However, limited data are available to evaluate the ergogenic potential of NO3− supplementation during other exercise modalities that mandate type II fiber recruitment, such as weightlifting exercise . In this systematic review, we examine the existing evidence basis for NO3− supplementation to improve muscular power, velocity of contraction, and muscular endurance during weightlifting exercise in healthy adults. We also discuss the potential mechanistic bases for any positive effects of NO3− supplementation on resistance exercise performance. Dialnet, Directory of Open Access Journals, Medline, Pubmed, Scielo, Scopus and SPORT Discus databases were searched for articles using the keywords: nitrate or beetroot and supplement or nut*r or diet and strength or “resistance exercise” or “resistance training” or “muscular power”. Four articles fulfilling the inclusion criteria were identified. Two of the four studies indicated that NO3− supplementation could increase aspects of upper body weightlifting exercise  performance (increases in mean power/velocity of contraction/number of repetitions to failure), whereas another study observed an increase in the number of repetitions to failure during lower limb weightlifting exercise . Although these preliminary observations are encouraging, further research is required for the ergogenic potential of NO3− supplementation on weightlifting exercise performance to be determined.Dietary nitrate (NO Weightlifting exercise is well established as an exercise modality of resistance exercise to improve skeletal muscle mass ,2, strenThere are different methods to assess muscle strength and power . Static 3−) ingestion, often administered as concentrated NO3−-rich beetroot juice (BR), has been reported to confer ergogenic effects in various exercise modalities  and [NO2−] following NO3− supplementation have been observed in skeletal muscle [2−] is dependent on methodological considerations, such as the supplementation regimen   increases dose-dependently after NO3− supplementation and is correlated with enhanced exercise capacity [3− dose administered in the study of Flanagan et al. [3− supplementation in that study. This interpretation is reinforced by Coggan et al. [3− ingestion was positively correlated with the increase in plasma [NO2−]. However, a limitation of all existing studies assessing the effect of NO3− supplementation on resistance exercise performance is the lack of plasma [NO2−] determination.Williams et al.  examined failure . In anot failure  observeds et al.  assesseds et al.  assesseds et al. ,121,122.s et al.  to assesined men . Therefon et al.  did not n et al.  was the n et al.  administs et al.   employed might also have contributed to the disparate findings across studies assessing the ergogenic potential of NOexercise ,116, when et al. , but then et al. . Given te fibers  and since fibers , this miividuals ,127. Accividuals  and Rancividuals , who rec3− supplementation can enhance weightlifting exercise performance by increasing muscle power production, velocity of contraction and muscular endurance in healthy resistance-trained adults. However, the results are incongruous with inconsistencies likely linked to differences in supplementation strategies and exercise modality. Therefore, further research is required to assess the weightlifting exercise settings and populations in which NO3− supplementation is more or less likely to be ergogenic. Moreover, while encouraging preliminary evidence suggests that dietary NO3− supplementation may enhance weightlifting training quality, further research is also required to assess whether this translates into greater adaptations to chronic resistance exercise training.Taken together, the existing, albeit limited, evidence suggests that acute and short-term dietary NO3− supplementation, enhanced skeletal muscle contractile function has been observed during electrically stimulated contractions  without altering force production at a supraphysiological PO2 [2+] concomitant with lower submaximal, but not maximal, force at a physiological PO2 [2 administration increased time to task failure by offsetting the reductions in Ca2+ pumping rate and Ca2+ sensitivity [3− and/or NO2− can modulate skeletal muscle contractility via changes in skeletal muscle Ca2+ handling, the findings from Whitfield et al. [3− supplementation in human skeletal muscle is linked to increased content of Ca2+-handling proteins. Specifically, these authors observed an increased force production and rate of force production during evoked isometric twitches in healthy humans without changes in skeletal muscle CASQ, DHPR or SERCA protein content following 7 days of BR supplementation.Using a mouse model, Hernández et al.  demonstrl muscle . There iical PO2 , or loweical PO2 , during sitivity . While td et al.  challeng3− supplementation is the post-translational modification of the skeletal muscle contractile or Ca2+-handling proteins [−) to form RSNO groups in a reversible process termed S-nitrosylation [2+ release could contribute to enhanced muscle contractility following NO3− supplementation [2+-handling proteins. An interesting observation by Flanagan et al. [3− supplementation despite no change in weightlifting exercise performance. However, other studies have not observed changes in EMG after NO3− supplementation [3− supplementation alters neural drive. Further research is required to evaluate how NO3− supplementation can modulate excitation–contraction coupling in human skeletal muscle.Another mechanism that could improve skeletal muscle contractile function after NOproteins . Indeed,sylation . S-nitrosylation . For exasylation . The potsylation , troponisylation , SERCA [sylation  and ryansylation ,144, andsylation . In addisylation , which sentation . Importan et al.  was thatentation ,103 and,3− supplementation has been reported to alter high-energy phosphate turnover and phosphorus metabolites in human skeletal muscle [3− supplementation has been reported to lower the high-energy phosphate cost of skeletal muscle contractile force production [3− supplementation has also been shown to increase muscle blood flow [In addition to potential changes to excitation–contraction coupling proteins, NOl muscle ,94. Specoduction ,94 and toduction , factorsoduction . Dietaryood flow , which mood flow ,108 and ood flow ,105,106.3− supplementation can improve skeletal muscle contractile function and might enhance weightlifting exercise performance in humans. Therefore, NO3− supplementation holds promise as an effective nutritional ergogenic aid for weightlifting exercise. The potential candidate mechanisms for improved weightlifting exercise performance after NO3− supplementation include enhanced excitation–contraction coupling, via modulation of Ca2+-handling and contractile proteins [3− supplementation. Furthermore, while NO3− supplementation appears to potentially enhance resistance training quality, it is unclear if this will translate into improved weightlifting training adaptations. Notably, although NO3− supplementation has been reported to enhance the adaptations to sprint interval training [2− has been reported to activate AMPK [3− supplementation impacts chronic adaptations to weightlifting exercise training.Taken together, the existing evidence suggests that NOproteins ,135,139;proteins ,94; and proteins . Howevertraining ,147, thetraining ,149. Forate AMPK , which iate AMPK ,149. The3− on various aspects of exercise performance, the number of high-quality studies  focused on weightlifting exercise is limited, which restricted the sample analyzed in the present systematic review. In addition, existing between-study differences in the supplementation dosage (from 32.5 mg NO3− to 6.4 mmol NO3−) and the period of supplementation (from acute to chronic over 6 days), along with differences regarding the type of exercise selected, prevented a firm conclusion on the ergogenic potential of NO3− supplementation on weightlifting exercise performance at this stage. Nevertheless, this systematic review is an important contribution to the literature as it highlights both the potential promise of NO3− supplementation as an ergogenc aid for weightlifting exercise performance and the necessity to conduct further studies to improve understanding on this topic.Although there are numerous studies analyzing the effect of NO3− supplementation holds promise as a nutritional intervention to enhance weightlifting performance in resistance-trained males. Indeed, NO3− supplementation can improve muscular power production, velocity of contraction, and the number of repetitions to failure during weightlifting exercise. Given the important athletic and clinical implications of improved weightlifting exercise performance, NO3− supplementation might offer potential as an ergogenic and therapeutic nutritional aid. The mechanistic bases responsible for the potential ergogenic effect of NO3− supplementation on weightlifting exercise performance may be linked to improvements in skeletal muscle excitation–contraction coupling, high-energy phosphate metabalism and perfusion. However, further research is required to resolve the putative underlying mechanisms for, and the conditions in which, NO3− supplementation might enhance weightlifting exercise performance, as well as its effects on chronic adaptations to weightlifting exercise training.In conclusion, the limited exisiting literature suggests that acute and short-term dietary NO


Supplemental Digital Content is available in the text In France, one in eight patients with acute ST-segment elevation myocardial infarction (STEMI) is admitted direct to an emergency department (ED) in a hospital without percutaneous coronary intervention (PCI) facilities. Guidelines recommend transfer to a PCI center, with a door-in to door-out (DI–DO) time of ≤30 min. We report DI–DO times and identify the main factors affecting them.RESURCOR is a French Northern Alps registry of patients with STEMI of <12 h duration. We focused on patients admitted direct, without prehospital medical care, to EDs in 19 non-PCI centers from 2012 to 2014. We divided DI–DO time into diagnostic time  and logistical time .P = .017) or transfer by air ambulance (P = .004); shorter distance from referring ED to PCI center (P < .001); shorter time from symptom onset to ED admission (P = .002); thrombolysis (P = .006); and extended myocardial infarction (P = .007).Among 2007 patients, 240 were admitted direct to EDs in non-PCI centers; 57.9% were treated with primary angioplasty and 32.9% received thrombolysis. Median (interquartile range) DI–DO time was 92.5 (67–143) min, with a diagnostic time of 41 (23–74) min and a logistical time of 47.5 (32–69) min. Five patients (2.1%) had a DI–DO time ≤30 min. Five variables were independently associated with a shorter DI–DO time: local transfer (mobile intensive care unit [MICU] team available at referring ED) (In view of longer-than-recommended DI–DO times, efforts are required to promote urgent local transfer and use of thrombolysis. Early reperfusion  has been shown to significantly improve both short- and long-term outcomes in meta-analyses.,7Acute ST-segment elevation myocardial infarction (STEMI) is a major cardiovascular emergency.–10 However, one-third of STEMI patients are admitted direct to an emergency department (ED) , and some of these hospitals lack PCI facilities.,11 The European Society of Cardiology (ESC) recommends urgent transfer to a PCI center, with a door-in to door-out (DI–DO) time ≤30 min . Thrombolysis must be used if reperfusion is delayed by >120 min.In France, contacting the emergency call center gives dedicated access to the prehospital Emergency Medical Service (EMS). When STEMI is diagnosed by the EMS, patients receive prehospital care in MICUs en route to centers with percutaneous coronary intervention (PCI) facilities. This immediate activation of reperfusion pathways reduces treatment delays and patient mortality.–19 in France, where prehospital organization is different, DI–DO times have not yet been published. We sought to report DI–DO times in a French registry and identify the main factors affecting them.Although US and Canadian studies have reported DI–DO times,2 with a reported accuracy of 84%. The registry listed initial characteristics  for all STEMI patients; the data were recorded by the treating physician. All patients provided oral consent. Patients were enrolled prospectively; additional data were recorded retrospectively from the STEMI QUAL project, which was approved by the French administrative regulatory body . The STEMI QUAL project was supported by a grant from the French Ministry of Health (PREPS 2014 14–0040). The registry area encompasses three main French administrative departments in the French Northern Alps, with 19 EDs without PCI facilities and five PCI centers ”.We analyzed data from patients with STEMI of duration <12 h (acute STEMI) enrolled in RESURCOR—an ongoing registry in the French Northern Alps started in 2002,We analyzed data from patients with acute STEMI who were admitted direct to 19 EDs of non-PCI centers (January 2012 to December 2014) and were subsequently transferred to a PCI center. These patients arrived at the ED by personal transport, regular ambulance, or with the fire service. We excluded patients who received prehospital medical care in a MICU, patients hospitalized for another condition and who subsequently developed STEMI, patients admitted direct to an intensive care unit, and patients who had an early cardiac arrest before ED admission. For this analysis, the MICU team  was distinguished from the ambulance (means of transport); the possibility of transfer from a non-PCI center to a PCI center depended on the availability of a MICU team and ambulance at each ED . Six air ambulances were available.DI–DO time was defined as the time from arrival at, to discharge from, the initial ED in the non-PCI center (the “referring ED”). DI–DO time was divided into “diagnostic time” (from admission to the ED to transfer decision) and “logistical time” (from transfer decision to discharge from the ED). The transfer decision time was taken as the time of the physician's call to the emergency call center. The transfer was then organized by the emergency call center. The transfer was “local” if the MICU team was available at the referring ED and “non-local” if they were from another center.2.1t test or the Wilcoxon test were used to compare continuous variables; the chi-square test was used to compare categorical variables. Identification of DI–DO time explanatory variables was done using multivariable analysis (linear regression) . All tests were two-tailed; P < .05 was considered statistically significant. Statistical analyses were performed using R software, version 3.2.2 .Continuous variables are expressed as median (interquartile range [IQR]) and categorical variables as count (percentage). Student's 3Among 2007 STEMI patients enrolled in the RESURCOR registry during 2012–2014, 425 (21.2%) were admitted direct to an ED, of which 240 (56.5%) were admitted to an ED without PCI facilities . Most of the remainder received prehospital care by a MICU team, that is, followed the EMS pathway (70.6%). The characteristics of patients admitted direct to an ED in a non-PCI center were generally similar to those admitted to an ED in a PCI center , 97 underwent a non-local transfer to a PCI center (40.4%), and 34 were transferred by air ambulance (14.2%) .3.2Only five patients (2.1%) had a DI–DO time of ≤30 min. Median (IQR) DI–DO time was 92.5 (67–143) min, divided into a diagnostic time of 41 (23–74) min and a logistical time of 47.5 (32–69) min.P < .001); transfer by air ambulance (P = .004) or local transfer (P = .017); shorter time from symptom onset to ED admission (P = .002); use of thrombolysis (P = .006); and extended myocardial infarction (P = .007). Supplemental Digital Content 6, contains details of all factors, whether associated or not.Five major factors were associated with a shorter DI–DO time Fig. : shorterP < .001 and P = .001, respectively) . A shorter time from symptom onset to ED admission decreased the DI–DO time by decreasing diagnostic time (P = .013), with a linear link. The use of thrombolysis decreased DI–DO time by reducing the diagnostic time (P = .001). A diagnosis of extended myocardial infarction decreased the DI–DO time by reducing diagnostic time (P = .01).Local transfer or transfer by air ambulance reduced the DI–DO time by improving logistical time  also exceeded those in published studies from the United States and Canada, which ranged from 51 to 74 min.–17 In two other US studies, median DI–DO times were reduced—from 83 to 68 min and from 97 to 58 min by making various changes to their protocols.Only 5/240 (2.1%) of our non-PCI center patients had a DI–DO time within the recommended ≤30 min.Among our non-PCI center patients, 57.9% underwent primary angioplasty, 32.4% within 120 min from first medical contact (admission to the ED) and the start of PCI . This is considerably less than in a 2009 Canadian study, where 66.6% of STEMI patients benefited from primary angioplasty, 92% within the recommended 120 min.Although our rate of thrombolysis was high, it may not have been enough in our mountainous region. Only 43.0% of thrombolysed STEMI patients had signs of reperfusion on arrival at the PCI center. Therefore, plans to transfer patients to a PCI hospital for potential rescue angioplasty should start as soon as thrombolysis is initiated, as an urgent transfer.P < .001). Similarly, US studies have reported that rural location significantly increased DI–DO times and that awaiting transport at the referring hospital was a common reason for delay. Transfer of the patient using an ambulance and MICU team provided by the local center is a critical point of care management to decrease transfer delays. However, less than half of the transfers were local. A shorter time from symptom onset to ED admission was also associated with reduced DI–DO (P = .002). Similar results have been reported in two studies from Canada.,13 Other studies have reported that various factors are linked with shorter DI–DO times, including emergency transport to the ED,,13,15 younger age,,15,17 and male sex.,14,15The type of transfer was one of the most influential factors on DI–DO time. Patients who underwent local transfer had significantly shorter logistical times than those who underwent non-local transfer . Also, we did not analyse clinical severity or instability, which can affect the time spent in the ED. The sample size also precluded us from calculating odds ratios for patients with DI–DO ≤30 min. The findings may not be entirely representative of patients with acute STEMI, due to the logistical difficulties related to the geographic location. These findings may not be generalizable to other countries due to the specificities of the French EMS system.5In our regional French registry, 12.0% of patients with acute STEMI did not follow the dedicated prehospital pathway, and were thus admitted direct to an ED without PCI facilities. DI–DO times were long, with both diagnostic and logistical times exceeding 30 min, and only 2.1% of patients had a DI–DO time within the recommended ≤30 min. As nearly three quarters of patients with STEMI benefited from primary angioplasty beyond the recommended 120 min, we should aim for shorter DI–DO times and faster transfers, using an urgent local transfer (or the nearest MICU if there is no dedicated team at the site), or promote thrombolysis for eligible patients. However, current recommendations of DI–DO ≤30 min as a goal may not be realistic in everyday clinical practice.The authors would like to thank the RENAU network for the practical implementation of this registry and statistical analysis.Conceptualization: Sandrine Clot, Thomas Rocher, Vincent Descotes-Genon, Loic Belle.Data curation: Sandrine Clot, Thomas Rocher, Mathieu Cardine, Mohamed Lotfi, Julien Turk, Pascal Usseglio, Vincent Descotes-Genon, Gerald Vanzetto, Dominique Savary, Guillaume Debaty, Loic Belle.Formal analysis: Claire Morvan.Funding acquisition: Sandrine Clot, Loic Belle.Methodology: Sandrine Clot, Thomas Rocher, Loic Belle, Claire Morvan.Project administration: Loic Belle.Resources: Claire Morvan, Sandrine Clot, Thomas Rocher.Software: Claire Morvan.Supervision: Loic Belle.Writing – original draft: Sandrine Clot, Thomas Rocher.Writing – review & editing: Sandrine Clot, Thomas Rocher, Mathieu Cardine, Mohamed Lotfi, Julien Turk, Pascal Usseglio, Vincent Descotes-Genon, Gerald Vanzetto, Dominique Savary, Guillaume Debaty, Loic Belle.


Candidatus Regiella insecticola” strain TUt, a facultative bacterial symbiont of the pea aphid Acyrthosiphon pisum, was analyzed. We determined a 2.5-Mb draft genome consisting of 14 contigs; this will contribute to the understanding of the symbiont, which underpins various ecologically adaptive traits of the host insect.The genome of “ Candidatus Regiella insecticola” strain TUt, a facultative bacterial symbiont of the pea aphid Acyrthosiphon pisum, was analyzed. We determined a 2.5-Mb draft genome consisting of 14 contigs; this will contribute to the understanding of the symbiont, which underpins various ecologically adaptive traits of the host insect.The genome of “ Candidatus Regiella insecticola” is a bacterial clade of facultative symbionts that is associated with the pea aphid Acyrthosiphon pisum and other aphid species and belongs to the Enterobacteriaceae family of gammaproteobacteria  and insufficient sequence depth (1× to 6×). A sequence homology search indicated that these contigs were derived from contaminating DNA or repeated sequences, and no plasmid sequence was observed. The remaining 14 contigs represent the genome of “Ca. Regiella insecticola” strain TUt, which consisted of 25,316 reads with 10× coverage. The total size of the contigs  was almost equivalent to the size of the draft genome sequence of “Ca. Regiella insecticola” strain LSR1  , 11 rRNA genes, 43 tRNA genes, and 408 pseudogenes.Here, we analyzed the genome of “t insect , 12. We d strain , 12. Surd strain , 14. We d strain  and 75 rla genes . The remla genes . Assemblf 40.1%) . Strain f 40.1%) , while if 40.1%)  and Prokf 40.1%) . The annf 40.1%) . In the Ca. Regiella insecticola” have been analyzed for another A. pisum-associated strain, LSR1  under accession no. BLXO01000001 to BLXO01000014 (draft genome) and DRA009060 (raw sequence reads).The genome sequence of “


Scientific Reports 10.1038/s41598-020-73297-5, published online 07 October 2020Correction to: The Acknowledgements section in this Article is incomplete.“We thank all children and their families and the three schools Maristas, San Antonio and San Pablo CeI for their help and support, specially to Mari Carmen Blaya, Jose Ignacio Peña and María José Martínez.”should read:“We thank all children and their families and the three schools Maristas, San Antonio and San Pablo CeI for their help and support, specially to Mari Carmen Blaya, Jose Ignacio Peña and María José Martínez. This work has been supported in part by The Spanish Government of Investigation, Development and Innovation (SAF2017-84135-R) including FEDER co-funding. The Autonomous Community of the Region of Murcia through the Seneca Foundation (20795/PI/18) and NIDDK R01DK105072 granted to M. Garaulet”


A comprehensive intuition of the systemic lupus erythematosus (SLE), as a complex and multifactorial disease, is a biological challenge. Dealing with this challenge needs employing sophisticated bioinformatics algorithms to discover the unknown aspects. This study aimed to underscore key molecular characteristics of SLE pathogenesis, which may serve as effective targets for therapeutic intervention.n = 6), generated by three platforms, which included SLE patients (n = 220) and healthy control samples (n = 135) were collected. Across each platform, we integrated the datasets by cross-platform normalization (CPN). Subsequently, through BNrich method, the structures of Bayesian networks (BNs) were extracted from KEGG-indexed SLE, TCR, and BCR signaling pathways; the values of the node (gene) and edge (intergenic relationships) parameters were estimated within each integrated datasets. Parameters with the FDR < 0.05 were considered significant. Finally, a mixture model was performed to decipher the signaling pathway alterations in the SLE patients compared to healthy controls.In the present study, the human peripheral blood mononuclear cell (PBMC) microarray datasets , (2) autoantigen presentation by MHCII , and (3) end-organ damage . As a remarkable finding, we demonstrated significant perturbation in CD80 and CD86 to CD28, CD40LG to CD40, C1QA and C1R to C2, and C1S to C4A edges. Moreover, we not only replicated previous studies regarding alterations of subnetworks involved in TCR and BCR signaling pathways  but also distinguished several significant edges between genes (PPP3 to NFATC gene families). Our findings unprecedentedly showed that different parameter values assign to the same node based on the pathway topology .In the SLE signaling pathway, we identified the dysregulation of several nodes involved in the (1) clearance mechanism (Applying the BNrich as a hybridized network construction method, we highlight under-appreciated systemic alterations of SLE, TCR, and BCR signaling pathways in SLE. Consequently, having such a systems biology approach opens new insights into the context of multifactorial disorders. Called as “the Great Imposter,” systemic lupus erythematosus (SLE) is a chronic and complex autoimmune disease with multisystem manifestations , 2. The Certain lines of evidence suggested that a break in central B cell tolerance results in autoreactive clones to reach the periphery , which fTowards a better intuition of molecular mechanisms involved in multifactorial immune-related diseases especially SLE, lots of researchers acknowledge the gene network approach which helps to prioritize main driver genes and pathways, to propose a better drug development . Bayesiacross-platform normalization (CPN) (1) increases sample sizes and improves gene signature selection [As a method that is widely used to integrate gene expression data, election –23, (2) election , 23. On election .With systems biology approach, we aimed to illustrate the superiority of using CPN and mixture model method and the BNrich to better understand new aspects of underlying molecular mechanisms in the pathogenesis of complex diseases such as SLE. Here, we concentrated on SLE, BCR, and TCR signaling pathways, which are among the most enriched pathways in SLE, to highlight significant alterations of those pathways in SLE patients compared to healthy controls. Besides the altered gene expression level, we demonstrated several significant intergenic relationships which can be proposed as effective targets for therapeutic intervention in SLE patients.n = 6) associated with SLE were downloaded and integrated by CPN method. Subsequently, we employed SLE, TCR, and BCR signaling pathways as structures of BNs and we determined significant node and edge parameters by BNrich method in each dataset independently. Consequently, significant parameters of all datasets were merged through mixture model method and key driver parameters in the selected pathways were defined  microarray datasets  samples, published or updated in 2010–2019, were downloaded from the Gene Expression Omnibus (GEO) database: GSE 17755 , GSE 123normalizeQuantiles function from R package limma [Reduce function in R to integrate each paired gene expression data emanated from the same platform. Finally, we have three major datasets from Affymetrix, Illumina, and Hitachisoft; each platform has a SLE patient group and a HC group. Afterwards, the empirical Bayes method (ComBat) from the R package sva was used for batch effect removal [Each expression dataset was preprocessed and normalized by ge limma . Then, wge limma  with Red removal .To reconstruct BN structures, the SLE (hsa:05322), TCR (hsa:04660), and BCR (hsa:04662) signaling pathways were implemented. All of the pathways were extracted directly from the KEGG database  .Y gene has X1, X2…Xp − 1 parents in the network, and Y for the SLE patient and HC datasets, respectively, they can be modeled as follows:In the parameter estimate step, the mean value of the expression for each gene (node) can be modeled as a linear regression of its parents’ (upstream) gene expression . When Y β(s/h)0 demonstrates the expression level of Y gene, known as a node parameter. The coefficients of X1, X2…Xp − 1 are the parameters of the edge in the trained BNs which reveal a biological relationship between two genes, Y and X. For example, the value of βs3 is the edge parameter between Y and X3 genes in the network structure related to SLE patient data and shows the value of the biological relationship between these two genes. The εc and εh are the residual values.In the SLE- and HC-associated trained BNs, the constant coefficient βs) with HCs (βh) by independent t test [βj)∗ which is the significant parameter based on false discovery rate (FDRj < 0.05) for Affymetrix (A), Illumina (I), and Hitachisoft (H) datasets.Using Eq. t t test  and gainThe estimated parameters of BNs have the following distribution :2\documAccordingly, the significant parameters had been distributed as follows:βA)∗∗∗ which are βf, have a normal distribution, and their mean and variance were defined by:By using the mixture model approach , we mergNA = 166, NI = 95, and NH = 94 were the number of subjects in Affymetrix, Illumina, and Hitachisoft. Since the range of the variances across final parameters is varied, their mean is divided by the standard deviation to achieve comparable parameters. Hence, we plotted the figures based on the following definition:where β∗(j) in each dataset was not significant, its mean and variance were considered to be zero in calculating the corresponding final parameter. When the βf was associated with the node parameters, the sign of μ∗(βf) indicated upregulation (+) or downregulation (−) of the gene and the absolute value of μ∗(βf) was used to measure the value of activation or inhibition. Where the βf was associated with the edge parameters, the sign of μ∗(βf) indicated increasing (+) or decreasing (−) biological function and the absolute value of μ∗(βf) was used to measure the value of these increased or decreased functions.If the parameter After downloading the six mentioned datasets Table , to reduWe did not observe any genes lost after performing CPN on Illumina and Hitachisoft related data, while in the Affymetrix platform, information of about 5.4% and 8.5% of genes in the datasets GSE 50772 and GSE 121239 were lost correspondingly. Considering the CPN benefits, we pursue of using all 166 samples of Affymetrix platform simultaneously instead of analyzing 81 or 85 individual samples  were determined using BNrich method. Table According to our results, 34 significant nodes and the C1QB → C2 edge were common among all datasets in the SLE pathway. Across datasets of all platforms, 53 nodes and 26 edges of the TCR signaling pathway and 36 nodes and 30 edges of the BCR signaling pathway were common. All significant parameters of different platforms, based on three examined signaling pathways, are presented in Additional file μ∗(βf) in all nodes and edges are represented in Table To distinguish the altered signaling pathway parameters in SLE patients compared to HCs, the identified significant parameters of all dataset merged by the mixture model method Eqs.  and 5, tμ∗(βf) in each desired pathway are illustrated in Table μ∗(βf) represented in Additional file The three nodes and edges that have the extreme values of TRIM21, H2AX, C1QA, C1QB, H2BC12, and H2BC12 were mostly upregulated. However, SNRPD3, SSB, MACROH2A2, GRIN2B, and C4B were mostly downregulated. In antigen presentation process mediated by MHCII, nodes such as HLA-DPB1, HLA-DMA, CD80, IL10, and TNF and three edges, CD80 and CD86 to CD28 and DD40LG to CD40, were found to be dysregulated.Among genes involved in the clearance mechanism of SLE pathway, FCGR1A, CTSG, ELANE, FCGR3B, FCGR3B, and C1QA were increased and the expression of C7, C3, C4B, C2, and C4A genes were decreased. Particularly, in that functional part of SLE pathway, we also found dysregulation of several important edges like C1R, C1S, and C1QB to C2  and 64 common dysregulated edges  between these two pathways  showed the significant reduction , which included three sections: (1) autoantigen clearance mechanism, (2) antigen presentation mediated by MHCII, and (3) tissue injury and end-organ damage. The uncleaned apoptotic components which recognized by both the innate and the adaptive immune systems can trigger the pathogenesis of SLE. Therefore, the dysregulation of effective genes involved in the clearance of apoptotic components has a fundamental role in SLE –43. In t22, whichn in SLE –46. In ln in SLE . The cleng agent . Despitee in SLE . Althougt of SLE . Our resnature.”  In a recCBLB, PPP3CC, NFKB1, CD3E, and ZAP70. Recently, a study by Matsuo et al. revealed that a genetic mutation in ZAP70 resulted in the TCR signaling defect which followed by T follicular helper (Tfh) cell development and the manifestation of lupus-like systemic autoimmunity [ZAP70 as a hub gene of the TCR signaling pathway implicated in SLE pathogenesis. Interestingly, the perturbation in TCR culminated to enhanced expression of CDK4, IL2, and IL10 genes. In the BCR signaling pathway, the most up- and downregulated genes were BCL10 and INPP5D, respectively. Furthermore, we also identified significant lower expression of CD79A and CD79B . In a meta-analysis of GWAS results, Julià et al. reported BCR signaling pathway as the most significant biological process and BCL10 and CD79A among top single markers associated with SLE [As outlined earlier, the fundamental importance of BCR and TCR signaling pathways in SLE pathogenesis has been well established via both clinical and experimental evidence –55. In timmunity . Having with SLE .PI3K/AKT signaling pathway and VAV gene homologs were altered; AKT1, AKT2 and VAV1, VAV2 expression levels were particularly reduced (Figs. AKT in SLE patients, which further support the major role of PI3K/Akt/TSC/mTOR signaling pathway in the disease [MAPK1, MAPK3, and PPP3CA genes. Similarly, the strong association of PPP3CA with SLE-like disease has been described as new [Interestingly, our results addressed the dysregulation of several noteworthy node and edge parameters shared by both TCR and BCR signaling pathways. Among them ed Figs.  and 5. P disease , 59. Accd as new . Moreoved as new .CD80 and CD86 → CD28, C1R and C1QA → C2, and C1S → C4B biological functions, PI3K/AKT signaling pathways, besides JUN gene as therapeutic targets in SLE disease. Moreover, analyzing the RNA-seq datasets could be better for distinguishing expression levels and biological relationships of genes’ isoforms in considered signaling pathways. It is necessary to examine the BNrich among some other autoimmune diseases in order to identify SLE signature genes more precisely.Further investigation will be required to evaluate the efficiency of In the present study, we exploited BNrich to hybridize intergenic relations inferred from signaling pathways and gene expression profiling to improve the insight about SLE as a complex disease. We figured out the significant genes and intergenic relationships and systemic alterations in SLE, TCR, and BCR signaling pathways. Through cross-platform normalization and meta-analysis, large-enough SLE patients were analyzed. Since multiple platforms were utilized, the effects of individual study-specific biases were reduced.Notably, the proposed computational methodology can be applied to a variety of complex diseases to illuminate new mechanistic insights in their pathogenesis.Additional file 1. Batch effect removal results. The box plots and PCA plots related to CPN results for Hitachisoft and Illumina platforms.Additional file 2. Significant and final parameters. The significant parameters of different platforms and the final parameters based on the mixture model in three examined signaling pathways.


Plasmodiophora brassicae Woronin, is one of the most important diseases of oilseed rape (Brassica napus L.). The rapid erosion of monogenic resistance in clubroot-resistant (CR) varieties underscores the need to diversify resistance sources controlling disease severity and traits related to pathogen fitness, such as resting spore production. The genetic control of disease index (DI) and resting spores per plant (RSP) was evaluated in a doubled haploid (DH) population consisting of 114 winter oilseed rape lines, obtained from the cross ‘Aviso’ × ‘Montego,’ inoculated with P. brassicae isolate “eH.” Linkage analysis allowed the identification of three quantitative trait loci (QTLs) controlling DI . A significant decrease in DI was observed when combining effects of the three resistance alleles at these QTLs. Only one QTL, PbBn_rsp_C03, was found to control RSP, reducing resting spore production by 40%. PbBn_rsp_C03 partially overlapped with PbBn_di_C03 in a nucleotide-binding leucine-rich repeat (NLR) gene-containing region. Consideration of both DI and RSP in breeding for clubroot resistance is recommended for the long-term management of this disease.Clubroot, caused by  Plasmodiophora brassicae Woronin, is one of the most important diseases of cruciferous crops worldwide, causing significant yield and quality losses in oilseed rape  and other Brassicas (P. brassicae consists of three main stages: (i) survival in the soil, (ii) root hair infection, and (iii) cortical infection. The pathogen survives in the soil as long-lived resting spores , Crr2, Crr3, Crr5A, CRa, CRb, CRd, CRs, Rcr1, Rcr2, Rcr3, Rcr5, and Rcr9WA (P. brassicae classified as per Rcr4) was found to control resistance to Canadian isolates of the pathogen representing pathotypes 2, 3, 5, 6, and 8, classified as per Rcr8 and Rcr9) controlled resistance to an isolate of pathotype 5X, as defined on the Canadian Clubroot Differential set  controlled resistance to non-pathotyped isolates of P. brassicae    and near strains . A seriee K92-16 . In addie K92-16 , one mode K92-16 , and nine K92-16 . Moreoverassicae , in an arassicae , and in rassicae .Previous studies have included very diverse genetic materials , whose use in breeding programs may be difficult given potential issues such as linkage drag . The chaP. brassicae resting spore dynamics in response to the cropping of CR B. napus cultivars indicated increases in soil inoculum loads  and fitness (measured as the reproductive rate of the pathogen), both of which are generally assumed to be positively correlated ; clubrooP. brassicae resting spore production and root gall size (disease severity), these variables are not necessarily correlated, and resting spore production can be affected by host resistance and environmental factors  and clubroot symptoms (DI) in a segregating DH population from the cross of ‘Aviso’ × ‘Montego,’ two modern winter oilseed rape cultivars partially resistant to clubroot with good agronomic characteristics. Second, it aimed to identify some recombinant lines of potential interest for breeding efforts, i.e., carrying favorable alleles at multiple QTLs to decrease DI and limit resting spore production.P. brassicae selection isolate “eH” (1 on the system of B. rapa subs. pekinensis) cv. Granaat  with four replicates (blocks) were established, with the experimental unit comprising six plants per genotype. Seeds of each genotype were sown in 4-cm-diameter pots (one seed per pot) filled with “Falienor 922016F3” potting mix , which consists of 65% Irish peat, 20% black peat, 15% perlite, and 2% clay (pH = 6.2). The greenhouse was maintained at temperatures between 19 and 22°C under a 16/8 day/night cycle. Plants were fertilized with “Liquoplant FD 134 hiver” nutrient solution  once or twice a week by sub-irrigation. Inoculations were conducted 7 days after sowing, by applying 1 ml of the Disease assessment was conducted 54 dai on a 0–3 scale following n0, n1,n2,n2 +, and n3 represent the number of plants in each severity class and N is the total number of plants evaluated. To confirm the pathotype designation of the isolate “eH” as P1, the isolate was inoculated on the differential hosts of B. napus cv. Nevin (ECD 06), B. napus cv. Wilhelmsburger (ECD 10), and B. napus cv. Brutor]. The Chinese cabbage ECD 05 was also included as a susceptible check in all of the experiments.Where After disease assessment, all roots in the experimental unit were pooled and stored at −20°C until processing. Resting spores in the pooled root samples were quantified by flow cytometry following 4 to 6 × 105 resting spores⋅ml–1. Particle counting was completed for each of the dilution points with the flow cytometer and an injection speed of 20. The measured resting spore concentrations were adjusted with a regression Eq. (2) obtained from the standard curve. The obtained regression curve had an R2 of 95%. Spore levels were expressed as RSP by dividing the total number of resting spores in the suspension by the number of plants in the pooled sample.A standard curve was generated from a suspension of resting spores, produced as described above for inoculum preparation, with the spore concentration estimated by counting in a Malassez cell. This solution was then diluted as needed to generate nine spore suspensions with concentrations ranging from 1 × 10RS is the adjusted resting spore count in the suspension and x is the particle count obtained with the flow cytometer.Where A population of 114 DH lines obtained from the cross of the winter oilseed rape cultivars ‘Aviso’ × ‘Montego’ was chosen for linkage analysis because previous experiments indicated that both parents are partially resistant to clubroot, and therefore the presence of new QTLs controlling the disease was suspected. The parental lines were used as controls in all experiments. The genetic map used for the analysis was described by nlme  observed in the jth year and in the kthblock; μ is the population mean; Gi is the effect of the ith genotype; Yj is the effect of thejth year; Bk(j) is the effect of the kth block nested with the jth year; GYik is the effect of the interaction between the ith genotype and the jth year; and εijk are the residuals. Bk(j) was considered as random.Where Broad sense heritability as defined by H2 is the broad sense heritability, σ2G is the variance of the genotype effect, σ2Gy is the variance of the genotype × year effect, y is the number of years, b is the number of blocks, and σ2e is the residual variance.Where lsmeans package  according to the ANOVA table were retained in the model. LOD values and R2 values for each QTL were also obtained by the fitqtl function. The confidence intervals of the QTLs were estimated with a LOD drop of one unit.Linkage analysis was conducted with the R/qtl package . At firsbzh’  (The frequency distribution for DI  and RSP < 0.001) , resulti7 to 1.4 × 108, with about 54.2% of the lines exhibiting a higher RSP than both parents. Only 30% of the lines produced fewer resting spores than the parental lines , while the remaining 29% had a DI between 50.7 and 98. RSP ranged from 5.6 × 10al lines .7 and 5.9 × 107 resting spores plant–1, respectively) , line 148 (DI 44.8 and 1.2 × 109 resting spores plant–1), line 120 (DI 36.3 and 9.2 × 108 resting spores plant–1), line 74 (DI 37.3 and 9.7 × 108 resting spores plant–1), line 168 (DI 44.8 and 9.3 × 108 resting spores plant–1), line 48 (DI 46.8 and 9.7 × 108 resting spores plant–1), and lines 109 and 39 (DI 48.9 and 9.2 × 108 resting spores plant–1). In contrast, some lines with an intermediate DI had a lower RSP, including lines 76 (DI 20.8 and 5.8 × 107 resting spores plant–1) and 164 (DI 32.8 and 5.7 × 107 resting spores plant–1)  but had significant resting spore production (RSP of 5.6 × 10ctively) . Similarplant–1) .P. brassicae and B. napus, respectively. The second part indicates the trait controlled by the QTL, DI, or RSP, in lowercase letters. The third part indicates the chromosome on which the QTL is located. SIM indicated the presence of two main QTLs controlling DI on chromosomes A04 and C03 . PbBn_di_A02 covered 661.48 kb in a region encoding 101 genes, while PbBn_di_C03 covered 788.420 kb in total, encoding 147 genes. Four genes related to plant resistance and defense were found in PbBn_di_A02 QTL. Two were annotated as hypersensitive-induced response protein (HIR), two as a cyclic nucleotide gated channel, and the fourth as a Mlo-like protein 6. In PbBn_di_C03, six genes related to disease resistance or defense were identified, including five TIR-NBS-LRR class disease resistance proteins and one LRR protein kinase-like protein.Only three of the four identified QTLs could be mapped on the The only QTL related to RSP, PbBn_rsp_C03, covered 1614.43 kb. In this QTL, 346 genes were encountered, of which 10 were related to disease resistance or defense, including five TIR-NBS-LRR class disease resistance proteins, three leucine-rich repeat (LRR) protein kinase-like proteins, one defensin-like protein 203, and one WRKY transcription factor 18. PbBn_rsp_C03 QTL overlapped with PbBn_di_C03 in the region from 28.10 to 29.01 cM . In the common region between PbBn_di_C03 and PbBn_rsp_C03 (4.09–4.50 Mb), three TIR-NBS-LRR class disease resistance proteins were found .8 resting spores plant–1) was 2.5 times greater than the lowest (5.6 × 107 resting spores plant–1). Such variation may be important from an epidemiological perspective, since doubling the number of resting spores released into the soil could result in faster and more significant inoculum increases for future crops.Research on clubroot resistance has focused mainly on disease severity expressed as a DI. Resting spore production in host genotypes has not been examined to the same extent, although this is an important measurement of pathogen fitness, affecting inoculum build up in infested fields. In the ‘Aviso’ × ‘Montego’ population, the highest observed RSP .Our results indicate that RSP is not as strongly correlated with DI (Spearman’s coefficient of 0.65) as might be expected based on the trade-off hypothesis, where within-host multiplication, within-host transmission, and virulence of pathogens are positively correlated traits ; and thu plant–1 . More reLinkage analysis showed that the genetic control of RSP and DI are related, since the QTLs PbBn_di_C03 and PbBn_rsp_C03 co-localized, suggesting that the C03 locus controls both traits. The importance of this genomic region is highlighted by the fact that PbBn_di_C03 explains the highest proportion of the variation for DI. The other loci, PbBn_di_A04 and PbBn_di_A02, only control DI and not RSP. We have reported this type of genetic architecture previously, where all QTLs controlling RSP also control DI, but not all DI QTLs control RSP . Co-locaP. brassicae isolate “eH” in another B. napus DH population , five QTLs controlling DI were located on chromosomes A05, A07, C02, C03, and C09  , and few resistance sources appear to be lines with good agronomic value. In the current study, the parents of the DH population were the cultivars ‘Aviso’ and ‘Montego,’ with the former released by Danisco seeds  in 2000 and the latter released by Limagrain  in 2002. ‘Aviso’ is a variety with good agronomic performance that shows resistance to other diseases such as blackleg (aculans) . These c7 and 1.45 × 108 resting spores plant–1. In contrast, in the ‘Darmor-bzh’ × ‘Yudal’ DH population, the RSP values ranged from 1 × 108 and 5 × 109  class disease resistance proteins, LRR protein kinase-like, defensin-like protein 203, and WRKY transcription factor 18 (CRa and Crr1) that have been molecularly resolved so far  is reduced.The ‘Aviso’ × ‘Montego’ population characterized in this study holds promise for the development of new CR oilseed rape cultivars, since the parental lines were recent varieties with good agronomic traits, and some of the tested lines presented very low DI accompanied by low RSP. While much of the emphasis in clubroot resistance breeding traditionally has been on selection based on low disease severity, resting spore production in plant material should also be considered. While breeding based on DI is critical for producing cultivars that perform well in The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.AB-R conducted some of the laboratory analyses, performed the statistical and linkage analyses, and wrote the manuscript. AL, AG, and MM-D contributed to development of the research concept and design of the study and reviewed multiple versions of the manuscript. AL and MJ designed and directed the execution of the experiments. SG and MJ helped to carry out the clubroot tests. SS provided project guidance and extensively edited the manuscript. All authors contributed to manuscript revision, and read and approved the submitted versionThe 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.


In population ecology, the concept of reproductive potential denotes the most vital indicator of chances to produce and sustain a healthy descendant until his/her reproductive maturity under the best conditions. This concept links quality of life and longevity of an individual with disease susceptibilities encoded by his/her genome. Female reproductive potential has been investigated deeply, widely, and comprehensively in the past, but the male one has not received an equal amount of attention. Therefore, here we focused on the human Y chromosome and found candidate single-nucleotide polymorphism (SNP) markers of male reproductive potential.p <  0.025) and r = 0.98 (p <  0.025), respectively. Next, we found that there are twofold fewer candidate SNP markers decreasing TBP–promoter affinity relative to those increasing it, whereas in the genome-wide norm, SNP-induced damage to TBP–promoter complexes is fourfold more frequent than SNP-induced improvement . This means natural selection against underexpression of these genes. Meanwhile, the numbers of candidate SNP markers of an increase and decrease in male reproductive potential were indistinguishably equal to each other (p <  0.05) as if male self-domestication could have happened, with its experimentally known disruptive natural selection. Because there is still not enough scientific evidence that this could have happened, we discuss the human diseases associated with candidate SNP markers of male reproductive potential that may correspond to domestication-related disorders in pets.Examining in silico  all 1206 unannotated SNPs within 70 bp proximal promoters of all 63 Y-linked genes, we found 261 possible male-reproductive-potential SNP markers that can significantly alter the binding affinity of TATA-binding protein (TBP) for these promoters. Among them, there are candidate SNP markers of spermatogenesis disorders , pediatric cancer  as well as male anxiety damaging family relationships and mother’s and children’s health . First of all, we selectively verified in vitro both absolute and relative values of the analyzed TBP–promoter affinity, whose Pearson’s coefficients of correlation between predicted and measured values were r = 0.84 (significance Overall, our findings seem to support a self-domestication syndrome with disruptive natural selection by male reproductive potential preventing Y-linked underexpression of a protein. In keeping with Royal Chapman’s  and EricTo find out how a person can increase one’s own and offspring’s quality of life and longevity, predictive-preventive personalized participatory medicine  uses theThe cornerstone of this postgenome medicine is the greatest twenty-first-century scientific project “1000 Genomes” , which hBecause a physician’s decision based on a patient’s individual genome affects health, quality of life, and longevity of this patient, only those biomedical SNP markers are suitable for this decision that are clinically proved by a comparison between cohorts of diseased and healthy people. Considering how much time, manual labor, and funding is required, this task actually seems impossible because each of the 10 billion human SNPs  may maniAs for accuracy of annotation, at present, the best one is achieved with SNPs in protein-coding regions of genes ; these SPreviously, we have created Web service SNP_TATA_Comparator  and usedBPY2, BPY2B, BPY2C, CDY1, CDY1B, CDY2B, DAZ1, DAZ2, DAZ3, DAZ4, DDX3Y, EIF1AY, HSFY1, HSFY2, PCDH11Y, PRKY, PRY, PRY2, RBMY1A1, RBMY1B, RBMY1D, RBMY1E, RBMY1F, RBMY1J, RPS4Y1, SRY, TGIF2LY, TSPY1, TSPY3, TSPY9P, and VCY1B) contain 75 unannotated SNPs that were studied here, but none of them was not predicted as a candidate SNP marker (data not shown).Using our public Web service SNP_TATA_Z-tester , we analThe humanSHOXgene encodes short stature homeobox (transcription factor). Figure SHOX gene containing the above-mentioned minor allele.D, nM,” namely: KD values of the equilibrium dissociation constant of complexes of TBP and one of the two entered alleles of the promoter under study and their standard errors in nanomoles per liter (nM). Additionally, this table shows a change (Δ) in gene expression and its Fisher’s Z-score with statistical significance α, as described elsewhere  as described within the subsection “In vitro verification” of the main section “Methods.” Figure As shown in this figure, TBP–DNA affinity decreased from 39 ± 7 nM for the synthetic oligonucleotide (ODN) identical to the wild-type allele of SNP rs1452787381 being tested  that corresponds to ZBED1 underexpression, whereas 11 SNPs  cause its overexpression, as calculated here  contains 13 SNPs  that can elevate the expression of this gene, as shown in Table SAKAP17A overexpression, our PubMed keyword search retrieved transcriptome data on azoospermia caused by testicular degeneration in Klinefelter syndrome , 100 μg/mL BSA, 0.01% of NP-40, and 5% of glycerol) at a fixed concentration (2 nM) of active TBP. The gels were dried, and Imaging Screen-K  was exposed to these gels for analysis on a Molecular Imager PharosFX Plus phosphorimager . The resulting autoradiographs were quantitated in the Quantity One 4.5.0 software (Bio-Rad) as displayed in Fig. graphpad-prism.software.informer.com/5.01), we calculated the equilibrium dissociation constant (KD). For details of our protocol for in vitro measurements of the equilibrium dissociation constant for TBP–ODN complexes, see ref. [The equilibrium dissociation constants .Additional file 1: Supplementary Results. Tables S1-S4. Candidate SNP markers of male reproductive potential within the protein-coding genes on the human Y chromosome.Additional file 2. Supplementary Method. A sequence-based statistical estimate of the SNP-caused alteration in the affinity of TATA box–binding protein (TBP) for 70 bp proximal promoters of a human gene containing an SNP under study.Additional file 3. Supplementary Keyword Search. Figure S. A flow chart of the keyword search for male reproductive potential components whose physiological markers correspond to alterations in the expression of human Y-linked protein-coding genes containing a given SNP under study.Additional file 4. Supplementary Electropherograms. The original, raw, unfiltered, uncropped, and unedited electropherograms used for Fig.


The camp resulted in a lower sleep duration , but an improved global sleep behavior score . During the camp, the performance support athletes received was not different to pre-lockdown (p > 0.05), but there was greater sports massage  and physiotherapy usage  (both p < 0.05). The adverse effects of lockdown were restored during the camp. A “quarantine” camp may offer comparable training experiences to pre-lockdown training, without inducing additional perceived stress. Coaches and sporting organizations may consider this approach as part of a virus mitigation strategy, whilst maintaining sport-specific training.Globally, COVID-19-related movement restrictions have caused significant disruption to athlete's training and sporting competitions. “Quarantine” camps are one approach to maintain sport-specific training, whilst minimizing the risk of COVID-19 transmission between athletes and society. This cross-sectional study investigated the effects of a “quarantine” training camp on athlete's routines and wellbeing, performance support, perceived stress and sleep behaviors. A survey was completed at the end of a 30-day “quarantine” camp, by 76 elite athletes (17–46 years), predominantly (~80%) Olympic/Paralympic and/or world championship representatives. Athletes described their experiences in comparison to; pre-lockdown training and/or training during “lockdown” (immediately prior to the “quarantine” camp). Compared to “lockdown,” the “quarantine” camp revealed improvements ( In light of the pandemic of coronavirus disease 2019 (COVID-19), the world's sporting calendar has experienced considerable disruption. A notable example is the postponement of the Tokyo Olympics 2020, with further postponements or cancellations affecting the World Athletics Indoor Championships, UEFA European Football Championship and Badminton Thomas Cup. Travel restrictions and closures of sporting facilities have halted the regular training practices of athletes. Many athletes have been unable to perform sport-specific training, due to movement restrictions  remained in place. This “quarantine” camp allowed coaches and athletes to return to sport-specific training, use regular training facilities and receive performance support from sports science/medical staff within the camp. It also allows athletes to focus on the quality of training, without having to worry about training facilities and nutritious food, less travel time to the training venue and external “distractions” , all of which will provide a conducive training environment. However, a “quarantine” camp necessitates stringent working and living procedures to maintain a minimized risk of virus transmission. Notably, athletes and staff must be tested upon entry and may not leave the camp throughout the duration of the camp. Therefore, despite the apparent benefit of resuming sport-specific training, a “quarantine” camp may also elicit a psychological strain on athletes, who are restricted in their movements and away from family/friends  and “normal” training conditions . We hypothesized that a “quarantine” camp would improve athlete's perceptions of training routines and wellbeing, access to performance support, stress and sleep, compared to the prior “lockdown” and “lockdown” period.A cross-sectional design was adopted, using questionnaires to assess athlete's perceptions of “before” lockdown, “during” lockdown, and “during” the “quarantine” camp. All data were recorded within a 5-day period following the completion of the 30-day camp and collected using a custom-made Google Form. All athletes attended the camp for at least 23 of the 30-day duration. Data collection occurred across three training venues within the country, all of which were subjected to the same operating procedures/restrictions and requiring COVID-19 testing upon entry and exit.n = 5), badminton (n = 16), diving (n = 10), gymnastics (n = 4), sailing (n = 9), swimming (3) and nine Paralympic sports; para archery (n = 4), para athletics (n = 3), para badminton (n = 6), boccia (n = 2), para cycling (n = 7), para powerlifting (n = 2), para swimming (n = 1), para table tennis (n = 2), and wheelchair tennis (n = 2). The total number of participants from able-bodied sports was 47 and 29 athletes were from Paralympic sports. Participants with intellectual disabilities were not recruited. The study was conducted according to the Declaration of Helsinki. Informed consent was obtained from all athletes, with data was processed anonymously. Ethical approval was not sought as these questionnaires were comparable to those they would routinely provide as part of their official duties as national athletes . All participants had between 8 and 22 years of competitive experience in their sport (15.5 ± 6.5 years). Of these athletes, ~80% have already qualified for the 2020 Olympics or have previously competed at the Olympics or World championships . Participants were from six able-bodied sports; archery  independent proofreading by two bilingual translators; (c) discussion and confirmation of the translation's accuracy by these two bilingual translators. A parallel comparison with the original English questionnaires was included for each question.The survey contained 6 sections; (i) athlete background and camp ratings, (ii) training routines and wellbeing, (iii) access to sport science support, (iv) perceived stress, (v) sleep behaviors, and (vi) lifestyle. Approximately 15–20 min was required to complete the full survey. To improve the response rate, the completion of parts 5 and 6 were optional, but 1–4 was compulsory. The original survey in English was translated into Malay to facilitate data collection among non-English speaking athletes. A three-step procedure was used to obtain the translation: (a) translation via negatively, in the following aspects.” Athletes were asked to respond to the statement on a 5-point Likert scale, with 1 and 5 representing strongly disagree and strongly agree ratings, respectively, 1-point increments. The scores of each question in Section 2 were subsequently reversed to obtain “positive scores,” to aid interpretation and reporting of these data . In Section 3, the utilization of performance support  was assessed using 10 custom-designed questions. The questions are related to training monitoring, recovery practices, mental wellbeing techniques and nutrition monitoring. Athletes indicated how frequently, on a monthly basis, they utilized performance support on a 5-point Likert scale of “0 or never,” “1–2 times,” “3–4 times,” “5–6 times,” and “more than 6 times.” Section 4 utilized the Perceived Stress Scale (PSS) , for duplication checking and identification of missing data. Normal distribution of data were determined using the Shapiro-Wilks test. This revealed the data not to be normally distributed, therefore non-parametric analysis was adopted. Friedman's ANOVA was used to identify differences across the three time-points; pre-lockdown, lockdown and “quarantine” camp, with Wilcoxon signed rank test used as a post-hoc to identify where differences occurred. For parts of the survey where only two time-points of data were collected , Wilcoxon signed rank test was again adopted to identify within-subject differences. Cohen's d effect sizes were calculated, with the following interpretation boundaries; <0.2 , 0.2 , 0.5 (moderate), and 0.8 (large) with the relevant citation  was observed for 7 out of 9 questions. Post hoc analysis found differences between lockdown (p < 0.05) compared to “normal” training, but these aspects were not different compared to pre-lockdown and the “quarantine” camp (p < 0.05). Differences between pre-lockdown and lockdown were identified for; access to gym facilities for strength training , access to sport-specific training , access to recovery services , food and meal choices , mental health , emotional health  and training motivation . Sleep quality and quantity did not change throughout the assessed periods .The effects of the “quarantine” camp on training routines and athlete wellbeing (Section 2) are shown in d = 0.39) and physiotherapy  usage increased during the “quarantine” camp (p < 0.05). The remaining aspects of performance support were not statistically different between pre-lockdown and “quarantine” camp.2 = 11.006, p = 0.004, Post hoc analysis indicated differences between pre-lockdown and lockdown , between pre-lockdown and “quarantine”  and between lockdown and “quarantine” . The PSS scores for pre-lockdown, lockdown, and the “quarantine” camp were 17.8 ± 4.3 pt, 18.6 ± 4.6 pt, and 17.2 ± 4.9 pt, respectively. The results indicate increased stress level during lockdown  which reduced during the “quarantine” camp  compared to pre-lockdown. Neither age, sex, nor competitive experience were correlated with the PSS total score for pre-lockdown, lockdown, and “quarantine” camp .A difference in the average PSS total score (Section 4) was observed between pre-lockdown, lockdown and “quarantine” camp . The mean change was from 8:08 ± 1:27 during lockdown, to 7:39 ± 0:44 h during the “quarantine” camp. There was a difference in the ABSQ global score between lockdown and the “quarantine” camp . A higher score during lockdown (37.0 ± 9.1) indicated impaired sleep behaviors during lockdown, relative to “quarantine” camp (34.9 ± 9.7). There were no correlations for age, sex, and competitive experience with the ABSQ scores during lockdown and “quarantine” camp .Sleep duration (Section 5) was higher during lockdown compared to the “quarantine” camp , “use social media” , and “online education” . However, all other aspects of weekly activities were not statistically different.small to moderate effects). However, our findings indicate that these detrimental effects were reversed during the “quarantine” camp . The “quarantine” camps allowed athletes to complete sport-specific training with other athletes, supported by coaches and performance support staff. Compared with lockdown, athlete's training routines, performance support, and perceived stress improved during the camp, along with sleep behaviors. These outcomes confirmed our hypotheses that the negative effects associated with COVID-19 pandemic lockdown would be reversed during a “quarantine” training camp.The current study is the first to report athlete perceptions during a “quarantine” training camp. During lockdown, we observed negative effects on athlete's access to training and recovery facilities, increased mental and emotional stress, fewer nutritional choices, reduced training motivation and increased perceived stress . This finding concurs with Pillay et al.  as a result of home confinement. Such negative states may be induced by nervousness, preoccupation and apprehension caused by an individual's perception of the situation  and emotional health (−12.8%), and training motivation (−18.7%) . This indicates that athlete's sleeping habits were not impacted on by the camp environment, whilst athletes could not sleep in their own bed.Sleep quality and quantity appeared unaffected during lockdown, compared to pre-lockdown. Sleep quantity during lockdown (>8 h) was higher than during “quarantine” camp (<8 h), although both remain within the recommended duration for athletes  Watson, . Similartrivial effect), , . Our dateffect),  and Italeffect), . The paneffect), . Event ceffect), . Howevereffect), . Individeffect), . In contElite athletes are routinely surrounded by a team of professionals dedicated to maximizing performance , for which we cannot demonstrate validity and reliability. Nevertheless, these questions were generic and we believe unlikely to be misinterpreted and as such, enhances our understanding of athlete's experiences within the camp.To sum up, we observed a variety of negative effects in elite athletes that are associated with home confinement. However, these effects were not maintained during the “quarantine” camp, which appeared to improve aspects of athlete's routines and wellbeing, perceived stress, performance support and sleep behavior. The outcomes of the current study have important implications for policy makers, governing bodies, coaches, sports scientists considering implementing “quarantine” camps in the months leading up to the delayed Tokyo 2020 Olympic Games.A “quarantine” training camp offers one option for athletes to maintain “normal” training practices, with access to coaching, sports science and medical staff, whilst minimizing the risk of COVID-19 transmission with wider society.Following a period of home confinement, athletes may utilize greater physiotherapy/masseur support than usual to assist with the training and recovery process.Caution should be taken when interpreting these findings in the context of planning camps that exceed a duration of 30 days.The raw data supporting the conclusions of this article will be made available by the corresponding author, upon reasonable request.The questionnaires were comparable to those the participants would routinely provide as part of their official duties as national athletes. This study was conducted in accordance with high ethical standards, and approved retrospectively by the institutional review board of Institut Sukan Negara (004/2020-005/2020). The participants provided their written informed consent to participate in the study.JW: original planning, overall contribution, study conceptual, study planning, data collection, data collation, data analysis, data interpretation, manuscript (first draft), manuscript editing, and critical revision of manuscript. SM: original planning, content contribution, study planning, data collection, manuscript (first draft), and manuscript editing. PL and CC: content contribution, study planning, data collection, manuscript (first draft), and manuscript editing. CJ: overall contribution, study conceptual, study planning, data collection, data interpretation, manuscript (first draft), manuscript editing, and critical revision of manuscript. All authors contributed to the article and approved the submitted version.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.


Elastin Microfibril Interfacer 2, in a cohort of gastric cancer patients. We discovered that EMILIN2 expression was consistently suppressed in gastric cancer and high expression levels of this glycoprotein were linked to abnormal vascular density. Furthermore, we found that EMILIN2 had a dual effect on gastric carcinoma cells: on one hand, it decreased tumor cell proliferation by triggering apoptosis, and on the other hand, it evoked the production of a number of cytokines involved in angiogenesis and inflammation, such as IL-8. Collectively, our findings posit EMILIN2 as an important onco-regulator exerting pleiotropic effects on the gastric cancer microenvironment.Gastric cancer is a frequent human tumor and often a lethal disease. Targeted therapy for gastric carcinomas is far behind vis-à-vis other solid tumors, primarily because of the paucity of cancer-driving mutations that could be efficiently and specifically targeted by current therapy. Thus, there is a need to discover actionable pathways/proteins and new diagnostic and prognostic biomarkers. In this study, we explored the role of the extracellular matrix glycoprotein EMILIN2,  •EMILIN2 is localized in the gastric lamina propria and its expression is down-regulated in gastric cancer.•High levels of EMILIN2 associate with elevated vascular density.•EMILIN2 impairs the proliferation of gastric cancer cells by evoking apoptosis.•Surprisingly, EMILIN2 triggers the expression of pro-angiogenic and pro-inflammatory cytokines. Gastric cancer (GC) is a relative common disease worldwide and the third leading cause of cancer-related deaths . Most ofRecent mutational analyses have prompted the employment of targeted drugs in patients displaying hyperactivation of epidermal growth factor receptor (EGFR), mTOR or c-Met. Unfortunately, clinical trials have shown that the GC patients do not benefit from these types of treatment . On the Another important target is angiogenesis, the development of new blood vessels from pre-existing vasculature. Angiogenesis is switched on during cancer development since vascularization is required to support tumor cell growth through the supply of oxygen and nutrients ,42. AntiAn increasing body of evidences highlights the crosstalk between angiogenesis and the immune response ,58, and,P < 0.001, We have previously shown abnormal expression of EMILIN2 in a relatively small cohort of GC patients . To corrNext, we verified if the expression of EMILIN2 would associate with the clinical GC stage (I-II or III-IV), histotype  or anatomical location  of the tumor. We found no significant correlation with any of these parameters C–E.P = 0.0116, Since we have recently demonstrated that EMILIN2 triggers angiogenesis by evoking IL-8 expression in endothelial cells and fibroblasts , we asseHaving established a correlation between EMILIN2 expression and GC, we hypothesized that EMILIN2 could directly impact the behavior of GC cells. To this end, we employed the GC cell lines AGS originally isolated from an adenocarcinoma of the stomach  and MKN74β1 and α9β1, the putative integrins engaged by EMILIN2, as assessed by FACS analyses .Next, we determined whether EMILIN2 could act as an adhesive substrate for AGS and MKN74 cells. To perform these studies, we used recombinant EMILIN2 protein purified in our laboratory . We employed recombinant fibronectin, collagen type I and IV, and laminin as positive controls. Notably, AGS and MKN74 cells did not adhere to EMILIN2 D. OverlaTo further dissect the role of the molecule in gastric cancer cell behavior, we challenged AGS and MKN74 cells with EMILIN2.P < 0.001, P < 0.01 and P < 0.03, respectively, Notably, EMILIN2 significantly impaired the proliferation of AGS and MKN74 cells  and increased that of insulin growth factor-binding protein 2 (IGFBP2) . Notably, EMILIN2 strongly elicited the expression of serine protease inhibitor1 (SERPINE1), vascular endothelial growth factor A (VEGFA) C. IntereCollectively, our findings demonstrate that EMILIN2 is a key component of the gastric parenchyma and tumor microenvironment, and that its loss may profoundly affect the outcome of GC patients acting at different levels of gastric cancer development and progression.In this study, we analyzed the expression of the ECM glycoprotein EMILIN2 in the gastric mucosa and gastric carcinomas from a cohort of well documented patients with GC. We found that EMILIN2 is expressed primarily and uniformly in the lamina propria of the gastric mucosa, although the expression levels are variable among patients. Based on the fact that EMILIN2 can have a dual effect on gastric cancer, i.e., it can have direct effects on tumor cells as well impinging on microenvironmental cues, we hypothesize that the expression of this important secreted glycoprotein could affect not only tumor onset but also tumor progression. Indeed, our findings show a strong correlation between EMILIN2 expression and blood vessel density, highlighting the prominent role of EMILIN2 in GC vascularization.We demonstrate that EMILIN2, despite not representing a substrate for GC cell adhesion, directly affects their viability. It is likely that the lower proliferation rate of AGS and MKN74 cells observed in the presence of EMILIN2 is due to the increased percentage of apoptotic cells observed under treatment with the soluble protein. In fact, in previous studies carried out using another cell model, we demonstrated that EMILIN2 can trigger tumor cell apoptosis by activating the extrinsic apoptotic pathway receptors DR4 and DR5 . AGS celWe further report for the first time that the direct action of EMILIN2 on GC cells could be the consequence of on an indirect impact on the tumor microenvironment. Our data point to EMILIN2 playing a role in the crosstalk among cancer, endothelial and inflammatory cells. This concept is supported and corroborated by an increase in cytokines in the gastric adenocarcinoma cells exposed to EMILIN2. Several of these cytokines are involved in tumor angiogenesis. For example, we find a marked increase in VEGFA, a master regulator of angiogenesis . InteresWe previously discovered that EMILIN2 induces IL-8 expression in endothelial cells and fibroblasts present in the microenvironment . In thisIncreased IL-8 expression was recently demonstrated to occur following the down-regulation of protein tyrosine phosphatase receptor delta (PTPRD) and to associate to increased angiogenesis and metastasis in GC . DespiteIn addition, IL-8 is also produced by mast cells, whose density is increased in GC and correlates with the tumor vascularization, the number of metastatic lymph nodes and the patients' survival . As to wThus, the multiple effects that EMILIN2 exerts in the tumor microenvironment are complex and the outcome is a balance between positive and negative regulators of tumor growth. It must be also pointed out that the intratumoral vessels developed in the presence of EMILIN2 are particularly inefficient and leaky . This maIn this perspective EMILIN2 may represent a key molecule regulating the interplay between angiogenesis and inflammation which represents a major field of investigation . Taken tFor this study, 19 patients with locally advanced gastric cancer were consecutively enrolled. The methodologies conformed to the standards set by the Declaration of Helsinki. This study was approved by the Institutional Board of the CRO-IRCCS, National Cancer Institute of Aviano (PN), Italy (IRB no. CRO-2014-03). The clinical evaluation is shown in 2 and verified to be free of mycoplasma contamination using the MycoAlert™ Mycoplasma Detection kit (LONZA). AGS cells were cultured in F-12 medium  containing 10% fetal bovine serum . Caco-2 and 293-EBNA cells were cultured in Dulbecco's modified eagle's medium (DMEM) with 10% fetal bovine serum ; for 293-EBNA cells 250 μg/ml G418 were added. The MNK74 cell line was cultured in RPMI 1640 medium with 10% fetal bovine serum . The polyclonal anti-EMILIN2 antibody was produced in our laboratories as previously described [The AGS, MNK74, 293-EBNA (E293), and Caco-2 cell lines were obtained from ATCC , maintained at 37° C under a humidified atmosphere containing 5% COescribed ,89. The escribed ,91. The w/v) of 1,4-diazabicyclo--octane (DABCO). Images were acquired with a Leica TCS SP8 Confocal system , using the Leica Confocal Software (LCS). Fluorescence intensity and quantification was evaluated by means of the Volocity software .For immunofluorescence analyses on tumor samples, serial cryostatic sections (7 μm) were collected on positively charged slides (BDH Superfrost Plus), air dried at room temperature (RT) and fixed with PFA for 15 min. After washing with phosphate-buffered saline (PBS), the slices were incubated with 0.5% Triton X-100 in PBS for 5 min at RT, saturated with 1% BSA, 10% normal goat serum (DAKO) in PBS for 1 h at RT, and stained ON at 4 °C with the appropriate antibodies. Next, the samples were washed with PBS and incubated with the appropriate secondary antibodies and TO-PRO3 to stain the nuclei for 1 h at RT. After washing with PBS, the slides were mounted in Mowiol containing 2.5% ; isotype control PE, anti-α4 PE and anti- α9β1 PE (Biolegend). Cells were analyzed using FACS LSR Fortessa (BD Biosciences) and data were analyzed using DIVA software (BD Biosciences).Cell were detached with 5 mM EDTA in PBS and stained with the following antibodies: isotype control APC, anti-αg for 5 min at 37 °C to synchronize the contact of the cells with the substrate, incubated for 20 min at 37 °C and subsequently mounted together with a similar miniplate (top unit) to create communicating chamber for a reverse centrifugation at 50 ×g. The relative number of cells bound to the substrate (i.e. remaining in the wells of the bottom miniplates) was estimated by top/bottom fluorescence detection in a computer-interfaced Infinite 1000 PRO microplate reader .The quantitative cell adhesion assay used in this study is based on centrifugation and has been extensively described . Six-welUpon treatment with 5 μg/ml of recombinant EMILIN2 or PBS for 24 h, or, in alternative, with conditioned media from mock or EMILIN-2-transfected E293 cells (EMILIN2 concentration ~5 μg/ml), total RNA was isolated from the cell lines with Trizol and reverse transcribed using AMV-RT . Semi-quantitative endpoint reactions were performed with GoTaq DNA polymerase  and Real-time PCRs with iQ™ SYBR® Green Supermix . The oligonucleotide sequences were: MCP-1 forward: ‘CAGAAGTGGGTTCAGGATTCC’, reverse: ‘ATTCTTGGGTTGTGGAGTGAG’; TIMP-1 forward: ‘CCTGCACCTGTGTCCCAC’, reverse: ‘TCTGGTTGACTTCTGGTGTCC’; Angiogenin forward: ‘AGTCAATTTTCCGTCGTCCG’, reverse: ‘AACAAAAGGTCCAGGTAGCTC’; CXCL-16 forward: ‘CCCATGGGTTCAGGAATTG’, reverse: ‘GGGGGCTGGTAGGAAGTAAA’; TSP1 forward: ‘CTCCCCTATGCTATCACAACG’, reverse: ‘AGGAACTGTGGCATTGGAG’; VEGFA forward: ‘AGTCCAACATCACCATGCAG’, reverse: ‘TTCCCTTTCCTCGAACTGATTT’; SERPINE1 forward: ‘CCAGCTGACAACAGGAGGAG’, reverse: ‘CTCCTTGTACAGATGCCGGA’; IGFBP2 forward: ‘ACATCCCCAACTGTGACAAG’, reverse: ‘ATCAGCTTCCCGGTGTTG’; IL-8 forward: ‘CATTGACCAAGGAAATCGGC’, reverse: ‘CACAGAGATAGTTACAGCCATACC’; GAPDH forward: ‘GAGAGACCCTCACTGCTG’, reverse: ‘GATGGTACATGACAAGGTGC’.cells were lysed in cold RIPA buffer  containing a protease inhibitor cocktail . Proteins were resolved in 4% to 20% Criterion Precast Gels  and transferred onto Hybond-ECL nitrocellulose membranes, blocked with 5% dry milk in TBS-T buffer, probed with the appropriate antibodies, and developed using enhanced chemiluminescence  or the Odyssey Infrared Imaging System .E293 cells were transfected using FuGene6 reagent . Cells transfected with the pCEP-Pu-EMILIN2 or the empty vector were then selected with 250 μg/ml G418 and 0.5 μg/ml puromycin. Confluent cells were then incubated in serum-free medium for 48 h and the conditioned medium collected. For Western immunoblotting, 2. For adhesion experiments, the E-plates 96 were precoated over night at 4 °C with the indicated molecules (10 μg/ml), and cells were then seeded at 50,000 cells/well in FCS-free medium. Cells were monitored once every 5 min for 2 h. Data analysis was performed using Real-Time Cell Analyzer software (version 1.2) supplied with the instrument.To quantitatively monitor cell behavior in real-time, we adopted the Real-Time Cell Analyzer dual plate instrument xCELLigence (Roche) which measures the electrical impedance caused by cell attachment and proliferation and expressed as the cell index, an arbitrary measurement defined as (Rn − Rb)/15, in which Rb is the background impedance of the well measured with medium alone, and Rn is the impedance of the well measured at any time (t) in the presence of cells. Thus, the cell index is a reflection of overall cell number, attachment quality, and cell morphology that change as a function of time. The Real-Time Cell Analyzer dual plate instrument was placed in a humidified incubator maintained at 37 °C 5% CO4 AGS and MKN74 cells were plated in 96 well plates, let adhere and incubated with 5 μg/ml of recombinant EMILIN2 or PBS for 24 h. In alternative, cells were treated with conditioned media from mock or EMILIN-2-transfected E293 cells (EMILIN2 concentration ~5 μg/ml). Next, cells were incubated for 3 h with 5 mg/ml MTT and absorbance detected at 560 nm. The results were reported as % of cell proliferation. The apoptotic rate was determined using the In Situ Cell Death Detection Kit, Fluorescein .10t-test for the comparisons between two groups. For all the evaluations reported in the manuscript the investigators were blinded. All the measurements were included for the statistical analyses and differences were considered statistically significant when P ≤ 0.05.Statistical analyses were performed with the SigmaPlot software and the values represent the mean ± SD. The statistical significance of the differences was determined by the two-sided Student's EA, AF, AC and EPo analysis of data, conducting and designing experiments, data interpretation, writing and editing of the manuscript.RC, MF, SM obtaining clinical samples.EPi conducting and designing experiments, editing of the manuscript.RP, AF conducting experiments.RD obtaining cryostat sections.MM and PS experimental supervision, data interpretation, editing of the manuscript.RVI critical revision and editing of the manuscript;MM writing of the manuscript.The collection of human materials was approved by the appropriate Institutional Board of the CRO-IRCCS of Aviano, Italy (IRB no. CRO-2014-03).The authors declare no competing interest in the design or interpretation of the experiments, or in the writing of the manuscript.


We characterized these phenolic/DDSQ hybrids using Fourier transform infrared spectroscopy; 1H, 13C, and 29Si nuclear magnetic resonance spectroscopy; X-ray photoelectron spectroscopy (XPS); and thermogravimetric analysis. The thermal decomposition temperature and char yield both increased significantly upon increasing the DDSQ content, with the DDSQ units providing an inorganic protection layer on the phenolic surface, as confirmed through XPS analyses. We obtained carbon/DDSQ hybrids from the phenolic/DDSQ hybrids after thermal curing and calcination at 900 °C; these carbon/DDSQ hybrids displayed electrochemical properties superior to those of previously reported counterparts.In this study we incorporated various amounts of a double-decker silsesquioxane (DDSQ) into phenolic/DDSQ hybrids, which we prepared from a bifunctionalized phenolic DDSQ derivative (DDSQ-4OH), phenol, and CH Compared with inorganic materials, organic polymers usually possess lower moduli and poorer thermal stabilities and mechanical properties. The incorporation of inorganic nanoparticles (NPs) nanotubes, or nanosheets , grap2O to obtain various phenolic/POSS hybrids, which also exhibited high thermal stability and low surface energies [Phenolic resin is a typical thermosetting resin that has many applications  because of its structural integrity and solvent-resistance. Nevertheless, phenolic resin has been neglected as a material for use in polymer nanocomposites because it has a three-dimensional structure even when it is not crosslinked. Several approaches have been developed, however, to overcome this problem to enhance thermal stability ,23,24,25Unfortunately, these phenolic/POSS nanocomposites prepared using multi-functionalized POSS NPs have led to insoluble cross-linked phenolic resins ,23,24,252O under basic conditions , charcoal, acetonitrile, ND, paraformaldehyde (CH2O), platinum divinyltetramethyldisiloxane complex (Pt (dvs)), 4-aminophenol, and phenol were purchased from Sigma–Aldrich . DDSQ-4OH  and 6.80 (k) ppm, confirming the formation of DDSQ-4OH. The 1H NMR spectrum of the pure PDDSQ (2 units: ArCH2Ar at 1.83 ppm (peak a), methylene ether (PhCH2OR) at 3.73 ppm (peak b), and methylol (OCH2OH) (peak c) at 4.82 ppm. Furthermore, the intensity of the signal of the aromatic protons from the 4-aminophenol unit at 6.24 ppm (l) had decreased significantly  and C=O (peak b) units appeared at 157.3 and 178.8 ppm, confirming the formation of DDSQ-4OH. In the spectrum of the pure PDDSQ (CH2Ar (peak c) units and at 68.1 ppm for the methylene ether (peak d) and methylol (peak e) units, along with the other signals from the DDSQ-4OH moieties, confirming the synthesis of the pure PDDSQ.DDSQ-4OH a featurere PDDSQ b, additi−1, corresponding to the Si–O–Si and C–O units, as well as a signal at 1265 cm−1 representing the Si–CH3 groups. The spectrum of the pure PDDSQ did not feature any obvious additional peaks when compared with that of DDSQ-4OH, with only the intensity of the signal representing the OH groups having increased in intensity after the reaction of CH2O to form the pure PDDSQ. 29Si NMR spectra of DDSQ-4OH and the pure PDDSQ. The spectrum of DDSQ-4OH featured three major peaks at –79.05, –64.17, and –21.34 ppm representing the SiO3R (T3), SiO2R2 (T2), and Si-C units, respectively (−1) was determined through GPC analysis, with a polydispersity index of 1.10  between the signals for the free and hydrogen-bonded OH groups can give a rough estimation of the average hydrogen bonding strength [ν = 95 cm−1) appeared to be weaker than that for the self-associative OH···OH bonding of the pure phenolic resin (∆ν = 150 cm−1). This result is consistent with our previous finding that the inter-association equilibrium constant of phenolic/POSS (KA = 38.7) is smaller than the self-association equilibrium constant of pure phenolic (KB = 52.3), based on the Painter–Coleman association model [1H spectra of pure phenolic resin, PDDSQ-50 and pure PDDSQ recorded at room temperature. The peak assignment is similar with Having successfully synthesized the pure PDDSQ, we prepared PDDSQ hybrids incorporating various amounts of DDSQ, as displayed in 02/A1510 . Furtherstrength . The aveon model . Further2 bridges, the second step at ca. 450 °C is due the broken of crosslinking network structure, and the third at ca. 500 oC comes from the C–OH units in phenol were broken and then form H2 gas. Td10) and char yields at 800 °C of these PDDSQ hybrids. The char yields and values of Td increased significantly upon increasing the DDSQ concentration, as expected. For example, the char yield and value of Td increased by 12.2 wt% and 79 °C, respectively, for the PDDSQ-80 hybrid when compared with those of the pure phenolic resin. This behavior can be explained by considering the nano-reinforcement effect of incorporating inorganic DDSQ NPs into a phenolic matrix. The DDSQ units were presumably dispersed well within the phenolic resin at the nanoscale, stabilized through hydrogen bonding and covalent bonding, and, thus, they could enhance its initial decomposition temperature. Furthermore, the DDSQ units could retard the thermal motion of the polymer through the covalent bonding, and could also increase the chain spacing, giving rise to the lower thermal conductivity. It has been proposed that DDSQ units prefer to be oriented toward the air side in related nanostructures, because the low surface free energy of the siloxane units would screen out the polar functional units  [Although we had detected only weak intermolecular interactions between the phenolic OH groups and the DDSQ siloxane units, these interactions did still enhance the thermal stability of the PDDSQ hybrids, as determined using thermal gravimetric analyzer . The TGA trace  of the prethane) ,39. As a2 atmosphere. −1 for the C=O groups and at 1432 cm−1 for the aliphatic CH2 units from the ND moieties both disappeared, and new peaks appeared at 1385 and 1612 cm−1 representing the carbon structure. Most importantly, the signals at 1090 cm−1 for the Si–O–Si units and at 1265 cm−1 for the Si–CH3 units were still present after thermal heating at 900 °C, suggesting that the DDSQ units remained dispersed within the carbon matrix in the carbon/DDSQ hybrids. Raman spectroscopy  revealed the local graphitic structure of this carbon/DDSQ hybrid , suggesting the presence of oxidized graphene after thermal treatment at 900 °C (1592 cm−1) or 1000 °C (1620 cm−1). Furthermore, the intensity ratio ID/IG can be used to roughly characterize the degree of graphitization. The ID/IG ratio (1.35 after treatment at 900 °C) decreased significantly to 1.01 after thermal treatment at 1000 °C, indicating a highly defected carbon structure containing Si, O, and N atoms in the former system, as confirmed by the Transmission electron microscope  images in q of 16.52 nm−1 (d = 0.38 nm) was due to the (002) plane of the carbon structure  [We used cyclic voltammetry (CV) and the galvanostatic charge/discharge (GCD) method in 1 M KOH as the aqueous electrolyte in a three-electrode system to investigate the electrochemical performance of our PDDSQ hybrids after thermal treatment at 900 °C. 5 mV s−1 d–f. The 5 mV s−1 d had a r5 mV s−1 f had a qs−1 (−1) ,46. A sis−1 (−1) . Furthers−1 (−1) ,49.−1. The GCD curves of the carbon material derived from pure phenolic were triangular in shape, suggesting EDLC characteristics, while those of the carbon materials derived from PDDSQ-50 and pure PDDSQ were triangular with a slight bend, implying the presence of both pseudocapacitance and EDLC [and EDLC . The dis−1, the specific capacitance of the carbon material derived from pure PDDSQ (689.5 F g−1) was much larger than those of the carbon materials derived from PDDQ-50 (258.8 F g−1) and pure phenolic (89.9 F g−1). The excellent performance of the carbon material derived from pure PDDSQ was due to its many electroactive centers  making it easier for the electrolyte to access the electrode surface [−1, all of the carbon/DDSQ hybrids displayed high cycling stability, retaining over 90% of their original capacitances (−1, it displayed close to 99.9% retention; thus, by adjusting the ratios of phenol and DDSQ-4OH during the copolymerization reaction we could balance the electrode materials to obtain both a large specific capacitance and high stability. We used Equation S1 to calculate the specific capacitances from the GCD curves . At a cucitances b. Althou−1 at a power density of 337.5 W kg−1; its energy density was maintained at 49.5 W h kg−1 even at a high power density of 13.5 kW kg−1. Compared with other materials prepared in previous studies, our present carbon/DDSQ hybrids displayed excellent properties and were much simpler to prepare, suggesting that they have potential for use as future electrode materials [aterials ,59,60,61We have synthesized PDDSQ hybrids incorporating various amounts of DDSQ and confirmed their chemical structures using FTIR and NMR spectroscopy. All of these PDDSQ hybrids had char yields and thermal stabilities higher than those of the pure phenolic resin, and they increased upon increasing the DDSQ content in the hybrid systems. The DDSQ units provided an inorganic protection layer on the phenolic surface, as confirmed through XPS analysis. We obtained carbon/DDSQ hybrids after thermal curing and calcination of our PDDSQ hybrids; these carbon materials displayed electrochemical properties superior to those of previously reported counterparts.