Schistosoma spp. that present limitations for culture maintenance. This strategy also has prompted the construction of multi-epitope based vaccines, with great immunological foreseen properties as well as being less prone to contamination, autoimmunity, and allergenic responses. Therefore, in this study we applied a robust immunoinformatics approach, targeting S. mansoni transmembrane proteins, in order to construct a chimeric antigen. Initially, the search for all hypothetical transmembrane proteins in GeneDB provided a total of 584 sequences. Using the PSORT II and CCTOP servers we reduced this to 37 plasma membrane proteins, from which extracellular domains were used for epitope prediction. Nineteen common MHC-I and MHC-II binding epitopes, from eight proteins, comprised the final multi-epitope construct, along with suitable adjuvants. The final chimeric multi-epitope vaccine was predicted as prone to induce B-cell and IFN-γ based immunity, as well as presented itself as stable and non-allergenic molecule. Finally, molecular docking and molecular dynamics foresee stable interactions between the putative antigen and the immune receptor TLR 4. Our results indicate that the multi-epitope vaccine might stimulate humoral and cellular immune responses and could be a potential vaccine candidate against schistosomiasis.Schistosomiasis remains a serious health issue nowadays for an estimated one billion people in 79 countries around the world. Great efforts have been made to identify good vaccine candidates during the last decades, but only three molecules reached clinical trials so far. The reverse vaccinology approach has become an attractive option for vaccine design, especially regarding parasites like  Schistosoma. The World Health Organization (WHO) classifies schistosomiasis as the second most importance socioeconomic disease in the world and as the third most relevant parasitic disease regarding public health ; Africa and South America (S. mansoni); China and the Philippines (S. japonicum)  .Strategies for schistosomiasis control and elimination have been implemented and reexamined in the last three decades –8. MajorSchistosoma has biological features that turn it into a tough task. Different parasite evolutionary phases and their remarkable ability to evade and subvert immunological mechanisms of elimination represent a significant bottleneck .Given the above, here we based our work on an immunoinformatics approach to identify all hypothetical S. mansoni available in this database were gathered. The PSORT II serverProteins sequences were obtained from the GeneDB databaseIn order to confirm PSORT II prediction and identify the extracellular domains from plasma membrane sequences, CCTOP topology predictor50 value <500 nM were selected for further analysis.The aforementioned extracellular domains were submitted to IEDBProteins extracellular domains were also used to predict linear B-cell epitopes, which was carried out by ABCpred serverInitially, CLTs and B-cell epitopes, provided by the same extracellular region, were aligned. CTLs fitting within B-cell sequences were then submitted to the analysis of immunogenicity. The chosen tool50 < 50 nM were selected. Subsequently, CTLs and HTLs epitopes overlapped with B-cell sequences were aligned with each other to select those matching. For all these alignments the MULTALIN tool was usedSame alignment approach was conducted for HTLs and B-cell epitopes. Only HTLs fitting within B-cell sequences and presenting ICThe ability of previously selected epitopes to act as an antigen was evaluated by the VaxiJen serverP9WHE3). Subsequent, the Pan DR epitope (PADRE – AKFVAAWTLKAAA) adjuvant was also fused to act as a T helper Lymphocyte stimulus. Then, CTLs epitopes were added followed by HTLs. Finally, the C-terminal portion included a 6xHis tag for further purification assays.In order to fuse the selected epitopes in a rationally immunogenic fashion we used suitable linkers and adjuvants. The well-described linkers EAAAK, GGGS, GPGPG, HEYGAEALERAG, and AAY were used to connect different components. The N-terminal region of the multi-epitope protein starts with HIV TAT peptide that plays a cell penetrating role. Next, to improve the immunogenicity, we choose the 50S ribosomal protein L7/L12 (Locus RL7_MYCTU) (Accession no. Schistosoma species (S. japonicum and S. haematobium) were evaluated using UniProt Knowledgebase dataset. This former analysis was conducted for the final eight proteins from which we have predicted our epitopes.To evaluate similarity with human proteins and therefore reduce autoimmunity possibilities, a BLASTConsidering the importance of the microbiota in maintaining the organism's homeostasis, we submited the multi-epitope protein against the proteome of commonly found gut microbiota, with the aid of Pipeline Builder for Identification of drug Targets (PBIT) serverTo assess the antigenicity of whole multi-epitope vaccine, VaxiJen serverThe presence of IFN-γ inducing epitopes in the multi-epitope vaccine sequence were predicted by IFNepitope serverConsidering that IFNepitope server was trained with MHC II epitopes, we further submitted all final MHC II epitopes selected before chimera assembly. This analysis was carried out in order to check whether the chimeric-MHC II epitopes we selected would be predicted as an IFN- γ inducing peptide. In this case, the server module used was the module “Predict,” which ranks the inserted peptides in positive or negative order, based on their potential to induce IFN- γ.6. The chosen method was the IEDB recommended and the proteasome type was the immunoproteasome. Epitopes were predicted based on HLA allele set mentioned earlier, in the CTL epitope prediction section.After multi-epitope vaccine conception, we decided to confirm if upon processing steps in the cell, our chimeric-MHC I epitopes would be generated. In order to perform this we applied the Proteasomal cleavage/TAP transport/MHC class I combined predictorin silico immune simulations were conducted by C-ImmSim serverTo further characterize the immunogenicity and the immune response of the multi-epitope vaccine, Escherichia coli, yeast, and mammal cells) and the instability index  structure was obtained using the Iterative Threading Assembly Refinement (I-TASSER) serverThe predicted vaccine 3D structure from I-TASSER was submitted to refinement in order to improve structure quality, both locally and globally. This task was carried out by GalaxyRefine serverDiscontinuous or conformational B-cell epitopes are formed as a result of protein folding that can bring into proximity distant residues and form it. It has been estimated that >90% of B-cell epitopes are discontinuous . In ordeSchistosoma antigens. Then, for docking analysis, the TLR 4 receptor structure was retrieved from the RCSB PDB database (PDB ID: 2Z63) and edited by Chimera softwareThe ability to trigger innate immune receptors is broadly described for To predict the 3D structure of multi-epitope vaccine and TLR 4 receptor complex, the SwarmDock server was used. The server has a flexible protein-protein docking algorithm, which initially pre-process and minimize input structures and performs docking through a hybrid particle swarm optimization/local search. Next, it minimizes, re-ranks and clusters docked poses. Lastly, democratic clustered structures are provided as PDB format . The anaMolecular dynamics (MD) simulation is applied to determine the stability of receptor-ligand complexes . GromacsE. coli (strain K12). The output also includes two more parameters; Codon Adaptation Index (CAI) and percentage of GC content. For CAI, the ideal value is 1.0 and the GC content should gather values between 30 and 70%. To carry the cloning into an expression vector, pET28a(+), SnapGene software was used. SalI and BamHI restriction sites were introduced flanking the multi-epitope sequence to ensure the suitable insertion into the plasmid.Finally, in order to provide a feasible plasmid construction harboring the multi-epitope sequence, Java Codon Adaptation Tool (JCat) server was used. This server provides a codon optimized version of the interest DNA sequence, based on a chosen organism . Here weGene DB search provided 584 hypothetical transmembrane protein sequences, from which 37 were predicted as most likely located at plasma membrane by PSORT II server. Out of it, CCTOP predictor allowed us to identify 54 extracellular domains ≥ 30 amino acids.50 < 500 nM, considering a set of 27 alleles. Diverse epitopes were predicted to more than one MHC alleles and they were considered as one in further analyses.IEDB server predicted promiscuous CTLs epitopes covering 108 alleles. It was found 13,981 epitopes presenting percentile rank <0.5, for all extracellular domains. In turn IEDB foreseen 6,893 HTLs epitopes presenting percentile rank <0.5 and ICThe ABCpred server predicted linear B-cell epitopes for all 54 extracellular domains. The analysis generated a total of 1,583 sequences.The alignment of multiple sequences aid to identify and select epitopes harboring cellular and humoral inducing ability. The first alignment was carried out between CTLs and B-cell epitopes, which generated a set of CTLs epitopes overlapping with B-cell sequences. Out of this, a second set of peptides were obtained through class I immunogenicity analysis. The outcome was a group of best CTL epitopes.50 < 50 nM. Finally, both best epitopes groups (CTLs and HTLs) were aligned to each other and yielded 55 matched sequences.Similarly, a set of HTLs epitopes overlapping with B-cell sequences was obtained and improved by selecting only those with ICThe best 55 epitopes were submitted to VaxiJen antigenicity analysis. Out of the total, 19 epitopes presented score >0.7  and therIn order to construct the multi-epitope vaccine, we have used suitable adjuvants and linkers. The final sequence is represented in Homo sapiens proteins did not reveal any match. The same result was observed when chimeric epitopes were individually submitted to a BLAST analysis. As expected, the search for similarity between multi-epitope protein and gut microbiome revealed considerable identity for the 50S ribosomal protein L7/L12, but non-homology for other components, considering server's cutoff , >20 h in yeast and >10 h in E. coli, both in vivo. Regarding solubility, SOLpro server provided good prediction (0.9010) upon overexpression in E. coli.Based on amino acid sequence, ProtParam predicted a set of physicochemical parameters for the multi-epitope vaccine. It has foreseen a size of 52 kDa and isoeletric point (pI) of 5.93. The total numbers of positive and negative charged residues were 38 and 46, respectively. The instability index (II) was computed to be 31.24, which classifies the multi-epitope vaccine as a stable molecule. This stability was reinforced by the aliphatic index, 86.06. The higher the aliphatic index, the more stable is a protein in a broad range of temperatures. During heterologous expression in bacteria or yeast is essential a long period of half-life. In this sense, our molecule presented an estimated half-life of 30 h in mammalian reticulocytes . Ten best templates (Z-score ranging from 1.01 to 5.59) were used to predict five potential tertiary structures of the multi-epitope vaccine. Each of these five models presented an individual C-score value: −0.41, −2.34, −2.59, −4.30 and −2.70. C-score range is typically between −5 and 2, with values >-1.5 indicating a correct global topology. Therefore, we selected the C-score −0.41 model as our multi-epitope tertiary structure.The I-TASSER server starts modeling process from structure templates identified in the PDB library. The server can generate tens of thousands of template alignments, but it only uses the ones of the highest significance, which is measured by the After choosing the best 3D model, a refinement process was carried out by GalaxyWeb server in order to improve structure quality. As a result, the server generated five refined models . To seleAccording to ElliPro server, a total of 253 residues were distributed in 10 conformational B-cell epitopes, with scores ranging from 0.60 to 0.877. The epitopes size ranged from four to 77 residues .The SwarmDock server and LIGPROT v2.2 software allowed us simulate and analyze possible interactions between the ligand (multi-epitope vaccine) and the immune receptor (TLR 4). The best docked structure was selected based on minimum energy value, −43.71. This complex exhibited 14 hydrogen bonds, 29 hydrophobic interactions for the ligand and 21 for the receptor, as demonstrated in In order to confirm the proper engagement of TLR 4 with the putative multi-epitope vaccine, a MD simulation was conducted. After energy minimization step, temperature and pressure equilibration phase were assessed during 100 ps. Temperature raised quickly and reached 300 K, which was kept throughout the interval analyzed . SimilarSubsequently, the final trajectory was used to analyze some essential parameters; the first one was the RMSD. This parameter measures reflect the stability between the receptor and ligand where mild fluctuations point to a stable interaction. The second parameter, RMSF, represents amino acids side chains fluctuations. Elevated fluctuations at the plot indicate highly flexible regions in the receptor-ligand complex while mild ones point to continuous interaction between receptor and ligand molecules. Most of our selected complex has presented uninterrupted interactions (0.5 nm), except for a specific region of highly flexibility (1.8 nm), as demonstrated in Jcat server generated an optimized DNA sequence presenting good parameters. The CAI value was 1.0, indicating high chance of increased expression. GC content reached 54.25%, which remains in the optimal range (30–70%). Expression vector pET 28a (+) carrying the multi-epitope vaccine insert is represented in the Historically, vaccine development has been considered the best cost-effective method for combating diseases caused by infectious pathogens . For decin silico analysis. As constituents of this putative antigen, authors selected T and B-cell epitopes found in seven major vaccine targets  described for the disease . Our selection criteria allowed us to reach a final set of epitopes, CTLs and HTLs, with great affinity scores for MHC molecules, and being potentially recognized by B-cells. This feature is essential for vaccine approach, given the relevance of neutralizing antibodies against the pathogen and the support of helper T cells in the prolonged antibodies production, as well as in maintaining the activation of CTL response . FurtherConnection between epitopes, as well as their merge with other chimera components, was achieved by using linkers. Studies of naturally-occurring multiple domain proteins have prompted the use of linker peptides for artificial protein fusion . In the Leishmania major  have been widely investigated as possible agents for vaccine delivery. The integration of these peptides with antigenic sequences aims to improve their uptake by Antigen Presenting Cells (APCs), thereby favoring antigen processing and presentation processes . Here, wia major , clearly+ T cell responses, and IFN-γ production, as outcomes closely related to protection against the parasite infection , which is also described as essential for combating Schistosoma infection , primarily through their secretion and excretion products . Durães E. coli (10h) and the molecule stability  propose the usage of this organism as a platform for heterologous expression. For this reason, we have optimized vaccine candidate codons, based on E. coli strain K12, and performed the in silico cloning in a common expression vector, pET28a (+). Moreover, predicted isoelectric point (5.93) indicates that under neutral pH conditions the protein will assume a negative global charge, which lean toward to aid affinity purification procedure, as those carried in Nickel (Ni2+) immobilized columns (The physicochemical features foreseen for the multi-epitope candidate strongly suggest heterologous expression and antigen purification as viable processes. The predicted half-life in  columns , 84. BesS. japonicum and S. haematobium (After decades searching for an effective antigen to be used as proper vaccine against schistosomiasis, few molecules reached clinical trials, keeping the pursuit for alternative approaches open . An attrmatobium . On the matobium . Smp_175matobium .in vitro and in vivo assays.In general, extracellular domains of proteins will always be an appreciable source of vaccine candidates for different pathogens. Likewise, studies of hypothetical proteins can translate into the discovery of potential candidates and stimulate their investigation/annotation. Therefore, we have combined these concepts into a multi-epitope protein that presented itself as a good putative antigen. Immunogenic, physicochemical and structural properties suggest that our vaccine candidate might yield promising results on near future The original contributions presented in the study are included in the article/RS, ST, VA, and DL designed the whole analysis. RS, LF, FO, ML, MP, EM, AT, and RK carried out most of the analysis. RS, ST, and DL wrote the manuscript. RS prepared figures. DL submitted this paper. All authors reviewed the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


This study shows that starch type can also be an important parameter to control this mesoporosity. Carbons with controlled mesoporosity (Vmeso from 0.1–0.7 cm3/g) have been produced by the pre-mixing of different starches using an ionic liquid (IL) followed by a modified Starbon® process. The results show that the use of starch from corn and maize  is the better combination to increase the mesopore volume. Moreover, “low-cost” mesoporous carbons have been obtained by the direct carbonization of the pre-treated starch mixtures with the IL. In all cases, the IL can be recovered and reused, as demonstrated by its recycling up to three times. Furthermore, and as a comparison, chitosan has been also used as a precursor to obtain N-doped mesoporous carbons (5.5 wt% N) with moderate mesoporosity (Vmeso = 0.43 cm3/g). The different mesoporous carbons have been tested as cathode components in Li-O2 batteries and it is shown that a higher carbon mesoporosity, produced from starch precursor, or the N-doping, produced from chitosan precursor, increase the final battery cell performance (specific capacity and cycling). Previous studies have demonstrated that the mesoporosity of carbon material obtained by the Starbon Every year, there is an increasing demand of energy due to growing populations. Most of the consumed energy still comes from fossil fuel . Their combustion is causing the greenhouse gas emissions , contributing to climate change. In order to mitigate global climate change, the transition from fossil fuel’s energy to zero-emission energy is necessary and is only possible with the development of sustainable materials and technologies. 2, steam) or chemical  methods [2RR) [The production of porous carbon materials from natural and biomass sources such as polysaccharides  and as  methods ,15, Carbds [2RR) ). Furtheds [2RR) , Na-ion ds [2RR) , Li-S [1ds [2RR) , and Li-ds [2RR) ).® process [In 2007, White, Clark, Budarin, and MacQuarrie, from the University of York, designed and patented a method called Starbon process  for the Starch is one of the major polysaccharide sources in nature, present in roots , tubers , cereals , and fruits . It is mainly composed by amylose and amylopectin, and proteins and lipids in minor quantities. Amylose and amylopectin are polysaccharides formed by glucose units joined by glycosidic bonds in a lineal and branched structure, respectively, depending on the source of the starch. The ratio amylose: amylopectin varies and so are the physico-chemical properties of the material. Several studies demonstrated that the higher the amylose content is in starch, the higher the porosity of the carbonized material is .Ionic liquids (ILs) are ionic and non-molecular salts that melt below 100 °C. Normally, they are composed by an organic cation and an organic or inorganic anion, and they are very interesting because of their distinctive properties, such as low vapor pressure, high thermal stability, recyclability, conductivity, and so on . Due to ® process (with ultrasonication instead of a microwave treatment at high temperatures). The mesoporosity is influenced not only by the amylose content but also by the starch type. In this way, the starch mixture of Maize starch  and Hylon VII starch  with 60 wt% of amylose results in a higher mesoporosity than the pure Hylon VII starch  with 70 wt% of amylose. At lower amylose content (less than 45 wt%), the higher mesoporosity is obtained with the mixture of Maize and Hylon V. The recovery and reuse of the IL is studied up to three times with minor changes in the final mesoporosity. Additionally, we show a low-cost process for the production of mesoporous carbons from starch by avoiding the need to perform gelation, retrogradation, and freeze-drying (a saving of four days) as required in the Starbon® process. Finally, we demonstrate that the performance of these carbons as cathode materials in Li-O2 batteries is greatly influenced by their mesoporosity, resulting in higher capacity and cycling for the carbons with a higher mesoporosity or N-doping [In this work, we demonstrate the preparation of carbon materials with controlled mesoporosity from polysaccharides using a process based on an IL pre-treatment (with recovery and reuse of the IL), which is followed by a modified StarbonN-doping ,26,27.® . Pure amylose, pure amylopectin, chitosan (MMW), potassium iodide (KI), dimethyl sulfoxide (DMSO) 99.9% v/v, and para-toluenesulfonic acid monohydrate (p-TSA) were purchased from Sigma-Aldrich . The ionic liquid (IL) 1-butyl-3-methylimidazolium chloride 99% (BMIMCl) was purchased from abcr . Ethanol 96% v/v, iodine (I2), and sodium hydroxide (pellets) were purchased from Scharlab . Tert-butanol (TBA) was obtained from Honeywell .The starches maize (M), Hylon V (HV), and Hylon VII (HVII) were obtained from IngredionThe amylose content in the commercially available starches was determined by a colorimetric method measuring the absorbance at 600 nm of a complex formed between amylose and iodine by UV-Vis spectrophotometry. The equipment used was Jasco V-630 UV-Vis spectrophotometer .−1 and a spectral range from 4000 to 500 cm−1. Proton nuclear magnetic resonance (1HNMR) spectroscopy was performed using a Bruker Avance DPX spectrometer  of 250 MHz (5.8 T) equipped with an automatic sample changer Bruker BACS-60 and a probe 1H/13C/31P.Fourier-Transform Infrared-Attenuated Total Reflection (FTIR-ATR) spectroscopy was performed using an Agilent Cary 630 FTIR , after 32 scans with a spectral resolution of 4 cmThermogravimetric analysis (TGA) was performed with a TA instrument Thermogravimetric Analyzer Q500 . The samples were loaded in flat platinum pans and the measurements were taken under a heating rate of 10 °C/min from room temperature to 900 °C.Gas Permeation Chromatography (GPC) was performed using an Agilent 1200 Infinity GPC  equipped with a 390-LC Multi Detector Suite.® Field Emission-Scanning Electron Microscopy (FE-SEM) microscope  equipped with a GEMINI® II column, and High-Resolution Transmission Electron Microscopy (HR-TEM), using a Jeol JEM-2011 scanning microscope .High Resolution Scanning Electron Microscopy (HR-SEM) was performed using a Carl Zeiss MERLINBJH/SABJH.Nitrogen adsorption and desorption isotherms were measured using a Nova 2200e Surface Area and Pore Analyzer from Quantachrome Instruments . The surface area was determined by the application of multi-point Brunauer-Emmett-Teller (BET) equation and the pore size was calculated according to the Barrett, Joyner, and Halenda (BJH) average pore diameter equation, D = 4VQuantitative Nitrogen determination was measured by an Elemental Analyzer Eurovector EuroEA3000 .The micro-Raman spectra were recorded using a 532 nm laser excitation line with a dispersive spectrometer Jobin-Yvon LabRam HR 800 , coupled to an optical microscope Olympus BXFM .2 2.5 mM in distilled water (this solution must be stored in darkness at 4 °C). Then, different starch solutions containing different ratios of pure amylose and amylopectin  were prepared by dissolving 20 mg of mixture into 8 mL of DMSO and heated for 15 min at 85 °C in a water bath. The solutions were allowed to cool and then diluted to 25 mL with distilled water. After, a solution was prepared by the addition of 40 mL of distilled water to 1 mL of the starch precursor solution, which was followed by the addition of 5 mL of the iodine precursor solution and, finally, diluted to 50 mL of distilled water. The formation of an amylose-iodine complex was measured by the Ultraviolet-Visible (UV-VIS) absorbance at 600 nm. The calibration graph obtained was used to calculate the real amylose content of the commercially available starches. For this, previously, the starches were defatted with a treatment with 120 mL 75% n-propanol at 85 °C for 7 h in a Soxhlet extractor to eliminate the lipids. Then, the determination of amylose in the commercial starches was carried out by the same procedure described above but adding 20 mg of the starch.In order to determine the real content of amylose of the obtained commercial starches ,29, a caThis process was carried out by dispersing a 10 wt% of the starch mixture with distilled water under strong magnetic stirring for 1 h. Then, the dispersion was ultrasonicated for 15 min (2200 J) using a tip  in a pulse mode of 1 s. After cooling at room temperature, the gel was maintained at 5 °C for 2 days to allow the retrogradation of the structure. After that time, 30 wt% of TBA with respect to the total water content was added to the gel and kept for 1 h in a rotatory mixer at lower speed rotation (5 rpm). Then, 1 wt% of p-TSA with respect to starch was added and kept for 1 h under rotation. The gel was then frozen with liquid nitrogen and dried for 2 days in a freeze-dryer  at 0.02 mbar and room temperature. After freeze-drying, an expanded dried gel was obtained. The carbonization process was carried out under inert atmosphere (argon) in a multi-stage process from room temperature to 800 °C for 24 h in a laboratory chamber furnace .−1. Finally, the precipitate was separated by vacuum filtration and dried overnight at 80 °C and 60 mbar, obtaining a completely white powder.In the first step of this process, the starches were dispersed in an IL. For this, BMIMCl was heated at 95 °C under mechanical stirring until completely melted. Then, 5 g of the starch or the mixture were added to the IL (20 wt% of starch) and stirred for 6 h at 95 °C by forming a viscous white dispersion. After that, the starch or the mixture was quantitatively separated from the IL using several washing steps. In the first step, 75 mL of distilled water was added dropwise, the dispersion was heated to 70 °C, and 150 mL of ethanol were added dropwise to precipitate the starch. The mechanical agitation was maintained overnight, and the next day the precipitate was separated by centrifugation at 4500 rpm for 10 min, which was followed by several washings with ethanol until complete elimination of IL, detected by FTIR at 1560 cm® based process was performed as explained before in In the following step, the StarbonThis process involves the previous preparation of the starch or mixture in the IL, as explained in After the starch separation in the process described in The preparation of N-doped mesoporous carbon from chitosan was carried out by dissolving 2 g of chitosan into 80 g of distilled water, acidified with 1 mL of acetic acid. The mixture was stirred for 1 h and then it was maintained for 1 day at 5 °C for the gelation process. Then the gel was precipitated into 100 mL of NaOH 3 M and washed with distilled water until complete neutralization. The precipitate was separated and 30 wt% TBA was added and kept under rotation for 1 h. The mixture was freeze-dried for 2 days before carbonization at 1000 °C under argon, following the same conditions.2-cathodes were prepared by bare coating of a slurry composed of the studied carbons, 5 wt% Polyvinylidene Fluoride  as a binder dispersed in 1-methyl-2-pyrrolidinone  and carbon black  as a conductive additive with a 8:1:1 final dried weight ratio. They were mixed in a glass vial and stirred with an electromagnetic stirring method for at least 4 h and, finally, mixed with a homogenizer  for at least 10 min. Afterward, the prepared ink was spread onto a gas diffusion layer . The wet thickness was 100 μm and coating was made with the bar coating method. The coated GDL was dried in an oven at 50 °C overnight under vacuum 0.05 Pa. Subsequently, disk-shaped electrodes were punched with a diameter of 18 mm and an active carbon electrode loading ranged from 3.2 to 4.3 mg/cm2. The cell was assembled in an argon-filled glovebox and then connected to oxygen gas circulation (purity grade 4.5–99.995%) through the cathode with a flow rate of 5 mL/min during the experiments.All electrochemical tests were carried out in the same reference battery cell system. An ECC-Air cell  was used for testing all active materials. In the test system, a metal lithium chip as an anode  and a glass fibre filter paper  as a separator, which was soaked with 1M LiTFSI-TEGDME electrolyte for 5 min, were used. The electrolyte was prepared mixing 1 M bis(trifluoromethane)sulfonimide lithium salt  in tetraethylene glycol dimethyl ether . O+ cut-off voltage was used for the galvanostatic cycling tests. The full discharge capacity was measured by discharging the cell at a constant current density of 100 mA/g of active carbon. Then another cell was assembled for the cyclability test and the performance of the material was studied by charging and discharging the battery cell at 50 mA/g with a limited capacity of 300 mAh/g. All electrochemical measurements were performed using a VMP3 and BCS810 multi-channel galvanostat-potentiostat . A 2.15 and 4.35 V vs. Li/LiA calibration curve was obtained from the absorbance measurements of the mixtures of pure amylose and amylopectin with iodine, as shown in ® process with some modifications. The results obtained with each of them are explained in the following sections. Mesoporous carbons have been prepared following two different processes based on the Starbon® based process. It is already known that some IL are good solvents for polysaccharides and can increase its solubilization in comparison with water [In an attempt to control the mesoporosity of the final carbons from the starting starch mixtures and to try to improve it, a pre-treatment of the starch and mixtures with an IL has been introduced before the modified Starbonth water ,30. Acco® based process. In order to generate carbons with different amylose contents than the commercial ones, various mixtures of Maize, Hylon V. and Hylon VII were prepared and the final nanoporosity was measured , which is higher than pure Hylon VII with a 70 wt% amylose (0.80 cm3/g and 77%). However, at lower amylose content (less than 50 wt%), the highest mesoporosity is obtained with the Maize Hylon V mixture, showing that the final mesoporosity not only depends on the amylose content but also on the type of starch. The pore size grows with increasing amylose content and is higher for the mixture of MHV vs. MHVII at the same amylose content.For this process and as explained in the previous section, the starch mixtures were first blended with an IL and then processed following a modified Starbonmeasured . An expemeasured . The higThe morphology of the carbons prepared from Maize and HVII with different contents of amylose was studied by HRSEM microscopy. The obtained images in To better understand the higher mesoporosity of some of the mixtures, GPC studies were performed to study the possible influence of molecular weight of the starch mixture on the porosity. For this, GPC analysis was performed on starch mixtures after their treatment with the IL. ® based process. According to the results  with the IL in the pre-treatment step to obtain the highest mesoporosity, different concentrations were studied. For this, mesoporous carbons were prepared following the previous described procedure with the starch mixture dispersion in IL and after the modified Starbon results , the dis® process consisting of a preparation of the mixture of starches in water. Then a gelation with sonication, a solvent elimination by freeze-drying, and finally the carbonization at 800 °C without the IL pre-treatment. The porosity .To study the IL chemical stability after its use, several analyses were performed . The mesoporosity increases for the MHVII samples as the amylose content increases, as expected. A high content of nitrogen (5.5 wt%) was measured for the chitosan-derived carbon. The reference sample  has a low mesoporosity.The nanoporosity data can be seen in D/IG . The full discharge capacity was measured by discharging the cell at 100 mA/g. Then the cyclability of the cell is studied by charging and discharging the battery cells at 50 mA/g with a limited capacity of 300 mAh/g. The results are shown in The mesoporous carbons produced in the previous section were tested in the Li-O® based process. In this sense, the starch with a higher amylose content, for instance Hylon VII with 70 wt% amylose, produced carbons with a higher mesoporosity. In this study, we demonstrated that, apart from amylose content, the type of starch mixture influences the final meso-porosity. For that, we applied a pre-treatment of the starch mixtures with an IL followed by a modified Starbon® process with sonication for the gelation step instead of a microwave treatment. We showed that, at high amylose content (higher than 50 wt%), the highest mesoporosity was obtained at 60 wt% amylose using a mixture of Maize and Hylon VII instead of the pure Hylon VII with a higher amylose content (70 wt% amylose). At lower amylose content (less than 50 wt%), the best mixture to increase the mesoporosity of the carbon was the mixture of Maize and Hylon V. This result was also confirmed by the HRSEM images of the mixtures Maize and Hylon VII, where the nanorod-like carbon structures forming the mesopores became smaller and more homogeneously distributed through the material when the amylose content was increased  with and without the IL pre-treatment , but a more cost effective process was achieved by direct carbonization of the IL-treated starches (0.33 cm3/g for 60% MHVII and 0.44 cm3/g for 70% HVII) with a higher mesoporosity than the carbons obtained directly by a modified Starbon® based process (0.25 cm3/g for 60% MHVII and 0.23 cm3/g for 70% HVII).In this work, we also demonstrate the effect of the process used on the final carbon mesoporosity. For this, we compared the modified Starbon2 battery cells were Chitosan and 60% MHVII with 4057 mAh/g and 3978 mAh/g specific capacity and 69 and 60 cycles, respectively, without any capacity lost. When compared to the reference material , we see that the capacity was doubled and the cyclability of the cell increased more than four times. From these results, it is demonstrated that, for Li-O2 technology, the total carbon mesoporosity contributed highly on the performance of the materials for both specific capacity and number of cycles. For the carbons obtained from MHVII, the one with the highest mesoporosity (60%MHVII) has the highest capacity and the one with the lowest mesoporosity (36%MHVII) has the lowest capacity. This shows the importance of the mesoporosity in Li-O2 technology, as it enables a better oxygen diffusion and lithium transfer [2O2 formation, which is directly related to the capacity. However, in the case of the carbon obtained from chitosan, although the mesoporosity is lower, the capacity was the highest achieved. In this case, the effect of the presence of nitrogen clearly improves the performance of the material, as already described in the literature [Best performing materials in Li-Otransfer  and, theterature ,32,33,34® based process, using sonication for the gelation and followed by retrogradation, freeze-drying, and carbonization under inert conditions. A low-cost process to produce mesoporous carbons directly from the carbonization of the IL-treated starches is also shown. Different starches, starch mixtures, and different concentrations were tested to find the optimal conditions that resulted in a carbon material with an increased mesoporosity. The highest mesoporosity was found for the mixture of starch from maize (Maize) and corn (Hylon VII) containing a 60 wt% amylose and prepared with a concentration of 20 wt% in the IL . For all these methods, it was demonstrated that the used IL can be recovered and reused without any effect on the final mesoporosity. Moreover, an N-doped mesoporous carbon was obtained from chitosan following a similar Starbon® based process.This work describes new processes to produce carbon materials from polysaccharides with controlled mesoporosity consisting in the IL pre-treatment of starch or starch mixtures. For the fabrication of high mesoporous carbons, this IL pre-treatment is followed by a modified Starbon2 batteries showing an increased capacity and cycling stability with the carbons with the higher mesoporosity. The doping with nitrogen, in the case of the chitosan-derived carbon, showed a further increased performance. These carbons were tested as cathode components in Li-OProcess for preparing mesoporous carbon material. LEITAT Submitted 28/02/2019 Patent nr. P20382135.0.


Advances in the health sciences rely on sharing research and data through publication. As information professionals are often asked to contribute their knowledge to assist clinicians and researchers in selecting journals for publication, the authors recognized an opportunity to build a decision support tool, SPI-Hub: Scholarly Publishing Information Hub™, to capture the team’s collective publishing industry knowledge, while carefully retaining the quality of service.SPI-Hub’s decision support functionality relies on a data framework that describes journal publication policies and practices through a newly designed metadata structure, the Knowledge Management Journal Record™. Metadata fields are populated through a semi-automated process that uses custom programming to access content from multiple sources. Each record includes 25 metadata fields representing best publishing practices. Currently, the database includes more than 24,000 health sciences journal records. To correctly capture the resources needed for both completion and future maintenance of the project, the team conducted an internal study to assess time requirements for completing records through different stages of automation.The journal decision support tool, SPI-Hub, provides an opportunity to assess publication practices by compiling data from a variety of sources in a single location. Automated and semi-automated approaches have effectively reduced the time needed for data collection. Through a comprehensive knowledge management framework and the incorporation of multiple quality points specific to each journal, SPI-Hub provides prospective users with both recommendations for publication and holistic assessment of the trustworthiness of journals in which to publish research and acquire trusted knowledge. Journal trustworthiness and rigor have been much discussed since the transition to electronic publishing in the 1990s . The broThe shift in the payment model, whereby funding often derives from author-paid article processing charges, has created an opportunity for players who are less driven by standards to earn a profit potentially using misleading or non-transparent practices , 14. TheLong-established bibliographic databases—such as Ulrichsweb, OCLC’s WorldCat, and the National Library of Medicine (NLM) Catalog –23—offerTo assist authors in making decisions about where to publish, several organizations have issued guidance and checklists, including: Think. Check. Submit. , the DirHowever, finding each component of information for multiple journals can be a time-consuming and daunting process. Several tools attempt to automate journal selection for potential authors and facilitate quick discovery of relevant journals for publication, based on scope and other elements such as impact metrics –41. In tThere is a clear challenge in evaluating quality and validity in the increasingly complex world of publishing. As has been our experience at the Center for Knowledge Management at Vanderbilt University Medical Center, information specialists are often called upon to assist clinicians and researchers in identifying where to publish their research and, in the process, steer them away from journals that lack rigor. As requests for assistance with journal investigation and selection have become more frequent, our team has recognized the need to scale the process through an innovative approach to automating its information specialists’ collective knowledge of the publishing industry. Making the process scalable allows us to provide this important service while freeing our team’s limited resources to address other challenges. Knowledge management approaches—such as knowledge curation, data organization, and content maintenance—can be directly applied to help guide clinicians and researchers through this dynamic landscape.Our team’s experience applying knowledge management strategies to inform clinical decision support , 43 was Our center’s approach to identifying where to publish manuscripts has traditionally involved manually capturing multiple data points to generate a snapshot of a journal’s scope and publishing policies. We have leveraged our manual process, combined with best practices from the aforementioned criteria and checklists, to define the metadata for the Knowledge Management Journal Record. The record focuses on multiple journal rigor and quality data points that can support informed decision-making for all journal types and avoids prescribing subjective assessments. The Knowledge Management Journal Record is intentionally designed to present users with impartial, current data about journals in as objective a manner as possible, allowing users to judge which elements of the journal record are most important to them.The current fields in the Knowledge Management Journal Record have been selected based on a comprehensive review of publication guidelines and standards and the ease with which fields could be captured in a structured and/or semi-automated manner. The fields are organized in four sections: “General Information” ; “Metrics & Indexing” ; “Publication Policies” ; and “Open Access” . Leveraging the Knowledge Management Journal Record as its core infrastructure, we created SPI-Hub as a decision support tool for journal identification and assessment. SPI-Hub currently includes three primary functionalities: “Search by Topic,” “Search by Journal,” and “Search by Author.” Links to journal suggestion and selection resources that are publicly available are also provided in a “Resources” tab for prospective users to consult as a complement to SPI-Hub’s information. A “Contact Us” page allows users to provide feedback and suggest journals to add to the SPI-Hub database. The “Contact Us” page has a tab with Frequently Asked Questions , which pTo help prospective authors find journals that publish works on a given topic, the tool offers a multifaceted search against PubMed. Using the Entrez Programming Utilities interface (E-utilities) provided by the National Center for Biotechnology Information (NCBI), we first recommend subject keywords from the PubMed autocompleter  for userThe ranking uses a weighted algorithm that is largely informed by the team’s expertise and knowledge of authorship rules and regulations, publication standards, and the publishing industry. Results are displayed in two columns, with one column showing the default results ranking and a second column with citation-based impact metrics added to the weighting, because impact metrics are often used by authors as a criterion for journal selection but have known limitations as indicators of journal quality –49. OnceThe journal name search allows users to find detailed information about a specific journal. This function searches across all names and alternate titles of a journal in our database and utilizes an autocompleter for convenience. Upon a successful match, SPI-Hub retrieves the Knowledge Management Journal Record entry for the journal .The author search functionality provides another mechanism for identifying potential publication venues by allowing the user to quickly view the scholarly journals in which colleagues working in similar areas of interest and research have published. The interface allows the user to input structured citation lists that may constitute an online curriculum vitae, including an ORCID identifier, an NCBI My Bibliography link, or the public uniform resource locator (URL) of a Zotero personal library or group –52. ThisTo initially populate the database, we downloaded the “List of All Journals Cited in PubMed”  and usedBased on patterns observed through manual completion of over 2,000 records, some fields are populated using information from publisher websites through semi-automated methods and manual review. When possible, publisher-level policies are established that apply to multiple journals. For example, a publisher may state that all of their journals undergo double-blind peer review, which enables us to create a standardized message for every journal produced by that publisher. Automated data accuracy and integrity checks are run periodically and used to update the database as new information becomes available from data sources.SPI-Hub includes approximately 24,000 currently active journal titles. Record completion is ongoing, with priority given to the journals in which our institution’s authors have most commonly published as well as the journals with the largest publishers, because those journals allow the record to be rapidly completed through automation of publisher-specific data. An internal study conducted to assess resource allocation for project completion and future maintenance, which detailed 4 distinct stages of automation, showed that the team was able each time to reduce the average time needed to fully complete journal records .The last stage of automation, which is applicable to journals from large publishers, requires manual intervention only for 3 of the 25 metadata fields, showing a time reduction from 12.16 to 1.62 minutes to complete an entire record. With the introduction of automation, variances and errors were also greatly reduced, resulting in less time needed for quality control during the data acquisition phase of the project. To date, more than 16,100 records are fully completed in our database, with the remainder at least 50% complete.SPI-Hub is a decision support application to aid researchers in identifying journals in which to publish and for reviewing journal transparency and rigor. A user can quickly review all information in a Knowledge Management Journal Record holistically to gain important insights into a single journal. By comparing two or more records, a user can identify and evaluate differences. For example, the integration into the record of data from different sources allows users to distinguish between a journal’s self-reported impact metrics and a verifiable JCR impact factor –60 and iFigures 1Our automation study demonstrates the efficiency of the automated and semi-automated techniques applied to SPI-Hub. Importantly, it shows that in our current work flow, the time needed to complete Knowledge Management Journal Records can be considerably shorter than with a manual process. These automation findings have important implications, as the significant reduction of the manual effort improves the feasibility of keeping the information current. This will be key to the long-term sustainability of SPI-Hub.As we further develop SPI-Hub, a robust maintenance strategy is also being implemented and will be key to the tool’s ongoing success and usefulness. This strategy includes periodic assessment of changes and updates to journal websites and third-party data sources alike. Maintenance, with all of its challenges, requires an ongoing, significant behind-the-scenes effort because journal details  change over time and sources change their data structure. For example, the Committee on Publication Ethics’s most recent change on how to search its membership data has already necessitated an update of our automated data gathering.While SPI-Hub offers clear benefit as a decision support aid for journal identification and selection, there are two important limitations to report. First, while much progress has been made in the automation of data collection, more work is needed to fully automate the process. Second, at this stage, no effort has been made by the team to collect feedback from papers’ authors and peer reviewers about their experience working with specific journals or publishers, which could provide additional information about whether the journal or publisher actually adheres to its stated policies.Our approach offers the benefit of an open and transparent process: SPI-Hub makes every effort to provide unbiased, factual information by which users can perform journal quality assessments. All stakeholders recognize how critical it is that researchers and individuals who are seeking journals in which to publish are able to assess their rigor and transparency. Through a comprehensive knowledge management framework and the incorporation of multiple quality points specific to each journal, SPI-Hub provides an opportunity for holistic assessment of the trustworthiness of journals in which to publish research and acquire trusted knowledge.Through a series of planned rollouts, beta testing, and collection of anonymous user feedback , SPI-HubAppendix AClick here for additional data file.Appendix BClick here for additional data file.Appendix CClick here for additional data file.Appendix DClick here for additional data file.Appendix EClick here for additional data file.Appendix FClick here for additional data file.Taneya Y. Koonce, MSLS, MPH, taneya.koonce@vumc.org, https://orcid.org/0000-0002-4014-467X, Associate Director for Research, Center for Knowledge Management, Strategy and Innovation, Vanderbilt University Medical Center, Nashville, TNMallory N. Blasingame, MA, MSIS, mallory.n.blasingame@vumc.org, https://orcid.org/0000-0003-0356-9481, Information Scientist, Center for Knowledge Management, Strategy and Innovation Vanderbilt University Medical Center, Nashville, TNJerry Zhao, MS, MLIS, jerry.zhao@vumc.org, Senior Application Developer, Center for Knowledge Management, Strategy and Innovation, Vanderbilt University Medical Center, Nashville, TNAnnette M. Williams, MLS, annette.williams@vumc.org, https://orcid.org/0000-0002-2526-3857, Senior Information Scientist, Center for Knowledge Management, Strategy and Innovation, Vanderbilt University Medical Center, Nashville, TNJing Su, MD, MS, jing.su@vumc.org, Information Scientist, Center for Knowledge Management, Strategy and Innovation, Vanderbilt University Medical Center, Nashville, TNSpencer J. DesAutels, MLIS, spencer.desautels@vumc.org, https://orcid.org/0000-0002-6120-2496, Information Scientist, Center for Knowledge Management, Strategy and Innovation, Vanderbilt University Medical Center, Nashville, TNDario A. Giuse, Dr. Ing., MS, FACMI, dario.giuse@vumc.org, Associate Professor, Department of Biomedical Informatics, Vanderbilt University School of Medicine and Vanderbilt University Medical Center, Nashville, TNJohn D. Clark, MS, john.clark@vumc.org, Senior Application Developer, Center for Knowledge Management, Strategy and Innovation, Vanderbilt University Medical Center, Nashville, TNZachary E. Fox, MSIS, zachary.e.fox@vumc.org, Associate Director for Information Services, Center for Knowledge Management, Strategy and Innovation, Vanderbilt University Medical Center, Nashville, TNNunzia Bettinsoli Giuse, MD, MLS, FACMI, FMLA, nunzia.giuse@vanderbilt.edu, https://orcid.org/0000-0002-7644-9803, Professor of Biomedical Informatics and Professor of Medicine; Vice President for Knowledge Management; and Director, Center for Knowledge Management, Strategy and Innovation, Vanderbilt University Medical Center, Nashville, TN


Optimal management of post-concussion symptoms (PCS) remains ill-defined but includes multimodal, symptom-guided plans of care. Osteopathic manipulative treatment (OMT) may be used as an adjunct treatment for PCS.We present a case of a motor vehicle collision victim whose PCS improved directly and progressively after OMT. To our knowledge, this is the first report of OMT utilized for PCS management after polytrauma and as part of an organized trauma system.Previous studies discuss potential benefits of OMT for patients with PCS after sports-related injuries, and none account for management of multiply injured patients as part of an organized trauma system. Further study of OMT for PCS is warranted and would benefit by recruiting patients from trauma centers in order to observe a range of mechanisms of injury that result in concussion. The Centers for Disease Control and Prevention estimates 3.2 to 5.3 million people in the United States live with concussion-related disability . OptimalOsteopathic manipulative treatment (OMT) is a set of hands-on techniques to diagnose and treat illness or injury. OMT generally involves movements of a patient’s muscles and joints using techniques that include stretching, gentle pressure and resistance . OMT in We present a multiply injured patient who suffered a motor vehicle collision (MVC) with soft tissue injury, skull and orthopedic fractures and persistent PCS. Her PCS improved directly and progressively after OMT sessions. To our knowledge, this is the first description of OMT utilized for PCS management after polytrauma and as part of an organized trauma system.A 66-year-old female with no significant past medical history presented as a trauma system activation after a high-speed MVC. Workup revealed left squamous temporal bone fracture with extension to the sphenoid sinus, associated intrasinus hemorrhage and minimal pneumocephalus, right frontal sinus fracture with extension into the right orbital roof, left frontal scalp hematoma, left supraorbital laceration and non-displaced left clavicle fracture. Computerized tomography did not reveal intracranial hemorrhage Figure .The laceration was repaired, and the clavicle fracture was treated with a sling. She was observed in hospital for two days for pain control and physical therapy, and then discharged to home in stable condition.First follow-up in the trauma clinic on post-injury day 9 revealed left parietal, neck and shoulder pain, anhedonia, agitation and insomnia for which watchful waiting and analgesics were prescribed. She completed follow-up with orthopedics and neurosurgery, who also recommended watchful waiting and analgesics. Five weeks later, she presented to trauma clinic complaining of worsening headaches to the left occipital, temporal and parietal areas and persistent decreased activity level. Osteopathic cranial examination revealed sphenobasilar synchondrosis compression, left occipitomastoid compression, left temporal bone external rotation, bilateral frontal compression, decreased cranial rhythmic impulse and suboccipital muscle spasm. The patient was advised of possible benefits from and literature on OMT after head injury.She returned to the office two weeks later for a planned OMT session. The headaches had become more intense and persistent, were associated with worsening left neck pain and interfered with concentration and sleep. OMT was performed and targeted cranial, cervical spine and bilateral upper extremity somatic dysfunctions. Gentle myofascial release, balanced ligamentous tension, paraspinal inhibition and OMT in the cranial field were performed over the course of 35 minutes with a focus on achieving sutural decompression and alignment, restoration of intracranial fluid motion and increase of locoregional vertebral and soft tissue mobility. This session resulted in immediate and progressive improvement of neck, head and left upper extremity pain and mobility. The head and neck pain ceased completely one day later, the first time she had been pain-free since the MVC. Her sleep quality significantly improved, allowing increased activity level. Her self-reported use of analgesics decreased. She returned one month later for planned OMT. The intense head and neck pain had not recurred. However, she reported left visual field central and lateral gaze blurriness since the MVC for which she had seen a neuro-ophthalmologist in the interim, who advised watchful waiting. This symptom had become more apparent and bothersome since relief of the previous pain syndrome. She also noted left periauricular paresthesia, mild dizziness and mild left temple pain localized to the intersection of the frontal and sphenoid sutures which had also become more apparent. Gentle OMT focused on the treatment of left sphenoid torsion, left rib exhalation dysfunction, diaphragmatic motion and soft tissue somatic dysfunction. She experienced immediate improvement of pain and periauricular paresthesia and transient improvement of dizziness and blurriness. Thereafter, she was able to return to driving in a limited fashion.She returned one month later complaining of improved but persistent left temple pain and visual field blurriness. An extended, 45-minute treatment with OMT in the cranial field resulted in immediate and progressive improvement in pain, dizziness and visual field blurriness. Thereafter, she was able to return to full athletic activity and had increased driving ability.Subsequent sessions of OMT consisted of reassessment and treatment of full-body somatic dysfunction and optimization of cranial function, including use of Fulford’s dural release performed as follows: the patient is asked to gaze in a right or left direction, and the physician rotates the head in the same direction up to the restrictive end point of rotation; the patient then gazes in the other direction as the physician rotates the head to the other end point of rotation; this is subsequently performed with rotation and gaze in opposing directions; each maneuver is repeated until fluid motion is achieved . She expThroughout her course, no aggressive OMT techniques  and no additional complementary and alternative medicine modalities were used, and no major manual therapy adverse events(e.g. neurovascular injury) occurred . No quan  A prospective case series describes an association between a focused, five-day craniosacral manipulation regimen and immediate and long-term PCS improvement ), and six sessions were required for return to near-full activity. In contrast, Castillo et al. provided six weekly OMT sessions and Yaodescribed five biweekly OMT sessions before return to functional activity level ,7. BasedSimilarly, optimal timing to start and frequency of OMT sessions for PCS have not been established as national osteopathic organizations do not provide guidance on the subject. OMT was first performed for our patient approximately six weeks post-injury in part to allow completion of other subspecialist follow-up, and fracture healing-acute fracture is a relative contraindication to OMT of the fracture . In contChronicity of PCS also does not seem to prohibit improvement after OMT, as Wetzler et al. describe former National Football League players who suffered years of head injuries experiencing significant improvement after five days (10 sessions) of focused craniosacral manipulation . WetzlerThe underlying pathophysiology of PCS is not well defined nor is the cause of benefit of OMT for PCS . Zwibel Given limited targeted treatment regimens for PCS and the reported profound benefits of OMT, further utilization and study of OMT for PCS are warranted. Future work should use a quantitative assessment tool to track patient progress and would benefit by recruiting patients from trauma centers in order to observe a range of mechanisms of injury that result in concussion.


Puccinia striiformis f. sp. tritici (Pst) CY32 and P. recondita f. sp. tritici (Pt) HnZU18-3, two destructive rust fungi in wheat, are sequenced and determined. Their circular mitogenomes are 102,521 bp and 77,894 bp in length and contain 24 and 23 tRNA genes for Pst and Pt, respectively, and both consist of 14 protein-coding genes located on the same strand. Phylogenetic analysis revealed the phylogenetic relationships of Pst and Pt based on mitogenomes conform to the previous identification in genomic level.The complete mitogenomes of  Puccinia striiformis f. sp. tritici (Pst) and Puccinia recondita f. sp. tritici (Pt), members of most prevalent plant pathogenic fungi, cause wheat stripe rust and leaf rust across the world  located on the same strand for Pst and Pt. 24 and 23 tRNA genes on the same strand were identified using tRNA scan-SE (Lowe and Chan Pst and Pt, respectively.The mitochondrial base composition for Pst and Pt based on mitogenomes, we constructed the maximum-likelihood tree based on concatenated amino acids using Mega-X (Kumar et al. Pst and Pt are consistent with theirs in genomic level (Zheng et al. The two mitogenomes are highly syntenic using mitogenome alignment. In order to infer the phylogenetic relationship of


The online environment brings both challenges and opportunities. The skills learned in journal clubs remain highly relevant where the ability to critique rapidly generated information and apply evidence to patient care is vital. Creativity and flexibility are needed to ensure that learners’ needs are met and efforts are made to involve those who may not be naturally drawn to online environments. This article explores how journal clubs have been approached in the past, both in person and more recently online, considers techniques for maintaining engagement in online teaching and proposes new approaches for future journal clubs. Attending and presenting at a journal club are recommended for all trainees, being a workplace-based assessment (WPBA) for core trainees within their curriculum and required for annual review of competency progression (ARCP).1 Monitoring of attendance and reinforcement of its importance are likely to vary within UK deaneries and internationally.The Royal College of Psychiatrist’s curriculum for core trainees requires the development of critical appraisal knowledge, skills and habits for practising evidence-based medicine (EBM).3 However, recent difficulties in engaging doctors in journal clubs have been discussed.2 In 2009, Agell wrote of a ‘journal club syndrome,’ in which he reported an observed correlation between reported ‘mishaps’ preventing trainees from engaging in the learning process and the timing of their scheduled presentations.4 This somewhat cynical view highlights the long-standing anxieties and struggles with engagement that trainees might face.Doctors assembling to review current literature has been recorded as far back as 1875.5 Other aspects, such as article choice, are also specified. In psychiatry, Swift built on these strategies, highlighting the importance of structure, clear aims, a social aspect for the group and discussing selection of the facilitator .6 Moving away from the traditional format by introducing ‘evidence-based journal clubs’ has been suggested.7 Such sessions would address a specific clinical problem by referring to available literature.In 2008, Deenadayalan et al recommended establishing an overarching goal for a journal club which is reviewed and agreed by participants, recording and expecting attendance, establishing sustainable leadership with access to a statistical expert as needed and providing incentives such as food.Although the COVID-19 pandemic resulted in a rapid move to online learning, journal clubs have existed in the virtual world for some time. These can take the form either of synchronous learning, when a teacher is present at the same time as students, or of asynchronous learning, when a teacher directs study but is not present when the learning takes place.8 Other e-sources are available for accessing mental health information, an example being the Mental Elf, which has used blogs, social media and gamification to incentivise users.9Many asynchronous options for journal review have been developed, supporting the face-to-face meetings. MacLaren et al have discussed the internet possibilities, using social media platforms such as Facebook, Twitter handles and Mentimeter as a means to widen potential journal club participation and support learning.13 in which participants contribute via tweets over a specified time period , providing free participation, open journal access, time efficiency and a diverse international forum for discussion. Instructions for those wishing to set up an asynchronous journal club can involve a number of steps, including incorporating other social media platforms and considering the most convenient time for its target audience, to make their establishment accessible to all.14 Plante et al note that content may be unregulated and that some might struggle to learn from reviewing past discussions, and suggest that Wiki Journal Club  overcomes these barriers by providing editorial and professional moderation, easily referenced and referred to.15 MacRae et al found a significant improvement in critical appraisal skills among participants of their surgical journal club, compared with a control group, but acknowledged that those involved in a Twitter journal club are likely to be more motivated than the general surgeon population.10There has been a recent surge of asynchronous Twitter journal clubs,16 More recently, the BJPsych has collaborated with university psychiatric societies to help successfully launch live virtual journal clubs, with student presentations followed by expert-panel discussion.17 A similar approach has been followed with dental trainees, highlighting the benefits of accessibility, ease of interaction and effective learning.18Synchronous online journal clubs have had less exploration. Among Lin & Sherbino's suggested steps for setting up an online journal club is the added possibility of ‘live’ video panel discussions using technology such as Google ‘Hangouts on Air’ and YouTube.19 but the exact prevalence of use among doctors, or more specifically trainees, is to our knowledge not reported. If medical education becomes dependent on social media platforms, the risk is that some will be alienated or simply not reached.Although additional online options may increase the potential for journal club participation, they are unlikely to suit all trainees. Twitter is the most popular form of social media communication among healthcare professionals,20 Additional considerations are needed  review describes techniques used by educators to maximise student engagement online, necessitated by COVID-19.22 These include the need for effective organisation and structure, and varied options for student interaction, such as using online chat features, polling, hand-raising and breakout rooms.22 Warren et al argue the need for flexibility in this process, incorporating different learning modalities, considering technical resources and support.23 Engagement can still be delayed unless a learner is willing to change habits and thus suspend their disbelief in a new approach, with additional support and time needed for them to do so.24Coppola et al proposed that the online instructor's role has three domains: cognitive, affective and managerial, achieving a balance between imparting information effectively, communicating approachability, enthusiasm and intimacy, and being an effective organiser and administrator.24 In the current climate, using cases or studies involving COVID-19 specifically might assist with this.Online learning is often more effective when activities are ‘authentic’, using complex clinical scenarios and tasks to increase the perceived level of relevance for the learner.25 Haggis proposes that teachers address student diversity by shifting their approach away from the struggling learners towards a more dynamic stance, considering the overall student–teacher interactive process.25 Problematic areas need to be addressed, including acceptance of the wide range of students’ experience, motives, interaction and communication preferences. The instructor could consider using diverse examples when setting problems/tasks and fostering social relationships within the group, while still respecting the student's own responsibility for driving their learning and seeking help.26 Forcing some students to contribute, when not part of a shared collaborative process, could endanger their autonomy and motivation.27One must consider all students rather than just the most vocal or communicative.In 2020, since the COVID-19 pandemic, psychiatry training has depended on online technology to replace all face-to-face journal club meetings. Within months, our use of platforms such as Zoom and Microsoft Teams became the ‘new normal’ and the only option for teaching and networking. Although some participants were already familiar with the array of online options, many may not have been and could still be struggling to keep up.Within South West London and St George's Mental Health NHS Trust, we have continued to run a weekly journal club using the virtual platform of Microsoft Teams, supported by a WhatsApp group to offer further discussion as needed. Efforts were made to boost the effectiveness of the journal club by recruiting additional higher trainees to support the sessions, increasing the level of support offered to the lead presenter and making particular efforts during journal club meetings to encourage all to contribute. Although the numbers in the virtual room have been noticeably higher than those in face-to-face meetings before March 2020 – up to two or three times as many – engagement levels have been difficult to measure, with many of those present in the virtual room not taking an active part. In the absence of visual cues, with most turning off their screen camera and not using the ‘chat’ function, we struggle to assess who is actively engaging in the group and who finds it useful. Several trainees have commented on missing the social aspect of the group.As it has become unclear how long COVID-19 restrictions will continue, morale has declined. The initial high level of discussion has not been maintained; not all trainees have appeared confident to engage, either by speaking openly, using the written chat/Q&A function or through the WhatsApp group. This experience is, of course, likely to vary between different hospitals, trusts and deaneries, depending on the confidence of trainees, as well as the ethos and culture of the organisation.Box 2Lessons learned from recent experienceAssessing and maintaining engagement in online forums is a challengeUncertainty about the future can be unsettling and disempowering for some traineesTrainees feel a loss of the social aspect of journal clubsLearning and training opportunities have been threatened by clinical workload, technological limitations and noisy surroundingsFacilitator encouragement, support and continuity are criticalTrainees appear to appreciate a flexible approach, such as presenting articles in pairs, structuring their session in varied ways and focusing on different aspects of the paper. Some are interested in statistical methods and details, whereas others prefer to focus on how the research relates to previous and future research or how they can make use of the data in their practiceReflections and feedback from trainees are continuously sought and contribute to future planning , but we 1 The current pandemic reminds us how critical evidence-based medicine is to our practice, with a flurry of new research on COVID-19, and the need to recognise how new research should be integrated into practice. The transition to online journal clubs now requires a new set of organiser and administrator skills, which is a challenge in some departments. Even before 2020, journal clubs were struggling with participant engagement.6 Online journal clubs might be embraced by those with research or education focus; but there is a high risk that others could feel excluded. Others might be starting to get online fatigue.Journal clubs continue to be a valued part of psychiatry training, offering a key opportunity to learn and practise critical appraisal skills, presenting skills, critical thinking and structured discussion. There is no current alternative within the RCPsych curriculum that offers these opportunities, and journal club remains a WPBA for core trainees.23 The use of clinical problems and specific tasks, similar to the previously introduced evidence-based journal clubs, may provide additional relevance and authenticity for learners.24 Considering the diverse range of learner needs and preferences is advised.25 The online platform potentially supports educators in managing diversity by improving accessibility, in particular for those shielding or working on a different clinical site. Publicising and encouraging the use of additional asynchronous learning resources9 such as Twitter journal clubs to support sessions14 is worthwhile.Considering techniques for effective engagement in online teaching can enhance our ability to facilitate journal clubs.22 Trainee uncertainty about the future and feelings of social loss need to be acknowledged, as does the consideration of variability in resources and skills, with limited availability of functioning computers for some. Trainees have fed back that they appreciate additional guidance on presentation style, paper choice and additional resources. They have expressed a preference for flexibility, not defining the style of presentation and allowing the option of multiple communication forms . A challenge of measuring active participation remains, as the administrative resources required to record contributions might not be feasible within all training programmes. Forcing contribution could endanger trainee autonomy,26 although it should be recognised that it is an expected and important part of the trainee curriculum.1When running future synchronous online journal clubs, instructors need to transform their approach, not only in offering appropriate guidance and knowledge, but also in helping to establish an approachable atmosphere, optimising the potential for discussion and collaboration.This article can only be the start of a more complex discussion and re-evaluation of the function, purpose and delivery of online journal clubs and the part they play in the psychiatry curriculum, as well as considering the level of support provided to trainees and by whom. Collaboration and discussion between professional educators in neighbouring training programmes will help explore and expand the wider networking potential of journal clubs.


Monolithic zirconia is able to achieve certain aesthetic, but its durability in resisting fracture has been questioned, as fractures often originate from margins of restoration. This study determined fracture resistance of highly translucent monolithic zirconia crowns with different margin designs in terms of marginal thickness and collar height. +) were selected for the fabrication of zirconia crowns according to different designs, including varying margin thicknesses  and collar heights , which resulted in CLNC, CLLC, CLHC, CHNC, CHLC, and CHHC groups (15 crowns each). The crowns were seated on a metal die and loaded vertically through round end punch (θ = 10 mm), contacting with inclined planes of cusp in a testing machine with crosshead speed of 0.2 mm/min until fracture. Videos with a rate of 50 frames/second were used to record fracture. Fracture load (N) and durable period (s) were compared for significant differences using ANOVA and Bonferroni test (α = 0.05).  Zirconia blanks . No significant impact of collar height was shown, either on load-bearing capacity or durable time (p > 0.05). No interaction between two factors was presented (p > 0.05).  The mean ± sd of fracture load (N) and durable time (s) were 3211 ± 778 and 212 ± 47 for C Heavy chamfer margin provided stronger zirconia crown than light chamfer, but both were capable of withstanding fracture load higher than maximum masticatory force. Neither presence nor absence of collar indicated any impact on strength. Fabrication of zirconia crowns with either heavy or light chamfer margin and either presence or absence of collar, with the consideration of emergence profile, should be considered. The zirconia frameworks were predominantly fabricated in the form of yttria-stabilized tetragonal zirconia polycrystalline (Y-TZP) [transformation toughening” process [Nowadays, all ceramic restorations have been widely used for fixed partial dentures because of their biocompatibility and aesthetics outcomes. Zirconium dioxide  , 2. Natu (Y-TZP) . Eventua (Y-TZP) . Upon ad (Y-TZP) . This re process , 6.Zirconia restorations have been gaining much popularity as a result of advances in the dental technology of restoration fabrication using computer-assisted design (CAD) and computer-assisted manufacturing (CAM) processes that were capable of producing precise restorations . Dental Recently, the development of translucent zirconia has been introduced into dental practices, challenging the natural aesthetic appearance of zirconia, as it can be used solely in the aesthetic zone as monolithic high translucency zirconia restoration. This material demonstrates high strength levels, and it has been used increasingly in dental practices, especially for chair-side fabrications of zirconia restorations. However, it has been detected that the origin of failure for the zirconia crown fracture during clinical usage was reported to have been generated from the crown margin . The marRegarding the fabrication of monolithic zirconia crowns, the marginal areas are areas of minimal thickness that frequently lead to the easy fracture of the crown . The souTherefore, the objective of this experimental study was to compare the fracture load and durable time of fracture for monolithic zirconia crowns with different geometric designs of margins upon compressive loading. The null hypothesis was that different marginal designs of HTMZ restorations had no significant effect on fracture load or durable time for fracture strength.L) with a width of 0.8 mm and another with a heavy chamfer margin (CH) with a width of 1.2 mm. They possessed an axial surface height of 5.5 mm with a 6-degree taper, leading to a total convergence angle of 12 degrees. The abutment possessed round occlusal and cervical line angles with diameters of 6.0 mm and 7.0 mm, respectively (Two sets of stainless-steel definitive dies were designed using the full-features of a three-dimension (3D) computer-assisted design application  and milled using a computer numerical control milling machine . Two stainless-steel dies were identically manufactured in a cylindrical shape to simulate molar abutment ; one witectively . The abuThe metal dies were scanned using a digital scanner . The 3D printer  was used to fabricate the light polymerization resin dies. Six resin dies were fabricated with the monomer, which had a structure based on acrylic esters  in order to replicate the definitive resin dies, which would be used in the process of the fabrication of zirconia crowns for each group.L: 0.8 mm thickness) and the heavy chamfer (CH: 1.2 mm thickness)—as well as the height of cervical collar—no collar (NC: 0.0 mm collar height), low collar (LC: 0.5 mm collar height), and high collar (HC: 1.0 mm collar height)—which resulted in six groups of zirconia crowns: CLNC, CHNC, CLLC, CHLC, CLHC, and CHHC groups as shown in Ninety zirconia crowns were fabricated for six testing groups, based on the margin designs of the restorations that were related to the designs for the emergence profile present in Figures + White; Amann Girrbach AG, Koblach, Austria). The pre-sintered milled zirconia restoration was performed in an oversize dimension for each model, in order to compensate for the dimensional shrinkage of 25–30%. They were then sintered in a furnace , according to the manufacturer's recommended firing parameters, at a temperature of 1450°C for a 120-minute holding period, which concluded with a sintering process that took a total of 7.5 hours for each zirconia crown. After the sintering process, the sintered restorations were tried, adjusted, and finished on the resin die until completely seated.Six resin dies for each group were digitally scanned using the digital scanner , and they were used to design the zirconia crowns with the computer-assisted designed (CAD) software . The digitally designed crowns for each group were confirmed as having an identical anatomy, contour, and emergence profile using the computerized control milling in CAD-wax . The digital files of each restoration were transferred to the CAM to be milled with the green stage zirconia blocks . This machine used a round-end (10 mm diameter) hard steel punch at a crosshead speed of 0.2 mm/min, in order for the circumferential hoop stress at the crown margin to develop, as presented in Upon the ultimate crack, the fractured specimens were retrieved and investigated with a stereomicroscope  at 50–100 magnification, using different light sources to identify the origins of the fracture, which were then further investigated for fracture characteristics under a scanning electron microscope . The fractured specimens  were mou++® statistics , in order to estimate the characteristic strength (σo) and the Weibull modulus (m).The mean and standard deviation (sd) of the compressive load (N) at the point of failure and the time until fracture (s) of each group of zirconia crowns were calculated and then further analyzed using a two-way analysis of variance (2-way ANOVA). This was calculated in conjunction with a post hoc Bonferroni multiple comparison, which used statistical software  to determine the significant differences between the fracture strength and the duration of the fracture among all groups. The Weibull analysis was then performed to determine the strength of reliability using WeibullHLC, followed by CHHC, CHNC, CLNC, CLLC, and CLHC. The mean ± sd values of the compressive load at the point of fracture (N) of the zirconia crowns with CL and CH designs were 3055 ± 1012 and 4362 ± 909, respectively. The mean ± sd values of the compressive load at the point of fracture (N) of zirconia crowns with NC, LC, and HC designs were 3718 ± 977, 3764 ± 1327, and 3644 ± 1187, respectively, as shown in p < 0.05). However, the varied marginal collar design and the interaction between the marginal thickness and marginal collar designs did not indicate a significant influence on the compressive load at the point of failure of the zirconia crown, as presented in T-tests demonstrated that the different marginal thickness showed significant differences (p < 0.05) in the compressive load at the point of fracture, as presented in p > 0.05) in the value of the compressive load at the point of fracture in different collar designs, as presented in p < 0.05), only between CLNC-CHLC, CLNC-CHHC, CLLC-CHNC, CLLC-CHLC, CLLC-CHHC, CLHC-CHNC, CLHC-CHLC, and CLHC-CHHC, as presented in m” from highest to lowest, as CHNC (5.04); CLNC (4.59); CLHC (3.81); CHHC (3.72); CHLC (3.40); and CLLC (2.31), which indicated the relative survival probability of fracture in each group, as shown in The mean, sd, 95% confidence interval, and Weibull modulus of fracture strength (N) for each group are illustrated in LNC; 188 ± 53 for CLLC; 192 ± 27 for CLHC; 245 ± 35 for CHNC; 228 ± 29 for CHLC; and 227 ± 37 for CHHC, as presented in HNC, followed by CHLC, CHHC, CLNC, CLHC, and CLLC, respectively. The mean ± sd values of the time until fracture (s) with CL and CH designs were 197 ± 44 and 233 ± 34, respectively. The mean ± sd values of the time until fracture (s) with NC, LC, and HC designs were 228 ± 44, 208 ± 47, and 210 ± 37, respectively, as shown in p < 0.05). However, the varied collar design and the interaction between marginal thicknesses and marginal collar designs did not significantly influence the amount of time until the fracture of the zirconia crowns (p > 0.05), as shown in T-test demonstrated that the different marginal thicknesses showed significant influence on the amount of time until the fracture (s) of the zirconia crowns (p < 0.05), as presented in p > 0.05), as presented in p < 0.05), only between the CLLC group and the CHNC group, as presented in The mean ± sd values of durable time until fracture (s) were 212 ± 47 for CThe fractographic analysis of the fracture surface, according to the SEM photomicrographs, indicated patterns of fracture clues in the monolithic zirconia crowns of each group, showing an area of mist, arrested lines, and hackle features, as presented in Figures H-margin over the CL-margin on the marginal endurance of the crown to resist fracture was possibly related to the marginal thickness, which enhanced the fracture resistance of the monolithic zirconia crowns. This seems to show a preference towards the heavy chamfer margin over the light chamfer margin for monolithic Y-TZP restoration. This finding is in accordance with other previous studies [This study has indicated that the thickness of the margin design significantly affects the load-bearing capacity and durable time until fracture of HTMZ crowns. However, the design of collar did not have any influence on the load-bearing capacity and time until fracture of HTMZ crowns. Thus, the null hypotheses were rejected for the thickness of the chamfer margin but accepted for the collar design. The impact of the C studies –21. Alth studies . Therefo studies . This st studies , 31.H-group) was capable of surviving for a longer period before fracturing upon bearing load than the light chamfer margin design (CL-group). Thus, the difference in marginal design significantly affected the duration that the crown is able to resist fracture. In this study, the absence of a collar design on the HTMZ crown did not exhibit any ability to resist fracture for a longer duration than those with a collar, regardless of whether the collar was high or low. The study clearly indicated that the presence of a collar design in HTMZ crowns was absolutely possible without any reduction to the period that the crown can withstand a fracture. Thus, the absence of a collar design tends to induce more biological disadvantages for periodontium than the presence of collar design. The absence of a collar tends to result in the overcontouring of the crown at the margin, and it does not fabricate the correct emergence profile for the HTMZ crown to simulate the natural contouring of the teeth [In terms of the duration of time until fracture, the study indicated that the heavy chamfer margin design  was within the range of customary dental ceramic materials used in dentistry [The ultimate load-bearing capacity of the HTMZ crowns has been quested for clinician for the purpose of using these crowns in practice, as the disadvantage of using all ceramic restorations is that fractures often occur, the majority of which start in the margin. The most regularly used margin for ceramic restorations is a chamfer margin with either a heavy or light chamfer. Thus, the study was designed to investigate the effect of margin design on the load-bearing capacity of the HTMZ in resisting fractures, and the period that the fracture load could be withstood. The heavy chamfer is generally used for conventional ceramic restorations of normal periodontal health, and the latter is often used in cases of a compromised periodontium. Yet, both designs need to be beneficial for periodontal heath in terms of the emergence profile. This study has clearly been created in order to answer these specific issues. By designing the investigation of load resistance to fracture, the zirconia crowns need to be supported by metal dies in order to be ensured of having a strong supporter to derive for ultimate fracture load for fracturing crown without any destroying on the supporter. All HTMZ crowns were seated on the metal die supported during testing without cementation owing to reach the objective of inducing the circumferential hoop stress on the margin to induce crack initiated from the margin and elimination of any confounding factors from the chemical and mechanical properties of cement that might affected fracture resistance , 32. Thiact area ; howeveract area , 30. In entistry . As suchCrown design must be resistant enough to fracture to endure mastication and ensure positive periodontal health. This study has clearly indicated that chamfer margin design, both light and heavy, fulfilled the requirement for the load-bearing capacity of HTMZ restorations to be satisfactorily used in clinical practice. The absence of a collar design did not enhance the fracture strength of the HTMZ crown, in comparison to the presence of collar, regardless of whether the collar was high or low. Thus, this study recommends that the collar design should be advocated for HTMZ crowns, so as to ensure good periodontal health and the natural simulation of tooth contours in an appropriate emergence profile, especially in clinical situations when using a zirconia restoration on a patient with a compromised periodontal health. Accordingly, only in the limited clinical situations in which the restoration is being carried out on a patient with a healthy periodontal situation should the absence of a collar be acceptable; however, it was suggested that the emergence profile of the restoration should always be considered, in order not to affect a patient's periodontal health.The study has indicated that the design of heavy chamfer margins provided a stronger and more durable zirconia crown than light chamfer margin. Nevertheless, both heavy and light chamfer margins were capable of withstanding a fracture load that is higher than the maximum masticatory force of humans. The design of a zirconia crown without a collar did not demonstrate a better load-bearing capacity or level of endurance, in comparison with those with a collar, regardless of whether it was a high or low collar. However, the presence of a collar seems to provide a favourable emergence profile, in comparison with the absence of collar, which suggests that the collar design should be routinely advocated for in HTMZ crowns. Using a zirconia crown with no collar should be limited to cases of healthy periodontal health.


International Journal of Cardiology Heart & Vasculature, and discuss potential factors which may explain this observation.The COVID-19 pandemic has a significant impact on our daily life due to national lockdown restrictions and measures to reduce social contacts to prevent the wider spread of the virus. As one component of these restrictions, most cardiology conferences were canceled or turned into virtual online meetings Following the issued lockdown restriction initiated in mid-March 2020, a transient but substantial increase in submission rate of scientific manuscripts to this journal was noted . DespiteSeveral factors might have contributed to this disproportional transient increase in submission rate. COVID-19 introduced a new research topic, which motivated several groups all over the world to investigate the putative interaction patterns with their respective research topic Although the lockdown restrictions during the COVID-19 pandemic were associated with a substantial increase in submission rate to the journal, it is unclear, whether the increase in submission rate will remain stable and ultimately translate into published manuscripts of better quality. Finally, we hope that this pandemic gets under control very quickly to allow face-to-face meetings and conferences, which created the basis of fruitful networking and productive collaborations for decades.The authors report no relationships that could be construed as a conflict of interest.