This enzyme is also known to function as a molecular chaperon to stabilize protein conformation during the folding process. In this study, we noted FK506 binding protein (FKBP)-type PPIase from a hyperthemophilic archaeon Thermococcus sp. strain KS-1 (PPIase KS−1) to improve the solubility of Pseudomonas putida aromatic amino acid decarboxylase (AADC) that is an indispensable enzyme for fermentative production of plant isoquinoline alkaloids. AADC fused N-terminally with the PPIase KS−1 (PPIase KS−1-AADC), which was synthesized utilizing Escherichia coli host, showed improved solubility and, consequently, the cell-free extract from the recombinant strain exhibited 2.6- to 3.4-fold elevated AADC activity than that from the control strain that expressed the AADC gene without PPIase KS−1. On the other hand, its thermostability was slightly decreased by fusing PPIase KS−1. The recombinant E. coli cells expressing the PPIase KS−1-AADC gene produced dopamine and phenylethylamine from L-dopa and phenylalanine by two- and threefold faster, respectively, as compared with the control strain. We further demonstrated that the efficacy of PPIase KS−1-AADC in solubility and activity enhancement was a little but obviously higher than that of AADC fused N-terminally with NusA protein, which has been assumed to be the most effective protein solubilizer. These results suggest that PPIase KS−1 can be used as one of the best choices for producing heterologous proteins as active forms in E. coli.Peptidyl-prolyl  Thermococcus sp. KS-1 as an effective molecular chaperon in E. coli.Use of FKBP-type PPIase from Solubility and activity enhancement in aromatic amino acid decarboxylase (AADC).Thermococcus sp. KS-1 PPIase gene as a powerful fusion counterpart for the AADC gene.Demonstration of the cis-trans isomerase  is an enzyme that catalyzes the racemization reaction of amino acid proline , and parvulin, and all of these have been shown to exhibit similar enzymatic activities    in E. coli  was used as a host for protein overproduction. The plasmids pET-3a and pET-43.1a  were used as cloning and expression vectors for the AADC and NusA-AADC structural genes respectively. The structural gene of AADC  was amplified from the genomic DNA of P. putida KT2440 by the polymerase chain reaction (PCR) using PrimeStar GXL DNA polymerase  with a primer pair 5′-AAACCCCATATGACCCCCGAACAATTCCG-3′ and 5′-AAAGGATCCTCAGCCCTTGATCACGTCCTG-3′ (The NdeI and BamHI restriction sites are underlined respectively). The amplified fragments (approximately 1.5 kbp) were treated with NdeI and BamHI and inserted into pET-3a digested by the same restriction enzymes. The 1.2 kbp fragment containing the lacI gene, amplified by PCR using pET-43.1a as a template and a primer pair 5′-GGCGCCATCTCCTTGGATCCCGGACACCATCGAAT-3′ and 5′-CCGCAAGGAATGGTGCTAGTCATGCCCCGCGCCCA-3′, was inserted into SphI site of the resulting plasmid by using In-Fusion® HD Cloning Kit (15 bp homologous nucleotides in primers used for recombination are underlined). The constructed plasmid (pAADC) was used for overproduction of wild-type P. putida AADC. The Thermococcus sp. KS-1 FKBP-type PPIase gene (GenBank accession no. AB012209.1) was fused to N-terminus of AADC by inserting the aadc gene into pFusion-F87V . After digestion with SacI and XhoI, the fragment was inserted into the similarly-cut pFusion-F87V to construct pPPIase KS− 1-AADC. The NusA-AADC-expressing plasmid (pNusA-AADC) was constructed by amplifying the aadc gene from pPPIase KS− 1-AADC by using a primer pair 5′-GCTACTAGTCTGGTTCCGCG-3′ and 5′-CCCCTCGAGTCAGCCCTTGATCACGTCCTG-3′ (The SpeI and XhoI restriction sites are underlined respectively), and the amplified fragments were inserted into pET-43.1a at the SpeI and XhoI sites. Summary of the expression construct of pPPIase KS− 1-AADC was shown in Fig. E. coli strains. When the production of dopamine and phenylethylamine was performed, M9-0.2 (w/v) % glucose minimal medium supplemented with 1 mM L-dopa or L-phenylalanine were used for cultivation. Ampicillin was added to media at 100 µg/ml to maintain the expression vectors in the E. coli cells. L-Dopa, dopamine, L-phenylalanine, and phenylethylamine were purchased from NakaraiTesque Co. .Luria-Bertani (LB) (Difco) was routinely used for cultivation of the E. coli BL21(DE3) derivatives carrying pAADC, pNusA-AADC, or pPPIase KS−1-AADC were cultivated in 50 mL LB at 25 °C with shaking at 140 rpm, and when the turbidity at 600 nm reached to 0.4−0.6, isopropyl-β-d-thiogalactopyranoside (IPTG) was added at the final concentration of 0.2 mM. Temperature was then changed to 18 °C, and the cultivation was continued with shaking for 18 h. The cells were harvested by centrifugation at 5000×g for 8 min, and were disrupted by ultrasonication in 50 mM potassium phosphate (pH 7.0) containing 4 mM 2-mercaptoethanol (2-ME) and 200 µM pyridoxal 5′-phosphate (PLP). Cell-free extract and insoluble cellular debris were separated by following centrifugation at 15,000×g for 10 min. The solubility of the proteins were analyzed with SDS-PAGE by applying cell-free extracts and cellular debris equivalent to 10 µg of the cells.The Cell-free extracts were evaluated for the AADC activity by using the methods we described in the previous study with some modifications  prior to the HPLC analysis. AccQ-tag amino acid analysis column (Waters) was used to separate the compounds, and fluorescence was detected with excitation wavelength at 250 nm and emission at 395 nm. Elution program was set according to the manufacturer’s instruction.The AADC- and PPIase E. coli BL21(DE3) carrying pAADC, pNusA-AADC, or pPPIase KS−1-AADC were cultivated in LB and the expression of target genes from the T7 promoter were induced by IPTG. In the PPIase KS−1-AADC fusion construct, a 14 amino acids linker (TSLVRRGSHMEFEL) was introduced between two structural genes of the proteins, since this length had been found to be effective for stabilizing the Bacillus megaterium P450 BM3 (variant F87V) when expressed in E. coli BL21(DE3) . These results indicated that PPIase KS−1 greatly improved the solubility of AADC, and the effectiveness of PPIase KS−1 was comparable to the NusA system, which is regarded as one of the most efficient protein solubilizer generally used among the E. coli overexpression constructs.KS−1-AADC were significantly higher than AADC without fusion partner, well reflecting the high solubility of the NusA and PPIase KS−1 systems. Although the thickness of protein bands were almost similar between NusA- and PPIase KS−1-fusions, the AADC activity toward L-dopa was significantly higher in PPIase KS−1-AADC as compared to NusA-AADC (1750 ± 240 vs. 1360 ± 70 nmole/min/mg protein respectively), thus indicating that the efficiency of the PPIase KS−1 system is comparable or even superior to NusA.The AADC activity was measured using these cell-free extracts, and the results were shown in Fig. B. megaterium P450 BM3 (variant F87V) protein was improved by the existence of PPIase KS−1 at N-terminus  than that of wild-type AADC-producing strain . The latter strain was capable of producing phenylethylamine not more than 0.6 mM even after 138 h from the start of cultivation. PPIase KS−1-AADC fusion thus elevated the production level of active AADC molecules, leading to the increment of dopamine and phenylethylamine accumulation.Next, we evaluated the production ability of aromatic amines (dopamine and phenylethylamine) of the strains expressing the wild-type AADC and PPIase P. putida-derived AADC linked to FKBP-type PPIase KS−1 at the N-terminus was significantly increased in recombinant E. coli cells, as compared with the non-fused protein. A similar result has been shown in P450 BM3 (variant F87V) from Bacillus megaterium , and its high solubilizing activity was confirmed in E. coli  instead of AADC (Misawa e al. KS−1, it is possible that denaturation or decomposition of the PPIase KS−1 and AADC fusion protein were promoted due to intense structural change provoked by heat treatment. This result is also conflicting with improvement of protein solubility and activity, and future follow-up survey is required.It was found that the thermal stability of AADC was slightly weakened by fusing with PPIase Thermococcus sp. KS-1 PPIase is sufficiently small, only 17.5 kDa, when compared to NusA that is a large protein of 54.9 kDa, so that the use of such a small PPIase KS−1 protein as a fusion counterpart is obviously advantageous and the size of a plasmid constructed can be reduced. In recent synthetic biology, it is necessary to simultaneously express a large number of heterologous genes in the same cell, therefore the compactness of PPIase KS−1 should surely be useful. P. putida AADC is a protein used in the synthetic-biological production of plant isoquinoline alkaloids using E. coli cells (Kim et al. Thermococcus sp. KS-1 PPIase can be a good choice for improving the production property of heterologous proteins, which contribute to an important issue in synthetic biology to efficiently provide starting materials or intermediates.However,


Larix sibirica Ledeb.—with sulfamic acid in 1,4-dioxane using different activators has been studied for the first time. The dynamics of the molecular weight of sulfated arabinogalactan upon variation in the temperature and time of sulfation of arabinogalactan with sulfamic acid in 1,4-dioxane has been investigated. It has been found that, as the sulfation time increases from 10 to 90 min, the molecular weights of the reaction products grow due to the introduction of sulfate groups without significant destruction of the initial polymer and sulfation products. Sulfation at 95 °C for 20 min yields the products with a higher molecular weight than in the case of sulfation at 85 °C, which is related to an increase in the sulfation rate; however, during the further process occurring under these conditions, sulfation is accompanied by the destruction and the molecular weight of the sulfated polymer decreases. The numerical optimization of arabinogalactan sulfation process has been performed. It has been shown that the optimal parameters for obtaining a product with a high sulfur content are a sulfamic acid amount of 20 mmol per 1 g of arabinogalactan, a process temperature of 85 °C, and a process time of 2.5 h.The process of sulfation of arabinogalactan—a natural polysaccharide from  The anticoagulant drug most widely used in modern medical practice is heparin, a natural hexosaminoglycan of animal origin. Heparin is used to improve the hemocompatibility of the surface of medical devices ,2,3. ThiL. sibirica Ledeb), Gmelina (L. gnielinii Rupr.), and Cajandera (L. cajanderi Mayr.) [A promising and available substance alternative to heparin is sulfated arabinogalactan ,24,25,26i Mayr.) . The quai Mayr.) . For gali Mayr.) ,28. The i Mayr.) .It was found that in AG macromolecules from western larch, arabinose units are located at the ends of lateral branches consisting of three or four monosaccharide residues . The conArabinogalactan exhibits a wide range of biological properties: the immunobiological, hepatoprotective, antimutagenic, mitogenic, gastroprotective, and membranotropic activity; the probiotic, mycogenic, hypolipidemic, and immunomodulatory characteristics; the dispersing effect; etc. ,29,30,313-dimethylformamide complex in dimethyl sulfoxide [At the laboratory of natural synthons and ligands of the Irkutsk Institute of Chemistry, Siberian Branch, of the Russian Academy of Sciences, sulfated arabinogalactan in the form of a potassium salt was obtained by sulfation with a SOulfoxide . The preulfoxide .As is known, the anticoagulant activity of sulfated polysaccharides directly depends on the sulfation method used, which affects the degree of sulfation, arrangement of sulfate groups, molecular weight, etc. ,19,20,21Current methods of AG sulfates producing are based on the use of aggressive and environmentally hazardous sulfating agents such as sulfuric anhydride and chlorosulfonic acid ,23,24,26In the continuation of the work , the aimAs is known, the process of sulfation with sulfamic acid is activated by organic bases ,36,37. SIn , the sulw value of 12,310 Da. An increase in the substituent chain length in the urea molecule leads to a decrease in the sulfur content in arabinogalactan sulfate; for ethyl urea, it becomes 8.0 wt %. It should be noted that the use of the ethyl urea and hydroxyethyl urea activators ensures the same sulfur content (8.0 wt %) and PD value (1.22) in the product, with slightly different Mw values . The use of biuret as an activator in the process of sulfation of arabinogalactan with sulfamic acid leads to the formation of a product with the lowest sulfur content (7.6 wt %).It was shown that, at the introduction of substituents into the urea molecule, their activating ability in the reaction of arabinogalactan sulfation with sulfamic acid decreases. In particular, the process activated by methylurea yields sulfated arabinogalactan with a sulfur content of 9.4 wt % and a MThe highest sulfur content (12.6 wt %) and molecular weight  in arabinogalactan sulfated with sulfamic acid were obtained with the urea activator. The results are consistent with the literature data .In this study, we exploited the effects of the process temperature and time and the amount of the sulfating complex on the sulfur content in the synthesized arabinogalactan sulfates.1 , sulfation time X2 , and sulfating complex amount X3 per 1 g of arabinogalactan . The output parameter of the sulfation process was sulfur content Y1 (wt %) in arabinogalactan sulfate. The Box‒Behnken experimental design (BBD) was used. Each experiment was carried out in duplicate. The designation of variables is given in The three factors included in the study as independent variables  were sulfation temperature XThe experimental results are shown in An increase in the process temperature should lead to an increase in the rate of sulfation, and the rate of depolymerization . It is oAccording to the data given in p < 0.05. For all independent variables in the area of the factor space, in our case, p < 0.0043 was observed . The straight shows the calculated Y1 values and dots correspond to the results of observations. The proximity of the dots to the line confirms the good predictive properties of Equation (1).The predictive properties of Equation (1) are illustrated in The dependence of the output parameter Y1 on the variable factors X1 and X2 is described by a slightly curved response surface and reaches a ‘plateau’ after the X2 value of 2.5 h . The depIn addition, the quality of the approximation is characterized by the coefficient of determination R2adj. In the problem under consideration, it is R2adj = 94.0%, which indicates the high quality of the approximation. Therefore, we can state that the data obtained using Equation (1) are consistent with the observation results and this equation can be used as a mathematical model of the investigated process.The calculated optimal parameters of the sulfation of arabinogalactan with sulfamic acid in dioxane in the presence of urea are a sulfamic acid amount of 15 mmol per 1 g of arabinogalactan, a temperature of 85 °C, and a time of 2.5 h.A classical method of purification of high molecular weight compounds from low molecular weight impurities is the dialysis process, in which low molecular weight components are removed through a semipermeable membrane. A membrane is conventionally a film based on cellulose derivatives with a pore size from 3 to 100 kDa. However, this polymer purification method is rather laborious. In view of this, we compared the molecular weight characteristics of the sulfated arabinogalactan samples obtained directly from the reaction medium and purified by dialysis. Chromatograms of the crude sulfated arabinogalactan samples are shown in It was found that the main arabinogalactan peak with a retention time of ~22 min shifts to the left with an increase in the sulfation time, which indicates an increase in the mass of polymer chains due to the included sulfate groups. The peaks with a retention time of more than 25 min correspond to the initial reagents  and to the low molecular weight reaction products.The peaks corresponding to the main product are somewhat broadened, which can be attributed to the inhomogeneity of the initial arabinogalactan molecules and the fact that the low- and high-molecular chains are sulfated differently.To remove the low molecular weight chains, the initial arabinogalactan was also dialyzed and then sulfated .At a sulfation process time of 60 min, the ‘low molecular weight’ shoulder appears at the main arabinogalactan peak. As the time increases to 120 min, arabinogalactan completely passes to the low molecular weight form. In this case, the product yield was lower than 30%. These changes may indicate that, during dialysis of the initial arabinogalactan, the stabilizing polysaccharide chains were removed, which intensified the glycosidic bond breaking and the formation of low molecular weight products during sulfation.The GPC data on the dialyzed sulfated arabinogalactan samples obtained at a temperature of 85 °C and the ratio of the amount of the complex to the amount of the polysaccharide corresponds to 1:15 (g:mmol) are presented in According to the GPC data, during the dialysis, low molecular weight impurities were removed almost completely, while the retention time and form of the main product remained unchanged, in contrast to the case of the crude samples. Thus, the GPC technique can be used to analyze the molecular weight characteristics of crude products of sulfation of arabinogalactan without the additional laborious dialysis process. The use of cellophane bags in dialysis, which have smaller pore sizes than those used, could lead to the appearance of the low molecular weight fraction peak on the chromatogram and a different molecular weight distribution.The GPC data on the MWD for the samples of the reaction mass of arabinogalactan sulfation at different process times and temperatures of 85 and 95 °C are given in The GPC data showed that, in contrast to the initial arabinogalactan, the number-average and weight-average molecular weights of sulfated arabinogalactan increase with the sulfation temperature already in the first 10–30 min of the process due to the introduction of sulfate groups; the degree of polydispersity changes insignificantly as compared with the value for the initial arabinogalactan and remains within 1.11–1.16 .Sulfation of arabinogalactan at a temperature of 85 °C and a process time of up to 90–120 min  proceedsThe chromatographic study of the molecular weight characteristics of the arabinogalactan sulfation products obtained at 95 °C yielded different results .p increases, most likely due to the introduction of sulfate groups into the macromolecule, while the Mw value decreases with an increase in polydispersity. Such a change in the molecular weight characteristics can result from the partial destruction of both the initial and sulfated arabinogalactan. To 120–180 min of the sulfation process, the molecular weights significantly decrease. A decrease in the molecular weight was previously observed during mechanochemical activation of arabinogalactan [Up to a 50-min process time, the maximum peak coordinate Mgalactan . As was galactan , during The obtained results of the study of the molecular weight characteristics of the samples of the reaction mass sulfated at 85 and the duration of the process 40–180 min show that under these conditions the formation of sulfated products with close values of the average molecular weight, molecular weight distribution, and sulfur content (~12%). In contrast, the results of the chromatographic study of the molecular weight characteristics of the samples of the reaction mass sulfated at 95 °C and a process duration of 20–60 min show the possibility of obtaining sulfated arabinogalactan of different average molecular weight, but close molecular weight distribution and sulfur content (~12%). The results obtained in the future can be useful in studying the effect of the preparation conditions and the value of the molecular weight of the sulfated biopolymer on its anticoagulant activity.Larix sibirica Ledeb.) produced by OOO “Khimiya drevesiny”  under the trade name “FibrolarS” was used as the initial material.AG from the wood of the Siberian larch , using the modified procedure, described in . 1,4-dioThe dynamics of the molecular weight characteristics of the reaction mass during the sulfation of arabinogalactan was studied by gel permeation chromatography (GPC). The reaction mass samples (100–200 mg) for the GPC investigations were taken after 10, 20, 30, 40, 50, 60, 90, 120, 150, and 180 min of the process, dissolved in 5 mL of distilled water, and neutralized with an aqueous solution of ammonia to pH 7.3 (pH = 7) as a mobile phase. The column was calibrated by the polyethylene glycol standards . The eluent flow rate was 1 mL/min and the sample volume was 100 μL. Before the analysis, the samples were dissolved in the mobile phase (1 mg/mL) and filtered through a 0.22 μm hydrophilic PTFE membrane filter (Agilent). Data collection and data processing were performed using the Agilent GPC/SEC MDS software.The molecular weight distribution (MWD) of the reaction mass samples was determined on an Agilent 1200 Infinity II multi-detector GPC/SEC system with triple detection: refractometer (PL), viscometer (VS) and light scattering (LS). The separation was made on two Aquagel-OH Mixed-M columns  using the aqueous solution of 0.1 M NaNOThe sulfur content was determined on a Thermo Quest Flash EA-1112 thermal analyzer . This section may be divided by subheadings. It should provide a concise and precise description of the experimental results, their interpretation, as well as the experimental conclusions that can be drawn.The software Statgraphics Centurion XVI V16.1.09, DOE block (design of experiment), was used for numerical optimization of the AG sulfation process .In this study, the dynamics of the molecular weight characteristics of sulfated arabinogalactan upon variation in the temperature and time of sulfation of arabinogalactan with sulfamic acid in 1,4-dioxane was investigated.It was found that, during sulfation of arabinogalactan at 85 °C with an increase in the process time from 10 to 90 min, the molecular weights of the reaction products increase due to the introduction of sulfate groups without significant destruction of the initial polymer and sulfation products. Sulfation at 95 °C for 20 min leads to the formation of the products with a higher molecular weight than at 85 °C, since the sulfation rate increases; however, in the further course of the process under these conditions, sulfation is accompanied by destruction and the molecular weight of the sulfated polymer decreases.The conditions for obtaining sulfated arabinogalactan with the desired molecular weight characteristics were established.The effect of urea-based activators on sulfation of arabinogalactan with sulfamic acid was examined. It was shown that an increase in the substituent chain length leads to a decrease in the sulfur content in arabinogalactan sulfate. It was found that urea exhibits the best activating ability among the investigated activators.The Box‒Behnken numerical optimization of the process of sulfation of arabinogalactan with sulfamic acid in 1,4-dioxane was performed. It was shown that the optimal parameters for obtaining arabinogalactan sulfate with the maximum sulfur content (12.8 wt %) are a sulfamic acid amount of 20 mmol per 1 g of arabinogalactan, a process temperature of 85 °C, and a process time of 2.5 h.


After attending this session, you will be able to:Recognize the molecular heterogeneity of large B-cell lymphomas and understand current molecular classifier.Learn how new treatment strategies can be developed based on identified target alterations.Learn which prognostic and predictive biomarkers are biologically meaningful, robust, reproducible, widely available and affordable to serve as companion diagnostics.Recognize the optimal treatment for your DLBCL patients, learn the most recent data on the integration of novel drugs to standard chemmoimmunotherapy and discuss how new combo strategies, based on molecular and biological DLBCL subsets, may improve the outcome of DLBCL.Diffuse Large B-cell lymphoma (DLBCL) is the most common lymphoid malignancy. Despite its high curable rate, 60%, with Rituximab-CHOP, 40% of the patients did not respond or relapse. To improve their outcome, we need to learn more about the heterogeneity and complexity of DLBCL. This educational session is focused on molecular and pathological subtyping of aggressive lymphomas and how treatment can be driven accordingly.Dr Chapuy will review genetic analysis of DLBCL. Gene recurrent mutations, somatic copy number alterations and low frequency alterations has been studied leading to the identification of five distinct molecular DLBCL subsets and other similar models. These genetic features provide new insights in the DLBCL pathogenesis, predicting the outcome and may lead the pathway to novel combination treatment approaches.Cell of origin (COO) has been extensively studied in DLBCL with activated B-cell-like subgroup carries the worst prognosis. The underlying genetic differences have been shown to differ between COO subgroups although with considerable overlap. Double hit lymphomas with MYC, BCL2 and/or BCL6 have a dismal prognosis and their recognition is mandatory because of different treatment strategy. The expression of predictive biomarkers as MYC, BCL2, CD30 and other is also important with several uncertainties. Dr de Jong will review all these histopatological findings.Based on these premises, many expectations have been placed on clinical trials using novel compounds as Ibrutinib, Lenalidomide, Venetoclax and others with R-CHOP, based on COO classification or biomarker expression to improve the results. Positive and negative results have been reported. Dr Trneny will critically discuss these findings, providing new insights for a possible future molecular driven DLBCL therapy.


Nowadays, gelatin, a molecular derivative of collagen, has gained increasing interest in tissue engineering applications due to excellent biocompatibility, biodegradability, availability, process simplicity, and low costs. In this study, we fabricated tannic acid-crosslinked gelatin nanofibers by electrospinning method. In order to increase the bio-functionality of scaffolds, they were exposed to the atmospheric air plasma. Several analytical tools were used for evaluation of nanofibers including scanning electron microscopy (SEM), atomic force microscopy (AFM), attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy, X-ray diffraction (XRD), and water contact angle equipment (CA) together with biocompatibility study using fibroblast cells. Results demonstrated that atmospheric air plasma is not only able to improve the hydrophilicity of nanofibers but it also improves the bio-functionality against human skin fibroblast cells. Hence, we recommend atmospheric air plasma pre-treatment approach for the surface functionalization of gelatin nanofibers for skin tissue engineering applications. Gelatin is a natural biopolymer obtained from collagen under controlled hydrolysis. It has well-known properties for drug delivery, wound healing, and tissue regeneration applications ,4,5,6,7.Electrospinning is a simple and effective method for producing long, uniform, and fine diameter nanofibers. Resultant scaffolds from different nanofibers have shown specific properties such as high surface-to-volume ratio, high porosity, and long lengths. Notably, they have been used in various products such as artificial tissues, biomedical products, electronics, sensors, and nanocomposites ,13,14,15w/w) was prepared by dissolving 2 g gelatin in 10 g acetic acid. Then, it was stirred at 30 °C for 4 h. The electrospinning process was conducted under the following conditions using the electrospinning device from Fanavaran nano-meghyas Co. : high voltage value of 15 kV, distance of tip of needle to collector of 15 cm, and feeding rates of 0.6 mL/h. Then, nanofibers were collected onto an aluminum (Al) sheet. In order to generate crosslinked nanofibers, gelatin scaffolds were treated in a solution containing 5% w/w tannic acid and finally placed in a vacuum oven at 45 °C for 3 h. Gelatin powder , tannic acid, and acetic acid (66%) were provided from Sigma Aldrich. According to the prior research, acetic acid was used as a solvent for gelatin electrospinning . The spiCrosslinked nanofiber scaffolds were treated in a PF-200 plasma DBD device  for 90 s in the presence of compressed atmospheric air. The voltage applied in this process was AC and up to 100 kV, the gas pressure was 2 L/min and the distance to nozzle was 3 mm. Plasma-treated samples were kept under vacuum prior to characterization.The morphology of gelatin nanofibers was illustrated with scanning electron microscope . In this context, samples were first coated with sputter coating (Au layer) under vacuum conditions. We applied a pumping system along with a coating thickness controller of MTM-20 at a sputtering power of 30 W. The coating thickness and the target-to-substrate distance were 10 and 50 mm, respectively. The SEM working distance was set at 9 mm for all samples and a magnification of ×10 K (K = 1000) was used. SEM device was operated at a 25 kV accelerating voltage.RMS) of nanofibers, as the most important parameter for roughness, was calculated according to Equation (1):n , and AFM was employed to evaluate the surface topography and the roughness of tissues on a non-contact mode . It had a single-beam Pt-coated fpN01S tip with a length of 130 µm, width of 35 µm, resonance frequency of 152 kHz, and force constant of 5.3 N/m. The root mean square . The spectra were collected after 64 scans per sample. X-ray diffraction studies of nanofibers were conducted from wide-angle X-ray diffractograms using a Philips X’Pert Pro Multipurpose X-ray Diffractometer operating at 40 mA. Ni-filtered Cu Ka radiation generated at 40 kV (k = 0.1542 nm) and the measured angle ranged from 4 to 70°, with the scan speed of 1°/min. Wettability of the scaffolds was assessed using a water contact angle system supported by a video camera equipment . To achieve the contact angle (CA), the water droplet size was set at 0.5 mL and three samples were evaluated for each test; the average value of CA was finally reported.2 condition. Then, final cells (fourth passage) were cultivated on gelatin films with a size of 10 mm × 10 mm, by using a microscope slide cover glass (22 mm × 22 mm). After 24 h, films were washed twice with PBS, then fixed using 2.5% glutaraldehyde for 1 h at 4 °C, dehydrated by graded ethanol, and allowed to air-dry overnight. The dried films were finally imaged using SEM and optical microscopy with a cell seeding density of 10,000 cells/cm2 of medium. We also studied the effect of atmospheric air plasma treatment of gelatin films on the human dermal fibroblast cell culture. HDF-1  cells were purchased from Royan Institute, Iran. In this regard, primary cells, derived from human skin fibroblasts were cultured in Dulbecco’s modified Eagle’s medium  prepared with 10% fetal bovine serum , 100 IU/mL penicillin, and 100 μg/mL streptomycin. The cells were remained at 37 °C in 5% COThe surface morphology of gelatin nanofiber scaffolds was assessed before and after treatment with air pressure plasma, using SEM and AFM methods. For an accurate surface topography evaluation of scaffolds, we led a complementary AFM analysis C,D. We a−1 due to N–H stretching of amide bond, C–H stretching at 2925 cm−1, C=O stretching at 1635–1651 cm−1, a band at 1449 cm−1 from C–C bond, and peaks at about 610-66 cm−1 related to C–H bond [−1 which was attributed to the random coil and alpha-helix conformations [−1 [C–H bond ,24. Seveions [−1 .−1. Moreover, a strong absorption band was exhibited at 3400–3500 cm−1 due to overlapping of N–H and O–H stretching vibrations. We also found the C=O stretching vibrations (amide I) at 1651 cm−1 [−1 was attributed to the C=O stretching (amide I) and N–H bending vibrations (amide II). The intensity of this band was changed after atmospheric-air plasma treatment. Furthermore, the intensity of bands at 1449 (C–C bond), 1241, and 1077 cm−1 (referred to C–O stretching bond) was also decreased after atmospheric-air plasma functionalization of gelatin nanofibers. For the atmospheric-air plasma-treated gelatin nanofibers, C–H stretching was observed at 3300 and 2947 cm651 cm−1 . In addiThe X-ray diffraction (XRD) test was conducted for the untreated and atmospheric-air plasma-treated gelatin nanofibers, and results are indicated in One of the most important properties of biomedical scaffolds is their hydrophilic/hydrophobic functionalities . For thiFibroblast cells were put on the scaffolds and analyzed with inverted optical microscope along with SEM after 24 h. As it can be seen from the inverted microscope B. AccordWe studied the effect of the atmospheric-air plasma on the physical, chemical, and bio-functional properties of gelatin nanofibers. Surface roughness of nanofibers was significantly increased after atmospheric-air plasma treatment due to ionization and chemical degradation. Furthermore, several chemical changes were proved by the ATR-FTIR spectroscopy. In contrast to our previous study on argon and argon–oxygen plasma-treated gelatin films, we did not observe any major changes in the XRD patterns of gelatin after atmospheric-air plasma treatment. However, the intensity of peaks was changed in comparison with the untreated gelatin. Furthermore, the water CA of gelatin was changed from 20.65° to 11.8° after atmospheric-air plasma treatment. This can be possibly due to the growth of new hydrophilic groups such as oxygen-containing polar domains on the surface of scaffolds. Notably, our process led to an improvement in the hydrophilicity, which was previously confirmed elsewhere ,36. Fina


The threat from antimicrobial resistance (AMR) continues to grow. Japan’s National Action Plan on Antimicrobial Resistance, which was formulated in 2016 and targets six areas, has already had a major impact on the countermeasures implemented against AMR. Particular advances have been made in AMR-related surveillance, and we now know the situation regarding antimicrobial use and antimicrobial-resistant bacteria in the country. Educational and awareness-raising activities for medical professionals and the general public have been actively implemented and seem to have contributed to a gradual move toward an appropriate use of antimicrobials. However, there is still insufficient understanding of the issue among the general public. Determining how to use surveillance results and implementing further awareness-raising activities are crucial to address this. Tasks for the future include both raising awareness and the promotion of AMR research and development and international cooperation. The government’s next Action Plan, which will detail future countermeasures against AMR based on the outcomes of and tasks identified in the current Action Plan, has been delayed due to the COVID-19 pandemic and is urgently awaited. Antimicrobial agents have made it possible to treat and prevent bacterial infections, thereby saving countless lives. However, the effectiveness of these agents is declining due to increase in antimicrobial resistance (AMR). It is estimated that AMR is responsible for at least 700,000 deaths a year worldwide . AMR is The World Health Organization (WHO) published the Global Action Plan on Antimicrobial Resistance  in 2015,Japan’s Action Plan consists of six areas, each with goals and detailed measures for achieving them . The strThe second major characteristic is that the Action Plan sets target figures (outcome indices) . For exaA new Action Plan was expected to start from 2021, continuing on from the first 5-year Action Plan (2016–2020), in order to continuously implement measures against AMR. However, the formulation of the next Action Plan was delayed due to the impact of the 2019 novel coronavirus (COVID-19) pandemic, and its contents have not yet been released as of this writing (October 2021). Against this backdrop, this article reviews the outcomes of the current Action Plan narratively and discusses potential future countermeasures against AMR in Japan. To our knowledge, this is the first article that summarizes the AMR-related activities and results under the National Action Plan in Japan.To effectively implement measures against AMR, it is essential to increase awareness of AMR and provide educational opportunities to both medical professionals  and the general public. In Japan, the Cabinet Secretariat has been holding the National Awareness Conference on Measures Against Antimicrobial Resistance every November since 2016 and presents awards for significant public awareness and educational activities on measures against AMR . The conThe AMR Clinical Reference Center at the NCGM is responsible for education and awareness-raising activities for medical professionals . It orgaA questionnaire survey on antimicrobial management for doctors working at clinics across Japan revealed that they have high levels of awareness  and suggHealth literacy is also lower in Japan than in Europe , which mExposure to information on antibiotics and AMR from reliable sources such as specialists or public organization websites has been shown to lead to good awareness and motivation for behavioral changes , indicatPharmaceutical companies have conducted campaigns to promote the proper use of antimicrobial agents among the general public, but they are relatively limited in scale and may not be very effective. The involvement of pharmaceutical companies is expected to have a great effect on physicians and other healthcare professionals.Surveillance of both antimicrobial-resistant organisms and the use of antimicrobials is essential to implement measures against AMR. In Europe, both types of surveillance have been conducted and are used as indicators for AMR control. In Japan, drug-resistant bacteria surveillance has been conducted since before the implementation of the Action Plan, when it was already known that the proportion of drug-resistant bacteria varied according to the type of bacteria compared with other countries . HoweverThe Clinical Laboratory Division of the Japan Nosocomial Infections Surveillance (JANIS) run by the Ministry of Health, Labor, and Welfare is responsible for surveillance of antimicrobial-resistant bacteria, and the results of surveillance were used when setting the outcome indices for the Action Plan. According to the 2019 annual report, the number of participating medical institutions was 2075, which was 24.8% of the total number of hospitals (8372) in Japan , su, su26], The Action Plan includes the promotion of basic and clinical research related to AMR control as well as research to clarify the socioeconomic impact of the AMR-related diseases. The Japanese government has allocated more funds to projects and research related to AMR. In the medical field, the Ministry of Health, Labor, and Welfare has established the Antimicrobial Resistance Research Center and the AMR Clinical Reference Center as well as a surveillance system to provide a platform for promoting research. The impact of AMR in Japan has become clear as a result of these efforts ,4. The MNew antimicrobials and therapeutic agents with novel concepts need to be developed to tackle AMR. Although the governments of various countries and a number of international organizations have funded such research and development, the profits achieved to date do not yet cover the development costs. As a result, development remains inadequate to control the spread of AMR. A system that ensures a sufficient return on investment in drug development is needed . The devAnother important issue is the stable supply of the antimicrobials currently used in clinical settings. A shortage of cefazolin  in 2019 caused increased demand for other antimicrobials and consequent instability in antimicrobial supply ,42. ManyNeedless to say, action on AMR is an important international issue. Cross-border movement of individuals is currently limited due to the COVID-19 pandemic, but the spread of AMR across borders will attract attention again after the pandemic. Sharing information with international organizations and other developed countries and promoting measures against AMR in developing countries will help with the implementation of measures against AMR within Japan.The Ministry of Health, Labor, and Welfare has organized the Tokyo AMR Health Conference four times since 2016. The number of participating countries was 12 in 2016, 10 in 2017, 17 in 2018, and 20 in 2021 (online). Each conference reaffirms the policy that Japan, in conjunction with the WHO Western Pacific Office, will promote measures against AMR in the Asia-Pacific region and proactively support each country’s actions ,46,47. AThese efforts are expected to lead a range of actions in future, although the outcomes might not be seen for a while.Streptococcus pneumoniae isolates and in the resistance rates to methicillin in Staphylococcus aureus isolates and carbapenems in Pseudomonas aeruginosa isolates. However, these are all still much higher than the corresponding outcome indices. The resistance rates to carbapenems in Escherichia coli and Klebsiella pneumoniae isolates were unchanged or slightly decreased, whereas the resistance rate to fluoroquinolones in E. coli isolates clearly increased. The development of antimicrobial resistance in Enterobacterales is of concern worldwide, and Japan is no exception. Even after the first 5-year Action Plan ended, the development of AMR had not abated. Thus, a sense of crisis may be needed in order to tackle this problem.The achievement in terms of the outcome indices is summarized in Antimicrobial use has decreased gradually since the Action Plan was published, with a marked drop in 2020 . BecauseAs described above, at present, most of the outcome indices have not yet been met. However, the gaps are closing for many of the indices, indicating that the implementation of the Action Plan is producing some results, albeit not fully satisfactory.Measures against AMR, especially the surveillance system that provides the basis of actions, have shown considerable advancements since the implementation of the Action Plan started. Although this article focuses on the human medical sector, various measures have also been taken in the veterinary and environmental sectors. Activities implemented in accordance with the Action Plan and achievements relative to the outcome indices need to be reviewed when considering future activities.The Japanese government plans to implement further measures against AMR in the next Action Plan after 2021. However, because of the COVID-19 pandemic, formulation of the next Action Plan has not yet been completed as of this writing (October 2021). Early formulation of the next Action Plan would help with the continuous implementation of measures against AMR.Staphylococcus aureus (MRSA) and fluoroquinolone-resistant E. coli [E. coli, is high [E. coli resistant to third-generation cephalosporins, which was added as the global indicator 3.d.2 of the Sustainable Development Goals (SDGs) [For the next Action Plan, outcome indices must be decided carefully. For example, simply reducing the total use of antimicrobials should not be the target. Any reduction would be pointless if the necessary use of antimicrobials was decreased as well. Thus, specific indices for the inappropriate use of antimicrobials need to be set based on reports from Japanese institutions. Moreover, while the decreases seen in the resistance rates to antimicrobials in bacterial isolates are favorable, the impact of these decreases will be lost if the number of cases of infectious diseases caused by these bacteria increases. Thus, more specific indices, such as the number of cases of bloodstream infections caused by antimicrobial resistance bacteria, will be more significant for clinical practice and public health. More than 8000 individuals die every year due to bloodstream infections by methicillin-resistant  E. coli . The bur is high . The pers (SDGs) , could bAwareness of the appropriate use of antimicrobials has risen in recent years. The importance of measures for preventing infections in the clinical settings as well as collaboration between governments and medical institutions has been recognized anew during the COVID-19 pandemic. The existing directions will be maintained by introducing actions based on new outcome indices in the new Action Plan. Furthermore, the One Health approach will continue to be followed to further promote action in the veterinary and environmental sectors, and new issues, such as a stable antimicrobial supply, will be taken into account.Measures against AMR are an important theme requiring constant, long-term efforts. It is worth emphasizing that, globally, implementation needs to be done within the context of the SDGs . This isThis study has several limitations. First, the Action Plan is extensive, and this study did not cover every aspect of it. For example, this study did not evaluate the AMR-related efforts being made by the agricultural and environmental sectors. In addition, this study is a narrative review and may not fully reflect all of the achievements of the Action Plan to date. Finally, the movement within the government toward the next Action Plan may not have been fully taken into account.A variety of measures have been implemented under the Action Plan for AMR Measures published by the Government of Japan in 2016. Novel approaches have been taken in various fields, including the promotion of the One-Health approach. In the field of surveillance in particular, significant progress has been made compared with the pre-Action Plan period, including the establishment of novel surveillance systems and the launch of a website summarizing AMR-surveillance results. Antimicrobial use has decreased and is approaching target levels, but there is a need for careful monitoring due to the impact of the COVID-19 pandemic. In addition, the target percentage of drug-resistant bacteria has not yet been reached. The next Action Plan should set targets and decide what measures to take in the future based on a careful consideration of the efforts made so far.


To the editors:Response to treatment: The authors cite an overall tumor response rate of 80% and a complete response (CR) rate of 13%. These response rates, and the CR rate in particular, are higher than would be expected even in highly resourced settings where the adverse socioeconomic factors the authors cite as reasons for their high mortality rate are not present. This raises concerns about the rigor with which KS response and progression-free survival were documented. Response of KS was measured infrequently , and response assessment was described as consisting of measurement of “sentinel KS lesions”, which is only one of several measures required for a full KS response assessment. In addition, the duration of response was not specified.We write to comment on the recent article by Coldiron and colleagues  that des2.Safety: Except for blood counts performed prior to each chemotherapy cycle, formal assessments of safety and adverse events were done infrequently during the course of treatment. Other than blood counts, it is not stated whether any other tests to monitor safety  were conducted routinely, and participants were apparently questioned about adverse events only at the times of their 3- to 6-monthly KS evaluations. Thus, it is not surprising that the reported rates of moderate to severe toxicities were lower than those reported from studies of PLD conducted in highly-resourced settings , 3. MoreOf study participants who achieved a response to PLD and whose chemotherapy was then discontinued, 28% required re-treatment after subsequent KS progression. In our experience, recurrence or progression of KS after apparently successful chemotherapy is not uncommon, nor is response to a subsequent course of treatment. We are, however, puzzled by the authors’ statement that “this phenomenon does not seem to be related directly to PLD”, as most agree that chemotherapy is not expected to cure KS, and that the cause of progressive KS is likely multifactorial. Nonetheless, it is disappointing that the investigators did not track ART adherence and virological failure, which are likely contributing factors to KS progression.In this context, we highlight the authors’ comment regarding a prospectively randomized trial of chemotherapy with antiretroviral therapy (ART) in advanced KS , which w3.Mortality: Overall mortality and, especially, loss to follow-up were high in this study. The loss to follow-up rate of 13% compares unfavorably with the < 1% we observed in our randomized trial , and at We have additional concerns about participant screening. Formal assessment of cardiac function was done rarely (5 of 116 participants), and echocardiography was only performed if symptoms of CHF were detected. While we appreciate that the risk of cardiotoxicity is lower with PLD than with conventional doxorubicin, symptoms may be an unreliable indicator of cardiac dysfunction in this vulnerable patient population. For example, as noted in a recent review [In summary, while we applaud these investigators for attempting to provide improved treatment for patients with advanced HIV-associated KS, we urge caution in interpreting the results presented. We suggest a cautious approach to patient screening prior to initiating chemotherapy, and careful attention to response assessment, safety monitoring and control of HIV and concomitant illnesses during therapy. Finally, we agree that a rigorously controlled, prospective, head-to-head comparison of PLD and paclitaxel in Africa is warranted. Such a trial is being planned, which will include not only clinical and safety endpoints, but also formal evaluations of quality of life as well as cost- and cost-effectiveness comparisons.Susan E. KrownLondon, United KingdomMargaret Z. BorokFaculty of Medicine and Health Sciences, University of Zimbabwe, Harare, Zimbabwe


The results indicated that a basis weight in excess of 0.4 g/m2 of PAN electrospun fibers yielded a filtration efficiency over 97%, with breathing resistance values that increased proportionally with the amount of basis weight added. With the aim of retaining filter efficiency whilst lowering breathing resistance, the basis weight of 0.4 g/m2 and 0.8 g/m2 of PAN electrospun fibers were strategically split up and stacked with SPP in different configurations. The results suggested that a symmetric structure based on SPP/PAN/PAN/SPP was the optimal structure, as it reduces SPP consumption while maintaining an FFP2-type of filtration efficiency, while reducing breathing resistance, specially at high air flow rates, such as those mimicking FFP2 exhalation conditions. The incorporation of zinc oxide (ZnO) nanoparticles within the electrospun nanofibers in the form of nanocomposites, retained the high filtration characteristics of the unfilled filter, while exhibiting a strong bactericidal capacity, even after short contact times. This study demonstrates the potential of using the symmetric splitting of the PAN nanofibers layer as a somewhat more efficient configuration in the design of filters for respirators.Electrospinning has been used to develop and upscale polyacrylonitrile (PAN) nanofibers as effective aerosol filtration materials for their potential use in respirators. The fibers were deposited onto non-woven spunbond polypropylene (SPP) and the basis weight (grammage, g/m The unexpected appearance and rapid spreading of the severe acute respiratory syndrome coronavirus SARS-CoV-2) at the end of 2019, has affected over 14 million people and caused over a million deaths all over the world as of November 2020  at the e. The SARIn light of this, there is an urgent need to generate highly efficient filtration materials that can help from a prophylactic view-point to fight the current pandemic ,5. CurreElectrospinning is becoming an emerging technology in the area of filtration to generate nanofiber-based filter materials. These fibrous materials at the nano scale have significant morphological advantages over current conventional filters, such as fiber diameters within the nanoscale, homogeneous and interconnected small pore size, good mechanical properties and a high specific surface area of the fibers. These characteristics favor a mechanical filtration mechanism, where interception and Brownian diffusion retention of small particles between 50–5000 nm result in highly efficient filtering materials ,13. Due Electrospun nanofibers are normally deposited onto a substrate, commonly a non-woven fabric . Once anSimilarly, filters of PVDF electrospun fibers were produced with different fiber diameters  and were also corona-discharged to enhance differences between mechanical and electrostatic capture . Filter Among the different types of polymers used in filtration, polyacrylonitrile (PAN) has become a good candidate due to its good mechanical properties and hydrophobicity, as well as both its thermal and chemical stability ,18,19. A2) . F. F2 were2 were used, as these resulted in high filtration materials as monolayers, and a pressure drop was deemed acceptable if extrapolated to 200 cm2 areas . Filter efficiency and, therefore, breathing resistance, can be tailored by either modifying the fiber diameter, packing density and/or the fiber morphology of the electrospun mat. In this case, increasing the deposited mass of nanofibers with the same fiber diameter increased filter efficiency, but also led to an increase in the packing density, which ultimately increased breathing resistance. Although most of the approaches for electrospun filters are based on single layers of nanofibers, the arrangement of the nanofiber layers have shown certain improvements in filtration and pressure drop ,11,20,402 of basis weight (distributed as 0.8 g/m2 for the monolayer and 0.4/0.4 g/m2 for the multilayer filters), a filtration efficiency over 99% was achieved, regardless of the layer distribution. For the distribution of 0.2/0.2 g/m2, the values were between 97–98%, with very similar values across the different stacked structures. The results in filtration efficiency of the multilayer structures can be seen in 2 (distributed as 0.4 g/m2 for the monolayer and 0.2/0.2 g/m2 for the multilayer filters). The use of the symmetric multilayer structure (SPP/PAN/PAN/SPP) reduced slightly the breathing resistance at 30, 95 L/min and 160 L/min when compared to the monolayer structure. On the other hand, the alternate structure of SPP/PAN/SPP/PAN/SPP 0.2/0.2 g/m2 had an overall mild increase in breathing resistance for all the air flows when compared to the monolayer structure. For the case of the 0.8 g/m2 structures , which could result in membrane dysfunction, interruption and blockage of transmembrane electron transport [The biocide capacity of the ZnO-loaded nanofibers was then analyzed. The antimicrobial effect of ZnO particles is exerted by different mechanisms of action such as the creation of reactive oxygen species (ROS) and ion release to the fiber surface  after 24 h. The samples were tested with the nanofibers with the base weight of 0.4 g/m2. The filter material containing 1 wt.% of ZnO showed a strong reduction (R ≥ 3) against both types of bacteria, indicating a strong antimicrobial activity. In the case of the PAN-ZnO at 3 wt.%, the S. aureus and E. coli showed an even higher reduction. For the sample with 3 wt.% loading against S. aureus, no counts were measured, so by default a R > 3 was considered. On the other hand, the filters containing ZnO nanoparticles at 10 and 20 wt.% showed a strong antimicrobial activity (R ≥ 3), but there was an evident decrease in growth inhibition when compared with the filters containing ZnO at 3 wt.%. Such a decrease can be ascribed with nanoparticle agglomeration within the fibers due to excessive filler loading in the fibers. In this regard, 3), a relatively good dispersion and distribution across the fibers was observed. However, as the concentration increases, the presence of agglomerates becomes more pronounced, especially for PAN-ZnO20. These observations can be correlated with the reduction in antimicrobial activity reported in 3 were selected and additionally measured at different time points of bacterial exposure , in order to determine the bactericidal kinetics of the nanofibers once microorganisms are stopped at the filter surface. Similar findings have been reported by other authors, concluding that at high concentrations of nanoparticles, aggregates may form, decreasing the ZnO activity against bacteria ,48,49. F3 electrospun nanofibers with a total basis weight of 0.4 g/m2 , and especially for the configuration of SPP/PAN/PAN-ZnO3/SPP. Factors such as small and homogeneous pore diameters and high porosity are key factors to obtain a low breathing resistance material [3 fibers, obtaining a higher breathing resistance when two split nanofiber layers of PAN-ZnO were considered  and, moreover, with a strong antibacterial activity.Overall, these results indicate that ZnO-loaded PAN nanofibers are an effective technology to generate a highly efficient antimicrobial filter media for respirators, with values of up to 99% in filter efficiency, and relatively low breathing resistance . This study demonstrates the feasibility to generate a high filtration efficiency filter media with antimicrobial properties. It also suggests that the symmetric splitting up of the nanofiber layer is the optimal method to constitute the most efficient filters with this technology.In this study, PAN electrospun nanofibers were deposited onto non-woven polypropylene as monolayer fibrous structures. A range from 0.1–1 g/mThe conceptualization of this work is based on the patent P202030319.


N = 1685) in the USA. In Study 1, we drafted, tested, and modified vignettes with reference to pilot data on the relevance of the scenarios and response options. In Study 2, regulatory flexibility of sustaining daily routines, as calculated in term of context sensitivity and responsiveness to feedback, was formulated correlations with self-reported instruments to demonstrate its concurrent validity, discriminant validity, and criterion-related validity. In Study 3, path analysis examined the associations of regulatory flexibility of sustaining daily routines with psychological distress and well-being, and the moderating effects of subjective financial strain on the associations. Results showed that the inverse associations of context sensitivity and responsiveness to feedback with depressive symptoms were stronger at medium/high levels relative to lower levels of perceived financial strain. The inverse association between context sensitivity and positive affect was significant only at higher levels of strain. Our findings could provide a feasible direction for developing scalable behavioral interventions for potential mental health problems, especially among those with a lower socioeconomic status. A vignette approach was adopted to investigate flexibility of sustaining daily routines and whether and how this ability was related to mental health at different levels of financial strain. Three separate studies were conducted with community-dwelling adults ( Despite the fact that the USA is one of the most affluent countries in the world, there are consistent unequal distributions of income and wealth, with the richest 5% of the population owning two-thirds of the wealth . At leasA growing body of evidence suggests a strong socioeconomic gradient in mental health. In an 8-wave prospective study in American working adults between 1999 and 2013, a decline in income was associated with higher levels of psychological distress . ControlA series of studies that were conducted in the USA or other more developed parts of the world have found a close association between financial strain and daily living. In many families, family time involves functional tasks such as cooking, hygiene, and childcare ,11. But In the current investigation, we aimed to examine the ability of sustaining daily routines from a regulatory flexibility perspective. In the sections that follow, first, the current conceptual and empirical literature on regulatory flexibility will be discussed. Then, we will introduce a new conceptualization on how to study daily routines in adaptation to different forms of stressors, followed by an application of the individual components in the regulatory flexibility perspective to explain the role of daily routines in adaptation.Everyday life contains a multitude of routines and interactions with different people in our social network. A recent model suggests that sustainment of daily routines is one of the basic processes for demonstrating psychological resilience over time ,19. DuriUpholding functional daily routines can be a natural, adaptive coping process in chronic financial strain despite restrictive contexts. Low-income families often strive to create a more resourceful environment for adaptation. Conversations and problem solving with regard to financial strain, chores, childcare, and spousal conflicts have been observed across studies ,28,29. TEffectiveness and efficacy of sustaining daily routines could be evaluated in terms of different components of regulatory flexibility . FlexiblThese components tend to be inter-related and have shown predictive relationships to adjustment and psychopathology. For example, a greater ability to detect the presence of cues for regulation correlates with a greater flexibility in coping and emotion regulation, whereas a higher ability to detect an absence of cues correlates with lower anxiety and depressive symptoms . Apart fIn the current investigation, we used a vignette approach to adapt the context sensitivity and responsiveness to feedback components to assess individuals’ ability to adjust to changes in daily living due to financial difficulties. A vignette is a brief description of a person or situation written with the aim of simulating specific realistic features of real-world settings . Based oIn the present study , vignettUnder immediate deprived conditions, such as financial difficulty due to job loss, mass conflicts, and natural disasters, disruption and termination of primary routines could lead directly to rapid breakdown of the overall regularized daily living, whereas disruption and termination of secondary routines could only reduce overall resourcefulness, leading to gradual breakdown of daily living ,22,41,42With the adaptive priority of (1) primary over secondary routines and (2) disruption over termination, the goal of Study 1 was to develop vignettes that simulate realistic changes in daily routines in the face of financial strain and realistic responses. The context sensitivity and responsiveness to feedback components of regulatory flexibility guided the development of each vignette, which encompassed three components, namely scenario, changes, and standardized responses. The scenario described the financial strain experienced by an individual. Two changes in daily routines were also included. Responses with respect to the changes were written. A pool of 24 scenarios were drafted to reflect financial strain, with 4 scenarios randomly assigned on each of the following combinations of routines changes: (1) primary-disrupted, primary-terminated; (2) primary-disrupted, secondary-disrupted; (3) primary-disrupted, secondary-terminated; (4) primary-terminated, secondary-disrupted; (5) primary-terminated, secondary-terminated; (6) secondary-terminated, secondary-disrupted. The actor in each scenario was described with a name, gender, age, full-time/part-time job, relationship, and with/without children. Based on a review of previous studies using a vignette task , all sce“Levy is a 46 years old employee of a car wash company. Levy’s partner is also employed. Though they have joint income, they struggle to make ends meet each month, and so, Levy is constantly on the search for more work hours or a second job.” As each of the 24 scenarios contained 2 daily routines changes, a pool of 48 daily routines changes were generated. Sample daily routines changes: “You used to sleep well at night and take naps during the day. Now, you take naps or gets good sleep at night only occasionally.” (Disrupted primary routines); “You used to do a lot with friends each month. Now, you cut back on the amount of time spent with friends.” (Disrupted secondary routines); “You used to regularly sleep well. Now, you often go sleepless.” (Terminated primary routines); and “You used to hang out with friends at the local bar/café/restaurant on a regular basis. Now, you can no longer meet friends there.” (Terminated secondary routines). After obtaining ethics approval from the Institutional Review Board for the Protection of Human Subjects at Teachers College, Columbia University (IRB number: 17-351), the survey session was advertised on Amazon’s Mechanical Turk (MTurk) service (MTurk) as “A survey about daily stress” and consisted of demographic items and the draft items for realistic changes of daily routines. MTurk facilitates experimental and survey data collection using crowdsourced convenience samples. There is evidence showing that data derived from MTurk participants is valid, representative , and relM = 34.5, SD = 10.9). To minimize multiple participations, we restricted each MTurk Worker ID to one participation only. Participants were paid US $3.10 for their participation. In the survey, the instructions read, “Below are several real-life changes some people go through. Rate to what extent these changes accurately describe the reality.” Participants rated each item on a 5-point Likert scale ranging from 0  to 4 (Very much). Ratings of the items were averaged. The 48 changes ranged from 2.35 to 2.84 with median score of 2.52 and mean score of 2.53 (SD = 0.11).The accuracy of the routines changes on describing the reality were examined by a sample of 356 people  aged between 18 and 73 years (I would work on showering more regularly. (Consolidate/replace disrupted primary routines)I would start meeting friends again. (Consolidate/replace terminated secondary routines)I would get more regular exercise. (Consolidate/replace unrelated routines)I would start spending more time on relaxation. (Add unrelated routines)I would not do anything.Following presentation of a scenario with two daily routines changes, participants would rank five standardized responses in reaction to the changes. Two responses were consolidation and/or replacement of the two daily routines changes. Two responses were consolidation and/or replacement and addition of routines that were not the two changes. The fifth response option indicated no action. For example, with disrupted hygiene and terminated social activity, the responses would be as follows:In this item, according to the predicted adaptive priority of primary routines and disruption/termination, consolidation of personal hygiene  is prioritized over replacement of social activity (terminated secondary), followed by consolidation of unrelated routines  and then addition of unrelated routines . The ranking indicated context sensitivity in each item.Monitoring feedback is about the efficacy of the regulatory strategy that has been enacted. Based on the feedback, individuals prioritize, again, consolidation, replacement, and addition of routines if necessary . If the Following development of the vignette task for assessing context sensitivity and responsiveness to feedback in sustaining daily routines in time of financial strain, Study 2 aimed to investigate the concurrent validity, discriminant validity, and criterion-related validity of the vignette task.Data were extracted from different measurements in two study samples (sample 2 and 3). A total of 791 participants  aged between 18 and 72 years completed the measures and were paid US $3.20 for their participation. Each MTurk Worker IDs was restricted to one participation and thus did not duplicate across the two surveys. The study protocol was approved by the Institutional Review Board for the Protection of Human Subjects at Teachers College, Columbia University (IRB number: 17-351).One item was randomly selected from the four scenarios in each of the six combinations. A total of 6 vignettes were used to assess context sensitivity and responsiveness to feedback, each of which was scored using rankings of actions towards daily routines changes during financial strain. In each item, the first ranking indicated context sensitivity, whereas the second ranking indicated responsiveness to feedback. The standard rankings for the vignettes were formulated based on the aforementioned adaptive priority. Spearman’s rank correlation coefficient assesses how well participants’ rankings matched with the standard rankings, ranging from −1 (perfect inverse correlation) to 1 (perfect positive correlation) with 0 indicating no correlation. Context sensitivity was indicated by averaging correlation coefficients in all the first rankings whereas responsiveness to feedback was indicated by averaging correlation coefficients in all second rankings. Higher Spearman’s rank correlation coefficients indicated higher levels of context sensitivity or responsiveness to feedback.The 20-item Context Sensitivity Index (CSI) assessed the general ability to perceive cues of contextual demands . It inclThe Social Problem-Solving Inventory-Revised Short-Form (SPSI-R: SF) was used to assess social problem-solving ability ,53. SPSISustainability of Living Inventory (SOLI) was utilized to measure regularity of daily routines . SOLI coThe 16-item Flexible Regulation of Emotional Expression (FREE) Scale assessed the ability to flexibly regulate emotional expression . The scaThe PCL-C contains 17 items addressing the three DSM-IV diagnostic criteria for PTSD , which aConcurrent validity was established by quantifying the correlations of regulatory flexibility in sustaining daily routines with CSI  and SPSIThe results on concurrent, discriminant, and criterion-related validity are summarized in Study 3 examined the association between sustainment of routines and mental health and the moderating effect of subjective financial strain in the association. Subjective financial strain, denoting individuals’ evaluation of the insufficiency of income for basic living, has been consistently associated with mental health across different populations ,7,8,9,56M = 35.2, SD = 10.9) completed the measures and were paid US $3.20 for their participation. The MTurk Worker IDs was restricted to one participation and did not duplicate those in Study 1 and Study 2.With ethics approval from the Institutional Review Board for the Protection of Human Subjects at Teachers College, Columbia University (IRB number: 17-351), data collection was conducted using the MTurk Service. Sample 4 was used in the current study. A total of 538 participants  aged between 18 and 79 years  was used to assess anxiety symptoms . ParticiThe 9-item Patient Health Questionnaire (PHQ-9)  was usedPositive affect was measured by the positive affect subscale of Positive and Negative Affect Scale (PANAS) . ParticiSatisfaction with Life Scale (SWLS) assessed participants’ well-being in terms of satisfaction and fulfillment of their current state . ParticiZero order correlation and Mann–Whitney U tests were utilized to identify potential confounding demographic variables. Path analysis was performed using a lavaan package in R  to test r = −0.019, p = 0.664).As shown in β = −0.22, 95% CI: −0.32, −0.12) and depressive symptoms . No significant association was found between responsiveness to feedback and outcomes. Perceived financial strain was positively associated with depressive symptoms .Descriptive statistics and correlations of the study variables are summarized in β = 0.23, 95% CI= 0.16, 0.30) and positive affect . Context sensitivity was significantly inversely associated with anxiety symptoms  and depressive symptoms . No significant association was observed between responsiveness to feedback and outcomes. Context sensitivity × perceived financial strain was significantly inversely associated with depressive symptoms  and positive affect . Responsiveness to feedback × perceived financial strain was significantly inversely associated with depressive symptoms . No significant association was found between the interaction terms and anxiety symptoms and life satisfaction. Age (β = −0.20 to −0.13), income level (β = −0.15 to 0.20), education level (β = 0.14 to 0.16), marital status (β = 0.16 to 0.24), and employment status (β = 0.07) were included as exogenous confounding variables in both models .Model fit remained acceptable after adding the two interaction terms: CFI = 0.96, TLI = 0.91, RMSEA = 0.06 , and SRMR = 0.06. Perceived financial strain was significantly associated with depressive symptoms , medium , and high  levels of financial strain, the association was stronger as the perceived financial strain increased  of perceived financial strain but not at lower and medium levels (ps > 0.106)  but not at lower and medium levels of perceived financial strain   b. The inctively) .In three studies, we developed and validated a vignette task for assessing regulatory flexibility of sustaining daily routines in the face of financial strain and tested its associations with psychological outcomes in separate samples of participants. The ability of sustaining daily routines was assessed in terms of context sensitivity and responsiveness to feedback detailed in the regulatory flexibility perspective . ConcurrThis study is among the first to develop and use a novel method, i.e., a vignette task, to assess people’s coping with financial strain as a realistic and ecologically valid stressor. Previous studies on the association of subjective financial strain with mental health outcomes and coping processes demonstrate interrelationships more than causal relationships ,7,8,9. PContext sensitivity and responsiveness to feedback of sustaining daily routines were moderately positively associated with problem-solving ability, indicating the emphasis on cognitive processes that are needed for adopting effective strategies to regularize daily routines. The ability to sustain daily routines during financial strain was furthermore distinguished by null or weak correlations with measures assessing the regularity of daily routines and the flexible regulation of emotion expression. The moderate positive associations of context sensitivity and responsiveness to feedback in sustaining daily routines with general ability of context sensitivity demonstrated an overlap between the current vignette measures of task-specific regulatory flexibility and general scenario-based general measures of the same construct. Specifically, context sensitivity and responsiveness to feedback in daily routines sustainment was more strongly associated with higher sensitivity to the absence of contextual cues relative to sensitivity to the presence of them. Several kinds of dysfunction  are characterized by insensitivity to cue absence ,66. MoreOur findings extend the current literature on context sensitivity and responsiveness to feedback to studying daily functioning during financial difficulties. Most if not all of the previous studies have investigated context sensitivity and responsiveness to feedback with regard to emotion regulation. Emotion context insensitivity has been found to be associated with several emotion disorders, most notably anxiety and depression ,67. PrevA significant inverse association was found between context sensitivity and positive affect at higher levels of perceived financial strain while such associations were non-significant at lower and medium levels of perceived financial strain. People perceiving higher levels of financial strain are more likely to be have a lower socioeconomic status and fewer coping resources . Even thn = 1685). There is supportive evidence on the representativeness, reliability, and validity of data collected on MTurk [Several limitations of this study should be noted. First, our findings were generated from four sets of MTurk crowdsourcing data (on MTurk ,50,51. Won MTurk ,3, as weon MTurk , suggestNotwithstanding the above limitations, our findings on regulating daily routines for mental health in financial difficulties provide a feasible cost-effective approach for developing scalable behavioral interventions that could be conducted by trained non-specialists, digital treatments, and guide self-help for the early management of potential mental health disorders . First,


NSMF gene, its origin and we will summarize the evidence for the existence of splice isoforms and their expression pattern.Pyramidal neurons exhibit a complex dendritic tree that is decorated by a huge number of spine synapses receiving excitatory input. Synaptic signals not only act locally but are also conveyed to the nucleus of the postsynaptic neuron to regulate gene expression. This raises the question of how the spatio-temporal integration of synaptic inputs is accomplished at the genomic level and which molecular mechanisms are involved. Protein transport from synapse to nucleus has been shown in several studies and has the potential to encode synaptic signals at the site of origin and decode them in the nucleus. In this review, we summarize the knowledge about the properties of the synapto-nuclear messenger protein Jacob with special emphasis on a putative role in hippocampal neuronal plasticity. We will elaborate on the interactome of Jacob, the signals that control synapto-nuclear trafficking, the mechanisms of transport, and the potential nuclear function. In addition, we will address the organization of the Jacob/ Activation of Ca2+ signaling in the neuronal soma includes modulation of calcium release from intracellular calcium stores, backpropagating action potentials, somatic propagation of dendritic Ca2+ spikes that are independent of action potentials  and L-type voltage-gated Catentials . Thus, n factors . HoweverLong-distance transport of macromolecular protein signaling complexes can potentially provide a more precise means of encoding and transducing different types of synaptic activation to the nucleus. The type of information might include for instance the localization and a number of activated NMDAR and published evidence suggests that synapto-nuclear protein messenger might convey this type of information to the nucleus where it is translated in distinct long-lasting changes in gene expression .The prevalent model of activity-induced protein transport from synapses to the nucleus implies the binding of a nuclear localization signal (NLS) in synapto-nuclear protein messengers with one of the importin-α family members in response to synaptic activation. This is followed by binding to importin-β and association with a dynein motor that eventually mediates the transport of the protein complex along microtubule toward the nucleus .NSMF gene, is a synapto-nuclear messenger that encodes and transduces NMDAR signals to the nucleus  and Huntington’s disease (HD) . Especia2+ sensor protein Caldendrin binds to a central IQ-like motif of Jacob and thereby masks a bipartite NLS involved in importin-α binding  via Ca2+/CaMKII-α (en route to the nucleus (CaCaMKII-α . This reCaMKII-α . Concomi nucleus .KPNA2 gene), which links cargo to importin-β1, directly interacts with the NLS of Jacob and this interaction is essential for transport , which indicates that this type of signaling requires a certain level of network maturation and substantial expression of GluN2B-containing NMDAR at extrasynaptic sites , a CREB coactivator, replaces LMO4 from the transcription factor complex and impairs its transcriptional activity . Furtheric sites .Although extensively studied in the context of neurodegenerative diseases, the extrasynaptic NMDAR pathway has also been shown to play an important role in classical synaptic memory mechanisms  and behaStudies on the function of Jacob as synapto-nuclear protein messenger have been mainly done in pyramidal neurons of the hippocampus and cortex . HoweverNSMF expression appears to be developmentally regulated with the highest mRNA levels during synaptogenesis, between the second and the third postnatal week  that is part of the 3‘UTR region of the transcript (in situ hybridization (MERFISH) revealed that in comparison to the cellular distribution of ∼4200 RNA species in hippocampal primary cultures Jacob/NSMF mRNA expression in distal dendrites belongs to the top 10% of transcripts showing the highest dendrite-to-soma transcript ratio  indicates 13 isoforms for mouse Jacob/NSMF  out of which 8 are validated. For the human Jacob/NSMF gene  5 out of 10 transcripts predicted by automated computational analysis are confirmed.The Jacob16 exons  and high spliced . The exiNSMF mRNA expression have shown that several Jacob/NSMF transcripts are present in the mouse, rat, and human brain ; NSMF gene is present in vertebrates comprising all groups of the taxon euteleostomi, namely Tetrapoda and Osteichthyes (bony fish). The recently updated NCBI database (09.2021) contains entries for NSMF genes in 280 species. Examples include mammals  but also birds , reptiles , amphibians , and fish  (NSMF gene sequence in a scaffold of the sea lamprey (P. marinus). Although the total gene size varies from 8.5 kb in mouse to approximately 80 kb in zebrafish the exon organization is highly conserved with an exception for the lowest vertebrate P. marinus where exon boundaries are shifted, and the number of Jacob coding regions is 17. It is striking that the homology in the Jacob amino acid sequence between R. norvegicus, G. gallus, D. rerio (NSMF a gene) and P. marinus is very high in its C-terminus, especially within the regions encoded by exons 10-16 where most species show sequence identity. The differences in amino acid sequence between different species concern largely parts of the protein encoded by exon 3 , as well as exons 8 and 9. Altogether, these findings suggest that Jacob is expressed throughout all vertebrates  of different exons of diverse species, Ensembl genome database project  . An exte. rerio)  search itebrates  with theIn recent years several motifs and binding partners of Jacob have been identified . The proRodent Jacob is a 532 amino acid (aa) protein  with no The binding of Jacob to Caldendrin relies on its central α-helix  that is NSMF gene encode the bipartite NLS  and CH-HC motif  in which two cysteine and two histidine residues coordinate zinc binding were predicted in Jacob sequence . These cSeveral studies support the idea that Jacob serves as a mobile signaling hub that by docking to nuclear targets might induce long-lasting changes in gene expression.At present, not much is known about how long-distance transport of other synapto-nuclear protein messengers might converge to CREB signaling. The most prominent candidate for such a role is probably the nuclear translocation of CREB-Regulated Transcriptional Co-activator (CRTC1). Interestingly, both CRTC1 and Jacob bind to the bZip domain of CREB . This opThe exon/intron structure and the amino acid sequence are highly conserved in mammalian species, although gene size varies due to different sizes of introns. The high conservation of the C-terminus compared to the N-terminus is striking. Since the majority of known Jacob functions are linked to its N-terminus, the role of splice isoforms lacking this protein part is hard to predict. Furthermore, we undertook the effort of describing the plethora of isoforms to stress that we only begin to understand the role of the gene and that there may be additional functions.Interestingly, the N-terminus is the region that has the lowest evolutionary conservation and disordered structure without clear domains. It is possible, that only phosphorylation of some of the numerous predicted sites and/or binding to interacting partners stabilizes the protein, which could be linked to specific signaling events. One might also speculate that many aspects of NMDAR signaling to the nucleus might have evolved relatively late, possibly with the evolution of spine synapses and regulated dynamics of NMDAR localization at synaptic and extrasynaptic sites.Another important aspect of synapto-nuclear communication is the retrograde transport from presynaptic sites along the axon. This has been well-documented for importins upon axonal injury , and alsKMG, JB, GMG, MRK, and AK wrote the manuscript. AK prepared the figures. All authors commented on 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.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Supercapacitors have been considered as one of the main energy storage devices. Recently, electrospun nanofibers have served as promising supercapacitor electrodes because of their high surface area, high porosity, flexibility, and resistance to aggregation. Here, we investigate the effects of electrospinning parameters and nickel precursors on the nanostructure of electrospun nickel oxide (NiO), as well as on their electrochemical performance as supercapacitor electrodes. In contrast to the case of using nickel nitrate, increasing the nickel acetate molar concentration maintains the flexible fibrous sheet morphology of the as-spun sample during the polycondensation and calcination of NiO. As a result, our flexible electrode of NiO nanofibers derived from nickel acetate (NiO-A) exhibits much better electrochemical performance values than that of nickel nitrate-derived NiO. To further improve the electrochemical storage performance, we combined NiO-A nanofibers with single-walled carbon nanotubes (CNTs) as a hybrid electrode. In both half-cell and full-cell configurations, the hybrid electrode displayed a higher and steadier areal capacitance than the NiO-A nanofibers because of the synergetic effect between the NiO-A nanofibers and CNTs. Altogether, this work demonstrates the potency of the hybrid electrodes combined with the electrospun NiO-A nanofibers and CNTs for supercapacitor applications. Currently, electrochemical energy storage devices (ESDs) have gained a huge amount of attention worldwide . It has 2SO4 electrolyte and compared the capacitance performance of hydrous ruthenium oxide (h-RuO2) and multi-walled carbon nanotube (MWCNT) within this device. At 10 V/s of scan rate, h-RuO2 displayed 3.2 mF cm−2 of areal capacitance, while the areal capacitance was only 0.2 mF cm−2 for MWCNT under the same condition. Amongst the electrospun nanofibers of transition metal oxides, nickel oxide (NiO) is one of the most widely studied electrodes, owing to its rich natural resources, high electrochemical capacitance, and low toxicity. For instance, Kolathodi et al. , (I) is the discharge current [A], (Δt) is discharge time [s], (A) is the electrode area [cm2], and (ΔV) is the potential drop during discharge in [V] unit  unit . The ele2 of carbon cloth substrate  by drop-by-drop at 60 °C on a hotplate. This conductive substrate is considered to be a flexible current collector to support the electrode. Afterward, the coated substrate was dried in the furnace for 30 min at 60 °C. In parallel, commercial single-walled carbon nanotubes (CNTs) were drop-casted on carbon cloth with PTFE using the same previous steps to fabricate CNTs electrodes. To produce a hybrid electrode, NiO-A, CNTs, and PTFE were mixed in a ratio of 2:2:1 on carbon cloth. In our work, we fabricated half-cells: PVA+N electrode, NiO-N electrode, NiO-A electrode, CNTs electrode, and NiO-A+CNTs hybrid electrode. Further, two full-cell supercapacitors were built up: NiO-A//CNTs device (where NiO-A is the positive electrode and CNTs is the negative electrode) and NiO-A+CNTs//CNTs device (where NiO-A+CNTs is the positive electrode and CNTs is the negative electrode).The Ni-based sample was mixed with polytetrafluoroethylene (PTFE) binder in a mass ratio of 4:1 and dispersed in ethanol by ultrasonication for 5 min. Then, the mixture was cast on 1 cm−1 to 100 mV s−1 and simultaneously measured the produced current. In CD experiments, we executed the areal current selected from 0.06 mA cm−2 to 10 mA cm−2 and simultaneously measured the charging and discharging potential and time. EIS experiments were performed at a direct current (DC) with 0 V bias and a sinusoidal potential signal of 5 mV amplitude in a frequency range from 0.01 Hz to 100 kHz. EIS data were analyzed using an intricate plane impedance plot called Nyquist plot where the real part of the impedance (Z′) is represented by the x-axis and the imaginary part of the impedance (Z″) is assigned at the y-axis [x-axis of the Nyquist plot is correlated with the electronic resistance of the electrode and the current collector substrate in addition to the electrolyte solution, as they are connected in series, so it is called equivalent series resistance (ESR). In the high-frequency region, the diameter of the semicircle corresponds to the charge transfer resistance (RCT). It depends on the electrochemical activity of the redox reactions between electrode and electrolyte. The impedance data slop in the low-frequency region is related to the electrolyte ions diffusion into the electrode.In CV experiments, we supplied potential at different scan rates selected from 10 mV se y-axis . The firThe synthesis process of NiO nanofibers by electrospinning of PVA and different NiO precursors is presented schematically in −1 (40.9 × 10−4 m2 S mol−1) is much smaller than the ionic conductivity of NO3−1 (71.42 × 10−4 m2 S mol−1) [Afterward, we applied the calcination technique to create NiO from electrospun PVA/NiN nanofibers. We selected 650 °C as a calcination temperature because it is higher than both the decomposition temperatures of PVA and NiN. Surprisingly, the calcination of the nanofibers resulted mainly in nanoparticles, while occasional ultrathin nanofibers can be observed b. This mS mol−1) . TherefoSEM images of the PVA/NiAc nanofibers electrospun from 0.2 mol/L of NiAc, before and after calcination, are illustrated in −1, which is assigned to vibration of the aliphatic C-H bond [−1, which corresponded to the symmetric NO3− stretch [−1 for NiAc, which corresponded to the C-H stretching vibration [−1 and 957.31 cm−1, which corresponded to the CH3 asymmetric bending and (C–C) vibrations, respectively. All these peaks clearly appear in the spectra of electrospun PVA/NiN and PVA/NiAc nanofibers, which implies homogeneous dispersion of PVA and nickel precursors in the nanofibers without deterioration or any chemical reactions. After calcination of the nanofibers, two main characteristic bands appeared at 498 and 1090 cm−1, which represented the first and second-order phonon scattering of NiO, respectively [The formation of NiO from the PVA/NiN nanofibers was confirmed by Raman spectra a,b. InteC-H bond . For NiN stretch . On the ibration . Meanwhiectively . The XRD spectra for the NiO resulting from PVA/NiN and PVA/NiAc are compared in 2 (x-axis than that of NiO-N (~80° vs. ~70°) and has smaller real impedance values than that of NiO-N (4–5 ohm vs. 5–6 ohm). These differences indicate that the ion diffusion and the drift current in NiO-A are faster on the interface of the NiO-A electrode-KOH electrolyte. In general, larger angles than 45° and small impedance are expected for supercapacitors because the electrical charge storage mechanism that occurs at the electrode-electrolyte interface is faster than that which occurs in the entire bulk electrode of batteries [The electrochemical performance of NiO-based electrodes was evaluated and compared in terms of CV, CD, and EIS. According to 2 b. As a r2 c. The deatteries . 2). It is well known that the chemical redox reactions, in general, may require a longer time than the formation time of the electrical double layer. Therefore, we hybridized NiO with CNTs into a hybrid electrode that could maintain the high capacitance in both slow and fast charging conditions at low and high applied currents, respectively.Overall, our comparative result suggests that the structural and morphological features of NiO greatly influence its electrochemical storage performance. NiO resulting from nickel acetate possesses numerous advantages, including nanofibrous morphology, defined crystalline phase, high electrochemically active surface area, good electrical conductivity, and resistance to aggregation and/or agglomeration. All these properties are greatly beneficial for the efficiency of the redox reaction occurring between the electrode and the electrolyte. On the other hand, we notice that both the areal capacitances of both NiO-A and NiO-A drop if their charging process was quick (in few seconds) by applying high areal currents  of NiO-A, CNT, and NiO-A+CNT electrodes are close to each other, but we found that the hybrid electrode has a more stable areal capacitance in a wider range of areal currents than both the NiO-A nanofibers and CNTs  of NiAc can be loaded within PVA solution to perform stable electrospinning. Consequently, we obtained smooth and uniform NiO nanofibers without beads or fusion after calcining the electrospun PVA/NiAc nanofibers. Compared with NiO-N, NiO-A nanofibers exhibited higher areal capacitance, longer CD time, and lower transfer resistance in a half-cell configuration. Next, we combined NiO-A nanofibers with CNTs to make a hybrid electrode. In both a half-cell and full-cell configuration, the hybrid electrode displayed a higher areal capacitance than NiO-A nanofibers. Moreover, the hybrid electrode also exhibited steady areal capacitance values at different current ranges with small transfer resistance. Overall, the hybrid electrode combined with the NiO-A nanofibers and CNTs could be one of the promising electrodes for the supercapacitor.


Fibrinogen is a unique precursor of fibrin and cannot be compensated for by other coagulation factors. If plasma fibrinogen concentrations are insufficient, hemostatic clots cannot be formed with the appropriate firmness. In severe trauma patients, plasma fibrinogen concentrations decrease earlier and more frequently than other coagulation factors, predicting massive bleeding and death. We review the mechanisms of plasma fibrinogen concentration decrease, which include coagulation activation-induced consumption, hyper-fibrino(geno)lysis-induced degradation, and dilution by infusion/transfusion. Understanding the mechanisms of plasma fibrinogen concentration decrease in severe trauma patients is crucial. Fibrinogen is a glycopeptide that facilitates the formation of blood clots. It is synthesized in hepatocytes, with a molecular weight of 340 kDa , 2. The In cases of severe trauma, depleted plasma fibrinogen levels are frequently observed before dilution by infusion –9. FurthAlthough decreased plasma fibrinogen levels on arrival at the emergency department are an important risk factor of poor outcomes, the plasma fibrinogen concentration threshold considered as critically low has not been well-established in trauma patients. A decade ago, guidelines suggested that plasma fibrinogen concentrations of 1.0 g/L represented the critical threshold in bleeding patients . HoweverSince, plasma fibrinogen concentrations decrease earlier and faster than other coagulation factors in severe trauma patients , 9, 13, Following trauma, and particularly blunt trauma complicated by severe tissue injury, massively injured tissues accelerate spontaneous thrombin generation, induced by pro-coagulants in plasma Fig.  14–17].–17.14–17Extracellular DNA and DNA-binding proteins are the principal DAMPs that comprise the pro-coagulants detected in severe trauma patients. Histone and histone-complexed DNA fragments have been detected in the systemic circulation just after trauma , 19. FurVarious cell-derived microparticles have been detected during the acute phase of severe trauma –32. PlatThese DAMPs and microparticles are released into the plasma from injured organs/tissues just after trauma and activate the coagulation cascade following the conversion of fibrinogen to fibrin. Furthermore, massive DAMPs and microparticles induce consumptive coagulopathy , 30, 31.The newly formed fibrin is subsequently converted to a D-dimer and fibrin/fibrinogen degradation products (FDP) via degradation by hyper-fibrino(geno)lysis, which is a combination of fibrinolysis and fibrinogenolysis , 16.Coagulopathy caused by severe trauma is known as disseminated intravascular coagulation with fibrinolytic phenotype and is characterized by hyper-fibrino(geno)lysis, which is a combination of fibrinolysis and fibrinogenolysis , 33–42. acute release of t-PA [Weibel-Palade bodies are storage granules found in systemic vascular endothelial cells and normally contain t-PA –45. The  of t-PA , 44. Thi of t-PA , 46, 47. of t-PA –53. Typi of t-PA , 51, 53.2 plasmin inhibitor complex levels are reported to result in hyper-fibrino(geno)lysis [2-plasmin inhibitor [2-plasmin inhibitor trigger the release of plasmin and induce hyper-fibrino(geno)lysis.In severe trauma, elevations in D-dimer and FDP levels are frequently observed and are complicated with coagulopathy, regardless of severe shock , 54–59. no)lysis –59 but ano)lysis , 12, 16.no)lysis . This tyno)lysis . Some reno)lysis , 23. In no)lysis . In seveno)lysis –17. Thisnhibitor , 41, 61.Severe trauma-related depletion of plasma fibrinogen levels is observed before and upon emergency department admission, and levels continue to decrease after blood infusion/transfusion therapy initiation –9. We shIn clinical settings, we usually evaluate the plasma fibrinogen level by conducting measurements in a laboratory. Although knowledge of plasma fibrinogen levels is required for prompt treatment of patients with severe trauma, the laboratory measurements of fibrinogen levels usually takes more than 30 min. Therefore, the early evaluation of fibrinogen levels is considered important , 53, 66.Early evaluation of hyper-fibrino(geno)lysis is difficult in clinical settings. Shock-induced hyper-fibrino(geno)lysis is diagnosed via thromboelastometry –53. HoweAlthough fibrinogen is an important factor in hemostasis, it is easily decreased to critical levels in severe trauma patients , 13, 62.