AMG 076. These observations suggest that different metabolic state of the animals may be critical in determining how antagonism of MCHR1 may contribute to body weight loss, through increasing metabolic rate or suppressing appetite, or both.Obesity has been shown to be a risk factor for type 2 diabetes and is known to exacerbate diabetes in humans (Chen et al. Rodents do not express MCHR2 (Tan et al. A number of MCHR1 antagonists have been developed as potential anti-obesity drugs (Borowsky et al.


We propose a quite concrete approach based on the notion of Köthe-Toeplitz duals with respect to the non-Newtonian calculus. Finally, we derive some inclusion relationships between Köthe space and solidness.The important point to note is that the non-Newtonian calculus is a self-contained system independent of any other system of calculus. Therefore the reader may be surprised to learn that there is a uniform relationship between the corresponding operators of this calculus and the classical calculus. Several basic concepts based on non-Newtonian calculus are presented by Grossman (1983), Grossman and Katz (1978), and Grossman (1979). Following Grossman and Katz, in the present paper, we introduce the sets of bounded, convergent, null series and  It is certainly not unusual to measure deviations by ratios rather than differences. For instance, during the Renaissance, many scholars, including Galileo, discussed the following problem. Two estimates, 10 and 1000, are proposed as the value of a horse, which estimates, if any, deviates more from the true value of 100? The scholars who maintained that deviations should be measured by differences concluded that the estimate of 10 was closer to the true value. However, Galileo eventually maintained that the deviations should be measured by ratios, and he concluded that two estimates deviated equally from the true value. From the story, the question comes out this way, what if we measure by ratios? The answer is the main idea of non-Newtonian calculus which consists of many calculuses such as the classical, geometric, anageometric, and bigeometric calculus.Bashirov et al. , 2 have bs*, cs*, and cs0* consisting of the sets of all bounded, convergent, null series based on the non-Newtonian calculus, as follows:ω* = {x = (xk) : xk ∈ C*for all  k ∈ N}. One can conclude that the sets bs*, cs*, and cs0* are complete non-Newtonian metric spaces with the metric d∞N defined byFollowing Tekin and Başar  we can cbv*, bvp*, and bv∞* of bounded variation sequences in the sense of non-Newtonian calculus, as follows:bv*, bvp*, and bv∞* are complete with corresponding metrics on the right-hand side with (Δx)k = xk ⊖ xk−1, x)k′ = xk ⊖ xk+1 for all k ∈ N.Secondly, we introduce several sets  generator is a one-to-one function whose domain is R and whose range is a subset of B⊆R, the set of real numbers. Each generator generates exactly one arithmetic, and conversely each arithmetic is generated by exactly one generator. As a generator, we choose the function β such that its basic algebraic operations are defined as follows:x, y ∈ R(N), where the non-Newtonian real field R(N)≔{β{x} : x ∈ R} as in [AR} as in .β-positive real numbers, denoted by R+(N), are the numbers x in R such that β-negative real numbers, denoted by R−(N), are those for which beta-zero, beta-one, β(0) and β(1). Further, β-integers turns out to be the following:The X be a nonempty set and let d* : X × X → R(N) be a function such that, for all x, y, z ∈ X, the following axioms hold:  x = y,(NM1)   d* = d*,(NM2)   (NM3)   Then, the pair  and d* are called a non-Newtonian metric space and a non-Newtonian metric on X, respectively.Let X =  be a non-Newtonian metric space. Then the basic notions can be defined as follows. x = (xk) is a function from the set N into the set R(N). The β-real number xk denotes the value of the function at k ∈ N and is called the kth term of the sequence.A sequence xn) in a metric space X =  is said to be ∗-convergent if for every given n0 = n0(ε) ∈ N and x ∈ X such that n > n0 and is denoted by *limn→∞xn = x or n → ∞.A sequence (xn) in X =  is said to be non-Newtonian Cauchy (∗-Cauchy) if for every n0 = n0(ε) ∈ N such that m, n > n0.A sequence  is defined as β(|β−1(x)|) and is denoted by |x|β. For each number x in B ⊂ R(N), x1 and x2 is defined by alpha-generator in  also are the ordered pair of arithmetics (β-arithmetic and α-arithmetic). The sets beta-generator generates beta-arithmetics, respectively. Definitions given for β-arithmetic are also valid for α-arithmetic.Suppose that α-arithmetic is used for arguments and β-arithmetic is used for values; in particular, changes in arguments and values are measured by α-differences and β-differences, respectively. The operators of this calculus type are applied only to functions with arguments in A and values in B. The ∗-limit of a function with two generators α and β is defined byf is ∗-continuous at a point a in A if and only if a is an argument of f and *limx→af(x) = f(a). When α and β are the identity function I, the concepts of ∗-limit and ∗-continuity are identical with those of classical limit and classical continuity.The important point to note here is that α-arithmetic to β-arithmetic is the unique function ι (iota) that possesses the following three properties:(i)ι is one to one.(ii)ι is from A onto B.(iii)u and v in A,For any numbers,  It turns out that ι(x) = β{α−1(x)} for every x in A and that n. Since, for example, α-arithmetic can readily be transformed into a statement in β-arithmetic.The isomorphism from C*; that is,Let C*, ⊕, ⊙) is a field. of ∗-complex numbers, the formal notation,N-series. Also, for integers, n ∈ N, the finite ∗-sums sn* = ∗∑∑k=0nzk* are called the partial sums of complex N-series. If the sequence ∗-converges to a complex number s*, then we say that the series ∗-converges and write s* = ∗∑∑k=0nzn*. The number s* is then called the ∗-sum of this series. If (sn)  ∗-diverges, we say that the series ∗-diverges or it is ∗-divergent.Given a sequence -triangle inequality).(ii)(iii)Let  and zk*, tk* ∈ C* for k ∈ {0,1, 2,3,…, n}. Then, For any Following Tekin and Başar , we can z* ∈ C* be an arbitrary element. The distance function d* is called ∗-norm of z* and is denoted by z1*, z2* ∈ C* we have d* is the induced metric from Let z* by z* but leaves the real part the same. ThusLet (i) Let α and β be the same generator functions and z* ∈ C*. Then the following condition holds(ii) Let C*, d*) is a complete metric space, where d* is defined by l∞*, c*, c0*, and lp*, p ≥ 1, are sequence spaces.The sets (b)λ* denote any of the spaces l∞*, c*, and c0* and z = (zk*), t = (tk*) ∈ λ*. Define d∞* on the space λ* by λ*, d∞*) is a complete metric space.Let (c)l∞*, c*, and c0* are Banach spaces with the norm ||z||∞* defined byThe spaces (d)lp* is Banach spaces with the norm ||z||p* defined byThe space The following statements hold.bs*, cs*, cs0* and bv*, bvp*, bv∞* consisting of all bounded, convergent, null series and the sets of bounded variation sequences in the sense of non-Newtonian calculus which correspond to the sets bs, cs, cs0 and bv, bvp, bv∞ over the complex field C, respectively.In the present section, we introduce the sets μ* denote any of the spaces bs*, cs*, and cs0* and z = (zk*), t = (tk*) ∈ μ*. Define d∞N on the space μ* byα, β operators and corresponding function ι = βα−1. Then,  is a complete metric space.Let cs* and cs0*, we prove the theorem only for the space bs*. Let the ∗-sums ∗∑∑k=0nzk*, ∗∑∑k=0ntk* ∈ C*, where z = (zk*), t = (tk*) ∈ C*. Then the following metric axioms in  (NM1) From  it can b d∞N = d∞N holds.(NM2) It is trivial that the condition  z = (zk*), t = (tk*), w = (wk*) ∈ C*. In fact by taking into account (NM3) We show that ∗triangle inequality in  Since the axioms (NM1)–(NM3) are satisfied,  is a non-Newtonian metric space. It remains to prove the completeness of the space bs*.Since the proof is similar to the spaces xm) be a ∗-Cauchy sequence in bs*, where xm = {x1m) = {xk(1), xk(2),…, xkm) and show that x ∈ bs*. From (r → ∞ and m > n0 we havexm) ∈ bs*, there exists k ∈ N. Thus, (m > n0k ∈ N whose right-hand side does not involve k. Hence (xk) is a bounded sequence of geometric complex numbers; that is, x = (xk) ∈ bs*. Also from (m > n0m → ∞ for an arbitrary ∗-Cauchy sequence (xm). Hence bs* is complete.Let , y = (yk) ∈ bv*, and (Δx)k′ = xk ⊖ xk+1. Then,  is a complete metric space. Similarly, one can conclude that the other bounded variation sets  and  are also complete.Let bv* is a Banach space with the norm ||x||bv* defined byThe space The idea of dual sequence space which plays an important role in the representation of linear functionals and the characterization of matrix transformations between sequence spaces was introduced by Köthe and Toeplitz , whose mλ*, μ* ∈ C*, the set S, defined byλ* and μ* for all k ∈ N. One can easily observe for a sequence space ν* of ∗-complex numbers that the inclusions S ⊂ S if ν* ⊂ λ* and S ⊂ S if μ* ⊂ ν* hold.In this section, we focus on the alpha-, beta-, and gamma-duals of the classical sequence spaces over non-Newtonian complex field. For λ* ⊂ ω* which are, respectively, denoted by {λ*}α, {λ*}β, and {λ*}γ, as follows:wk⊙zk) is the coordinatewise product of ∗-complex numbers w and z for all k ∈ N. Then {λ*}β is called beta-dual of λ* or the set of all convergence factor sequences of λ* in cs*. Firstly, we give a remark concerning the ∗-convergence factor sequences.Firstly, we define the alpha-, beta-, and gamma-duals of a set Throughout the text, we also use the notation “<” for a non-Newtonian linear subspace which was created in .∅ ≠ λ* ⊂ ω*. Then the following statements are valid. λ*}β is a sequence space if φ* < {λ*}β < ω*, where {λ* ⊂ μ* ⊂ ω*, then {μ*}β < {λ*}β.If λ* ⊂ {λ*}ββ : = ({λ*}β)β.φ*}β = ω* and {ω*}β = φ*.{Let w = (wk), m = (mk), and n = (nk)∈{λ*}β. (a)λ*}β < ω* holds from the hypothesis. We show that m ⊕ n ∈ {λ*}β for m, n ∈ {λ*}β. Suppose that l ∈ λ*. Then (mk⊙lk) ∈ cs* and (nk⊙lk) ∈ cs* for all l ∈ λ*. We can deduce thatIt is trivial that { m ⊕ n ∈ {λ*}β. Now, we show that t⊙w ∈ {λ*}β for any t ∈ C* and w = (wk)∈{λ*}β, since (wk⊙lk) ∈ cs* for all l ∈ λ*. Combining this with ((tk⊙wk)⊙lk) = tk⊙(wk⊙lk) ∈ cs* for all l ∈ λ* we get t⊙w ∈ {λ*}β. Therefore, we have proved that {λ*}β is a subspace of the space ω*.Hence, (d)ω*}β ⊂ φ*. Suppose that w = (wn)∈{ω*}β and z = (zn) are given with ∗-division by zn⊙wn = 1* if wn ≠ 0* and zn : = 1* otherwise. By taking into account the set φ* from inclusion (a), then there exists an integer N ∈ N for all n ≥ N such that Using (a) we need only to show { w⊙z ∈ cs* implies that w ∈ φ*. The rest is an immediate consequence of this part and we omitted the details.Further, Since the proof is trivial for the conditions (b) and (c), we prove only (a) and (d). Let c0*}β = {c*}β = {l∞*}β = l1*.{l1*}β = l∞*.{The following statements hold: l∞*}β ⊂ {c*}β ⊂ {c0*}β by l1* ⊂ {l∞*}β and {c0*}β ⊂ l1*. Now, consider that w = (wk) ∈ l1* and z = (zk) ∈ l∞* are given. By taking into account the cases (c)-(d) of w⊙z ∈ cs*. So the condition l1* ⊂ {l∞*}β holds.(a) Obviously {y = (yk) ∈ ω*∖l1*, we prove the existence of an x ∈ c0* with y⊙x ∉ cs*. According to y ∉ l1* we may confirm an index sequence (np) which is strictly increasing with x = (xk) ∈ c0* by xk : = (sgn*yk⊘p), the non-Newtonian complex signum function is defined byy = (yk) ∈ C*. Finally, by using α = β, we getnp−1 ≤ k < np. Therefore y⊙x ∉ cs* and thus y ∉ {c0*}β. Hence {c0*}β ⊂ l1*.Conversely, for a given l∞* ⊂ ({l∞*}β)β = {l1*}β since {l∞*}β = l1*. Now we assume the existence of a w = (wn)∈{l1*}β∖l∞*. Since w is unbounded there exists a subsequence (wnk) of (wn) and we can find a real number (k + 1)2 such that k ∈ N1. The sequence (xn) is defined by xn : = (sgn*(wnk)⊘(k + 1)2) if n = nk and x ∈ l1*. However,w ∉ {l1*}β, which contradicts our assumption and {l1*}β ⊂ l∞*. This step completes the proof.(b) From the condition (c) of ζ-duals .In addition to the statements in ∅ ≠ λ* ⊂ ω*. Then the following statements are valid:φ* < {λ*}α < {λ*}β < {λ*}γ < ω*; in particular, {λ*}ζ is a sequence space over C*;λ* < μ* < ω*, then {μ*}ζ < {λ*}ζ;if I is an index set, if λi* are sequence spaces, and if λ* : = ⋃i∈Iλi*, then 〈λ*〉ζ = ⋂i∈I{λi*}ζ, where the notation "〈〉" stands for the span of linear subspace over C*;λ* ⊂ {λ*}ζζ : = ({λ*}ζ)ζ.Let l∞* < cs* < bs*. We only show the cases (c) and (d) taking ζ = alpha. The rest of the parts can be obtained in a similar way.(c)λi* ⊂ 〈λ*〉 the following 〈λ*〉α ⊂ {λi*}α and 〈λ*〉α ⊂ ⋂i∈I{λi*}α hold by (b). On the other hand, if y ∈ ⋂i∈I{λi*}α, that is, y ∈ {λi*}α, then x⊙y ∈ l1* for all x ∈ λ* and therefore y ∈ {λ*}α ⊂ 〈λ*〉α.Now, as an immediate consequence (d)λ* ⊂ {λ*}ζζ. Let w ∈ λ*; then w⊙z ∈ l1* for all z ∈ {λ*}α; thus w ∈ {λ*}ζζ and λ* ⊂ {λ*}ζζ by (a).We can deduce The case (b) obviously is true, and (a) follows from λ* ≠ {λ*}ζζ as we get from ζ = β and λ* : = c0*. We have {c0*}ββ = l∞* ≠ c0*. This remark gives rise to the following definition.Here ζ ∈ {α, β, γ} and let λ* be a sequence space over the field C*. λ* is called ζ-space if λ* = {λ*}ζζ. Further, an α-space is also called a Köthe space or perfect sequence space.Let From λ* is a sequence space over the field C* and , then {λ*}ζ is a ζ-space; that is, {λ*}ζ = {λ*}ζζζ.If λ*}α = {λ*}β = {λ*}γ. This gives rise to the notion of solidity.Now we look for sufficient conditions for {X be a sequence space. Then X is called solid ifLet λ* be a sequence space over the field C*. Then λ* is solid if and only ifLet u = (uk) ∈ ω* and x = (xk) ∈ λ* with ξk, ηk ∈ R. Obviously the condition ψk, δk ∈ R. We obtain ψk2 + δk2 ≤ ξk2 + ηk2 since k ∈ N. We may choose a real number γk ∈ R with ψk2 + γk2 = ξk2 + ηk2 and consider w = (wk) defined by w ∈ λ* and ζk ∈ R such that ζk2 + δk2 = ξk2 + ηk2 it follows similarly that Let λ* < ω* is any sequence space over the field C*; then the following statements hold. λ* is a Köthe space, then λ* is solid.If λ* is solid, then {λ*}α = {λ*}β = {λ*}γ.If λ* is a Köthe space, then λ* is a ζ-space.If Consider that λ* < ω* be a sequence space over the field C*.(a)λ* is a Köthe space and u ∈ ω*, then u ∈ {λ*}αα if and only if the condition u⊙z ∈ l1* holds for all z ∈ {λ*}α. Besides this we obtain u = (uk) ∈ λ* and v = (vk) ∈ ω* and the statementIf  z ∈ {λ*}α. Therefore v⊙z ∈ l1*. Hence w ∈ λ* and λ* is solid over the field C*.holds for each (b)λ* is a solid sequence space over C*. To show {λ*}α = {λ*}β = {λ*}γ, it is sufficient condition to verify {λ*}γ < {λ*}α in v = (vk) ∈ {λ*}γ; that is,Consider that  λ*, for z = (zk) ∈ λ*, the condition u = (uk) ∈ λ* for all k ∈ N. Therefore by combining this with the inclusion (By taking into account solidness of nclusion  we deduc u⊙v ∈ l1* < cs*. Hence v ∈ {λ*}α and {λ*}γ < {λ*}α.holds and (c)This is an obvious consequence of Let φ*, ω*, lp*, c0*, and l∞* of sequences are solid.The sets c* and bv* of sequences are not solid; therefore none of them is a Köthe space.The sets ζ ∈ {α, β, γ}, thenFor each (i)l1*}ζ = l∞* and {l∞*}ζ = l1*;{(ii)ω*}ζ = φ* and {φ*}ζ = ω*.{ζ ∈ {α, β, γ} and c0* < μ* < l∞*, then {μ*}ζ = l1* and μ* ⊂ {μ*}ζζ = l∞*. In particular, {c0*}ζ = {c*}ζ = l1*, and each of c*, c0* is not a ζ-space.If The following statements hold. c0* and l∞* are solid, we know that {c0*}ζ = {l∞*}ζ = l1*. So the statements in (d) are obtained from That the specified spaces in cases (a-b) are solid is an immediate consequence of their definition. Additionally, the cases (i-ii) of (c) can be obtained ζ-duals of the spaces cs*, bs*, bv*, and bv0*. We will find that none of these sequence spaces is solid; in particular, none of them is a Köthe space.Next, we determine the cs*}α = {bv*}α = {bv0*}α = l1*; {bv0*} = bv*∩c0*,{cs*}β = bv*, {bv*}β = cs*, {bv0*}β = bs*, {bs*}β = bv0*;{cs*}γ = bv*, {bv*}γ = bs*, {bv0*}γ = bs*, {bs*}γ = bv*.{The following statements hold: cs*, bs*, bv*, and bv0* of sequences are β-spaces (ζ = beta), but they are not Köthe spaces. Moreover, the sets bs* and bv* of sequences are γ-spaces (ζ = gamma), whereas both cs* and bv0* are not γ-spaces. None of the spaces cs*, bs*, bv*, and bv0* is solid.In particular, the sets cs*}ζ, ζ ∈ {α, β, γ}, and the proofs of all other cases are quite similar.We prove the cases for the spaces {x = (xk) ∈ cs* and y = (yk) ∈ l1*. Then,x||cs* is defined by (y ∈ {cs*}α which gives the fact that l1* ⊂ {cs*}α.(a) Let fined by . Therefoy = (yk) ∈ {cs*}α∖l1*. Then to every natural number p we can construct an index sequence (np) with np < np+1 and p ∈ N. Define x = (xk) byβ-division β{(−1)k2p−} for np < k ≤ np+1. Then x = (xk) ∈ cs*. According to the choice of np, the inequalitiesp2p diverges. Thus, x⊙y ∉ l1*, which implies y ∉ {cs*}α. This contradicts the fact that y ∈ {cs*}α. Therefore {cs*}α ⊂ l1*.Conversely, suppose that bv0*}α ⊂ l1* holds as well if we take the sequence (xk) byThe condition {u = (uk) ∈ {cs*}β and w = (wk) ∈ c0*. Define the sequence v = (vk) ∈ cs* by vk = (wk ⊖ wk+1) for all k ∈ N. Therefore, ∗∑∑k⊙  vk  ∗-converges, butl1* ⊂ cs* yields (uk) ∈ {cs*}β ⊂ {l1*}β = l∞*. Then we derive by passing to the ∗-limit in  ∈ {c0*}β = {c0*}α = l1*; that is, u ∈ bv*. Therefore, {cs*}β⊆bv*.(b) Let ges, but∑∗ k=0 n u = (uk) ∈ bv*. Then, (uk ⊖ uk−1) ∈ l1*. Further, if v = (vk) ∈ cs*, the sequence (wn) defined by wn = ∗∑∑k=0nvk for all k ∈ N is an element of the space c*. Since {c*}α = l1*, the N-series ∗∑∑kwk⊙(uk ⊖ uk−1) is ∗-convergent. Also, we havewn) ∈ c* and (uk) ∈ bv* ⊂ c*, the right-hand side of inequality ⊙uk or ∗∑∑k=0∞uk⊙vk  ∗-converges. Hence bv*⊆{cs*}β.Conversely, suppose that  we have∑∗ k=m n ∈ {cs*}γ and v = (vn) ∈ c0*. Then, for the sequence (wn) ∈ cs* defined by wn = (vn ⊖ vn+1) for all n ∈ N, we can find a non-Newtonian real number n ∈ N. Since (vn) ∈ c0* and (un)∈{cs*}γ ⊂ l∞*, there exists a non-Newtonian real number n ∈ N. Therefore,uk ⊖ uk−1) ∈ {c0*}γ = {c0*}α = l1*; that is, (un) ∈ bv*. Therefore, since the inclusion {cs*}γ ⊂ bv* holds, we conclude that {cs*}γ = bv*, as desired.(c) By using (a), it is known that Although all arithmetics are isomorphic, only by distinguishing among them, we do obtain suitable tools for constructing all the non-Newtonian calculi. But the usefulness of arithmetics is not limited to the construction of calculi; we believe there is a more fundamental reason for considering alternative arithmetics; they may also be helpful in developing and understanding new systems of measurement that could yield simpler physical laws. average speed. The definition “distance traveled per unit time” is incomplete because it fails to provide a method of determining the average speed of an accelerated particle. The definition “distance divided by time,” though not incorrect, is a gross oversimplification that fails to reveal the underlying issues. Fortunately there is a completely satisfactory definition, which undoubtedly was known to Galileo. Then we isolate a constant in each given uniform motion by defining speed to be the distance traveled in any unit time-interval. Finally, for a particle that moves nonuniformly in a distance d in time t, we define the average speed to be the speed that a particle in uniform motion must have in order to travel a distance d in time t. In our opinion, neither the simplicity nor the obviousness of the answer, d/t, justifies its use as the definition of average speed .Of course, we can only speculate as to future applications of the non-Newtonian calculi. Perhaps they can be used to define new scientific concepts, to yield new or simpler scientific laws, to solve heretofore unsolved problems, or to formulate and solve new problems. For example, one constructs non-Newtonian calculi of functions of two or more real variables by choosing an arithmetic for each axis. It might even be profitable to seek deeper connections among the corresponding operators of the calculi. Like as, some of them are postponed to our future works.


Brain-Derived Neurotrophic Factor (BDNF) is a neurotrophin that generally promotes neuronal proliferation, survival and plasticity. These effects occur through cellular secretion and subsequent binding of BDNF to its cognate receptor, TrkB, which promotes paracrine and autocrine signaling cascades. A BDNF/TrkB autocrine loop has been implicated in the survival of cells representing several human cancers and is associated with poor prognosis. The HTLV-1 protein, HBZ, is localized to the nucleus where it is capable of deregulating gene expression. In this study, we found that HBZ increases expression of BDNF. There are a variety of transcriptional variants of BDNF, and HBZ increased expression of several of these. Furthermore, multiple transcriptional variants of BDNF were upregulated in HTLV-1-infected cells compared to normal CD4 cells. Expression of the TrkB receptor was also higher in HTLV-1-infected cells than in normal CD4 cells. This trend similarly occurred when comparing ATL patient and healthy donor specimens, with patient cells exhibiting higher levels of BDNF and TrkB expression. TrkB appeared to localize to the cytoplasmic membrane of HTLV-1-infected cells, suggesting a BDNF/TrkB loop contributes to the survival of these cells. This hypothesis was confirmed by chemically blocking the binding of BDNF to TrkB or by inhibiting TrkB signaling, as both treatments led to a significant increase in apoptosis. Together these results suggest that HBZ activates a BDNF/TrkB autocrine loop that enhances survival of HTLV-1-infected cells. This mechanism may ultimately facilitate the survival of ATL cells.