Group I introns are mobile genetic elements encoding self-splicing ribozymes. Group I introns in nuclear genes are restricted to ribosomal DNA of eukaryotic microorganisms. For example, the myxomycetes, which represent a distinct protist phylum with a unique life strategy, are rich in nucleolar group I introns. We analyzed and compared 75 group I introns at position 516 in the small subunit ribosomal DNA from diverse and distantly related myxomycete taxa. A consensus secondary structure revealed a conserved group IC1 ribozyme core, but with a surprising RNA sequence complexity in the peripheral regions. Five S516 group I introns possess a twintron organization, where a His-Cys homing endonuclease gene insertion was interrupted by a small spliceosomal intron. Eleven S516 introns contained direct repeat arrays with varying lengths of the repeated motif, a varying copy number, and different structural organizations. Phylogenetic analyses of S516 introns and the corresponding host genes revealed a complex inheritance pattern, with both vertical and horizontal transfers. Finally, we reconstructed the evolutionary history of S516 nucleolar group I introns from insertion of mobile-type introns at unoccupied cognate sites, through homing endonuclease gene degradation and loss, and finally to the complete loss of introns. We conclude that myxomycete S516 introns represent a family of genetic elements with surprisingly dynamic structures despite a common function in RNA self-splicing. E. coli rRNA gene numbering system [Group I introns are self-splicing and mobile elements sporadically distributed within genomes of bacteria, bacteriophages, chloroplasts, mitochondria, eukaryotic viruses, and the nuclei of eukaryotes ,2. Nucleg system .Group I introns catalyze their own splicing due to the intron-encoded ribozyme. The splicing mechanism is well described and involves two succeeding transesterification reactions initiated by an attack of an exogenous guanosine (exoG) cofactor ,13. The Nuclear group I introns may also carry additional genes and sequences. Most common are homing endonuclease genes (HEGs), which are found in approximately 5–10% of known nuclear group I introns and code for site-specific endonucleases involved in intron mobility ,20,21,22Mucilago crustacea that contained 34 copies of the intron internal guide sequence [Several examples of direct repeat (DR) arrays within nuclear group I introns have been reported ,11,32,33Naegleria species and in Allovahlkamfia and harbored complex insertions in the P6 segment corresponding to HEGs and lariat-capping ribozymes [In a previous study, we reported self-splicing and structural organization of nuclear S516 group I introns  and haveibozymes ,38,39.In the original S516 intron study [Didymium alpinum isolate Fr-K12, were retrieved from the NCBI database . Spores were germinated and approximately 108 amoeba cells were harvested from DS/2 agar plates, total DNA extracted, and rDNA sequenced as described previously [All rDNA sequences included in this work, except that of ® 2022.0.1 . Multiple sequence alignments were generated using MAFFT version 7.450 [® software program.SSU rDNA sequences (with introns removed) were assembled and aligned in the software program Geneious primeon 7.450 ,41 with on 7.450 ,43, exclon 7.450 ,8,11 usiThe tree-building methods of neighbor joining (NJ), maximum likelihood (ML), maximum parsimony (MP), and minimal evolution (ME), interpreted in MEGA X , were usD. alpinum (Fr-K12) identified a 534-bp group I intron  at position S516 in the nuclear SSU rRNA gene. An RNA secondary structure diagram is presented in Tetrahymena intron [Ribosomal DNA sequence analysis of the myxomycete a intron ,46,47, wD. alpinum Fr-K12) was provided in this study. To assess structural variations among the S516 introns, 143 nucleotide positions in the ribozyme core region common to all 75 introns were strictly aligned according to secondary structure features  tree  is in geL. columbinum isolates cluster together in the SSU rDNA analysis to a well-defined clade (blue box), the corresponding introns are found scattered on the tree, e.g., the L. columbinum (HQ687198) and L. columbinum (HQ687197) introns are distantly related to the other L. columbinum introns. (ii) SSU rDNA analysis strongly support that M. aggregatum and M. cribrariodes are two closely related taxa from the same genus (green box), whereas intron analysis of the core region indicates that the S516 introns are distantly related. This is further supported by structural comparisons , but their corresponding S516 introns suggest a distant relationship. In summary, our phylogenetic analyses suggest that myxomycete S516 introns share a complex evolutionary history, with several clear examples of both vertical and horizontal transfers.Most introns, however, showed a relationship that is not consistent with stable vertical inheritance. To better resolve the evolutionary relationship between S516 introns, we analyzed a smaller dataset consisting of 14 selected taxa , and froparisons , where PL. pseudomaculatum (JQ031985)  (T. varia (KM494993/4/5)  lacking a HEG insertion  , 190 amiY842033) , and 210993/4/5) . Furthernsertion . Interesnsertion A. A nuclnsertion B.Porphyra  [Allovahlkampfia (amoeba-flagellate) [Naegleria (amoeba-flagellate) [Ppo I from the myxomycete Physarum polycephalum [A hallmark of nuclear homing endonucleases is a His-Cys box motif consisting of two zinc coordinating structures (Zn-I and Zn-II) and two catalytic amino acid residues. d algae) , Allovahgellate) , and Naegellate) , as wellcephalum ,52.Elaeomyxa, Comatricha, Colloderma, Lamproderma, Licea, and Trichia), and all except two (Licea and Trichia) classified to the myxomycete order Stemonitales  harbors the largest motif (ca. 220 nt), which is located in P2 and repeated twice. (ii) C. nigricapillitia (AY643824) harbors the smallest motif (only 4 nt) in P2 and is repeated four times. (iii) C. pseudoalpina (DQ903673) harbors DRs in two different extension regions (P1 and P5). While the P1-DR motif is large in size (ca. 110 nt) and repeated four times, the P5-DR is composed of an approximately 60-nt motif and repeated twice. (iv) L. columbinum (HQ687204) also harbors DRs in two extension regions, but in P2 and P9. Two DR motifs are located in P2 (ca. 20 nt and ca. 40 nt), in 5 and 3 copies, respectively, and the DR motif in P9 is 28 nt in size and repeated twice. (v) E. cerifera (JQ031967) harbors three different DR motifs in P6 of 11 nt, 10 nt, and ca. 80 nt in sizes, repeated 4, 6, and 2 times, respectively. (vi) One each of two closely related T. varia introns contains  or lacks  a HEG. Intriguingly, the shorter variant contains a DR, including a HEG remnant sequence, at the proposed HEG deletion site (L. parasitica (JX481297) harbors a complex array in P2. Specifically, the intron folds into a typical group IC1 intron structure (Some isolates showed unique and characteristic DR features : (i) L. ion site . (vii) Ftructure A, and thtructure B. Seven tructure C.D. alpinum Fr-K12 intron. Extension sequences such as HEGs or DR arrays are all located in peripheral segments that do not interfere with the catalytic ribozyme core. HEGs located as insertions into segment P2 in five taxa are interrupted by small spliceosomal introns. Moreover, DR arrays were found in 11 taxa at six peripheral segments, and the L. parasitica intron was found to harbor a highly structured DR motif in 25 copies. Intron evolution analyses indicate a complex inheritance pattern that apparently includes both vertical and horizontal transfers.We report analyses of structural organization and molecular evolution of nucleolar group I intron at position S516 in myxomycetes. All 75 introns belong to the group IC1 subtype and fold at the RNA level into a highly conserved catalytic core, exemplified by the Porphyra red algae [Allovahlkampfia and Naegleria amoeba-flagellates [Naegleria homing endonucleases have been studied in more detail and found to recognize a 19 bp partially symmetrical sequence at the intron-less allele close to the S516 insertion site and to generate a pentanucleotide 3′ overhang at the DNA cleavage site [All HEGs in myxomycete S516 introns encode His-Cys homing endonucleases with a characteristic set of conserved amino acid residues involved in zinc coordination and catalysis. This His-Cys box was found to be highly conserved among S516 HEGs, also outside the myxomycete phylum. These include the ed algae  and the gellates ,39, whicage site ,53. The age site ,29,30,31age site .C. nigricapillitia to 23-nt motif in 25 copies in L. parasitica. A common feature to most arrays was non-identical sequence repeat motifs, which argues against slippage-like mechanisms during replication [Fuligo septica L1949 [M. crustacea S1389 [L. parasitica DR array may suggest a selective structural feature at the RNA level, but no function has currently been assigned.Extension sequences containing DR arrays were noted in six peripheral segments, including P2. The arrays varied from small insertions (4-nt motif in 4 copies) in lication ,55 as a lication ,9,11,33.ca L1949  and M. cea S1389 . The higLamproderma, Echinostelium, and Trichia, but also from Naegleria [Allovhalkampfia [Porphyra [Naegleria martinezi [Bangia fuscopurpurea [Acanthamoeba sp. Isolate [D. alpinum, as well as 93% of the myxomycete S516 introns. HEG deletion was further supported in one of the Trichia S516 introns and suggests a link between deletion of HEG and generation of DR. (4) The complete intron became lost, probably due to homologous recombination with an intron-less allele resulting in SSU rDNA lacking an S516 intron, as noted in the approximately 80% of myxomycetes assessed.The S516 group IC1 intron in myxomycetes and other eukaryotic microorganisms constitute a phylogenetic distinct family with a common origin . While iaegleria , Allovhalkampfia , and PorPorphyra . (2) To artinezi , Bangia purpurea , and an  Isolate . (3) TheThe myxomycete protists represent a unique model system for studying the evolutionary history of self-splicing introns due to an exceptionally high content of nucleolar group I introns. We analyzed 75 group I introns at position 516 in the SSU rDNA of myxomycetes representing five distantly related taxonomic orders. The result reveals a conserved group IC1 ribozyme core, but highly variable peripheral RNA domains containing direct repeat arrays and homing endonuclease genes in 20% of the taxa. All HEGs were interrupted by small spliceosomal introns, which probably facilitate the expression of protein-coding genes embedded in nuclear rDNA. Phylogenetic analysis identified a complex inheritance pattern that can be explained by repeatedly intron gain and loss during the evolution. The current study expands our understanding of the structural organization of group IC1 introns and their evolution.


Conventional psychiatric services are not always acceptable to indigenous communities and people.To present successful models of interactions of psychiatrists with indigenous patients and communities based upon our work with five communities in Maine.We reviewed the strategies that worked for community interaction from our project for supporting indigenous communities to implement medication-assisted treatment and we reviewed the literature to see what other strategies are reported successful.Psychiatrists working in these communities may need to share more personal details than to what they are usually accustomed to be accepted. They may need to acknowledge local culture and spirituality and work with traditional knowledge holders to create collaborative healing approaches. As part of this, a narrative approach appeared to work best in which the psychiatrist worked within the stories and beliefs of the community which required taking the time in dialogue to learn those stories and beliefs. Specifically, we address the challenges of flying into northern, rural, and remote communities, of academic physicians consulting to tribal-based opiate treatment programs, of modifying usual counseling techniques such as motivational interviewing to an indigenous population, and of the changes made in practice styles when taking into account the critiques made by indigenous people about medicine in general and psychiatry in particular.We propose that participatory action-based approaches can improve service delivery to indigenous people. Indigenous cultures share a collectivist mindset in which the needs of the group supersede the needs of individuals, a reliance upon stories, and commitment to a biopsychosocial and spiritual approach.


Here we reportan order-of-magnitude acceleration of the interfacial water dissociationreaction under visible-light illumination. This process is accompaniedby spatial separation of protons and hydroxide ions across one-atom-thickgraphene and enhanced by strong interfacial electric fields. The foundphotoeffect is attributed to the combination of graphene’sperfect selectivity with respect to protons, which prevents proton–hydroxiderecombination, and to proton transport acceleration by the Wien effect,which occurs in synchrony with the water dissociation reaction. Ourfindings provide fundamental insights into ion dynamics near atomicallythin proton-selective interfaces and suggest that strong interfacialfields can enhance and tune very fast ionic processes, which is ofrelevance for applications in photocatalysis and designing reconfigurablematerials.Recent experiments demonstrated that interfacial waterdissociation(H The proton currents were found to be exponentially acceleratedwith increasing E (that reached above 108 V m–1), a phenomenon known as the Wien effect.In particular, graphene’s perfect selectivity with respectto protons and its atomic thickness were crucial to observe the Wieneffect. These properties enable the intense interfacial E to separate protons from OH– ions across the atomicallythin barrier that prevents their recombination, thus yielding notableproton currents. The time scale of the involved separation processshould be extremely fast—as a first approximation, comparableto the time scale of proton transport and proton–OH– recombination in water, which is in the subpicosecond range.13 On the other hand, previous experiments showedthat proton transport through graphene electrodes is strongly enhancedunder illumination via a hot-electron-mediated mechanism, the so-calledphotoproton effect.14 Hot electrons ingraphene have a lifetime of ∼1 ps.16 If the proton-hydroxide ion separation across graphene is comparativelyfast, then in principle the photoproton effect should also acceleratethe transport of protons generated by interfacial water dissociation.In this work, we report such an acceleration.A unique combination of propertiesin graphene allows using it as a proton-permeable electrode. One-atom-thickgraphene exhibits high in-plane electric conductivity14 In brief, the crystals were suspendedover holes (10 μm in diameter) etched in silicon-nitride substrates.2 The resulting graphene film was electricallyconnected to allow for its use as an electrode . One side of the suspended graphene was decorated with Pt nanoparticlesdeposited via electron beam evaporation, which served to increasegraphene’s proton conductivity.2 The opposite side of the suspended graphene electrode faced a 1M KCl electrolyte with alkaline pH solution . Thehigh KCl concentration ensures that the electrolyte resistivity isnegligible, whereas the alkaline pH ensured that water is the onlysource of protons in the system. Hence, in this setup all proton currentsarise from the water dissociation reaction, as demonstrated previously.12 The inner side of the devices was also coatedwith an anion-exchange polymer (FAA FumaTech) which is an excellentOH– ion conductor.17 Thepolymer coating was not essential for the described experiments; however,it provided additional mechanical support for the membrane, improvingthe devices’ reliability (“Device fabrication”in Supporting Information). For electricalmeasurements, the devices were connected in an electrical circuitas shown in Supporting Information).Proton-permeablegraphene electrode devices were fabricated usingmonocrystalline graphene obtained by mechanical exfoliation, as reportedpreviously.lectrode S1. One s2O ⇆ H+ + OH–) withthesedevices was demonstrated in ref (+ + e– → H*@Pt) that flow into graphene through theelectrical circuit. These adsorbed protons eventually escape as hydrogenmolecules through the discontinuous Pt film.2 Notethat the μm-sized electrodes ensure that proton transfer throughgraphene dominates the resistivity in the circuit, with negligiblecontributions from the bulk electrolyte and counter electrode19 . On the other hand, the photoeffectin proton transport through graphene electrodes was demonstrated inref response of the devices with alkaline pH electrolytewas measured both in dark conditions and under solar-simulated illuminationof 100 mW cm–2 intensity (Oriel Sol3A light source).We found that the potential at zero current, φ, was negative,in agreement with the previous work12 . For small applied biases V around this potential, the I–V response was linear, which allowed extraction of the devices’proton conductivity, G = I/ΔV, where ΔV = V –φ. I–V response ofthe devices measured at pH 11 under illumination. Surprisingly, G increased by an order of magnitude with respect to thedark case. The inset of Figure S3). The absolute value of G in dark conditions changedwith pH, in agreement with the previous report.12 In all cases, we have observed a strong increase in G under illumination.The currentdensity vs voltage  dependence could be described by the empirical relation I ∝ P0.2, which is consistentwith the dependence found for the photoproton effect reported in ref . Besides graphene,this leaves only the silicon/silicon-nitride substrate as an alternativephotoabsorber. It is unlikely that the robust photoeffect we observedis due to silicon, since it is insulated from the electrical contactsby a thick 500 nm nitride layer . Moreover, we previously demonstrated that the photoeffect in thesedevices can be entirely suppressed if metals like Au or Ag were usedinstead of Pt nanoparticles,14 which wouldbe inconsistent with silicon as the photoabsorber. Nevertheless, wecharacterized the photoeffect using a 1310 nm light source. Such along wavelength cannot be absorbed by the ∼1.2 eV bandgap insilicon but is readily absorbed by graphene.20 We also studied these devices using an acidic pH polymer (Nafion)to study the role of this long wavelength on proton transport. To rule out anypossible artifact, we performed additional measurements.Neither the polymer support nor Pt nanoparticles could yield the photoresponsereported here S6. Besid12 of 108 V m–1, dissociateswater molecules into protons and hydroxide ions (“Wien effect”in Supporting Information). The same fielddrives protons through graphene and hydroxide ions into the electrolytebulk, separating the generated ion pairs across the proton-selectiveinterface that prevents their recombination. The transported protonthen adsorbs on the Pt nanoparticles on the opposite side of grapheneby acquiring an electron. The role of illumination can be understoodusing the following two observations. First, while the absolute valueof I depends on pH, the photoeffect always enhances I by a factor of ∼10 with respect to the dark case.This is the same enhancement reported in ref (I(P), reported here is the same as in ref (Supporting Information).Our results can be understood as follows. The intense electricfield at the graphene–water interface, which is of the orderd in ref  for devis in ref . These o2O ⇆ H+ + OH–) eventually leads to full electrolysis (H2O →H2 + 1/2O2), producing hydrogenand oxygen gas. The gas evolution rates are much slower than the dissociationstep21 . However, in our devicesthese reactions take place in the large Pt nanoparticle film (H2 evolution) and the Pt counter-electrode (O2 evolution).These catalytically active areas are several orders of magnitude largerin size than the graphene electrode and hence effectively behave asdrain reservoirs for the ions. Because of this, gas evolution reactionsare not a limiting factor in our devices,14 and therefore, we also expect to observe an acceleration of thesereactions under illumination. To confirm this, we measured rates ofH2 and O2 production directly, both in darkconditions and under illumination. For hydrogen measurements, thegraphene electrode faced a vacuum chamber connected to a mass spectrometer,whereas an oxygen-concentration sensor (Clark microelectrode) placedinside the electrolyte solution monitored oxygen production . For zero or positive voltagesapplied to graphene, no H2 could be detected by the spectrometer,in agreement with the previous work.14 For the negativepolarity, both H2 flux and electric current were detectedsimultaneously. 2 molecule was detected by the spectrometer.This charge-to-mass conservation is described by Faraday’slaw of electrolysis: ΦH2 = I/2F, with ΦH2 the hydrogen flux and F the Faraday constant. For O2 gas, the area-normalizedderivative of the oxygen concentration versus time, d(O2)/dt = ΦO, was also described byFaraday’s law, ΦO = I/4F  that was also consistent with Faraday’s lawof electrolysis. The found relations show that H2 and O2 molecules were generated in a 2:1 ratio with 100% Faradaicefficiency, which again shows that the measured currents in our devicesare due to water dissociation.Waterdissociation  and thelifetime of hot electrons in graphene . Wehave also shown that strong electric fields acting across atomicallythin and highly selective interfaces can enable ultrafast ion-chargeseparation. The fundamental insights gained here could be of interestfor development of photocatalysts, for which charge separation isa central consideration. Another emerging possibility is the use ofprotons to reversibly modify the electronic properties of materials.This has been explored in low power memory storage,22 plasmonic materials,23 and neuromorphichardware.24 The protonation dynamics inthese applications is important as it can control the response timein write/read or potentiation/depotentiation cycles.24 In addition, our work suggests that atomically thin interfaces cancontrol ion-charge separation dynamics at time scales comparable tothose in optoelectronics, which could open new opportunities in theseapplications.Our work reports a strong photoeffect in the interfacialwaterdissociation reaction using one-atom-thick graphene electrodes. Theobservation is attributed to acceleration of proton transport whichhappens in synchrony with the dissociation reaction. The findingsare consistent with both the fast rate of proton transport and proton–OH


The exceptional properties of two-dimensional (2D) solids have motivated extensive research, which revealed the possibility of controlling many characteristics of these materials through strain. For instance, previous investigations demonstrated that compressive deformation could be used to direct the chemisorption of atomic hydrogen and oxygen. Still, to our knowledge, there is no work detailing how strain affects the adsorption isotherms of 2D materials and the adsorption properties of materials such as the graphynes, which are monolayers composed of sp and sp The graphynes, which are carbon monolayers composed of atoms with sp and sp5, but the first member of this family, the graphdiyne, was only synthesized in 20106. Since then, the synthesis of 7, and other experimental works proposed using 8 and supercapacitor applications9. Regarding the electronic properties of the graphynes, theoretical investigations revealed that some are semiconductors with moderate gaps10, while others feature Dirac cones in their electronic band structure11. And, in the case of 11. Other simulations also characterized the mechanical properties of various graphynes, particularly under uniaxial strain.The isolation of graphene12. Deformation can also control the bandgap of other TMDs and phosphorene13. However, the magnitude of the bandgap change depends on the type of strain considered 13. Furthermore, the magnetic14 and thermal properties15 of certain 2D materials can be controlled through deformation.We mentioned above that strain can modify the electronic properties of 16. Similarly, biaxial strain increases the size of graphenylene pores, allowing the separation of different gas mixtures at particular pore sizes17. And by manipulating pore size using biaxial strain, it is possible to control water permeability in porous 2D materials while blocking the diffusion of salt ions dissolved in the water19. Finally, it is possible through deformation to change the adhesion in a covalent organic framework, and through this effect, it is possible to pick up and drop off other 2D materials20. This control of a material’s properties using strain has been termed strain engineering13, and here we investigate the effect of deformation on the adsorption properties of graphene and Of relevance to the present work, strain in porous 2D materials can also modify the size and shape of their pores. For instance, by applying uniaxial deformation, it is possible to control diffusion rates in porous graphene, with potential applications in gas separation21. At the same time, theoretical studies have determined the most stable configuration for the adsorbed H22, determined its binding energy curve23, and the adsorption isotherms24. Regarding the adsorption of CO21, while combined experimental and theoretical investigations examined the adsorption of CO25. Concerning methane, its adsorption on graphene was investigated experimentally at temperatures ranging from 253.15 to 293.15 K and pressures ranging from 0 to 8 MPa26. The authors found similar adsorption characteristics for CH27. Finally, note that various studies have also considered doping or decorating graphene with other elements or molecules to (1) improve its H29 and (2) CO31.The adsorption of the gas molecules considered here on graphene has been examined experimentally and theoretically. Experimental investigations on the adsorption of H32. Graphtriyne sheets have also been considered, and the results indicate that single layers of this solid could be used to separate CO33. Meanwhile, investigations on graphynes decorated with other elements are more common, aiming to improve: (1) hydrogen storage38, (2) methane storage39, and (3) CO43.On the other hand, the adsorption of gases on graphyne sheets has received less attention, and to our knowledge, there is no experimental work on this topic. However, theoretical studies have obtained the adsorption energies, preferential adsorption sites, and the change transfer for H44. Next, a DFT study proposed applying compressive strain to create ripples in graphene, creating convex regions where hydrogen preferentially adsorbs, allowing for the control of the bandgap45. Experimental studies then confirmed the preferential adsorption of atomic hydrogen on convexly curved areas in graphene grown on SiC and showed that this uneven adsorption does not occur in graphene bilayers47. Following simulation studies investigated the structure and electronic properties of pristine and hydrogenated corrugated graphene, intended to mimic graphene on SiC48. Regarding oxygen chemisorption, the simulation proposal was to use strain to introduce ripples, leading to preferential O adsorption on regions with high local curvature, followed by oxidative cutting of the resulting structures to produce graphene nanoribbons49. The effect of strain on adsorption properties has also been considered for transition metal dichalcogenides (TMDs) monolayers. Theoretical studies have introduced biaxial deformation to control the interaction of different TMDs with hydrogen50, NO52, and CO53. Sensor applications have been proposed for this effect52.Some studies have already considered applying strain to graphene layers to alter their adsorption properties, but these have focused on atomic hydrogen or oxygen chemisorption. Initial theoretical proposals introduced out-of-plane deformation in graphene to control the hydrogen-graphene binding energy. The authors discovered that convex regions are favorable for atomic hydrogen adsorptionHowever, to our knowledge, there is presently no work detailing how strain affects the adsorption isotherms of 2D materials and no study examining how deformation modifies the adsorption properties of the graphynes. Here, we investigate the adsorption properties of three gases on relaxed and strained monolayers of graphene and graphyne. For each combination of solid and gas, we calculate adsorption isotherms and enthalpies of adsorption. We also examined the structural changes and obtained the mechanical properties of all considered monolayers under biaxial strain.The methodology used to determine the effect of strain on the adsorption properties of mentclass2pt{minim57. Table 58 and Zhang et al. in the case of 59. Overall, graphene is the stiffest and strongest material, with a biaxial modulus of 1020 GPa and enduring stress values of up to 75.8 GPa. On the other hand, the Let us begin the discussion of the results by examining the mechanical properties of graphene and the graphynes. Figure mentclass2pt{minim60. Two of those studies considered the variation of these properties with the temperature. They found that the biaxial modulus55, the ultimate strain, and the ultimate strength of graphene decrease as the temperature increases57, in agreement with our findings.To avoid thermal perturbation on the stress–strain curve and to compare with values reported in the literature, we have also applied equal biaxial deformation at cryogenic temperatures  varies continuously with the deformation, intermediate values might be useful to separate hydrocarbon gases. For instance, at Inspecting the obtained pore diameters, we find that the triangular pores of see Fig. . On the One final note regarding the passage of gases through the graphyne membranes is that we compared energy profiles obtained using classical potentials with profiles obtained using Density Functional Methods in the Supplementary Information Figs. , S24. Wh63 and CCSD64. From Fig. The material deformation may also affect their affinities with gases. To investigate that, we performed grand-canonical Monte Carlo simulations to predict the adsorption isotherms of COon Figs. , S3.Furthermore, we compute the adsorption enthalpy at zero-loading, which is a measure of the heat released during the molecule adsorption and therefore provides a guide to the energy interaction between guest and host. Comparing the enthalpy values in Table entclass1pt{minimasee Fig. . A similFor HFor CHExcept for CHNow, let us consider the effect of the material’s deformation on the adsorption of COon Figs. , S5. To We have presented all isotherms up to now in units of mol of gas per kg of material, and there is a methodology issue behind this choice. Consider Fig. zLet us now carefully analyze the gas-monolayer interaction and the adsorption mechanism. Figure z < 2 Å . These probability values do not affect the results. In Fig. 78 and the enthalpy of adsorption at infinite dilution is given by 79.The COz-direction by computing the histograms of the height position of the adsorbed molecules. The histograms were constructed using bin width equal 1 Å and slider width equal 0.1 Å. To increase the number of adsorbed molecules and improve the statistics, we performed GCMC simulations at a higher pressure of 50 bar for the histogram calculations. We also mapped the interaction between a gas molecule at a given height and a monolayer using the LAMMPS code. To achieve this, we fixed the z position of a gas molecule and varied its x and y positions to scan the surface of the 2D material. And we recorded the interaction energy as we changed the molecule’s position. For each interaction energy map, we scanned an area with dimensions 20 Å For post-simulations analyses we obtained the density profile along Supplementary Information.


In recent years, the field of conductive fabrics has been challenged by the increasing popularity of these materials in the production of conductive, flexible and lightweight textiles, so-called smart textiles, which make our lives easier. These electronic textiles can be used in a wide range of human applications, from medical devices to consumer products. Recently, several scientific results on smart textiles have been published, focusing on the key factors that affect the performance of smart textiles, such as the type of substrate, the type of conductive materials, and the manufacturing method to use them in the appropriate application. Smart textiles have already been fabricated from various fabrics and different conductive materials, such as metallic nanoparticles, conductive polymers, and carbon-based materials. In this review, we study the fabrication of conductive fabrics based on carbon materials, especially carbon nanotubes and graphene, which represent a growing class of high-performance materials for conductive textiles and provide them with superior electrical, thermal, and mechanical properties. Therefore, this paper comprehensively describes conductive fabrics based on single-walled carbon nanotubes, multi-walled carbon nanotubes, and graphene. The fabrication process, physical properties, and their increasing importance in the field of electronic devices are discussed. Traditional textiles were created to protect people from the elements, such as cold and rain, and to serve as covering material. The two most important qualities associated with clothing are their ability to provide protection and their aesthetics. Throughout history, advances in smart materials and electronics have contributed to a unique potential that has led to the emergence of a new field called “smart textiles”. Smart textiles, also called intelligent textiles or e-textiles, are a type of intelligent materials that can detect and respond to changes in their environment . The stiAlthough recent advances in the field of smart textiles are extremely interesting, several obstacles still need to be overcome to make them suitable for commercial and economic use . UnfortuThis article is about the fabrication of smart textiles using carbon-based materials, especially SWCNTs, MWCNTs, and graphene, and is organized as follows: The smart materials, discussing the structure, physical properties, and potential applications of carbon nanotubes, SWCNTs and MWCNTs, and graphene. Then, there are three major sections focusing on the smart textiles fabricated with SWCNTs, MWCNTs and graphene, respectively. Meanwhile, the fabrication method, physical properties, especially electrical properties, factors affecting these properties, and potential applications of smart textiles are described.Materials that are described as “smart” or “functional” are usually part of a “smart system” that can sense and respond to its environment. If they are truly intelligent, they have a significant impact on the performance of smart textiles . In thisCarbon nanotubes (CNTs) belong to the fullerene family, which includes carbon allotropes whose atoms are connected in cage-like configurations, such as a hollow sphere, an ellipsoid, or a cylinder ,20,21,22CNTs are generally classified by the number of carbon layers into single-walled (SWNTs) or multi-walled (MWNTs) carbon nanotubes, as shown in CNTs were used in wide range of applications due to its small and lightweight, which makes them suitable for a . They caGraphene is a flat monolayer of carbon atoms densely condensed into a two-dimensional (2D) honeycomb crystal structure ,59,60,61The exceptional properties of graphene can be exploited in numerous applications, including biomedicine , membranConductive fabrics are usually made from various substrates, such as cotton , polyest2 stretching cycles with a minimum sheet resistance of 65 Ω/□ at 35% tensile load. Recently , conduct−1 was prepared by immersing cotton leaves in SWCNT ink [−1. This approach is characterized by its speed, simplicity, robustness, low cost, and easy scalability. The fabrication of conductive Lycra fabric has also been investigated using the conductive materials SWCNTs and polyaniline, using the immersion drying process [In another study, SWCNT ink was printed on a stretchable substrate using inkjet printing technique . It was ng study , a piezoWCNT ink . The resWCNT ink . The resWCNT ink  fabricat process . The res process , conduct process . The res−1 with a sheet resistance of 1.67 kΩ/□ after four times of immersion. In addition, the thermal conductivity of the fabric was also increased by 70%. MWCNTs dispersed in DMF were used in the recent study presented by Alamer et al. [−2. The results also showed that the conductive cotton treated with HNO3 resulted in a reduction of sheet resistance to 1.5 kΩ.cm−2 which was attributed to the interaction between MWCNTs and cellulose through glycosidic bonds.As discussed in the previous section, conductive fabrics made of SWCNTs have excellent electrical properties; however, SWCNTs are expensive, purification is difficult, and dispersion in liquid is also difficult. Therefore, many researchers focused on fabricating conductive fabrics using MWCNTs instead of SWCNTs because they are cheaper, can be produced in large quantities, and are more stable compared to SWCNTs. Rahman et al.  fabricatr et al.  to prepar et al. . The amo−2, had a specific capacitance of 8.01 F g−1, a high energy density of 6.30 Wh kg−1, and a cyclability of 5000. In the study presented by Nafeiea et al. [−3 S cm−1, which is ten times higher than the conductivity of the wool fabric treated with MWCNTs. In the study presented by Kowalczyk et al. [−1, and the sample exhibited good stability up to 3000 cycles. The conductive yarns based on MWCNTs were fabricated in the study presented by Abbas et al. [The conventional dyeing method was used to deposit synthetic MWCNTs on the surface of cotton fabrics . The depa et al. , conductk et al. , MWCNTs k et al. , where tk et al.  fabricats et al. ,124, in −1 at a graphene concentration of 14% by weight and under continuous mechanical stress.Incorporating graphene into textiles not only imparts conductivity to the textiles, but also enables the production of multifunctional textiles due to the excellent physical properties of graphene, as we discussed in −1 by using graphene oxide nanosheets as a conductive material and a coating reduction method. Briefly, the untreated silk fabric was immersed in bovine serum albumin, which generates a positive charge on the surface of the fabric and increases the absorption of the conductive material when the fabric is immersed in the graphene oxide nanosheet solution. Then, a hydrazine vapor reduction method was used to reduce the graphene oxide on the fabric. In the study presented by Zulan et al. [−1. In another study [−1 was prepared using reduced graphene oxide and a dip coating and reduction method. The results also showed that the conductive cotton yarn exhibited mechanical stability up to 1000 cycles and absorbed UV irradiation of about 1.0 mA/W under bending deformation. Yarns from Calotropis gigantean [−1 at high concentration and was shown to be resistant to washing, which was due to the hydrogen bonding formed between the fiber and graphene during the dyeing process.Lu et al.  fabricatn et al. , the coner study , a flexiigantean , which h2 for the untreated fabric to 23.15 Ω. cm2 for the fabric coated with three layers of reduced graphene oxide. In another study [Molina et al.  fabricater study , the knier study  were use−1 at a graphene concentration of 6 wt.%. Another interesting method to improve the bonding between graphene and cotton fabric and to fabricate a scalable conductive fabric with a length of 150 m and a sheet resistance of about 11.9 Ω/□ was presented in the study by Afroj et al. [4 S m−1. The biocompatible conductive fabric sensor was fabricated using graphene nanoplatelets dispersed in a water-based ink and screen-printed onto the fabric surface [−1 at a graphene concentration of 3.8 wt%.A conductive stretch-sensitive fabric was fabricated using graphene oxide nanosheets and a reducing deposition method . Brieflya et al.  found a j et al. . The conj et al.  was usedj et al. , and the surface . The res−1, and this value was stable up to five washing cycles. In the study by Sahito et al. [−1, which is not affected by bending and washing cycles. In another process [In addition, Yapici et al.  also fabo et al. , the suro et al. . A new m process , UV lighConductive fabrics based on SWCNTs have been prepared by various fabrication methods: chemical vapor deposition, filtration technique, dyeing and drying method, inkjet printing method, dipping and drying method, and drop-casting method. It has been shown that the electrical conductivity of conductive fabrics and sheet resistance has a wide range from low to high values, and depends on various factors, such as: deposition times, dopants, content of SWCNTs in the fabric, stretching cycles, number of coating layers, treatment of the fabric with plasma, and mixing of SWCNTs with other carbon-based materials, such as MWCNTS and graphene, or with conductive polymers, such as polyaniline and PEDOT:PSS. These SWCNTs based fabrics have been used in various applications such as: UV light shielding, piezoresistive sensor, supercapacitor, antenna, and metal thread.Conductive fabrics based on MWCNTs were prepared by various fabrication methods: Dipping and drying, drop casting and drying, dipping, impregnation and drying, dipping and drying, and tape casting. The fabricated conductive fabrics exhibited a wide range of electrical conductivity, which was influenced by several factors: Size of MWCNTs, number of dipping operations, type of organic solvents, content of MWCNTs, temperature, dopants, repetition of the fabrication process, use of anionic and cationic surfactants, and use of enzymes. These conductive fabrics based on MWCNTs have been used as electrodes in supercapacitors, and as electrodes in dye-sensitised solar cells and block UV light.Conductive graphene-based fabrics have been fabricated using graphene, graphene nanoribbons, graphene oxide, graphene nanosheets, natural graphite flakes, and graphene ink. Various methods were used in the fabrication process: wet coating, trapping method, chemical reduction method, vacuum filtration—hot press method, dipping and gamma ray reduction method, cationization—dip coating, coating and reduction method, dyeing and reduction method, dipping and reduction method, reducing deposition method, brush coating, pad dry curing method, screen printing method, chemical vapor reduction method, electrostatic self-assembly—low temperature reduction process, UV reduction method. The electrical properties of graphene-based materials are influenced by several factors: repetition of the manufacturing process, graphene content, graphene oxide concentration, tensile stress, cationization process, number of coating layers, number of dipping cycles, exfoliation agent, and number of printing cycles. Thus, the conductive fabrics produced have been used in various applications: electrode for a dye-sensitive solar cell, strain sensor, portable supercapacitors, UV blockers, heat-resistant gloves, electrocardiogram electrodes, and portable heaters. Finally, the field of smart textile fabrication from carbon-based materials is developing rapidly but needs further development to bring these applications from small scales (in research laboratories) to large scales . Therefore, more basic research is needed to enable the next wave of smart textile products.This review article summarizes the method of designing and fabricating electrical, flexible, and lightweight conductive fabrics with embedded SWCNTs, MWCNTs, and graphene, and their applications in the field of smart textiles. The development of smart textiles was carried out in three stages: the first stage is to impart conductivity to the textiles, the second stage is to fabricate the smart textiles, and the final stage is to functionalize the conductive yarns. Carbon-based materials, particularly SWCNTs, MWCNTs, and graphene, are discussed from their structure, physical properties, and potential applications to their use in the design and fabrication of conductive fabrics with a wide range of electrical conductivity and interesting physical properties that make them suitable for various wearable electronic applications. We come to the following conclusions:


Τ/τP-181 combination. Hence, TDP-43 combined with tau proteins could be a useful tool for the diagnosis of genetic FTD with TDP-43 underling histopathology, supplementing clinical, neuropsychological and imaging data.Cerebrospinal fluid (CSF) biomarkers, namely total tau, phospho-tau and amyloid beta peptides, have received much attention specifically regarding Alzheimer’s disease (AD), since they can detect the biochemical fingerprint of AD and serve as a diagnostic tool for accurate and early diagnosis during life. In the same way, biomarkers for other neurodegenerative disease pathologies are also needed. We present a case series of six patients with genetic frontotemporal dementia (FTD), with TDP-43 underlying proteinopathy, in an attempt to assess TDP-43 as a novel biomarker alone and in combination with established AD biomarkers for this specific patient group, based on the principles of personalized and precision medicine. Our results indicate that genetic TDP-43-FTD is characterized by increased CSF TPD-43 and increased TDP-43 × τ A common neuropathological feature of neurodegenerative diseases is cytoplasmic protein aggregates. Cytoplasmic mislocalization and aggregation of TAR-DNA binding protein 43 (TDP-43) is found in the vast majority of patients with amyotrophic lateral sclerosis (ALS), and in approximately half of patients with frontotemporal dementia (FTD) with ubiquitin inclusions .C9orf72 gene involving a hexanucleotide repeat expansion (GGGGCC) in the first non-coding intron of the gene, responsible for both familial and sporadic FTD and ALS, but more frequently for the combined FTD-ALS phenotype [Recently, great progress in the field of neurogenetics has resulted in the recognition of several gene mutations related to the FTD-ALS spectrum, especially with ubiquitin-TDP-43 predominant histopathology, thus highlighting a new brain proteinopathy. The most recent, but more common discovery, is a genetic mutation in the henotype .GRN) can also lead to FTD with TDP-43 inclusions [VCP) gene constitute another rare cause of FTD, which has been recognized to present with the clinical syndrome of “IBMPFD” , often in combination with ALS in certain families. [TARDBP) are also an extremely rare cause of the FTD phenotype [Heterozygous loss of function mutations in the progranulin gene (clusions . GRN mutamilies. . Mutatiohenotype . T) and phosphorylated (τP), as well as amyloid-β peptides (Aβ) expressing the pathological features of the disease, have been recognized as biomarkers with high diagnostic accuracy and are now included in research diagnostic criteria for AD [T, tau phosphorylated at the threonine 181 site (τP-181) and decreased Aβ with 42 amino acids (Aβ42) in the CSF, has currently been established as the biochemical signature of the disease in vivo [In recent years, cerebrospinal fluid (CSF) has proven to be an excellent material for biomarker discovery for neurodegenerative diseases . Due to a for AD ,13,14,15 in vivo . Τ/τP-181 may increase sensitivity and specificity (>80%) [P-181/τT ratio has been proposed as an indirect indicator of a non-tauopathic pathology [Accordingly, the detection of TDP-43 could be a specific biomarker for TDP-43 proteinopathy. Attempts to quantify TDP-43 in CSF have been successful ,18,19,20athology ,23.Since almost all the aforementioned studies lack pathologic confirmation, we isolated and present herein a case series of six patients with genetic FTD and a presumed TDP-43 underlying proteinopathy. We aimed to investigate how the above-mentioned biomarkers perform in a personalized setting, based on the principles of a precision medicine clinical approach. We also include the clinical and imaging data of this group of patients.A total of six patients were included in this case series study. All patients were hospitalized at the 1st Department of Neurology of the National and Kapodistrian University of Athens at Eginition Hospital during the years 2011–2018. The study was performed in accordance with the ethical guidelines of the Declaration of Helsinki, and had the approval of the local Ethical and Deontology committee of our hospital (244/5-7-2010). All study subjects and/or their relatives gave informed consent for inclusion in the study. Patients underwent detailed clinical, neuropsychological, biochemical and neuroimaging examination, to exclude secondary causes of dementia and establish an FTD diagnosis, based on the criteria of either Rascovsky et al.  for probT, τP-181 and Aβ42 and Aβ40 were measured in duplicate by commercially available enzyme-linked immunosorbent assay (ELISA) kits  according to manufacturer’s instructions, and blind to clinical diagnosis. The above biomarkers are measured routinely in the Neurochemistry and Biomarker Unit of our department, as part of an everyday diagnostic approach, as described elsewhere [The CSF levels of τlsewhere . TDP-43 T = 511.6 pg/mL, normal τP-181 = 28.7 pg/mL, decreased Aβ42 = 504.2 pg/mL, but normal Aβ42/Aβ40 = 0.1 and decreased τP-181/τT = 0.056 ratio. TDP-43 = 4.46 pg/mL and the formula TDP-43 × τΤ/τP-181 = 79.5 were increased. Genetic testing revealed a c.463-2A > G splice site variant—heterozygous mutation in the GRN gene. The patient also had more than a 2-fold decrease in plasma progranulin levels, as described elsewhere [A 61-year-old female patient was admitted to our department for progressive language impairment over the past 1–2 years. She had word-finding difficulty, with intact comprehension, an absence of phonological speech or grammar errors, no other significant cognitive/behavioral disturbances, and preserved activities of daily living. Neuropsychological assessment revealed mild cognitive impairment, with an MMSE score of 25/30 and a FAB score of 12/18. Her writing skills and phrase and sentence repetition were preserved, too. On magnetic resonance imaging (MRI), moderate cortical atrophy in the frontal and temporal lobes, with left predominance and relative preservation of the hippocampus, was apparent . CSF bioT = 565.12 pg/mL, normal τP-181 = 41.9 pg/mL, Aβ42 = 640 pg/m and Aβ42/Aβ40 = 0.11, while τP-181/τT = 0.074, was decreased. TDP-43 = 5.92 pg/mL and formula TDP-43 × τΤ/τP-181 = 79.9 were increased. Genetic testing revealed a c.934-1G > A splice site variant—heterozygous mutation on the GRN gene, and similarly, a decrease in plasma progranulin levels, as in case 1.A 60-year-old male patient, with a family history of dementia on his mother and maternal grandmother’s side, presented with language impairment over the last 10 years. Initially, he had difficulty naming objects and writing. Three years later he developed effortful speech with sound errors, decreased fluency, and memory complaints and disorientation. At the age of 55, a dysexecutive syndrome as well as behavioral disturbances (overeating and irritability) appeared. Gradually, he became almost mute, while bradykinesia and tremor on the right hand appeared. At the time of his admission, he was completely mute, and unable to eat. Neurological examination revealed diffuse muscle atrophies without fasciculations, bilateral Parkinsonian syndrome (more severe in the right limbs), pyramidal syndrome with spasticity, hyperreflexia, and Babinski sign bilaterally. Neuropsychological assessment was impossible to perform. MRI performed at the age of 57 revealed significant cortical atrophy in the frontal and temporal lobes with left predominance A–C, withT = 316.16 pg/mL and τP-181 = 27.2 pg/mL, decreased Aβ42 = 501.51 pg/mL but normal Aβ42/Aβ40 = 0.14 and low τP-181/τT = 0.086 ratio. TDP-43 levels were 5.74 pg/mL and TDP-43 × τΤ/τP-181 = 66.7, both increased. The genetic testing revealed a pathologic hexanucleotide repeat expansion in the C9orf72 gene.A 72-year-old male patient, with a family history of motor neuron disease (brother), gradually developed language  and memory impairment, bradykinesia and stereotypical movements of the right hand over the last two years. Three months before admission, his symptoms aggravated. Neurological examination revealed bilateral akinetic-rigid Parkinsonian syndrome, mild pyramidal signs (mainly on the right lower limb with hyperreflexia and extensor plantar response) and apraxia. Neuropsychological assessment revealed severe diffuse cognitive deficits . On MRI, severe cortical atrophy in frontal and temporal lobes was observed . CSF bioT = 250.3 pg/mL, τP-181 = 35.45 pg/mL and Aβ42 = 935.95 pg/mL, while τP-181/τT ratio was 0.14. The levels of TDP-43 = 2.48 pg/mL and TDP-43 × τΤ/τP-181 = 17.5 were also normal. Genetic testing, however, revealed a hexanucleotide repeat expansion in the C9orf72 gene.A 54-year-old female patient, with a medical history of depression with psychotic features was referred for evaluation. She had a family history of dementia (mother) and symptoms of motor neuron disease . The patient presented with apathy, inertia, disinhibition, loss of empathy and sympathy, unusual stereotyped and perseverative behaviors, and hyperorality with overeating of carbohydrates during the last two and a half years. One and a half year before her admission, she also developed inappropriate crying and a language impairment. Neurological examination revealed severely impaired non-fluent speech with single-word answers, severe disorientation in time and space, and frontal gait disorder. Neuropsychological assessment revealed moderate cognitive deficits with an MMSE score of 15/30 and a FAB score of 6/18. On MRI, some T2 hyperintense lesions attributed to chronic small vessel ischemia (Fazekas II), and mild cortical atrophy of the temporal lobes was observed . CSF bioVCP-associated syndrome was raised. Genetic testing  revealed a pathogenic heterozygous missense mutation p.R159H (c.476G > A) in the VCP gene, confirming the diagnosis of IBMPFD [A 62-year-old male patient, with a medical history of arterial hypertension and an operation for a herniated lumbar disc at the L4-L5 level, was referred for evaluation of neuropsychiatric symptoms. He had a family history of “presenile dementia” in his mother, ALS of his older brother, who died at the age of 53, and behavioral symptoms of another brother. The patient presented with progressively deteriorating steppage gait 20 years ago. A year before admission, the patient was unable to walk independently. Additionally, he developed obsessive behavior, aggression, palilalia and apathy. His neuropsychological evaluation revealed severe cognitive impairment, suggestive of predominant frontal lobe dysfunction, MMSE: 2 and FAB: 0 (not applicable). Brain MRI showed mild frontal lobe atrophy . EMG shof IBMPFD .T = 356.75 pg/mL, τP-181 = 32.77 pg/mL, low Aβ42 = 431 pg/mL, but normal Aβ42/Aβ40 = 0.1 and reduced ratio τP-181/τT = 0.091, while TDP-43 levels = 5.27 pg/mL, and TDP-43 × τΤ/τP-181 = 57.3 were increased.Lumbar puncture and CSF biomarker analysie were performed one year later and revealed normal τT = 328.05 pg/mL, τP-181 = 32.07 pg/mL, decreased Aβ42 = 397.1 pg/mL, with normal ratio Aβ42/Aβ40 = 0.17 and decreased τP-181/τT = 0.098, while TDP-43 levels 5.74 pg/mL and the formula TDP-43 × τΤ/τP-181 = 57.66 were increased. Genetic testing finally revealed a hexanucleotide repeat expansion in the C9orf72 gene.A 41-year-old male patient, with a medical history of dyslipidemia was admitted to the neurology department for delusions of religious content over the past 12 years. Moreover, apathy, a change in eating habits with carbohydrate craving and weight gain gradually developed, while dysarthria, executive deficits, stereotypical behaviors and inappropriate laughter appeared over the last two years. Finally, difficulty in swallowing liquids was noticed six months before admission. Neurological examination revealed pseudobulbar dysarthria, an absence of pharyngeal reflex, bilateral pyramidal syndrome and fasciculations in tongue and back muscles. Neuropsychological assessment revealed mild to moderate frontal cognitive deficits with an MMSE score 28/30, and a FAB score 15/18. Neurophysiological examination could not confirm lower motor neuron involvement. On MRI, significant cortical atrophy in frontal, parietal and temporal lobes was observed . CSF bioP-181 and only two out of six cases had increased τT. Four out of six cases had decreased Aβ42; however, all had a normal Aβ42/Aβ40 ratio. Five out of six patients had increased TDP-43 and formula (TDP-43 × τΤ/τP-181) values, while τP-181/τT was abnormal in all.C9orf72 gene; two had a novel mutation in the GRN gene which led to more than a 2-fold decrease of plasma progranulin levels, as expected [VCP gene. Although small in number, this group of patients is indicative of the most common mutations found in the Greek population [Herein, we present six patients with clinical, neuropsychological and imaging data fulfilling the criteria of Rascovsky et al.  and Gornexpected ,30; meanpulation ,31.T, τP-181, Aβ42 and Aβ40) and a novel emerging (TDP-43) biomarker were analyzed. Combinations of these biomarkers as the formula TDP-43 × τΤ/τP-181 and the ratio τP-181/τΤ were also calculated, as previously suggested [In this group of well-characterized genetic FTLD patients, established CSF biomarkers for AD diagnosis  system proposed by the National Institute of Aging and Alzheimer’s Association (NIA–AA) Research Framework group, AD biomarkers are divided into those with molecular specificity for AD, namely τent case . All pre42 was reduced, but the use of Aβ42/Aβ40 ratio restored normality, ruling out amyloidopathy (A). The Aβ42/Aβ40 ratio has been suggested to better reflect the presence or absence of amyloid pathology than Aβ42 alone. Furthermore, the Aβ42/Aβ40 ratio correlates better with amyloid load in PET (positron emission tomography) [In four out of the six presented cases, Aβography) , as it hography) ,34,35,36Τ/τP-181 formula. TDP-43 is a protein, which may be suggestive and specific of TDP-43 proteinopathy. Additionally, the τP-181/τT ratio was decreased, which in non-AD (FTD) patients could be considered as an indirect indication for a non-tauopathic pathology [The main findings of our study, however, were the increase of CSF TDP-43 levels  in five out of six cases, and the increased values of the TDP-43 × τathology ,23.C9orf72, GRN, VCP, and TARDBP genes for FTLD and the FTD-ALS spectrum [TDP-43 aggregation appears in approximately 50% of all FTLD cases  and in 97% of all ALS cases . Mutatiospectrum . TDP-43 spectrum . Abnormaspectrum .Although thought to exist as an intracellular/intranuclear protein, recent studies using ELISA and Western blot techniques suggest that TDP-43 can also be detected in extracellular fluids such as plasma and CSF .C9orf72 pathological repeat expansion, represents an exception. Nonetheless, the proposed mechanisms through which C9orf72 mutations cause neurodegenerative changes are three: C9orf72 loss-of-function through haploinsufficiency, RNA toxic gain of function, and gain of function through the aggregation of toxic dipeptide repeat proteins [The results of the present study indicate that CSF TDP-43 levels were increased in five out of six patients with TDP-43 histopathology-associated mutations. Case 4, with a proteins . The firproteins . As a reproteins . These Dproteins ,45. The TDP-43 abnormalities have also been observed in other neurodegenerative diseases as well, mainly in ALS, the other end of the ALS-FTD spectrum, but also in AD, limbic-predominant age-related TDP-43 encephalopathy (LATE), cerebral age-related TDP-43 with sclerosis (CARTS), and some other even rarer conditions ,46,47. RThere are also other biomarkers that can be used to differentiate FTD patients from patients with other neurodegenerative diseases, such as NfL . HoweverThe current study has certain limitations. TDP-43 pathology can only be presumed from genetic mutations, since it is an ante-mortem study. Another disadvantage is the small number of patients, coming from a single-center study that included only patients of Greek origin; however, the mutations that the patients harbored are indicative of the most commonly observed in TDP-43-related FTLD found in the Greek population ,31.Our study also has some important advantages. We describe patients that have undergone extended clinical, neuropsychological and imaging examinations, fulfilling the criteria of Rascovsky et al.  and GornBased on the results of the present personalized case series study, which is nonetheless indicative of the mutational profile of the population of Greece, the following conclusions can be drawn:Established AD biomarkers are reliable indicators for the exclusion of AD pathology in genetic FTLD.P-181/τΤ ratio may be useful as an indirect marker for non-tauopathic pathology.A decreased τTDP-43 alone and combined with tau proteins, could be a useful tool for the diagnosis of genetic FTD patients with TDP-43 underlying pathology.Future studies with larger and well characterized patient cohorts are needed to further explore the diagnostic potential of these biomarkers in differentiating TDP-43 proteinopathy from non-TDP-43-related pathologies.


Group II introns are extraordinarily versatile self-splicing ribozymes and retrotransposable elements widespread in bacteria and in bacterial-derived organelles (mitochondria and chloroplasts) of fungi, algae, plants, and of some early-branching metazoans . These lDespite their bacterial origin, it is now widely accepted that group II introns have been repurposed by natural evolution into the eukaryotic spliceosomal introns and the spliceosome . Both grIn bacteria, most group II introns encode a multifunctional reverse transcriptase (RT) and invade genomes through “retrohoming”, a highly site-specific mobility process operated both by the ribozyme and its encoded RT enzyme . RetrohoNatural group II introns are remarkably diverse regarding the structural organization of their ribozyme moiety, self-splicing pathways and the strategies they use to invade genomes. Here, I discuss recent findings regarding the splicing and mobility activities of group II introns that further emphasize the structural plasticity of these elements and their ability to be remodeled by molecular evolution into unique RNA-based machines, including the eukaryotic pre-mRNA splicing system.Group II introns possess a single catalytic center that positions two metal ions responsible for catalysis of the transesterification reactions . This orThe small terminal domain VI harboring the unpaired branchpoint adenosine , has lonRecent crystal structures of an intron lariat captured just after splicing completion revealed the crucial role of the 2′-5′ branch structure in promoting proper assembling of the second-step active site . UnexpecMore recently, cryo-electron microscopy (cryo-EM) structures of a bacterial subgroup IIB1 intron in complex with its reverse transcriptase and a DNA target allowed visualization of the large-scale toggling undertaken by domain VI between the two splicing steps . InteresBacillus genus that contain a large insertion of about 53/56 nucleotides (sometimes longer) located precisely between the end of domain VI and the intron 3′ splice site  is much more flexible than previously recognized and this plasticity could have an impact on 3′ splice site selection. Interestingly, some natural group II introns that deviate from the typical 6-nt/7-nt length of the bpA-3′ss spacer support this hypothesis. A remarkable example is provided by a number of closely related IIB introns found in bacteria of the ice site . These iice site . In anotirst one . Both 3′The ability to directly invade DNA by reverse splicing and the high specificity of target recognition make mobile group II introns unique retrotransposons. The DNA target is primarily recognized by extensive arrays of Watson-Crick pairings (∼12–14 bp) between the intron EBS sites and the complementary IBS sequences lying on the top strand of the target . ImportaThe studies discussed above highlight the flexibility of group II introns and their ability to acquire novel structural features with specific functionalities in splicing and mobility. Over the course of evolution, this remarkable plasticity not only allowed the great diversification of group II introns, but it certainly played a key role in the molecular process that repurposed group II intron self-splicing into one of the most complex cellular function of eukaryotes: spliceosomal pre-mRNA splicing. It is hypothesized that during this evolutionary pathway, fragmentation of ancestral group II intron RNPs gave rise to the small nuclear RNAs (snRNAs) U2, U5 and U6 that constitute the active core of the extant spliceosome . Concomi


This paper explores the capabilities that contribute to community transformation and the common pathways followed by communities in the 100 Million Healthier Lives SCALE  initiative in their transformation journeys towards a “Culture of Health”.Funded by the Robert Wood Johnson Foundation (RWJF), from 2016 to 2020, between 18 to 24 community coalitions nationwide participated in SCALE, the goal of which was to co‐design, implement, test, and scale up a model called the Community of Solutions (COS) Framework, that built community capacity around a set of skills and behaviors to advance culture change and create sustainable improvement in health, well‐being, and equity. We adapted and applied two qualitative research techniques, meta‐ethnography and participatory action synthesis, to evaluate SCALE initiative data.Eight concepts emerged that represent the knowledge, capabilities and practices commonly acquired and utilized across the communities. Overall, these concepts emphasize individual and team leadership, quality improvement skills, an intentional focus on equity, and partnerships for spread and sustainment. Concepts were linked into lines of arguments which were unique storylines explaining the transformation pathways. Three stories of the transformation process emerged from the data. Causal Loop Diagrams (CLDs) were created to represent non‐linear system relationships and visually capture some of the most important dynamics of the process of transformation. Even with vast heterogeneity among the SCALE communities and the diversity of activities that the communities undertook, our analysis showed there were a few basic principles that undergirded the process of building capability for transformation.The knowledge from our findings should be useful to expand further research and practice in community learning systems. The need for systems changes at the community level, and the urgent imperative to focus on equity has never been more apparent than during the devastation from COVID‐19, where a thousand different versions of the pandemic have been manifested in a thousand different communities. Public health analysis and responses to COVID‐19 have not adequately considered the effect of inequities arising from the intersection of race, gender, and class on an individual or community's experience of the pandemic.The COVID‐19 pandemic has merely reinforced what has been known for a long time. Health and well‐being are intimately connected to community level contexts and require collaborative, transformational solutions that are built on communities' capacity to learn, in real‐time, how to improve what matters most to them.1.2Community of Solutions (COS) Framework that built community capacity around a set of skills and behaviors to advance culture change and create sustainable improvement in health, well‐being, and equity.Convened by the Institute for Healthcare Improvement (IHI), 100MLives was a global movement and collaboration from 2014 to 2020 in support of Robert Wood Johnson Foundation's Culture of Health (COH) Strategy.1.3What are the key contributors to achieving capability for community transformation?What knowledge, capabilities, practices, and relationships did communities acquire and utilize?What common pathways did communities follow in their transformation journey?What are the mechanisms through which communities brought about change?Our description of SCALE in this paper focuses on the following four questions:22.1The theory of change for SCALE is the COS model shown in Figure 2.2The primary activities based on the COS model is the development—co‐designed with communities—of skills of community coalition members in five areas  through intense training, coaching and support activities. The key activities are shown in Table 2.3processes of change in the complex environments within which the communities operated, rather than attempting to assess the extent to which these processes resulted in the achievement of specific outcomes. The evaluation was intentionally designed as a participatory partnership between the funder, the implementation partners, the communities, and a multidisciplinary evaluation team led by the Universities of North and South Carolina. Recognizing the heterogeneity in coalition structures, community contexts, and areas of improvement emphasis, the evaluation approach was designed to be flexible, adaptive, and process focused. The evaluation also needed to be sensitive to the fact that much of the knowledge about the implementation was generated by and existed within the communities, and that their interpretation of the data was an important part of the knowledge synthesis process. The approach was related to learning or developmental evaluationThe primary intent of the evaluation was to advance knowledge about the 2.4inquiry, observation, and reflection were used to obtain data about the implementation to capture as much contextual information as possible. Together, these methods provided a multifaceted view of the SCALE implementation process that was synthesized and periodically shared with the implementation team and the communities to stimulate improvements to the program design and its execution.The formative evaluation approach is described in detail in Scott et al.2.5The summative evaluation was designed in the latter half of the project after the theory of change and the primary components of the program design had stabilized. This evaluation focused on the behaviors component of the theory of change through a longitudinal analysis of each community's traversal through SCALE. Since one of the goals of the summative evaluation was to generate insights from a plurality of voices, a collaborative bottom‐up methodology called ToAST (Telling our Amazing Stories Together) was developed.http://www.communitycommons.org/entities/ac745b94-1b33-4143-acc3-6df3e0217b3cDetails of the ToAST approach are described in Reed et al.improvement science and measurement, COS skills , spread and scale up, race, racism and equity and engaging people with lived experience. Across communities, the topical sub groups identified similar ideas that were categorized into themes. Subsequently, the subgroups contextualized the topic‐specific themes into community‐specific concepts representing the workgroup's interpretation of the particular elements of each community's journey through SCALE. For communities that had complete data on all the topical areas, the community‐level concepts were then assembled into overarching concepts representing the most common mechanisms employed by all communities to bring about change. Descriptions were developed for each of these concepts and were sent back to the coalition members to validate and revise if needed. Communities were also invited to create lines of arguments. In meta‐ethnography, these are new story lines or overarching explanations arising from the synthesis of the date. To create lines of argument, communities linked the overarching concepts visually into transformation pathways. The evaluation team used these to develop causal loop diagrams (CLDs) to highlight the nonlinear nature of the transformation process. Table To implement the ToAST approach, a synthesis workgroup consisting of volunteer participants from the SCALE communities and selected implementation and evaluation team members was assembled. The synthesis workgroup systematically worked through the seven phases of meta‐ethnography, working in four topical areas that roughly aligned with the skill areas in the theory of change: 3We describe results from both the formative and summative evaluations, organized by the questions of interest presented earlier in this paper.3.1robust theory of change that could serve as a common roadmap for all communities despite heterogeneity in contexts and areas of improvement focus; (b) a structured improvement process using a well‐known quality improvement method called the Model for Improvementimplementation support system for coaching communities through the process of improvement and change; and (d) an evaluation approach that provided ongoing data for learning and improvement. The existence of these areas was necessary but not sufficient. There needed to be a process that could facilitate the adaptation of these areas in response to changes in context or to new and emergent priorities. The process established for feedback of the formative evaluation results provided the mechanism to do this. An example of change to each area based on recommendations from the formative evaluation is shown in Table The formative evaluation revealed four key areas that were critical drivers for achieving the goals of SCALE: (a) a 3.2individual and team leadership, quality improvement skills, an intentional focus on equity, and partnerships for spread and sustainment. The first two concepts (nos. 1 and 2) related to individual leadership skills that gave the SCALE coalition members the confidence to approach complex problems and to engage in sensitive and difficult conversations. The next two (nos. 3 and 4) pertained to capabilities related to team leadership and practices that brought together those with local context specific knowledge needed to bring about change. Concepts 5 and 6 involved the acquisition of technical skills that were needed to complement local knowledge. A systematic approach to improvement based on a change theory, and accessing specialists  were practices utilized by the communities. Finally, as indicated by concepts 7 and 8, each community made equity and racism a central focus of their work and built deep relationships with other SCALE communities to support and sustain their work.The overarching concepts from the ToAST process represent the knowledge, capabilities, and practices commonly acquired and utilized across the SCALE communities. These are shown in Table These eight cross community concepts emerged in Phase 6 of the ToAST process as the output of multiple levels of synthesis shown in Table 3.3The transformation pathways shown in Figure It took longer than expected to develop the infrastructure of the “Quarterback Organization” and to provide engagement opportunities with activities. [Our SCALE community] was focused on fixing the lake and not fixing the fish, which involves more long‐term changes.The “build the core” story was reported by the coalitions that had strong existing connections with people with lived experience in their communities but first focused on strengthening their internal capacity by learning and internalizing improvement and relationship building tools and then used these to expand and deepen their relationships with the community to spearhead honest and authentic conversations about race and inequity. A core team member of a SCALE community that followed this pathway stated:We had a discussion on racism at every coalition and staff meeting. We created a safe and open space to discuss personal &/or patient experiences with racism or current events dealing with racism.The “create a shared language” story was followed by community organizations that began with using community partners with deep connections within the community to develop a shared language that facilitated mutual communication and dialog. Initial conversations to strengthen the ability to communicate was seen as especially important by these communities as was the need to create safe spaces for open and honest discussions and sharing experiences. These efforts accelerated trust and relationships in these communities. A coalition member stated:While all coalitions had a community champion from the outset, the “integrate people with lived experience (PLE) early” story emphasized developing community leaders as the first component of building the core team's ability to engage with the wider community. They also used the power of the local social networks to build relationships to build relationships quickly to advance equity work.A focus on authentic engagement of partners, building trust and relationships was foundational to the ability to scale up and sustain work. SCALE helped us offer more leadership development opportunities for community champions and staff members. This was valuable for building local capacity to improve authentic engagement and peer mentorship opportunities to lead civic engagement and service leadership.As one core team member remarked:3.4While the lines of argument are a convenient visual representation of how change happens, their sequential structure does not capture the complexity of the change process. CLDs allow us to better represent nonlinear system relationships. Using the data from Figure input into the system is the available external funding that influences the extent to which SCALE activities can be supported. The output from the system is the SCALE goal, that is, the capability for community transformation. As seen from the picture, there are three direct drivers of the output represented by the lines leading to the community transformation box: (a) the ability of communities to create collaborations with broad multisector leadership (including community members); (b) creating opportunities for dialog about addressing racism and equity and (c) engaging people with lived experience.In Figure SCALE program activities were intended to strengthen these drivers. The two places in the system where SCALE interventions were focused (leverage points to change the system) are indicated by the two arrows L1 and L2. L1 indicates that all SCALE training and support activities revolved around strengthening collaboration within the community. This enabled communities both to create leaders from within the community (loop R1) and facilitate dialog around difficult questions about inequity and race, without which true change is not possible. At the same time, to ensure that collaboration did not exclude those who needed to be at the center of transformation activities, SCALE required the intentional engagement of people with lived experience in leadership roles (leverage point L2), which strengthened the capability for transformation in multiple ways.As SCALE progressed and the community coalitions took on more complex projects, and attempted to spread what they had learned across the region, there was an increasing need for implementation support and technical assistance as indicated by the loops R2 and R3. These required increased resources, which were limited, by the available resources as shown in loop B1 . SCALE communities needed to do their best with fixed resources that were stretched thin, and these served as a limit to what the communities could ultimately achieve.4Even with vast heterogeneity among the SCALE communities and the diversity of activities that the communities undertook, our analysis showed there were a few basic principles that undergirded the process of building capability for transformation. A planned approach to transformation based on a theory of change, devoting time and attention to skill and relationship building and centering equity, ensuring the active engagement and leadership of community members and creating space for dialog and conversation were common ingredients across all communities. Trust within and across SCALE communities, and the ability to interact and learn were vital. Fostering collaboration and trust needed sustained coaching and technical assistance resources, both to build leadership and improvement skills and to recruit and engage community champions. The reliance on grant funding to support these activities was a limitation.4.1What does it mean to be in a community system that is truly engaged in learning to advance health, well‐being, and equity in sustainable ways? Results from our evaluation align with the existing literature related to multisector coalitions. Well‐known frameworks such as Collective Impactprocesses through which these capacities are developed. Even though these pathways were inductively constructed from the unique experiences of individual communities, we discovered enough commonality across communities to have confidence that Figures Our evaluation results also advance the knowledge about how community coalitions can build capacity to bring about change. The CLD and the transformation pathways extend the literature beyond just the factors that influence community capacity for change to hypotheses about the 4.2Our findings from SCALE about the dynamics that affect community change also allow us to set a direction for future research. Future efforts must focus on supporting community coalitions to reduce their reliance on external funding and technical assistance for their transformation initiatives. One possibility could be through strengthening regional hubs of excellence with local expertise that enable communities to learn from and support each other. SCALE began to build the infrastructure for regional change by requiring communities to spread the capabilities they had acquired, but in an environment where the communities had competing priorities, limited resources for implementation and staff turnover, there were limitations to what could be achieved. Part of the national and research conversation about shifting power to advance equity and dismantle racism should include ways to shift research and implementation resources to communities.Future research should investigate how to convert the constraints shown in Figure 5Our evaluation approach was designed to make the best use of existing project data, which was recorded in a variety of systems with varying degrees of completion. Consistent availability of high‐quality data to meaningfully document progress and learning was an ongoing challenge. The SCALE implementation teams introduced multiple measurement plans and mechanisms for data collection over the duration of the initiative, but in the end, there were still gaps in data quality and completeness. The ToAST process used the collective knowledge of the various stakeholder groups and triangulated data from various sources to develop the concepts and transformation pathways, but there were gaps in data that might affect the final results and conclusions. Also, our results have demonstrated progress in skills and behaviors, which are the first two components of theory of change. Given the range and complexity of the projects undertaken by the SCALE communities, there was not enough time for communities to demonstrate how this progress resulted in the achievement of the COH outcomes related to health, well‐being and equity before the funding ran out. This is not unusual, and our inability to evaluate whether the capabilities acquired by the communities resulted in improved health and well‐being is a serious limitation. The second CLD we have presented describes the scenario where communities have sustained financing to continue their transformation efforts. If this scenario were realized, it would be possible to follow communities over time to evaluate the impact of their work. Financial sustainability should be emphasized as a required component for future grants to facilitate this scenario.6The United States has invested trillions of dollars in health, well‐being and equity with little to show for these efforts. Simply investing more money and resources will not likely result in measurable improvement. Complexity‐informed approaches that approach well‐being through the lens of learning rather than through the lens of technical program implementation are needed. SCALE is an example of such an approach. Communities learned how answers to complex questions about change are not apparent at the start of the work, and that practices of learning and testing with meaningful measures to guide progress will help uncover solutions. Learning in a context of complexity requires a willingness to “fail forward,” in an environment where failures are opportunities for learning and growth rather than for judgment. Our results validate the relevance of the elements of the SCALE COS Theory of Change as the key components of skills and behaviors needed to bring about change. The extent to which the acquisition of these skills and behaviors drive sustainable health, well‐being and equity outcomes they aim to achieve should be a priority for future research.The authors have no conflicts of interest to declare.