Fusarium crown rot (FCR) is a chronic disease of cereals worldwide. Compared with tetraploid wheat, hexaploid wheat is more resistant to FCR infection. The underlying reasons for the differences are still not clear. In this study, we compared FCR responses of 10 synthetic hexaploid wheats (SHWs) and their tetraploid and diploid parents. We then performed transcriptome analysis to uncover the molecular mechanism of FCR on these SHWs and their parents.We observed higher levels of FCR resistance in the SHWs compared with their tetraploid parents. The transcriptome analysis suggested that multiple defense pathways responsive to FCR infection were upregulated in the SHWs. Notably, phenylalanine ammonia lyase (PAL) genes, involved in lignin and salicylic acid (SA) biosynthesis, exhibited a higher level of expression to FCR infection in the SHWs. Physiological and biochemical analysis validated that PAL activity and SA and lignin contents of the stem bases were higher in SHWs than in their tetraploid parents.Overall, these findings imply that improved FCR resistance in SHWs compared with their tetraploid parents is probably related to higher levels of response on PAL-mediated lignin and SA biosynthesis pathways.The online version contains supplementary material available at 10.1186/s12864-023-09268-7. Triticum aestivum, AABBDD), is the youngest allohexaploid formed nearly 10,000 years ago as a hybrid between an early-cultivated tetraploid wheat  and diploid wild goatgrass  [Hexaploid wheat, or common wheat  . Comparehii, DD) . Hexaplohii, DD) . Owing thii, DD) , 5.Fusarium pseudograminearum, is a soil-borne disease that seriously impacts wheat and barley production. This disease was first reported in Australia and now, it has become a prevalent disease, threating cereal production worldwide [Fusarium crown rot (FCR), primarily caused by orldwide , 7. Signorldwide , 9. Accoorldwide . FCR hasorldwide  reportedorldwide , currentF. pseudograminearum can provide new strategies to develop varieties with better resistance to FCR. In wheat, several transcriptome analyses have been performed to investigate defense responses in wheat after FCR infection [F. pseudograminearum infection are very similar to those induced by F. graminearum and F. culmorum [A better understanding of the mechanisms of the host defenses against nfection –15. For nfection . In a confection . More reculmorum .An interesting phenomenon is that hexaploid wheat is generally more resistant to FCR infection than tetraploid wheat. Differences in FCR resistance between tetraploid and hexaploid wheat have been uncovered by several studies. For instance, Daniel et al. (2008) reported that FCR inoculation reduced the yield of durum wheat by 58%, whereas the average yield loss in bread wheat was only 25% across different soil types . Liu et T. turgidum (AABB) and Ae. tauschii (DD). We performed transcriptome analyses of two SHWs and their tetraploid and diploid parents following FCR infection to identify genes and pathways associated with improved FCR resistance of SHWs. We found evidence that defense pathways regulated by phenylalanine ammonia lyase (PAL) genes are altered in SHWs compared with their tetraploid and diploid parents. We also measured six physiological parameters related to FCR resistance. Our findings revealed the genetic basis of improved FCR resistance in SHWs, which may help to develop highly FCR-resistant varieties in future.Recent studies have shown that reprogramming of transcriptomes, which occurs in newly formed hexaploid wheat, has distinct effects on growth vigor and growth adaptation following allohexaploidization , 19–21. T. turgidum ssp. dicoccoides, T. turgidum ssp. turgidum and T. turgidum ssp. dicoccon) and diploid parents (including Ae. tauschii ssp. tauschii and Ae. tauschii ssp. strangulata) were used (Table T. turgidum (AABB) and diploid goat grass Ae. tauschii (DD) were used as parental lines to generate SHWs [T. turgidum was used as a maternal parent and pollinated with Ae.tauschii anthers. S1 seeds were obtained from triploid F1 hybrids (ABD) after spontaneous chromosome doubling and were grown as the first generation of allohexaploid plants (AABBDD) [T. turgidum and 14 from Ae. tauschii) were used for further experimentation. Spontaneous chromosome doubling resulted from the union of un-reduced gametes [1 hybrid. All materials were preserved and provided by Triticeae Research Institute of Sichuan Agricultural University.In the study, 10 sets of SHWs and their tetraploid parents (including ate SHWs . T. turg(AABBDD) . Only pl gametes . No embrFusarium pseudograminearum isolate (NLYY), collected from Henan Province of China, was provided by Henan Institute of Science and Technology. The inoculum preparation, inoculation and FCR assessment were performed as described previously [6 spores/mL and then used directly for inoculation or stored at − 20 °C until needed. Tween 20 was added (0.1% v/v) to the spore suspension prior to use.A eviously . Briefy,Fusarium pseudograminearum isolate (Fp inoculation) or distilled water (mock inoculation) following the protocol described previously [Fp inoculation) or in distilled water (mock inoculation) for 1 min and seedlings were planted into the 50-well plastic trays containing sterilised nutrient soil. The plastic trays were arranged in the incubator and the settings for the incubator were: 25/16 (± 1) °C day/night temperature and 65%/85% day/night relative humidity.Seedlings at 4 days post-germination, were inoculated with eviously . Ten seeX/5 N)×100, where X is the scale value of each plant, n is the number of plants in the category, and N is the total number of plants assessed for each material.FCR severity was assessed on a 0–5 scale at fourth weeks after inoculation, where “0” represents no symptom and “5” represents whole seedling completely necrotic . A diseaT. turgidum ssp. turgidum and Ae. tauschii ssp. tauschii, and SHW-2 from the cross of T. turgidum ssp. dicoccoides and Ae. tauschii ssp. strangulata) and their tetraploid and diploid parents were selected for RNA-seq. The experimental design for each material contained two treatments (mock and Fp inoculation), one time point (3 days post inoculation) and three biological replicates. And a total of ten samples for each biological replicate were used for RNA extraction. Samples were harvested by cutting the stem bases (1 cm) at 3 days post inoculation and snap-frozen in liquid nitrogen and kept at − 80 °C until processed. Total RNA was isolated using a HiPure HP Plant RNA Mini Kit  according to the manufacturer’s instructions.Two SHWs (SHW-1 from the cross of https://ngdc.cncb.ac.cn/search/). Sequence quality assessment, trimming and RNA-seq analysis was performed using CLC Genomics 12.0 software with default parameters. The processed RNA-seq data in SHWs were aligned to complete mRNA (cDNA) sequences of Chinese Spring Reference genome IWGSCv1.1 . While RNA-seq data in tetraploid parents and diploid parents were respectively aligned to mRNA sequences of AABB sub-genomes  and DD sub-genome  from Chinese Spring Reference genome IWGSCv1.1. The expression level of each gene was calculated using TPM (Transcripts Per Kilobase of exon model per Million mapped reads ). Differentially Expressed Genes (DEGs) were restricted with the absolute value of log2 (fold Change) ≥ 1 and False Discovery Rate (FDR) < 0.05 as the threshold by performing pairwise comparisons of Fp treated and corresponding mock samples. DEGs were separated into up-regulated and down-regulated genes and used as individual test sets for GO enrichment analysis. GO term enrichment analysis was done using Triticeae-GeneTribe (http://wheat.cau.edu.cn/TGT/) [A total of 36 libraries were sequenced in the 150 bp paired-end mode using Illumina NovaSeq6000 platform at Berry Genomics Corporation . Sequence files were deposited at the National Genomics Data Center (NGDC) Genome Sequence Archive under BioProject ID PRJCA011781 (cn/TGT/) .http://www.ncbi.nlm.nih.gov/tools/primer-blast/) and listed in Table −∆∆CT). Genes with Ct values > 40 cycles were regarded as having no expression value.Thirteen DEGs were randomly selected and assessed using real-time quantitative PCR (RT-qPCR). Primers were designed using Primer-BLAST tool (Fp inoculation), one time point (3 days post inoculation) and three biological replicates. And a total of ten samples for each biological replicate were used. Samples were harvested by cutting the stem bases and snap-frozen in liquid nitrogen and kept at − 80 °C until processed.PAL activity, chitinase activity, salicylic acid (SA) content, jasmonic acid (JA) content and abscisic acid (ABA) content of 5 SHWs  and their tetraploid and diploid parents were determined using the ELISA method at FANKEL Industrial Corporation . The experimental design for each line contained one treatment . To measure basal lignin levels in stem bases, the experimental design for each line contained two time points (seedling and heading stage) and three biological replicates. And a total of ten samples for each biological replicate were used. Samples were harvested by cutting the stem bases and dried to constant weight.p < 0.05.Paired samples T-test was performed in Excel 2019 and significant differences was claimed at p < 0.01) infection than their parents  values than their diploid parents, whereas all SHWs had lower DI values than their tetraploid parents  randomly selected from the DEGs of two SHWs  enrichment analysis was performed to classify the functions of DEGs. This analysis revealed that 90, 48, and 26 terms related to biological processes were enriched (FDR < 0.05) in up-regulated DEGs of SHW-1 and its tetraploid and diploid parents, respectively , whereas 24, 36, and 17 functional biological process categories were enriched in down-regulated DEGs . In regard to SHW-2 and its tetraploid and diploid parents, 62, 45, and 20 functional categories under biological process were respectively enriched in up-regulated DEGs , whereas 33, 31, and 14 categories were enriched in down-regulated DEGs . GO terms enriched in up-regulated DEGs of SHWs were more abundant than those of their parents. Several defense-related pathways were commonly enriched in up-regulated DEGs of SHWs and their tetraploid and diploid parents, including cinnamic acid biosynthetic process (GO:0009800), chitin catabolic process (GO:0006032), L-phenylalanine catabolic process (GO:0006559), regulation of defense response (GO:0031347), glutathione metabolic process (GO:0006749), response to oxidative stress (GO:0006979)  SA contents of SHWs compared with their tetraploid parents  and heading (p < 0.01) stages  were significantly higher in SHWs than in their tetraploid parents (0.0992 U/g FW) and diploid parents (0.0933 U/g FW) , thereby exhibiting transgressive performance over both diploid and tetraploid parents.FCR has dramatically increased worldwide as a result of drier seasonal conditions and the adoption of moisture-preserving cultural practices, such as minimum tillage and stubble retention , 7. TetrTranscriptome reprogramming due to the introduction of the D genome occurred in SHWs, leading in turn to global changes in gene expression . These gAs an entry-point enzyme in the phenylpropanoid biosynthesis pathway, PAL converts phenylalanine to trans-cinnamic acid and free ammonia, thereby playing an important role in lignin and salicylic acid biosynthesis. Many studies have shown that PAL acts as a positive regulator of resistance to various plant biotic stresses –36. In tTaDIR-B1 improves resistance to FCR in wheat by increasing the accumulation of lignin [Lignin, an important component of the plant cell wall, provides an effective barrier for plants to resist pathogen infections and plays a crucial role in plant disease resistance . A recenf lignin , thus inThe importance of SA in plant defense against pathogen attack has been thoroughly documented , 39–42. Higher levels of response on PAL-mediated lignin and SA biosynthesis pathways may contribute to improved FCR resistance in SHWs compared with their tetraploid parents Fig. . HoweverPrevious transcriptome analyses have revealed various genes and pathways associated with FCR resistance in wheat, such as anti-microbial proteins, oxidative stress-related proteins, signaling molecules, primary and secondary metabolism, pathogen sensing and signaling, cellular transport and detoxification, and benzoxazinoid biosynthesis –15. HoweT. turgidum and Ae. tauschii. We confirmed improved FCR resistance in the SHWs compared with their parental genotypes. The transcriptome analysis of two SHWs and their tetraploid and diploid parents suggested that multiple defense pathways responsive to FCR infection were stronger in the SHWs than in the parents. Notably, dozens of PAL genes, involved in lignin and SA biosynthesis, exhibited a higher level of expression to FCR infection in the SHWs. Physiological and biochemical analysis validated that PAL activity and SA and lignin contents of the stem bases were higher in SHWs than their tetraploid parents. In summary, this study suggested that improved FCR resistance in SHWs compared with their tetraploid parents is probably related to higher levels of response on PAL-mediated lignin and SA biosynthesis pathways.In this study, we compared the FCR responses of 10 SHWs and their parental genotypes, Below is the link to the electronic supplementary material.Table S1. Primers for real-time quantitative PCR.Additional file 1: Table S2. RNA-seq read information and mapping statistics of 2 SHWs and their tetraploid and diploid parents.Table S3. Differentially expressed gene (DEGs) of 2 SHWs and their tetraploid and diploid parents.Table S4. GO terms enriched by up-regulated DEGs of 2 SHWs and their tetraploid and diploid parents.Table S5. GO terms enriched by down-regulated DEGs of 2 SHWs and their tetraploid and diploid parents.Table S6. PAL activity, chitinase activity, SA content, JA content and ABA content of the stem base in 5 SHWs and their parents following FCR infection.Table S7. Total lignin content of the stem base of 10 SHWs and their tetraploid parents at the seedling and heading stage.Figure S1. Validation of data of RNA-seq by RT-qPCRAdditional file 2:


The current review aims to provide an overview of the most recent research in the last 10 years on the potentials of graphene in the dental surgery field, focusing on the potential of graphene oxide (GO) applied to implant surfaces and prosthetic abutment surfaces, as well as to the membranes and scaffolds used in Guided Bone Regeneration (GBR) procedures. “Graphene oxide” and “dental surgery” and “dentistry” were the search terms utilized on the databases Scopus, Web of Science, and Pubmed, with the Boolean operator “AND” and “OR”. Reviewers worked in pairs to select studies based on specific inclusion and exclusion criteria. They included animal studies, clinical studies, or case reports, and in vitro and in vivo studies. However, they excluded systematic reviews, narrative reviews, and meta-analyses. Results: Of these 293 studies, 19 publications were included in this review. The field of graphene-based engineered nanomaterials in dentistry is expanding. Aside from its superior mechanical properties, electrical conductivity, and thermal stability, graphene and its derivatives may be functionalized with a variety of bioactive compounds, allowing them to be introduced into and improved upon various scaffolds used in regenerative dentistry. This review presents state-of-the-art graphene-based dental surgery applications. Even if further studies and investigations are still needed, the GO coating could improve clinical results in the examined dental surgery fields. Better osseointegration, as well as increased antibacterial and cytocompatible qualities, can benefit GO-coated implant surgery. On bacterially contaminated implant abutment surfaces, the CO coating may provide the optimum prospects for soft tissue sealing to occur. GBR proves to be a safe and stable material, improving both bone regeneration when using GO-enhanced graft materials as well as biocompatibility and mechanical properties of GO-incorporated membranes. The landscape of modern healthcare has been continuously shaped by groundbreaking advancements in materials science and technology . One sucPromising uses for graphene include lots of biomedical fields . GraphenOral surgery encompasses a spectrum of procedures, ranging from routine tooth extractions to complex maxillofacial reconstructions ,15,16,17At the heart of graphene’s allure lies its remarkable physical properties. Structurally, graphene consists of a single layer of carbon atoms arranged in a hexagonal lattice . This arGraphene’s electrical conductivity is equally intriguing. Its high electron mobility opens the door to applications involving electrical stimulation and biosensing. In the context of oral surgery, this property could lead to the development of implantable devices capable of monitoring healing processes in real time, thereby enabling timely interventions in case of complications . AdditioA cornerstone of successful oral surgery is the interaction between surgical materials and the complex biological environment . In-deptMoreover, graphene’s interactions with immune cells have raised intriguing possibilities for modulating immune responses during surgical interventions ,29. ThisThe prevention of post-operative infections remains a critical challenge in oral surgery . GraphenWhile the potential applications of graphene in oral surgery are undeniably exciting, several challenges must be addressed to ensure safe and effective clinical implementation. The scalable synthesis of graphene materials suitable for surgical use, long-term biocompatibility assessments, and regulatory approvals are among the foremost challenges. Additionally, the development of standardized surgical protocols and techniques for incorporating graphene into existing procedures is essential to ensure seamless integration and optimal outcomes ,38.As the frontiers of materials science and oral surgery intersect, graphene emerges as a transformative force poised to reshape the landscape of surgical practices . Its extThis systematic review was conducted by the standards of the Preferred Reporting Items for Systematic Reviews and Meta-analysis (PRISMA), and it was submitted to PROSPERO with number ID 453609.Graphene oxide, dental surgery, and dentistry were the search terms utilized on the databases  to select the papers under evaluation, with the Boolean operators “AND” and “OR”.The search was restricted to just items released in English during the previous ten years (July 2013–July 2023).The reviewers, who worked in pairs, chose works that satisfied the following criteria for inclusion: (1) animal studies; (2) clinical studies or case reports; and (3) in vitro and in vivo studies.Exclusion criteria were systematic reviews, narrative reviews, and meta-analyses.The screening procedure, which was carried out by reading the article titles and abstracts chosen in the earlier identification step, allowed for the exclusion of any publications that varied from the themes looked at.The complete text of publications that had been determined to match the predetermined inclusion criteria was then read.Reviewer disagreements on the choice of the article were discussed and settled.The quality of the included papers was assessed by two reviewers, RF and EI, using the reputable Cochrane risk-of-bias assessment for randomized trials (RoB 2). The following six areas of possible bias are evaluated by this tool: random sequence generation, allocation concealment, participant and staff blinding, outcome assessment blinding, inadequate outcome data, and selective reporting. A third reviewer (FI) was consulted in the event of a disagreement until an agreement was reached.Keyword searches of the Web of Science (55), Scopus (38), and Pubmed (200) databases yielded a total of 293 articles.The subsequent elimination of duplicates (61) resulted in the inclusion of 232 articles.Of these 232 studies, 213 were excluded because they deviated from the previously defined inclusion criteria.The screening phase ended with selecting 19 publications for this work.The results of each study are reported in The study data was selected by analyzing the study design, number of patients, intervention, and outcomes .This review discusses the potential of graphene oxide (GO) as a promising nanomaterial with exceptional physical and chemical properties. Recent research has focused on its applications in biomedical fields such as tissue engineering, antimicrobial materials, and implants . The revIn order to manage infection and stop bone loss, peri-implantitis therapy must remove polymicrobial biofilms from the implant site and lessen tissue invasion. Brushing and GO at high concentrations effectively decontaminate biofilms from exposed titanium surfaces, as shown by Qin et al. .The study by Ren et al. utilized GO as a coating on titanium foils to deliver drugs and enhance the growth and differentiation of rat bone mesenchymal stem cells (rBMSCs) into osteoblasts  46]. Th. Th46]. This study focuses on enhancing the antibacterial and cytocompatible attributes of titanium alloy implants by employing electrophoretic deposition to create bioglass (BG)-GO composites . The resThe physiochemical properties of GO-carbon fibers (CF)-PEEK on titanium implants were analyzed by Qin et al., who revealed that these coatings might greatly reduce the coefficient of friction of alloy and improve wear resistance . In addiS. mutans and P. gingivalis adhesion compared to the non-irradiated group [Photothermal therapy (PTT), an alternative antibacterial treatment, has a substantial impact on deactivating oral microbiota. The study by Park et al. involved applying graphene possessing photothermal characteristics onto a zirconia surface using atmospheric pressure plasma, followed by an assessment of its antibacterial effects against oral bacteria . To coated group . This reed group .Cheng et al. analyzed a new type of coating using a combination of GO and the antimicrobial peptide (AMP) Nal-P-113 on a smooth titanium surface . The stuS. mutans bacterial attachment and growth were significantly reduced on the Zr-GO surface, while the attachment of MC3T3-1 cells remained similar, but their growth and specialization improved on Zr-GO compared to Zr [The study by Jang et al. aimed to investigate the impact of applying GO onto a zirconia (Zr) surface on bacterial bonding and osteoblast activation . Two groed to Zr .S. mutans bacterial attachment and promoted osteoblast growth and specialization, suggesting a potential prevention of peri-implantitis by deterring bacterial adhesion and enhancing implant success by improving bone attachment [Conclusively, GO-coated zirconia hindered tachment .Recently, scientific research on graphene has focused on regenerative techniques in oral surgery, going on to investigate the efficacy of graphene oxide added to membranes or scaffolds compared with conventional methods with the hope that the results, combined with the potential of stem cells, will lead to a new class of nanomaterials with unique properties and a significant impact in the field of nanotechnology and oral health.For example, the evaluation of a bone graft material consisting of biphasic calcium phosphate (BCP) coated with rGO was the focus of the investigation by Jeong-Woo Kim et al. Osteoblast viability decreased as the concentration of rGO nanoplates increased in terms of cytotoxicity, with significant decreases at higher concentrations, while new bone production dramatically increased compared with the control group in in vivo tests using rat calvarial lesions. In fact, according to micro-CT and histomorphometric evaluations, rGO-coated BCP groups had higher volumes and percentages of new bone. The rGO4 group (with a 4:1000 ratio of rGO to BCP) showed the highest bone volume, demonstrating that the concentration of rGO in the composite material is important for bone regeneration .In contrast, Erika Nishida et al. investigated the effects of adding a GO monolayer solution to a three-dimensional collagen scaffold for possible use in bone tissue engineering . A speciThe research results of Jong Ho Lee et al. suggest that composite nanoparticles of hydroxyapatite (HAp) and rGO have extraordinary potential in enhancing the proliferation and osteogenic differentiation of pre-osteoblastic MC3T3-E1 cells.In particular, extracellular calcium deposition in MC3T3-E1 cells was significantly enhanced by rGO/HAp composite nanoparticles, and clearly, calcium accumulation is a key sign of bone tissue creation and extracellular mineralization, two crucial steps of bone regeneration. This implies that composite nanoparticles could create a favorable environment for bone tissue development, thereby promoting osteogenesis. In addition, the enzymatic activity of alkaline phosphatase (ALP), an early indicator of osteogenic differentiation, was significantly elevated in the presence of composite nanoparticles. This shows that composite nanoparticles can accelerate the differentiation process of pre-osteoblastic cells.The presence of composite nanoparticles also had a favorable impact on the deposition of osteogenic proteins, including osteopontin (OPN) and osteocalcin (OCN), which are essential markers of osteogenic cell development, suggesting a favorable impact on the expression of proteins important for bone growth. Therefore, these results also appear to be on the same wavelength as the other studies mentioned .The work of Izumi Kanayama et al.  investigS. aureus) and Gram-negative (E. coli) bacteria were analyzed; the antibacterial action of the composite was significant, and a larger clean zone was observed for higher concentrations of GO in the composite. The biocompatibility of the GO/HAp/PCL composite was also evaluated using MC3T3-E1 preosteoblast cells. Cell viability studies showed that the cytotoxic effects of the composite were minimal at lower concentrations: the cytotoxicity caused by HAp seemed to be attenuated by the addition of GO to the composite structure. It is also possible that GO has the ability to promote cell development, as it increases cell attachment and proliferation when added to the composite. Ultimately, the work showed that it was possible to successfully synthesize a GO/HAp/PCL composite with promising antibacterial qualities and biocompatibility [In the work of Chingis Daulbayev et al. , a compotibility ,71,72.In recently published work by Milena Radunovic et al. , the effThe study concluded that GO-coated collagen membranes limit inflammatory processes and promote attachment, proliferation, and osteogenic differentiation of DPSCs, emphasizing that efficacy is dose-dependent (the 10 g/mL concentration of GO produces the best results) .The study presented by Letizia Ferroni et al.  investigIn the very recent study by Elham-Sadat Motiee et al.  in 2023,The objective of the study by Alana P C Souza et al.  was to dThe risk of bias in the included studies is reported in The field of graphene-based engineered nanomaterials in dentistry is expanding due to their superior mechanical properties, electrical conductivity, and thermal stability. Graphene and its derivatives can be functionalized with bioactive compounds and added to dental materials, enhancing their properties. These materials stimulate tissue regeneration, cell differentiation, and proliferation while being biocompatible and low in toxicity. Dental implants and abutments with graphene coatings exhibit improved cytocompatibility, antibacterial properties, and osteoblast growth.Graphene-enhanced scaffolds and membranes for guided bone regeneration (GBR) also have improved physical properties, leading to enhanced bone formation. Laboratory tests indicate increased secretion of osteogenic markers and reduced inflammatory markers on graphene-coated materials. However, there are concerns about the safety of graphene and its derivatives, necessitating further research to understand their long-term effects. Overall, graphene materials hold great potential for improving oral surgery procedures in the future.


Conductive Ag electrodes were formed on PET substrates by inkjet printing. A thin film of GO-MoTe2 was deposited on the Ag electrode used for adsorbing humidity. The experiment’s results demonstrate that MoTe2 are attached to GO nanosheets uniformly and tightly. The capacitive output of the sensors with various ratios of GO/MoTe2 has been tested for different levels of humidity (11.3–97.3%RH) at room temperature (25 °C). As a consequence, the obtained hybrid film exhibits superior sensitivity (94.12 pF/%RH). The structural integrity and interaction of different components were discussed to afford the prominent humidity sensitivity performance. Under the bending condition, the output curve of the sensor has no obvious fluctuation. This work provides a low-cost way to build flexible humidity sensors with high-performance in environmental monitoring and healthcare.This study focuses on a novel humidity sensor composed of graphene-oxide (GO)-supported MoTe Humidity monitoring plays an essential role in the fields of aerospace, industry, agriculture, and even our daily lives. Resonance ,2, optic2 (molybdenum ditelluride) has attracted considerable attention in numerous fields, such as energy storage and optoelectronics, due to its high surface-to-volume ratio and favorable surface energy level [2 is used as a new semiconductor material for gas and pressure sensors. Wu et al. [2 gas sensor for NO2 detection with greatly enhanced sensitivity and recovery rates under ultraviolet (UV) illumination. Seunghyun et al. [2 engineered by strain. Meanwhile, graphene oxide (GO), one of the most popular materials around the world, is an important kind of material for preparing flexible sensors, especially humidity sensors, due to its dispersibility [2/mSiO2) via screen printing. The fabric humidity sensor exhibited high sensitivity (14.8 MΩ/% relative humidity (RH)) and low hysteresis (2.71 %RH) at a humidity interval from 23% to 97%RH. Chi et al. [Recently, transition metal dichalcogenides (TMDs) have excited many researchers owing to their excellent physicochemical properties and wide electronic applications ,22,23. Agy level ,25. Curru et al.  reportedn et al.  demonstrsibility , hydrophsibility , and larsibility . Zhu et sibility  designedi et al.  preparedi et al.  fabricat2 nanosheets for humidity detection was firstly demonstrated. Herein, we fabricated Ag electrodes by inkjet printing technology on a PET substrate. Then, a GO/MoTe2 sensing material was deposited on the electrodes by a drop-casting method. The morphology and nanostructure of the GO/MoTe2 nanofilm were confirmed by means of FTIR, Raman, SEM, and TEM. As a result, the sensor exhibited high response, good repeatability, and stability. At length, the underlying sensing mechanism of GO/MoTe2 toward humidity is also discussed.In this work, a printable and flexible capacitance sensor based on GO-supported MoTe2 powder was offered by Muke Nanotechnology Co., Ltd. . All other chemicals were at least of analytical grade and utilized without further purification.Graphene oxide  was supplied by XFNANO Co., Ltd. . The MoTe2 dispersion solution was synthesized by mixing different ratios of MoTe2 powder into 10 mL GO solution (2 mg/mL), and then the mixture was treated with ultrasound for 1 h at 100 w. Finally, three kinds of MoTe2/GO nanocomposite solutions with mass ratios of 5:1, 2:1, 1:1, 1:2, and 1:5 were obtained. The obtained dispersion solution was utilized for modifying as-prepared electrodes by means of a drop casting approach. In total, 2 droplets (1 μL single droplet volume) of the mixture solution were deposited by microinjector on the working electrode area of as-prepared flexible substrate. As control groups, pure GO and MoTe2 were, respectively, deposited on as-prepared electrodes by the same method.The GO/MoTeSEM images were examined by FEI Inspect F50 (SEM), and TEM images were inspected by FEI Talos F200S (TEM). The Fourier transform infrared spectroscopy (FTIR) was analyzed by Nicolet IS 10. Raman measurements were performed by Horiba Scientific LabRAM HR Evolution.2 sensor in various saturated salt solutions with different relative humidity (RH) levels [The humidity sensing experiments were carried out by placing the GO/MoTe) levels . The cap2/GO nano-hybrid are shown in 2 are confirmed. The wide absorption peak at 3458 cm−1 is attributed to the O-H stretching vibrations between the C-OH groups and water molecules [−1 is assigned to C=O stretching of carbonyl moiety functional groups. The sharp, tiny absorption peak at 1410 cm−1 is attributed to CO- carboxylic. The absorption peak at 663 cm−1 in nanocomposite can be assigned to MoTe2 [FT-IR spectrums of the graphene oxide and MoTeolecules . The shato MoTe2 .2 film. The measurements were conducted using a 532 nm laser. Combined with the spectra of MoTe2, the spectra of MoTe2/GO illustrated the four peaks at 120 (Bu), 145 (Bu), 197 (Bg), and 276 (Ag) cm−1 for the film blended with MoTe2, affirming the crystalline layered structure of our film. The positions and relative intensities of the four peaks match well with the previous literature reports [−1 and ~1583 cm−1 are defined as the D-band and G-band of graphene, respectively. The intensity ratio (ID/IG) of the two bands equals 1.5 and 1.19 for GO and MoTe2/GO, respectively. The ID/IG ratio can be seen as the crystallinity of graphene and a measure of defects in the skeleton structure of graphene. When the hydroxyl content is larger, the regularity of the material is worse. The GO used in this work may contain a lot of hydroxyl groups, which makes the material more hydrophilic. reports ,39,40. T2, and GO/MoTe2. From 2 exfoliated from bulk ones observed by SEM and TEM. 2 was well-dispersed in GO film. By comparing SEM and TEM images, it reveals that MoTe2 is doped between the GO sheets. In 2 (1 1 2) plane are observed with an 0.275 nm interfacial spacing.2/GO composites with different ratios were prepared. The humidity properties of the capacitive MoTe2/GO sensors at 500 Hz were measured and investigated as follows, as shown in 2 as the sensing material had only a 323 pF capacitance change from 11.3 %RH to 97.3 %RH. The response of the sensor with pure MoTe2 was remarkably lower than that of the sensor based on pure GO at different humidity conditions; however, the response of the sensor coated with MoTe2/GO (1:5) toward relative humidity reached 4386 pF in the whole RH range, which is about 13.5 times that of the sensor with pure MoTe2 and 1.3 times that of the sensor based on pure GO, respectively. These results show that the humidity sensitivity of the sensing film could be improved by doping MoTe2 in GO film.Humidity sensors based on MoTe2—too much is as bad as too little. At the outset, with the increase in the amounts of MoTe2 in GO film, the humidity response of sensor increases. When the mass ratio of both materials in the MoTe2/GO composite is 1:2, the humidity response of the sensor obtains the maximum value (capacitance change: 8093 pF), which is 25 times and 2.2 times those of the sensor with pure MoTe2 and the sensor with pure GO, respectively. Therefore, in the following further research, the sensor films are all in this ratio. Unfortunately, when the mass ratio of the two materials is further magnified (MoTe2/GO (1:1), MoTe2/GO (2:1), and MoTe2/GO (5:1)), the humidity response of sensor would actually decrease. It might be that excess MoTe2 could cover up not only the hydrophobic region of GO, but also part of the hydrophilic groups of GO, thus inhibiting the sensing performance improvement of the humidity sensor. It can be found that the addition of MoTe2 in GO thin film has little effect on its hydrophilicity under low humidity. However, under moderate and high humidity conditions, doping an appropriate amount of MoTe2 into GO thin films can significantly improve its hydrophilicity.The degree of improvement, however, depends on different addition amounts of MoTeIn To evaluate the accuracy and efficiency of the humidity sensor, the hysteresis characteristic is also a significant parameter. The dynamic hysteresis loop curve is shown in The repeatability of the as-fabricated sensor under both flat and bending state was studied between 11.3%RH and 97%RH. The response and recovery time is one of the important performance indicators to evaluate the sensor. 2, CH4, ethanol (C2H5OH), NH3, and NO2, have been tested to evaluate the cross-sensitivity of the sensor. The capacitance of the sensor to these gases (at 5000 ppm) was measured, as shown in Various gases, such as CO2/GO film is considered as a potential humidity sensing material. The combination of MoTe2 nanoparticles and GO nanosheets greatly enhances the ability of humidity sensing. MoTe2 with a sphere nanostructure can be embedded between nanosheets to play a supporting role, resulting in a great favor to the absorption and diffusion of water molecules [2/GO film can offer a higher proportion of available active sites for water molecule absorption and can accelerate the process of adsorption and desorption on the film surface owing to the larger surface specific area compared with GO [The above experimental results show that the sensor possesses excellent humidity sensing performance. Therefore, the MoTeolecules . Moreove with GO .−1, and the cycle of one breath is approximately 3–5 s [−1) and bradypnea (respiratory rate < 12 times min−1). As shown in −1, and the mouth breathing rate is 15 times min−1. Compared with a normal breathing (by nose) response, the capacitance variation range is larger due to the wetter and stronger airflow from the tester’s mouth. We simulated two kinds of abnormal breathing (tachypnea and bradypnea) and carried out a breathing experiment, as shown in In the field of health monitoring, with the development of wearable devices, new requirements are put forward for flexible sensors, especially in respiratory monitoring. Taking into account the response and recovery characteristics of the prepared sensor, it has ability to perform real-time respiratory monitoring in a non-contact manner. The normal respiratory rate of adults in a calm state is about 12–20 times minly 3–5 s . AbnormaWe also tested the non-contact skin humidity sensing performance, and the test method is shown in 2 nanosheets were deposited on the conductive Ag electrodes formed on PET substrates by inkjet printing to detect humidity. The SEM and TEM graphs demonstrate that MoTe2 are well-dispersed to GO nanosheets uniformly and tightly. By comparing the response of each sensor, the optimal composite ratio of materials is found out. The capacitive performance of the sensor has been tested at different states . The obtained hybrid film exhibits excellent sensitivity. Moreover, after bending, the response of the sensor does not change significantly, which is beneficial to its application in wearable devices. Due to its good response and recovery characteristics, the sensor also has a good performance in respiratory monitoring and non-contact measurement. The structural integrity and interaction of different components were discussed to afford the prominent humidity sensitive performance. This work proposed a low-cost and easy-to-operate preparation method of a flexible humidity sensor with high performance in environmental monitoring and medical care.In this paper, graphene-oxide (GO)-supported GO-MoTe


FUNDC1 is co-expressed with TDP-43, and both TDP-43 and FUNDC1 expression is correlated with genes associated with mitochondrial protein import pathway in brain samples of patients diagnosed with TDP-43 proteinopathy. FUNDC1 promotes mitochondrial translocation of TDP-43 possibly by promoting TDP-43-TOM70 and DNAJA2-TOM70 interactions, which is independent of the LC3 interacting region of FUNDC1 in cellular experiments. In the transgenic fly model of TDP-43 proteinopathy, overexpressing FUNDC1 enhances TDP-43 induced mitochondrial damage, whereas down-regulating FUNDC1 reverses TDP-43 induced mitochondrial damage. FUNDC1 regulates mitochondria-mediated TDP-43 degradation not only by regulating mitochondrial TDP-43 import, but also by increasing LONP1 level and by activating mitophagy, which plays important roles in cytosolic TDP-43 clearance. Together, this study not only uncovers the mechanism of mitochondrial TDP-43 import, but also unravels the active role played by mitochondria in regulating TDP-43 homeostasis.Though TDP-43 protein can be translocated into mitochondria and causes mitochondrial damage in TDP-43 proteinopathy, little is known about how TDP-43 is imported into mitochondria. In addition, whether mitochondrial damage is caused by mitochondrial mislocalization of TDP-43 or a side effect of mitochondria-mediated TDP-43 degradation remains to be investigated. Here, our bioinformatical analyses reveal that mitophagy receptor gene  TDP-43 gene have been identified only in a fraction of familial ALS-TDP and FLTD-TDP patients, but not in the majority of TDP-43 proteinopathy cases. Dysregulation of TDP-43 expression and protein homeostasis may contribute to the pathogenesis in most cases of TDP-43 proteinopathy.TDP-43 proteinopathy is a group of neurodegenerative diseases characterized neuropathologically by the presence of protein inclusions containing phosphorylated TDP-43 protein . This isMitochondrial damage is an early event and an important aspect of the pathogenesis of TDP-43 proteinopathy . A rangemt), and TDP-43 can be degraded by mitochondrial protease LONP1   for 30 min at 37 °C. To stop the reaction, 20 mM Tris-HCl, pH 7.8 was added at room temperature for 15 min. Mitochondria fractions prepared from DSP-treated cells were then used for Co-IP as described above.E. coli Rosetta strain . Bacteria were grown at 37 °C in LB medium with 100 μg/ml ampicillin and induced with 0.5 mM isopropyl-D-thiogalactopyranoside (IPTG) for 18 h at 16 °C. The cells were harvested and suspended in buffer A  and French pressed on ice at 1200 bar for 4 times. The lysate was centrifuged, and the pellet was resuspended with NTA buffer  containing 8 M Urea and loaded onto a Ni-NTA column . The column was washed with NTA buffer containing 8 M Urea and then TDP-43 bound to the column was eluted with NTA buffer containing 0.25 M imidazole and 8 M Urea. The collected fraction was either flash-freeze or dialyzed against 20 mM Tris, pH 8.0, 0.2 M NaCl for the following experiment. Human TDP-43 protein with a myc-his-tag was purified using Ni-Sepharose  following Tet-induction (1 μg/mL tetracycline) of the inducible HEK293 cells for 36 h. Purified human TDP-43 was analyzed by SDS-PAGE followed by Coomassie Brilliant Blue staining or by immunoblotting with anti-TDP-43 or anti-myc antibody.A cDNA encoding human full-length TDP-43 protein was cloned into vector pET32M3C, expressed as a fusion protein carrying amino-terminal thioredoxin and 6×his-tag and purified from the 2, 3% BSA, 4 mM ATP, 4 mM NADH), and 10 μg purified TDP-43 was then added to start the import assay. Unless specified otherwise, the solution was incubated for 30 min at room temperature. The mitochondria were collected following centrifugation at 8000 × g for 5 min at 4 °C. The mitochondria pellets were washed twice with 1 ml wash buffer at 8000 × g for 5 min at 4 °C, followed by “Proteinase K protection assay”.Mitochondria were purified according to “Biochemical fractionation experiments”. Mitochondria were suspended in 500 μL import buffer , the cells were harvested and mitochondria were purified, followed by “Proteinase K protection assay” and immunoblotting analysis. In different groups, the dosage and transfection time for the negative control siRNA (NC) were the same as the siRNAs targeting corresponding genes. The target sequences of siRNAs were listed as follows: DNAJA2: 5’-GGAAGAAGGTGATTAAAGA-3’; HSPA8: 5’-GCTGGTCTCAATGTACTTA-3’; TOM70: 5’-GAGGCAGCATGTACATGCA-3’.The siRNA was from RIBOBIO . Corresponding siRNA  was incubated with 30 μL lipofectamineRNA-seq datasets of human frontal cortex samples were downloaded from GEO database, including those from control subjects , or Two-way ANOVA followed by a post hoc test. For enumeration data, differences between different groups were analyzed using the Chi-square test or Fisher’s exact test, and Bonferroni correction was used for multiple comparisons. Measurement data are presented as mean ± standard error of the mean (SEM). Significance is indicated by asterisks: *P < 0.05; **P < 0.01; ***P < 0.001. The correlation coefficient was calculated using the Spearman method. Graphs were prepared using the R package “ggplot2” (v2.0.0) [Quantification of immunoblotting, TEM, and fluorescence data was performed using ImageJ2 (v2.3.0). Data were collected in Excel (Microsoft) and analyzed using the R program (v3.3.3). Differences between different groups were analyzed using a one-sided Student’s (v2.0.0) .Reproducibility checklistSupplementary informationOriginal Data File


N = 76), and whether it varied depending on maternal involvement in childcare and the perceived quality of past experiences of care.Attentional bias toward infant faces is associated with parental sensitivity and supports the infant-caregiver attachment relationship, ultimately fostering child health outcomes. However, experience-related determinants of parents' attentional bias to infant faces have been poorly investigated. We examined attentional bias to infant versus adult faces in a sample of same-sex mothers (A Go/no-Go attentional task was used to compare the effects of infant and adult faces in retaining attention. Maternal involvement in childcare was measured using items addressing nurturing behaviors. Memories of past experiences of care were collected using the short-form version of the Parental Acceptance-Rejection scale.Results confirmed that infant faces induced greater attentional bias compared to adult faces. More involved mothers were more biased, in terms of attention, to infant versus adult faces. Attentional bias to infant versus adult faces increased as mothers felt more rejected by their own fathers during childhood.Our findings suggested that attentional bias to infant faces might be associated with past experiences of care and direct commitment in childcare in same-sex mothers. Robust and accurate empirical findings on same-sex parent families are essential to inform social policies supporting these families’ well being. Parent–infant interactions rely on the ability to express signals through facial expressions . In partBehavioral tasks assessing attentional bias in parents typically present infant cues as distractors during conflict tasks . The attentional bias index refers to the difference in attention captured by distinct stimuli , which suggests a greater cognitive engagement to a particular stimulus. Using an attentional Go/no-Go task, Psychological characteristics of mothers have been found to modulate their attentional bias to infant faces . PearsonThe attentional bias toward infant faces might be also linked to the perceived quality of care received during childhood. According to Bowlby's theoretical work (1969/1982), repeated interactions with attachment figures are schematized in the form of Internal Working Models (IWM) that synthesize the main self-other interactive dynamics and are automatically activated by individuals when processing new situations. Similarly, in the theoretical framework of the Interpersonal Acceptance-Rejection Theory  confirm that infant faces retain more attention compared to adult faces; ii) investigate whether the attentional bias to infant versus adult faces is associated with mothers’ involvement with childcare; iii) investigate whether individual variations in past experiences of care with mothers' own caregivers are associated with the attentional bias to infant versus adult faces. First (i), we expect that infant faces interfere with the task performance more than adult faces, slowing RTs to peripheral stimuli in Go conditions. Given that an attentional bias to infant distressed faces has been specifically found in previous research ; 2013, wN = 76 mothers being in a same-sex couple participated in the study. Contact with most mothers was made through the Italian Association “Famiglie Arcobaleno” , which sent an invitation to participate in the study to all its members through the mailing list. A snowball sampling method was also used, such that mothers who participated in the study were asked to forward the study invitation to other same-sex mother families who might be interested in joining the study. To be included in the study sample, i) the age of mothers’ only or youngest child should range between 2 and 36 months; ii) mothers raised their child since birth. Both members of each same-sex couple were first invited to participate in the study together. However, whilst 70 mothers (92 %) participated in the experiment with their partner, 6 mothers (8 %) participated alone. Only participants who completed the whole experimental procedure (n = 73 mothers) were included in the final analyses; whilst two mothers did not complete the task, one mother was excluded from the final sample for technical issues while performing the task. The majority of participants were Italian (95.8 %), but 3 of them (4.2 %) reported to be of a different nationality . The Socio-Economic Status (SES) was calculated according to A group of Sociodemographic data was collected. Mothers were asked to answer all the questions by referring to the youngest child in case of multiple children.not at all; 2 = rarely; 3 = a few times a week; 4 = About once a day; 5 = more than once a day) were summed, with a higher score reflecting a higher degree of involvement. Cronbach's alpha was satisfactory (α = 0.81). The McDonald's ω total was 0.87.To assess parental involvement in childcare, mothers completed 10 items  adapted from the Parental primary caregiving role Structured Interview . The orialmost always true to 1 = almost never true). In this study, the two total scores had a good reliability .Mothers completed the Italian validated short-form version  of the PMothers completed a modified Go/no-Go task derived from an established paradigm  to measuTo overcome geographical barriers in recruiting the sample, all the experimental procedures were conducted online. Self-reports were administered through Qualtrics . The experimental task was run on JATOS . The tast-tests were run to check for potential differences within the sample. Correlation analyses were implemented. Missing data in self-reports was not replaced. Analyses were conducted for evaluating the number of correct answers for different blocks. The overall accuracy for Go trials was 96.6 %, which confirmed the ability of participants to complete the task as instructed. The percentage of false alarms  were 3.9 %. Response accuracy was analyzed using a generalized mixed-effects model. RTs were analyzed via linear mixed-effects models . Pearson correlation analyses did not show any significant associations between maternal involvement and PARQmother or PARQfather. PARQmother and PARQfather showed a low positive correlation .β = −0.015, SE = 0.002, t = −9.070, p < .001), as infants slowed RTs to a greater extent compared to adult faces. This effect remained statistically significant after increasing the complexity of the random effect structure by varying all the slopes  and only the slope of the main effects . Model 1 also evidenced a main effect of emotional valence , as mothers allocated greater attention to sad faces compared to happy and neutral faces. However, this effect was not statistically significant in all the subsequent models. The interaction between face age and emotional valence was not significant. In Model 2 . It highlighted a two-way interaction effect between face age and maternal involvement . Interestingly, more involved mothers were more biased, in terms of attention, toward infant versus adult faces . The interaction effect between face age and PARQmother was not significant. Model 4 confirmed the main effect of face age  and highlighted a two-way interaction between face age and PARQfather . That is, those mothers who felt more rejected by their own father were more biased, in terms of attention, by infant versus adult faces  has been considered an optimum point of reference for assessing caregiving involvement , we deciM = 46) was greater than the ones previously reported in other studies  = 35.81, M(wave2) = 34.00 and M(wave3) = 33.52; From the attachment theory perspective , the intContrary to our hypothesis, we did not find that attentional prioritization of infant over adult faces varied in relation to early care experiences with one's own mother. Compared to previous studies , it shouSome limitations of our study should point toward future directions. Future research might clarify the role of individual mental health as a potential covariate in relation to the behavioral findings e.g., . In non-In a considerable sample of same-sex mothers, we examined the contribution of past and current experiences of care on the attentional bias toward infant faces. Taken together, mental representations of care built during childhood and direct commitment in childcare were associated with same-sex mothers’ attentional bias toward infant faces. Apart from having a value on its own, inclusive research of different family contexts is needed conceptually and methodologically, as this might allow researchers to move beyond sex-related differences in families and consider different roles and arrangements . OverallThe authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.


Interactions between the programmed cell death receptor 1 (PD-1) and its ligand (PD-L1) lead to immune evasion in various tumors and are associated with poor prognosis in patients with pancreatic cancer; however, the roles of PD-L1-containing exosomes in pancreatic cancer is poorly understood. Here, we investigated the correlation between circulating exosomal PD-L1 (exoPD-L1) and PD-L1 expression in tumor tissue, and survival outcomes in patients with advanced PDAC.Exosomes were derived from pre-treatment serum samples isolated using ExoQuick kit from 77 patients with advanced pancreatic cancer. Exosomal PD-L1 (exoPD-L1) was detected by enzyme-linked immunosorbent assay, and matched tumor tissues PD-L1 expression were evaluated by PD-L1 immunohistochemistry (22C3) assay, described with combined positive score. Cutoff value of exoPD-L1 for survival was assessed with receiver operating characteristic curve analysis. Kaplan-Meier analysis was performed to obtain median overall survival (OS), and hazard ratio was estimated using a stratified Cox regression model.p = 0.023). There was a significantly higher proportion of elevated exoPD-L1 levels in patients with positive PD-L1 expression compared to patients with negative PD-L1 expression (p = 0.001). Patients were classified into groups with low and high exoPD-L1 levels using ROC curve-derived cutoffs . At a median follow-up of 8.39 months, the median OS was 13.2  and 6.36 months  in the low and high exoPD-L1 groups, respectively . ExoPD-L1 levels did not affect the proportion of CD8+CD69+ effector cytotoxic T cells in either of the groups (p = 0.166).The median exoPD-L1 serum concentration was 0.16 pg/mg, with undetected levels in seven patients. ExoPD-L1 levels were significantly higher in patients with systemic disease than in those with locally advanced disease (The serum-derived exoPD-L1 levels were higher in metastatic pancreatic cancer than locally advanced disease. Collectively, higher serum exoPD-L1 levels in patients with advanced pancreatic cancer suggested worse survival outcomes and may have clinical implications.The online version contains supplementary material available at 10.1186/s12885-023-10811-8. Pancreatic ductal adenocarcinoma (PDAC) is a lethal disease with a dismal 5-year overall survival (OS) rate of only 9% . AlthougIn many solid tumors, T-cell activation is inhibited by the interaction between the immune checkpoint programmed cell death ligand 1 (PD-L1) and programmed cell death receptor 1 (PD-1), which is an inhibitory receptor of activated T-cells. Suppression of T cell activation facilitates immune evasion by cancer cells and is associated with poor prognosis in patients with cancer . Thus, tThe TME of PDAC comprises regulatory T cells (Tregs), myeloid-derived suppressive cells (MDSCs), and tumor-associated macrophages (TAMs) that inhibit effector CD8 + T cells . PD-L1 oIn the present study, we investigated the correlation between PD-L1 expression in tumor tissue and circulating exoPD-L1 in patients with advanced PDAC. Furthermore, we evaluated whether circulating exoPD-L1 is associated with survival outcomes in patients with advanced PDAC. We also examined the correlation between CD8 + T cell counts and circulating exoPD-L1 levels.We enrolled 77 patients with advanced PDAC diagnosed at the Catholic University of Korea, Seoul St. Mary’s Hospital, between 2017 and 2021. Patients with available serum samples at the time of diagnosis for advanced disease were eligible according to the following criteria: (1) histologically confirmed PDAC; (2) locally advanced or metastatic disease; (3) survival confirmed at the time of data collection; (4) Eastern Cooperative Oncology Group (ECOG) performance status (PS) from 0 to 2. Peripheral blood samples were collected from patients before initiating systemic chemotherapy in a first-line palliative setting. Blood samples were delivered to the laboratory, immediately centrifuged at 2,500 × g for 10 min and stored in 2–4 mL aliquots at − 80 °C until use. Tumor tissue specimens were obtained from patients with PDAC who had undergone surgery or core needle biopsy. The study protocol was approved by the Institutional Review Board (IRB) of The Catholic University of Korea, Seoul St. Mary’s Hospital . Informed consent was obtained from all the patients. All procedures were conducted in accordance with Korean regulations and the Declaration of Helsinki.Serum exosomes were isolated using the ExoQuick exosome precipitation solution  as per manufacturer’s instructions. Thawed serum samples (500 µL) were centrifuged at 12,000 × g for 15 min to remove cells and cell debris. The supernatants were transferred to a sterile tube, mixed with ExoQuick reagent 120 µL) and incubated for 1 h at 4 °C, and then centrifuged at 1,500 × g for 30 min. The supernatant was removed, and the precipitate was centrifuged at 1,500 × g for 5 min to remove the residual ExoQuick solution and obtain precipitated exosome in the pellet. After discarding the supernatant, the pellets were resuspended in 250 µL sterile phosphate-buffered saline (PBS) and stored at − 80 °C until required. The NanoSight NS300 instrument with nanoparticle tracking analysis 2.3 software  was used to analyze exosomal size distribution . Exosome µL and iBriefly, 50 µg exosomes were resolved by 12% SDS-PAGE and transferred to nitrocellulose membranes  for western blot analysis. The membranes were incubated with primary antibodies against TSG101  and CD63 . Membranes were incubated with appropriate horseradish peroxidase-conjugated secondary antibodies , and the bands were visualized using a chemiluminescent western blot reagent .The expression of PD-L1 in serum-derived exosomes was detected using human PD-L1 DuoSet ELISA kits  according to the manufacturer’s protocols. The optical density was measured at 450 nm using a VersaMax Microplate Reader , and protein levels were calculated according to standard curves. The protein concentration of the isolated exosomes was determined using a BCA protein assay kit  according to the manufacturer’s instructions. The concentration of exoPD-L1 was standardized per mg of exosomal protein. For the detection of cytokines in the plasma, commercially available ELISA kits for transforming growth factor beta 1 (TGF-β1) (#DY240-05), TGF-β2 (#DY302), granzyme B (#DY2906-05), interferon-gamma (IFN-γ) (#DY285B-05), and interleukin (IL)-1β  were used according to the manufacturer’s protocol.Higher levels of sPD-L1 are associated with a poor prognosis in patients with various solid tumors . HoweverFormalin-fixed, paraffin-embedded tissues (4-µm-thick sections) from patients with advanced PDAC were tested for PD-L1 expression using the PD-L1 immunohistochemistry (IHC) 22C3 pharmDx assay  on an Autostainer Link 48 system with an EnVision DAB Detection System  following the manufacturer’s instructions. PD-L1 immunoreactivity was evaluated by two pathologists using a multihead microscope. PD-L1 expression in the specimen was described using a combined positive score (CPS), which is defined as the number of PD-L1 stained cells  divided by the total number of viable tumor cells and multiplied by 100 Peripheral blood mononuclear cells were isolated using Ficoll-Paque PLUS  density-gradient centrifugation and stored in liquid nitrogen until further analysis. Thawed peripheral blood mononuclear cells were incubated with anti-CD3 , anti-CD45 , anti-CD8 , and anti-CD69  antibodies for 20 min at 4 °C in the dark. The stained cells were analyzed using flow cytometry , and data were analyzed using FlowJo software .Descriptive statistics are reported as proportions and medians with ranges or interquartile ranges (IQR). A time-dependent ROC curve was used to assess the predictive performance of the clinical factors associated with survival outcomes. The cutoff values of the clinical factors were calculated based on Youden’s index. Differences between groups and associations between clinicopathological features and exoPD-L1 were determined using chi-squared and Fisher’s exact tests for categorical data and an unpaired t-test with Welch’s correction for continuous data. Pearson’s correlation coefficient was used to evaluate the correlation between exoPD-L1 and tumor PD-L1 expression. OS was estimated from the date of diagnosis of advanced disease to the time of last follow-up or death. Survival curves were estimated using the Kaplan–Meier method, and survival differences were compared using a two-tailed log-rank test. Cox proportional hazards regression models were used to identify the effects of the clinical factors on survival. The hazard ratio (HR) and 95% confidence intervals (CIs) were estimated for each factor. Results with two-sided p-values < 0.05 were considered statistically significant. Statistical analyses were performed using SPSS for Windows  and GraphPad Prism version 8.0 .The clinicopathologic characteristics of the 77 patients with advanced PDAC are summarized in Table Rs = 0.788; p < 0.0001; Fig. p = 0.023; Table ExoPD-L1 was undetectable or below the lower detection limit in seven of the 77 patients (9.0%). The median value of exoPD-L1 was 5.87 , and the median value of exoPD-L1 normalized with exosomal protein was 0.16 . Standardized exoPD-L1 concentrations were associated with corresponding exoPD-L1 levels .Among the 77 patients, PD-L1 expression in the tumor tissue was evaluated in 68 patients, and the correlation between tissue PD-L1 and serum exoPD-L1 levels was assessed. The median CPS of tumor PD-L1 was 3.0 IQR, 0–20), and 19 patients (27.9%) had negative tumor PD-L1 expression, defined as CPS < 1% . No linear correlation was observed between the tissue PD-L1 CPS and serum exoPD-L1 values in patients with advanced PDAC  with 52.6% sensitivity and 69.2% specificity. According to the cutoff value, the entire cohort was classified into low exoPD-L1 (45 patients) and high exoPD-L1 (32 patients) groups. The median OS was 13.2 months  in the low exoPD-L1 group, as compared with 6.36 months  in the high exoPD-L1 group , 56 patients (72.7%) had died. Among these deaths, 51 (92.7%) were related to disease progression, and five (8.9%) were attributed to treatment-related adverse events. The median follow-up was 8.39 months . In the entire cohort, the median OS was 9.28 months , and OS rates at 6 months, 9 months, and 1 year were 62.3%, 50.1%, and 40.6%, respectively. The optimal cutoff value that discriminated survivors at nine months was determined using the ROC curve. The optimal cutoff value of exoPD-L1 was 0.165 pg/mg, and the area under the curve was 0.617  and higher baseline neutrophil-to-lymphocyte ratio (NLR)  were significantly associated with worse OS. In addition, patients who received systemic chemotherapy had longer OS  than those who did not  and effector CD8 + T cell (p = 0.166) proportions were comparable between the two groups according to the exoPD-L1 level (Fig. p = 0.861; Fig. p = 0.031; Fig. The proportion of CD3 + CD45 + CD8 + T cells was measured in each patient, and the fraction of CD69 + CD8 + T cells was assessed to investigate cytotoxic T cell activity. The total cytotoxic T cell (Activation of the PD-1/PD-L1 signaling pathway can suppress T cell responses in various cancers, thereby leading to immune evasion . To the In a recent meta-analysis, elevated tumor PD-L1 expression was correlated with advanced stage and poor OS in patients with PDAC. However, the IHC methods for PD-L1 expression were not validated . High plIn this study, exoPD-L1 levels were not significantly correlated with clinicopathological features, except for the disease status. Patients with systemic disease had elevated exoPD-L1 levels compared to those with locally advanced disease, which was consistent with previous reports wherein exoPD-L1 levels correlated with advanced disease stage in patients with head and neck and lung cancer , 26. TheWe assessed the correlation between tumor PD-L1 IHC status and serum exoPD-L1 levels in patients with pancreatic cancer for the first time. Serum exoPD-L1 levels were higher in the positive than in the negative PD-L1 expression group. Consistently, in a previous study, tumor PD-L1 expression was associated with a higher percentage of PD-L1-expressing exosomes in patients with non-small cell lung cancer . In contSimilar to sPD-L1, serum-derived exoPD-L1 may be elevated in patients with increased systemic inflammation provoked by the inflammatory tumor type of pancreatic cancer . PancreaCirculating exosomes can interact with immune cells through various mechanisms. PD-L1-containing exosomes can suppress CD8 + T-cell activity by downregulating CD69 expression on the surface of activated T cells . AdditioSeveral limitations of our study need to be considered. First, the association between treatment resistance and exoPD-L1 was not evaluated because exoPD-L1 was not serially measured after treatment. Second, none of the patients in this study received anti-PD-1/PD-L1 blockade immunotherapy; therefore, we could not investigate the impact of exoPD-L1 on the response to anti-PD-1/PD-L1 blocking therapy. Third, tissue evaluation in relapsed patients was based on surgically resected specimens rather than relapsed tissues. Finally, the relationship between exoPD-L1 and tumor-infiltrating CD8 + T cells in the TME could not be investigated, as only serum-derived exosomes were evaluated and not exosomes from the tumor tissue.In conclusion, we found that exoPD-L1 was higher in PDAC patients with systemic disease than in those with locally advanced disease. Serum-derived exoPD-L1 levels correlated with PD-L1 expression in tumor tissue. Additionally, patients with PDAC and high exoPD-L1 levels had worse survival outcomes than those with low exoPD-L1 levels. Further studies exploring the association between PD-L1 expressing exosomes derived from tumor tissue and cytotoxic or immunosuppressive T cells in the TME are required to validate the clinical importance of exoPD-L1 in patients with pancreatic cancer.Below is the link to the electronic supplementary material.Supplementary Material 1


Magnaporthe oryzae is the causal agent of rice blast disease and can infect over fifty grass species. We demonstrate that rice blast fungi encode two diterpene synthases that produce normal pimara-8,15-diene and manoyl oxide scaffolds. Phylogenetic analysis of diterpene synthases among rice blast pathotypes showed functional conservation of these two core diterpene synthases amongst all pathotypes and suggests further expansion in those infecting select grass species. These insights into the blast fungal terpenome may inform efforts to counteract deleterious phytopathogens in crucial food crops.Plant-pathogenic fungi harbor various specialized metabolites including diterpenoids that function as hormones and virulence factors. The fungus  Ascomycota produce a diverse array of specialized metabolites with versatile roles that confer a competitive advantage in nature regarding virulence, interspecies communication and competition, and biotic and abiotic stress protection -geranylgeranyl diphosphate (GGPP) , also known as rice blast fungus, destroys 10–30% of total rice yield annually worldwide. Other than rice, the fungus infects over fifty grass species, the most alarming being its recent discovery in wheat  to shortlist bifunctional diterpene synthases. Class II cyclases are characterized by the DxDD motif involved in protonation-initiated cyclization. Class I active site has the characteristic Dxx motif that facilitates ionization initiated cyclization of the class II intermediate  and primers in Synthetic codon-optimized diterpene synthase genes for heterologous expression in E. coli cells pre-transformed with pIRS (to increase yield) for recombinant expression of each diterpene synthase. The resulting recombinant strains were grown at 37°C overnight in 5 ml of TB media (supplemented with 100 mM phosphate buffer pH 7.5 and appropriate antibiotics). One ml of overnight culture was used to inoculate 50 ml of TB media and the cultures were incubated at 37°C and 160 rpm till mid-log phase. At this point, the temperature was dropped to 16°C for 1 h and then 1 mM isopropylthiogalactoside (IPTG) was added for induction and 50 mM sodium pyruvate was added as a carbon supplement. The induced culture was further incubated for 3 days at 16°C and 160 rpm to allow product formation. The resulting products were extracted adding equal volume of hexane and the organic layer was separated, dried and re-dissolved in 1 ml of n-hexane for product analysis.The diterpene synthase products were determined using a modular metabolic engineering system  into yeast ZXB competent cells using previously described  values in the range from 60 to 600 starting from 9 min after sample injection until the end of the run. For identification of enzymatic products, the retention time and mass spectra of products were compared to authentic standards  on Agilent 7890/7000C GC-QQQMS using an Agilent HP-5MS column with 1.2 ml/min helium flow rate. Samples (1 ul) were injected in splitless mode using an Agilent 7693 autosampler. The following temperature program was used: the oven temperature started at 70°C, which was maintained for 1 min, and then increased at a rate of 20°C/min to 300°C, where it was held for another 2 min. The mass spectrum was recorded by mass-to-charge ratio  in M. oryzae reference genome (Mo70_15) to produce diterpenes. Using BLAST analysis, and presence of characteristic protein domains for published genome analysis we identified two putative diterpene synthases  among M. oryzae pathotypes that infect different host plants. Utilizing the diterpene synthase mining approach described in the Methods section and depicted in M. oryzae proteomes covering 14 pathotypes , brome grass (Bromus), and fingermillet (Eleusine) compared to rice (Oryza) pathotype. MoDiTPS6 is uniquely present in the pathotypes from two grasses –Triticum and Bromus. The observed expansion of DiTPS enzyme family aligns with several studies that have highlighted the fluidity of fungal genomes and expansion of various gene families  common ancestral origin of ent-kaurene synthases and MoDiTPS1, 4 and 6 clade. MoDiTPS2 and 5 have same node of divergence and do not group with any characterized DiTPS. However, DiTPS3 associates with a known phyllocladanol synthase , these were biochemically characterized using a previously described modular metabolic engineering system  or normal specific abietadiene synthase (AgAS) ,15-diene, indicating that this is relevant stereoisomer of CPP produced by its class II active site. The DiTPS2 class I active site also reacts with syn-CPP resulting in unidentified products but does not form any products with ent-CPP . Subsequent co-expression of the class I mutants with GGPP resulted in production of the expected intermediate 8α-hydroxy-CPP by DiTPS1 and CPP for DiTPS2 . To probs of CPP . To asseroxy-CPP ; MoDiTPS ent-CPP . These r ent-CPP .B71 and MZ5-1-6 may have variable regulator genes. The putative DiTPS2 cluster shows the presence of a complete and conserved biosynthetic gene cluster among M. oryzae pathotypes with precursor GGPP synthase (MGG_00026), oxidases and/or cytochromes P450 , dehydrogenase (MGG_00017); transcription factor (MGG_00032), and transporter (MGG_00040) , oxidases/cytochromes P450 , ligase (MGG_01951), and could possibly include a UDP glycosyl transferase (MGG_01961) . The synG_00040) . The biothotypes . These rMagnaporthe pathotypes infecting rice and other grass species. From the literature, the two DiTPSs were transcriptionally upregulated under various conditions in rice and fingermillet  are phylogenetically conserved across Magnaporthe pathotypes demonstrates an expansion of diterpene synthases that could be involved in adaptation of pathotypes for infection of other grasses. In particular, via intra-species duplication generated from the two core diterpene synthases (Fusarium species where terpene synthases were conserved across species, independent of host variety (Wiemann et al., F. fujikuroi. Nevertheless, this research provides a strong basis for further studies to investigate if the observed phylogenetic expansion is limited to duplication, but generally results in pseudogenes rather than neofunctionalization or whether the observed inactivity is an artifact of the experimental process. Our results also highlight the need for production of both pimara-8,15-diene and ent-manoyl oxide activity, as well as hinting at a relationship to pathogenicity. Thus, our study defines the diterpene biosynthetic capability in Magnaporthe pathotypes and provides impetus to further investigate the importance of chemical diversification in evolution and adaptation of plant pathogenic fungi. As terpenes mediate diverse functional roles, the conserved production of at least these two core diterpenes suggests a biological role for these diterpenes in the rice blast fungus.Interestingly, the phylogenetic analysis across ynthases . Terpeneynthases . Our resM. oryzae (Peters, ent-manoyl oxide, which are distinct from the diterpene scaffolds produced by rice (M. oryzae pathotypes. The BGC for MoDiTPS1 is more variable amongst the isolates which is consistent to the demonstrated variability in our biochemical analysis. The conservation of backbone formation and BGC suggest a conserved role of MoDiTPS2 in M. oryzae pathotypes. It would be worthwhile to confirm the BGC through experimental expression in heterologous hosts such as Aspergillus and yeast to not only identify the bioactive diterpenoids but understand the variability in the clusters.From the host plant perspective there is a strong diterpene-based chemical defense response in rice after infection by  Peters, . We show by rice . These dM. oryzae, the biochemical characterization described here provides information to enable experimental differentiation at the host-pathogen interface.As diterpenoids are essential for defense and resistance of rice to M. oryzae contains two diterpenes synthases that are evolutionarily conserved across different pathotypes, suggesting important roles for the resulting diterpenes. These labdane-related diterpenes produced by Magnaporthe are functionally distinct from those produced by rice. This research provides a strong basis for further studies of in vivo confirmation of gene function including testing whether the observed phylogenetic diversification confers an evolutionary advantage. Our study defines the diterpene biosynthetic capability in Magnaporthe pathotypes and provides insight to the importance of chemical diversification in evolution and adaptation.In conclusion, it is now evident that The original contributions presented in the study are included in the article/AS performed all the experiments. AS and HY performed the computational analysis. AS and SM analyzed the data and wrote the article. SM designed the research and supervised the project. All authors commented on the results and manuscript. All authors contributed to the article and approved the submitted version.Financial support was provided by the Department of Biochemistry and Molecular Biology, UMass Amherst and Richard and Susan Smith Family Foundation, Newton MA to SM. AS was supported by Lotta M. Crabtree Fellowship.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.


In this paper, an ultra-wideband and polarization-insensitive frequency-selective surface absorber is presented with oblique incident stable behavior. Different from conventional absorbers, the absorption behavior is much less deteriorated with the increase in the incidence angle. Two hybrid resonators, which are realized by symmetrical graphene patterns, are employed to obtain the desired broadband and polarization-insensitive absorption performance. The optimal impedance-matching behavior is designed at the oblique incidence of electromagnetic waves, and an equivalent circuit model is used to analyze and facilitate the mechanism of the proposed absorber. The results indicate that the absorber can maintain a stable absorption performance with a fractional bandwidth (FWB) of 136.4% up to 40°. With these performances, the proposed UWB absorber could be more competitive in aerospace applications. A frequency-selective surface (FSS), also known as an artificial electromagnetic metasurface, is a kind of periodic electromagnetic modulation structure ,2. BecauIn the available literature, most of the FSS absorbers are composed of multilayer structures and their loss modules are usually resistors, resistive inks, or resistive sheets. These absorbers can be categorized as single-band or multiband according to the resonance of the structures ,10. The To solve this problem, designing the optimal impedance matching effect at the oblique incident angle is proposed in this paper. Furthermore, using graphene coating can also obtain better broadband absorption and angular stability on this basis. The results of the research show that the performance of the absorber designed by this method will not deteriorate with the change in the incident angle within the range of 0–40°. On the contrary, its absorption band coverage is improved. The proposed ultra-wideband absorber with angular stability and polarization insensitivity may have a high application potential in the field of stealth.So far, there has been a common problem with conventional absorbers that the absorption performance worsens with the increase in the incident angle. As shown in The absorption mechanism of the absorber is the impedance matching principle. For the presented structure, the input impedance of the absorber was close to 377 Ω in a wide band, and the imaginary part was close to zero under oblique incidences, which matched the wave impedance of the air conjugate. At this time, the electromagnetic energy irradiated in the absorber hardly reflected and most entered the absorber. Graphene was used as the lossy material, so the electromagnetic energy entering the absorber was quickly lost to achieve perfect absorption.ε = 4.3). The purpose of using a two-layer lossy structure was to obtain a large bandwidth. Both the lossy layer I and lossy layer II had an absorption band. Combining the two lossy layers obtained an ultrawide absorption band. The symmetric graphene coating structure made the absorber insensitive to different incident polarization. To ensure zero transmission, a ground plane was provided on the bottom of another FR-4 substrate of the same thickness. After calculation and simulation, the optimized geometrical design parameters are shown in h1 represents from the lower part of the bottom plate to the lower surface of lossy layer I, and h2 represents from the lower surface of lossy layer I to the lower surface of lossy layer II.The unit cell geometry of the proposed FSS absorber is shown in T is the temperature fixed to 300 K.The graphene was composed of graphene flakes, carbon black, solvents, related binders, and dispersants. To ensure the carbon black and graphene flakes were uniformly dispersed, they were mixed and ultrasonically dispersed in a planetary mixer/deaerator  and an ultrasonic cleaner. Then, the graphene ink was printed on the prepared FR-4 substrate and cured at a high temperature . The comgd is the thickness of the graphene layer. The permittivity of the graphene was 2.3 on the research frequencies [Then, the value of the permittivity of graphene was calculated by using the formula :(2)ε=1+iquencies .To further facilitate this research, the absorption performances of different finite-size structures were modeled and simulated through the FSS FULL STRUCTURE function of CST Microwave Studio, as presented in d). Here, Z0 = 377 Ω represents the free space wave impedance, and As shown in C1, C2, and C3) and inductors  in a is the period of the unit structure, and l and d are the length and width of the graphene structure, respectively.To better predict the frequency response of the proposed FSS by ECM, the capacitors  and (4), it can be seen that the increasing incident angle mainly influenced the value of d and the period along the 11) of the lossy layers are shown in The ECM and unit cell geometry of the proposed FSS absorber were simulated using Keysight Advanced Design System (ADS) software and CST Studio Suite (CST) software, respectively. The simulated reflection coefficients (SMoreover, the input impedance of the absorber was extracted through the simulated S parameter, to quantitatively explain the working mechanism of the absorber. The real and imaginary parts of the impedance at 0° and 45° are shown in The performance of the design under different oblique angles of incidence was also studied to determine the suitability of the absorber for applications demanding stable performance at large oblique angles. To show more clearly the absorption performance of the presented structure, the absorptivity results are shown in In z-axis are shown in To verify the absorption performance of the proposed structure, the electric field distributions along the +R1 and R2 both in the range of 70–350 Ω/sq. As shown in R1 sheet resistance mainly affected the lower frequency absorption peak. In R2 sheet resistance mainly affected the absorbing bandwidth.Then, the sheet resistance of the graphene flake was analyzed. R1 and R2 directly influenced the real part and imaginary part of the impedance. As shown in R1 mainly affected the real part at the low-frequency absorption band and slightly affected the high-frequency absorption band. R2 influenced the real part at both the low-frequency and high-frequency absorption peaks. Meanwhile, the frequency point near zero of the imaginary part of the impedance moved, which means the range of the absorption band may change. The analysis of the real and imaginary parts of the impedance corresponded well with the absorption performance in The changes in the sheet resistance The comparison with FSS absorbers in other references is shown in In this article, a graphene-coating ultra-wideband FSS-based absorber with angular stability and polarization insensitivity performances was proposed. To mitigate the absorption performance degradation with the increasing incident angle, the optimal impedance matching effect was designed at the oblique incident angle. At the same time, graphene coating was used to obtain better broadband absorption and angular stability. As a result, the presented structure had a stable absorption performance up to 50°. It is worth pointing out that the FWB of the presented absorber maintained 136.4% up to 40°. Then, the ECM method was used to analyze the mechanism of the unit structure. According to these results, the proposed absorber may have application potential in the field of stealth.


Ketamine has been increasingly used to treat mental health conditions yet there is a lack of safety data on intramuscular (IM) and sublingual (SL) dosing in a community setting. The Roots to Thrive Ketamine assisted Therapy (RTT-KaT) program is a 12-week program with 12 Community of Practice (CoP) group therapy sessions and three ketamine sessions.To provide preliminary data on RTT-KAT adverse events to subsequently inform safe use of IM and SL ketamine for the treatment of psychiatric disorders.Retrospective chart review of the RTT-KaT Program on four cohorts (n=128) between September 2020 to December 2021. Eligible patients include those with post-traumatic stress disorder, depression, generalized anxiety, burnout/adjustment disorder, substance use disorder, obsessive compulsive disorder, disordered eating, and disordered sleep. Baseline characteristics and adverse events were captured including medication administration before, during, and after RTT-KaT sessions. Chi-squared test with Yates’ continuity correction was used to assess side effects in subgroups from ketamine administration.RTT-KaT was well tolerated with no loss to follow up. There were 351 IM (mean dose = 102.553mg) and 96 SL (mean dose = 276.667mg) sessions of ketamine. Of the 448 sessions, the prevalence of elevated blood pressure increased by 12.31% from baseline (36.85%), with all post-treatment elevations being transient. The prevalence of elevated blood pressure post-KaT session was also similar between IM (+11.69% from 37.71% baseline) and SL (+15.12% from 32.98% baseline) administration. Regarding adverse effects, 12.05% of sessions experienced nausea , 2.52% had an episode of vomiting , 3.35% had a headache , and seven sessions experienced dizziness. The incidence of adverse events was not significantly associated with past psychedelic experiences , nor past psychiatric diagnosis . . There was no significant association between administration route and incidence of nausea, which was the most common side effect. Male gender was also significantly associated with lower incidence of nausea .The group therapy model described provides a comprehensive approach and presents a promising model for operating a KaT program outside of a clinical trial setting.These findings suggest good safety and acceptability for RTT-KaT among individuals seeking treatment for mental health issues. Majority of participants did not experience adverse reactions and the adverse events that were recorded involved transient symptoms that were resolved with rest and/or medications.None Declared


C9orf72 or a mutation in progranulin (PRGN) or have a sporadic cause with no family history of dementia [C9orf72 mutation, dipeptide pathology was present in the retina in the absence of TDP-43 pathology [Frontotemporal dementia (FTD) is a clinically heterogeneous disease, characterized by behavioural, language and movement symptoms. The underlying pathology is termed frontotemporal lobar degeneration (FTLD) and consists of distinct underlying pathologies including aggregation of transactive response DNA-binding protein 43 (TDP-43), tau and fused-in-sarcoma (FUS) . Approxidementia , 10, 12.PRGN mutation, retinal changes have been observed and include thinning and formation of fluorescent lesions, which precede symptoms of FTD [PRGN mouse model, neurodegeneration and nuclear depletion of TDP-43 was reported without TDP-43 aggregation [There are limited data on retinal changes in FTLD. In donors with FTLD due to s of FTD , 14. In C9orf72 repeat expansion, one PRGN mutation and two were genetically sporadic . Pathologically, donors presented with FTLD-TDP types A (PRGN), B (C9orf72), C (sporadic) and E (sporadic) [n = 5), FTLD-FUS (n = 1), AD with limbic-predominant age-related TDP-43 depositions (LATE) (n = 2) and without (n = 1), donors with ALS due to TDP-43 (ALS-TDP (n = 2)) and neurologically healthy controls (n = 6). Retinal TDP-43 pathological burden in FTLD-TDP donors was scored into none, few, frequent and abundant inclusions. Details and pTDP-43 scores of the donors are listed in Supplementary File 1. Immunohistochemical stainings were performed for (pan)TDP-43, phosphorylated TDP-43 (pTDP-43), p62 (SQSTM1), and the dipeptide repeat proteins (polyGA and polyGP). Double-fluorescence with calbindin D28K and calretinin was performed to identify horizontal cells and amacrine cells, respectively (for detailed procedures see Supplementary File 1).Post-mortem retina tissue was obtained from 7 donors with FTLD-TDP, of which four had a poradic) . We alsoC9orf72 and PRGN mutation carriers, as well as the sporadic FTLD-TDP donors, TDP-43 aggregates were observed in the outer plexiform layer of the retina. No colocalization was found with markers for horizontal or amacrine cells. No TDP-43 inclusions were observed in the retina of FTLD-tau, ALS, AD, and neurologically healthy control donors. Interestingly, all C9orf72 mutation carriers showed abundant presence of p62 and display aggregation of dipeptides linked to the hexanucleotide repeat expansion in the inner nuclear layer of the retina [In e retina . The FTLC9orf72 hexanucleotide repeat expansion and showed no retinal or cortical TDP-43/dipeptide pathology. Recently, an increased signal for pTDP-43 in the retina was reported in 6 ALS donors [C9orf72 mutation carriers. This suggests that genetic factors play a role in the occurrence of TDP-43 aggregates. On the other hand, we also observed TDP-43 aggregation in sporadic FTLD-TDP cases. Interestingly, TDP-43 inclusions in the retinal internuclear layer have been reported in subjects with chronic traumatic encephalopathy (CTE) [The ALS patients included in the current study did not carry the S donors . We obsehy (CTE) , 4. AddiC9orf72 mutations, dipeptide pathology is also present in the retina. In this study, we observed no TDP-43 inclusions in the retina of 2 ALS donors without cortical TDP-43 depositions or AD patients with limbic TDP-43 pathology, suggesting that retinal TDP-43 pathology reflects the cortical involvement of TDP-43 aggregation. These findings provide opportunities for retinal TDP-43 aggregation as a biomarker for FTLD-TDP, using non-invasive retinal imaging techniques with the purpose of diagnosing and monitoring progression of FTLD-TDP.To summarize, we report that manifestations of pathological TDP-43 are present in the retina of donors with FTLD-TDP. In donors with Supplementary file1 (DOCX 25 KB)Below is the link to the electronic supplementary material.