To investigate the dynamical brain network changes before and after an absence seizure episode in absence epilepsy (AE).21 AE patients with a current high frequency of seizures and 21 sex- and age-matched health control (HC) who reported no history of neurological or psychiatric disorders and visited the hospital for routine physical examinations were included. Each included subject underwent a 2-h and 19-channel video EEG examination. For AE patients, five epochs of 10-s EEG data in inter-ictal, pre-ictal, and post-ictal states were collected. For the HC group, five 10-s resting-state EEG epochs were extracted. Functional independent components analysis (ICA) was carried out using the LORETA KEY tool.Compared with the resting-state EEG data of the HC group, the EEG data from AE patients during inter-ictal periods showed decreased alpha oscillations in regions involving the superior frontal gyrus (SFG) (BA11). From inter-ictal to pre-ictal, SFG (BA10) showed maximum decreased delta oscillations. Additionally, from pre-ictal to post-ictal, superior temporal gyrus (STG) (BA 22) presented maximum increased neural activity in the alpha band. Moreover, compared with inter-ictal EEG, post-ictal EEG showed significantly decreased theta activity in SFG (BA8).The changes in SFG alpha oscillations are the key brain network differences between inter-ictal EEG of AE patients and resting-state EEG of HCs. The brain networks of EEG oscillatory during peri-ictal episodes are mainly involving SFG and STG. Our study suggests that altered EEG brain networks dynamics exist between inter-ictal EEG of AE patients and resting-state EEG of HCs and between pre- and post-ictal EEG in AE patients. Childhood absence epilepsy (AE) accounts for 10–17% of all cases of childhood-onset epilepsy, making it the most common type of epilepsy in childhood . The cliOver the past decades, advances in neuroimaging and EEG techniques have led to a better understanding of the brain networks involved in AE. Some studies have found that the characteristic generalized spike-and-wave discharges are associated with “activations” and “deactivations” within a network of cortical or subcortical brain regions , 7. InteA widely used brain network analysis method, independent component analysis (ICA) is a classical electroencephalogram (EEG) signal analysis technique, which separates scalp electrical signals into their additive independent or source components , 11. AllIn the present study, we hypothesized that the occurrence and termination of absence episodes are accompanied by the change of brain functional networks. We aimed to investigate whether and how brain functional networks changed under three conditions, including inter-ictal, pre-ictal, and post-ictal states, in AE patients.A total of 21 patients with typical childhood AE were recruited from January 2016 until January 2019 in the neurology department of the Affiliated Hospital of Southwest Medical University. Inclusion criteria: (a) clinical diagnosis of childhood AE was established according to the International League against Epilepsy (ILAE) for a revised classification of epilepsy and epileptic syndromes ; (b) EEGSubjects washed and dried their hair before EEG data collection. The scalp EEG was acquired in a quiet, relaxed state in a semi-isolated room.For both patients and controls, video EEG was recorded from a nineteen-channel analog recorder (GALIEO EB Neuro with a camera) placed according to the 10–20 system for 2 h. EEG and video data were recorded in a time-locked pattern. The impedances of the electrodes were kept at <10 kΩ. The sample rate was 256 Hz for all the recordings.At the EEG monitoring start, the participants were told to keep awake, remain relaxed, keep their eyes closed and not to think of anything in particular for resting-state EEG data acquisition. Then the EEG-video recording lasted for up to 2 h. Following the EEG recording, the EEG patterns were evaluated by two reviewers. Reference montages and average montages were used as well as bipolar montages to assess epileptiform discharges and epileptic seizures. Those patients with frequent typical generalized ca. 3-Hz spike-and-wave complexes were included. The absence seizures were defined as generalized ca. 3-Hz spike-and-wave complexes discharges lasting more than 5 and <15 s. According to previous study (http://germangh.com). Those EEG matrices with low fractal dimensions would be recognized as artifact components and be removed automatically. Then, the EEG was recomputed to the common average reference. After preprocessing, the EEG of interest was cut out and translated to text format for further ICA analysis. For each subject in AE patients, we extracted five 10 s epoch EEG data for inter-ictal, pre-ictal (pre-seizure), and post-ictal (post-seizure) states. Five 10 s epoch EEG data in the resting state with eyes closed and for each subject in controls. inter-ictal EEG was defined as EEG epochs with eyes closed standing in the time points that there was no seizures onset before and after it within 1 min. Data preprocessing was carried out using MATLAB  toolbox EEGlab  format. The EEG was bandpass filtered between 1 and 45 Hz with a Hamming window after the FIR filter. Then the electromyogram artifacts and muscle artifacts were completely automated removed using blind source separation. Both electrooculogram artifact and muscle artifact corrections were carried out using a fast ICA algorithm. These procedures were done using an automated EEGlab plugin-in AAR  functional ICA method (eLORETA-ICA) was employed. eLORETA is a widely used method to localize multiple distributed cortical sources of EEG data in three-dimensional space . The LORZ scores higher than or equal to “3” were considered statistically significant and selected for later comparisons between two conditions.In short, the EEG signals of each subject were transformed to cross-spectral EEG matrix for each frequency band using the discrete Fourier transform. Then, these transformed files were used for computing the spectral density for each cortical gray voxel  and for each frequency band. Thus, we obtained six eLORETA images of cortical spectral density five frequency bands . These data correspond to a “function” of space  and frequency. After that, the data from each subject in two groups is concatenated. Thus, a matrix including two dimensions were created, in which one dimension corresponds to different subjects, and the other dimension corresponds to space-frequency. This process was known as functional data analysis. When independent component analysis is used to this matrix, general networks are found. This method is defined as functional ICA. Each functional network is composed of five images, one for each frequency band . Those cIn present study, to achieve an optimal number of components. We used sphericity test  to compuP < 0.05) in the ANOVA test among the four conditions was the variable considered in further two-by-two comparisons. For group comparisons between AE group and HC group, we used independent sample t-test. To compare the network coefficient in different states within the AE group, we used paired t-test. The null distribution between two conditions was evaluated by Monte-Carlo simulation with a repetition of 1,000,000 iterations. Only when all tests with P < 0.05 were considered statistically significant. A significant difference between two groups or states means they use the same network in a different way.According to ICA, we obtained the coefficient of each network component for each subject. One-way ANOVA was used to conduct statistical analysis on the overall difference in coefficients among groups. Only when one variable showed significant differences  code.Twenty-one AE patients (aged 9.6 ± 2.9 years) and 21 age- and sex-matched healthy subjects (aged 9.7 ± 2.6 years) were included in the study . ArtifacAccording to the sphericity test, we obtained 9 components in each comparison between two groups or conditions. We found that the most striking differences between the inter-ictal EEG of AE patients and resting-state EEG of HC subjects was in component one which showed that AE patients showed decreased alpha oscillations in regions involving superior frontal gyrus (SFG) (BA11) , 2.It was noted that from the pattern of functional networks changed in different state EEG of AE patients. SFG and superior temporal gyrus (STG) were the key brain regions in the changes of functional network pattern in AE patients. We found that in component one, SFG (BA10) showed maximum decreased brain activity in delta band in pre-ictal state, when comparing with inter-ictal state in AE patients. In addition, compared with pre-ictal state, superior temporal gyrus (STG) BA 22) in post-ictal state presented maximum increased neural activity in alpha band, which has been displayed in component three. Moreover, from components six, we noted that compared with inter-ictal EEG, post-ictal EEG in AE patients showed significantly decreased theta activity in SFG (BA8) , 2.2 in postPrevious reports on AE using MEG/EEG methods typically focused on ca. 3 Hz ictal spike-wave discharges , 23. TheIn clinical practice, normal EEG rhythm often been found in inter-ictal epochs of children with AE, similar to resting-state EEG of healthy controls, which means that it was hard to find potential abnormalities in inter-ictal EEG without spike-wave discharges of AE patients using visual inspection. However, when inter-ictal stage EEG from AE patients was compared with resting-state EEG from healthy controls, there is a significance reduction of alpha activity over SFG (BA11) in AE, which indicated that the altered EEG pattern in AE patients, could not be found by visual inspection, could be obtained by ICA. In addition, the alterations in alpha band in AE patients might suggest that they may possibly showed decreased cognitive function, considering that the modulations of alpha oscillations have been found to be involved in various cognitive domain function such as memory and attention –26. In fFrom inter-ictal to pre-ictal to post-ictal states, we observed a dynamic evolution of brain activity. Notably, EEG oscillations in SFG showed a prominent issue during the phase transitions, which is in good agreement with earlier functional MRI Studies , 31. AltThe relationship between absence seizures and the default mode network has been widely detected . The SFGIt is not unexpected that alpha oscillations in STG increased from pre-ictal to post-ictal, which is consistent with clinical observations that the normal background EEG rhythm will recovery after the terminations of absence episodes in AE. Nevertheless, we noted that frequency bands with maximum alterations from inter-ictal to pre-ictal, from post-ictal to inter-ictal were decreased delta oscillations, and increased theta activity, respectively. One reasonable explanation for these findings was that the cortico-thalamo-cortical network contribute to the onset and termination of seizure in AE –40. The Over the past years, EEG dynamics in the pre-seizure state have been widely studied for predicting epileptic seizures –47. ManyThe source assignment method used in this study, eLORETA has been identified showing good concordance with resected zone or intracranial EEG in epilepsy patients . This meStudy limitations include a relatively small sample. The used source localization method eLORETA may neglect some information from deep cortical and subcortical regions during the peri-seizure periods due to its low space resolution. The activity of cortico-thalamo-cortical network in peri-ictal period was not detailed investigated using combined EEG-fMRI to acquire an accurate location. In future research, we will continuously collect cases and overcome these limitations for a more comprehensive understanding of children AE.Our study provides evidences that the differences between inter-ictal EEG of AE patients and resting-state EEGs of healthy controls can be detected by eLORETA. The EEG networks of epileptic patients before and after absence seizures are significantly changed, which may involve the cortico-thalamo-cortical circuits. Overall, our study has advanced our knowledge about the dynamic process of brain networks for peri-ictal states in AE patients, which provide a better understanding of the onset and termination mechanisms of seizures from electrophysiological view.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by the Medical Ethics Committee of the Affiliated Hospital of Southwest Medical University. Written informed consent to participate in this study was provided by the participants' legal guardian/next of kin.All authors made a significant contribution to the work reported, whether that is in the conception, study design, execution, acquisition of data, analysis and interpretation, or in all these areas, took part in drafting, revising or critically reviewing the article, gave final approval of the version to be published, have agreed on the journal to which the article has been submitted, and agree to be accountable for all aspects of the work.This work was supported by the Youth Program of the National Natural Science Foundation of China (81804198) and Youth Fund of Southwest Medical University (2018-ZRQN-003).We thank the Special Training Program for Young Scientific and Technological Talents of Southwest Medical University 2020-2022 for sponsorship of JR.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


IJMS Special Issue “Perception, Transduction and Crosstalk of Auxin and Cytokinin Signals” (2021–2022).Auxins and cytokinins are considered the most important plant hormones, responsible for fundamental traits of the plant organism . These hReconstitution of the Cytokinin Signaling Pathway. Cytokinin signaling proteins are encoded by multigene families; therefore, determining the contribution of each component to a signaling pathway is challenging. This was addressed by researchers from the National Institute of Agricultural Sciences , who succeeded in reconstituting the cytokinin signaling pathway by the transient expression of genes encoding major players in the signal transduction chain in rice protoplasts [toplasts . Therebytoplasts . This netoplasts , fine-tuEvolution of the Cytokinin Perception Apparatus. Although the cytokinin signaling in typical model plants, such as Arabidopsis or rice, has been studied in-depth, the investigation of its evolution remains in its infancy. Hormone perception is based on sensing receptor proteins, so receptors are key components of signaling cascades. Lomin et al. (2021) [Physcomitrium patens, lycophyte Selaginella moellendorffii, and gymnosperm Picea abies. All the receptors or their individual sensory modules were analyzed in terms of their cytokinin-binding affinities, their ligand specificity, and the pH-dependencies of hormone binding. Through this integrative approach, new trends in the evolution of land-plant cytokinin receptors were identified. In particular, moss receptors were shown to strongly prefer isopentenyladenine (iP) over other forms of cytokinin nucleobases, whereas spruce receptors bound trans-zeatin (tZ) with much higher affinity than iP. These trends are consistent with the parallel evolution of active cytokinin forms which, in primitive plants, are mainly represented by iP, whereas in vascular plants starting from gymnosperms, specific enzymes (CYP735-like) for the direct synthesis of tZ from iP-type precursors appear.. (2021) , from thHow Do Cytokinins Get to Their Receptors? One of the recurring problems in cytokinin signaling research is the subcellular distribution of cytokinins between different cellular compartments. It is fundamentally important for any hormone, such as a cytokinin, to have access to a cognate receptor. The main cellular targets of cytokinins are currently thought to be the lumens of the endoplasmic reticulum (ER), where the sensory modules of the receptors are exposed [ exposed ,7. For o exposed , presentEnvironmental Cues Modulate Cytokinin Signaling. Plants’ survival in different climatic zones requires effective adaptation. The restructuring of the cytokinin regulatory system can play an important role in this [cis-zeatin signaling in apices. Since the publication of the article [ in this . A team  in this . They fo article , this scSynthetic Drugs with Unique Cytokinin or Anticytokinin Properties. The development of specialized methods for the analysis of cytokinin–receptor interactions [N6-benzyladenine with individual Arabidopsis receptors was studied. Three synthetic cytokinin S-enantiomers specific for AHK3 receptors were discovered: N6-((S)-α-methylbenzyl)adenine (S-MBA), 2-fluoro-S-MBA (S-FMBA) and 2-chloro-S-MBA (S-CMBA). Two other S-enantiomers were found to exhibit receptor-specific and chirality-dependent anticytokinin properties. These anticytokinins appear to act as allosteric regulators of the receptor activity. ractions ,12 creatractions , the intCytokinins + Auxins: Synergistic Morphogenic Effects. One of the most striking effects of cytokinins, observed almost immediately after the discovery of these hormones in 1955, is the ability, together with auxins, to induce organogenesis in undifferentiated callus tissue [s tissue . A high s tissue . The uniCytokinins + Auxins: Antagonistic Growth Effects. Despite the true synergistic effects in stimulating cell proliferation, auxin and cytokinin most often act as an antagonistic hormone pair [one pair . The crione pair . As a tyCytokinins + Auxins: Adaptation to Photoperiod Stress. Another example of a stress effect under the influence of the auxin–cytokinin duo was described in an article by a joint German–Czech group headed by Prof. Schmülling from the Free University of Berlin (Germany) [ahk2,3 mutants with loss of cytokinin receptor function, or gain-of-function YUC1 auxin synthesis mutants exhibited increased stress responses, while mutations causing the loss of the function of the auxin receptors TIR1, AFB2 and AFB3, in contrast, reduced stress levels. Thus, cytokinin signaling has been shown to play a protective role in this particular type of light stress, whereas auxin signaling enhanced the stress effect. This finding was in good agreement with earlier data on transgenic PHYB potatoes, which showed that cytokinin (kinetin) could reduce plant sensitivity to photoperiod [Germany) . In thistoperiod . Cytokinins + Auxins: Signaling Crosstalk at the Molecular Level. In the aforementioned research on the concerted action of cytokinins and auxins, the main topics were somehow indirectly related to the interplay between auxin and cytokinin signaling. Unlike these studies, the main purpose of the work by Kolachevskaya et al. [a et al.  from thea et al.  point toMicroRNA Fine-Tunes Auxin Signaling. miRNAs are short, single-stranded nucleic acid molecules that repress target gene expression, regulating plant growth and stress responses [TIR1/AFBs, ARF and AUX/IAA; (ii) the modulation of the free auxin content via the indirect regulation of auxin biosynthesis, metabolism and transport. The authors consider the latest studies on the miRNA-mediated regulation of the auxin pathway in the dicot Arabidopsis and the monocot rice, as well as new findings in other species, in order to provide a comprehensive picture of the important roles that miRNAs play in auxin signaling. The combination of certain miRNAs with the genes involved in auxin signaling creates specific regulatory modules that fine-tune development and stress responses in the plant kingdom. Some of these modules function as potent regulatory nodes to integrate auxin, cytokinin and other signaling pathways. Such regulation appears in a wide range of developmental processes in plants. The challenges remaining to be addressed are highlighted. esponses . The genesponses , the miRConcluding remarks. This Special Issue, “Perception, Transduction and Crosstalk of Auxin and Cytokinin Signals”, was aimed at bringing together researchers studying various aspects of the molecular basis of auxin and/or cytokinin perception and signal transduction (sduction . Indeed,


Systemic embolism was associated with mortality, and N-terminal prohormone of B-type natriuretic peptide (NT-proBNP) had predictive power for mortality. Conclusion: If diagnostic standard tools remain inconclusive, we suggest employing novel cut-off values to increase diagnostic accuracy and accelerate diagnosis. Patients with embolism or elevated NT-proBNP deserve a closer follow-up.Background: Diagnosis of infective endocarditis (IE) often is challenging, and mortality is high in such patients. Our goal was to characterize common diagnostic tools to enable a rapid and accurate diagnosis and to correlate these tools with mortality outcomes. Methods: Because of the possibility of including perioperative diagnostics, only surgically treated patients with suspected left-sided IE were included in this retrospective, monocentric study. A clinical committee confirmed the diagnosis of IE. Results: 201 consecutive patients  were finally diagnosed with IE, and 14 patients turned out IE-negative. Preoperative tests with the highest sensitivity for IE were positive blood cultures (89.0%) and transesophageal echocardiography (87.5%). In receiver operating characteristics, vegetation size revealed high predictive power for IE (AUC 0.800,  In our analysis, NT-proBNP had predictive power for the 30-day and the 1-year mortality with values < 3990 pg/mL and <2323 pg/mL favoring survival. Vegetation size failed to do so, although clearly associated with predictive power for the diagnosis of IE. The latter could somehow be surprising at first glance, but it is in line with a large study comparing surgical vs. medical-only therapy in patients with a vegetation size of <10 mm vs. >10 mm: Fosbol et al. showed that in 1006 patients with left-sided IE, medically managed patients with a vegetation size of >10 mm had a lower probability of survival, whereas surgically treated patients were indifferent [In our study, embolic events were relatively common, with an event rate of 70%. In the literature, the overall risk of peripheral embolism is described as between 20–50%, and cerebral embolism occurs in up to 60% of IE patients . A meta-This study recruited retrospectively; thus, not every patient had the entire palette of diagnostic tests. Further, we found that there were 10 HP-negative patients in our collective who nonetheless had positive blood cultures with pathogens that were compatible with an IE. In four of these patients, the duration of preoperative antibiotic treatment was >20 days. Thus, the affected valve may have healed by the time of surgery. An alternative site of infection could be identified in those patients (Supplemental In addition to previous knowledge and utilization of the cut-off value for vegetation size, we could add information that a vegetation size below our calculated cut-off value of 11.5 mm unlikely favors endocarditis. In combination with the cut-off values for inflammatory markers, clinicians have a gain in diagnostic arguments for decision-making. In terms of mortality, the presence of systemic embolism showed a significant association, which argues in favor of early surgical therapy. NT-proBNP, as an indicator of mortality, can help to identify patients at risk in order to recommend and utilize a tighter follow-up scheme.


Cardiovascular disease (CVD) is the leading cause of death in the United States with heart failure (HF) being the highest reason for hospital admission. Despite improved therapies for CVD patients, the 5-year mortality rate after HF hospitalization remains around 40% . Preclinhttps://www.frontiersin.org/research-topics/18185/role-of-molecular-modulators-in-combatting-cardiac-injury-and-disease-prevention-repair-and-regenera#overview), authors identify possible mechanisms for prevention, repair, and cardiac regeneration. Here, we summarize the major findings of interest to the readership and provide a frame of reference for future studies.In the current digest topic , a prevalent pathology with no specific therapy . Recent Rech and Rainer describe emerging evidence of the therapeutic potential for the innate immune DNA sensor cyclic GMP-AMP synthase (cGAS) and stimulator of interferon genes (STING) pathway in CVD. Many of the risk factors associated with CVD including smoking, obesity, and aging are accompanied by alterations in cGAS-STING signaling  is associated with beneficial outcomes such as decreased blood pressure and aortic stiffness, improved hypercholesterolemia, and enhanced cardiac mitochondrial function  is an essential cellular substrate critical for energy production. A decrease in NADnd aging , 14. The02921659 , the autfunction –18. JahaWang et al. thoroughly explored the role of heart stiffness, ECM proteins, and the combination of these factors on the cardiac regenerative response in juvenile mice. The investigators administered β-aminopropionitrile (BAPN) or genipin to alter tissue stiffness before subjecting mice to myocardial infarction (MI) at postnatal day 5. After MI, mice were given decellularized ECM (dECM) derived from either fetal or adult pigs. Consistent with published literature, fetal heart dECM produced pro-regenerative phenotypes including improved ejection fraction, reduced scarring, and increased cardiomyocyte cell cycle activity post-MI. Of particular novelty, the effects of fetal dECM were substantially accentuated when tissue stiffness was decreased by BAPN administration, suggesting an interaction between bioactivity and biomechanics in cardiac repair. Future studies identifying the specific ECM factors mediating biomechanical transduction pathways and cardiac regeneration will pave the way for new therapeutic approaches in post-MI patients.The ECM from neonatal hearts has pro-regenerative properties compared to that of the adult heart –22. DissKisling et al. report for the first time the existence of an intrinsic cardiac dopaminergic system as demonstrated by both D1R and D3R expression in murine cardiac tissue and fibroblasts. Mice with dysfunctional D3R displayed limited fibroblast proliferation and migration, reduced viability, and increased expression of collagen type 3. These phenotypes were recapitulated using a non-ergot pharmacological inhibitor of D3R. While a large body of evidence describes roles for dopamine and its receptors in the neuro and renal-vascular systems , HL145817 (KD-P), HL156022 (CO'M), HL141159 (CO'M), and HL152297 (LdCB), the Biomedical Laboratory Research and Development Service of the Veterans Affairs Office of Research and Development under Award Number BX003922 (KD-P), Advancing a Healthier Wisconsin Endowment (AHW) #5520561 (CO'M), American Heart Association IPA35260039 (KD-P), and the Department of Physiology startup funds from Augusta University (DI).The content is solely the responsibility of the authors and does not necessarily represent the official views of any of the funding agencies.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Background: This study was performed to identify key regulatory network biomarkers including transcription factors (TFs), miRNAs and lncRNAs that may affect the oncogenesis of EBV positive PTCL-U.Methods: GSE34143 dataset was downloaded and analyzed to identify differentially expressed genes (DEGs) between EBV positive PTCL-U and normal samples. Gene ontology and pathway enrichment analyses were performed to illustrate the potential function of the DEGs. Then, key regulators including TFs, miRNAs and lncRNAs involved in EBV positive PTCL-U were identified by constructing TF–mRNA, lncRNA–miRNA–mRNA, and EBV encoded miRNA–mRNA regulatory networks.Results: A total of 96 DEGs were identified between EBV positive PTCL-U and normal tissues, which were related to immune responses, B cell receptor signaling pathway, chemokine activity. Pathway analysis indicated that the DEGs were mainly enriched in cytokine-cytokine receptor interaction and chemokine signaling pathway. Based on the TF network, hub TFs were identified regulate the target DEGs. Afterwards, a ceRNA network was constructed, in which miR-181(a/b/c/d) and lncRNA LINC01744 were found. According to the EBV-related miRNA regulatory network, CXCL10 and CXCL11 were found to be regulated by EBV-miR-BART1-3p and EBV-miR-BHRF1-3, respectively. By integrating the three networks, some key regulators were found and may serve as potential network biomarkers in the regulation of EBV positive PTCL-U.Conclusion: The network-based approach of the present study identified potential biomarkers including transcription factors, miRNAs, lncRNAs and EBV-related miRNAs involved in EBV positive PTCL-U, assisting us in understanding the molecular mechanisms that underlie the carcinogenesis and progression of EBV positive PTCL-U. Peripheral T cell lymphoma, unspecified (PTCL−U) is one of the most common subtypes of mature post-thymic T cell neoplasms, which consists of variable immunophenotypes and morphological changes . PTCL-U Gene expression regulation is one of the key mechanisms for controlling biological processes in the organism . TranscrRecently, bioinformatics analysis as an interdisciplinary field has been regarded as a reliable technique for revealing molecular markers and therapeutic targets that may be implicated in the pathogenesis of cancer or other diseases . In the In this study, some key regulators including TFs, miRNAs and lncRNAs involved in EBV positive PTCL-U were identified and integrated them into a strong theoretical framework through constructing TF–mRNA, lncRNA–miRNA–mRNA, and EBV encoded miRNA–mRNA regulatory networks. The present study may offer new insights into the molecular mechanism of EBV positive PTCL-U and provide potential network biomarkers for EBV positive PTCL-U. The flowchart diagram for this study is represented in The microarray data for GSE34143 in the GEO database were obtained on the GPL6480 and GPL14550 platform. The expression profiles are provided for nine samples, including 3 EBV positive PTCL-U and 6 normal reactive lymph nodes.p-value < 0.05 were considered statistically significant. The analysis results were presented by volcano map.GEO2R is an interactive web tool that allows users to compare two or more sets of samples in the GEO Series and contributes to filtrating DEGs through the Bioconductor’s GEO Query and Limma R software . We usedp-value < 0.05 was considered statistically significant.To further evaluate the function of the DEGs, KEGG pathway and GO enrichment analyses were conducted. The GO functional enrichment analysis was applied to annotate genes and gene products, and also accomplish characteristic biological function originating from genomic or transcript omic data at three different levels including biological process (BP), molecular function (MF), and cellular component (CC) . KEGG haSearch Tool for the Retrieval of Interacting Genes (STRING) is a database resource that can investigate complex PPI information . We retrNetwork analysis was carried out to identify the key hub mRNAs with high degrees in the initial network. Module analysis was applied to screen the significant modules active in the network with the Molecular Complex Detection (MCODE) plugin in Cytoscape . Then, whttp://www.rna-society.org/virbase) is an online tool that provide a resource for efficient browsing and visualization of interaction networks in virus-host non-coding RNAs (ncRNAs) associated interactions  PTCL with normal/reactive tissue is not going to reveal the significance of EBV but that of the neoplastic process itself. Second, the sample size for the rare disease is too small while investigating a larger sample size in EBV positive PTCL-U would help to confirm our data. Third, the data in the current study were identified by bioinformatics analysis, and the findings are warranted to be validated by further investigations. Thus, further biological experiments are needed to explore the biomarker value of the identified network regulators and the underlying mechanisms. Last, we should also pay attention to the quality control of the assay and data generation letter analysis process, and the combination of NGS, GEP, CNA and other multi-omics analysis, which is beneficial to the molecular typing of PTCL-U and the search for new therapeutic targets.In summary, a network-based approach to identify potential biomarkers involved in EBV positive PTCL-U was developed by integrating the regulatory networks, including TF-mRNA, lncRNA-miRNA-mRNA, EBV-encoded miRNA-mRNA. Two transcription factors, including PAX5 and SPIB, were identified that were both associated with the tumorigenesis of EBV positive PTCL-U through regulating their target genes. Four hub mRNAs in this network, including CXCL10, CXCL11, CCL7, and CCL8, were identified which have also been demonstrated to influence the tumor microenvironment and infection status. Four miRNAs  and one unreported lncRNA, LINC01744, were identified in the ceRNA regulation network. In addition, EBV related miRNAs EBV-miR-BART1-3p and EBV-miR-BHRF1-3 were found to regulate CXCL10 and CXCL11 in the network. This study may provide a new avenue for investigating the regulatory mechanisms of EBV positive PTCL-U, and further molecular experiments are required to confirm these findings.


Alcohol use disorder (AUD) is characterized by enhanced cue-reactivity and the opposing control processes being insufficient. The ability to inhibit reactions to alcohol-related cues, alcohol-specific inhibition, is thus crucial to AUD; and trainings strengthening this ability might increase treatment outcome. The present study investigated whether neurophysiological correlates of alcohol-specific inhibition (I) vary with craving, (II) predict drinking outcome in AUD and (III) are modulated by alcohol-specific inhibition training. A total of 45 recently abstinent patients with AUD and 25 controls participated in this study. All participants underwent functional magnetic resonance imaging (fMRI) during a Go-NoGo task with alcohol-related as well as neutral conditions. Patients with AUD additionally participated in a double-blind RCT, where they were randomized to either an alcohol-specific inhibition training or an active control condition (non-specific inhibition training). After the training, patients participated in a second fMRI measurement where the Go-NoGo task was repeated. Percentage of days abstinent was assessed as drinking outcome 3 months after discharge from residential treatment. Whole brain analyses indicated that in the right inferior frontal gyrus (rIFG), activation related to alcohol-specific inhibition varied with craving and predicted drinking outcome at 3-months follow-up. This neurophysiological correlate of alcohol-specific inhibition was however not modulated by the training version. Our results suggest that enhanced rIFG activation during alcohol-specific  inhibition (I) is needed to inhibit responses when craving is high and (II) fosters sustained abstinence in patients with AUD. As alcohol-specific rIFG activation was not affected by the training, future research might investigate whether potential training effects on neurophysiology are better detectable with other methodological approaches. Alcohol use disorder (AUD) is a leading cause for societal and individual burden of disease , 2 and tOn a behavioral level, patients with AUD displayed inhibitory deficits –10 when Dysregulations in the inhibitory control network in AUD and other addictive disorders have been repeatedly observed , 17, 18 Following the logic that response inhibition is especially crucial in the context of alcohol-related cues, which may trigger strong appetitive processes , 39–41, Taken together, neuroscientific and experimental research in AUD suggests that inhibition is crucial, and probably also particularly difficult, when it must be exerted in an alcohol-related context which provokes craving.via the improvement of alcohol-specific inhibitory capacities. Such a working mechanism would potentially also induce changes in the neurophysiological correlates of alcohol-specific inhibition.Following a translational approach, such research led to the development of an alcohol-specific inhibition training (Alc-IT), which was designed to improve patients' inhibitory control over their responses to alcohol-related stimuli , 46. AlcThe presented study thus aims to (I) replicate earlier reports indicating that neuronal activation related to alcohol-specific inhibition increases with craving; (II) test whether neuronal activation related to alcohol-specific inhibition is related to drinking outcomes and (III) investigate whether the neurophysiological signature of alcohol-specific inhibition can be altered through alcohol-specific inhibition training.In the context of a randomized-controlled, double-blind, clinical trial investigating the effects of alcohol-specific inhibition training on drinking outcomes , the pret−value ≤ 63] as well as non-problematic drinking behavior , right-s, AUDIT  < 8; Alcs, AUDIT  < 2] wers, AUDIT  < 8 per n et al. . Three pAt baseline, a trained study member verified AUD diagnosis with the Diagnostic Expert System for Psychiatric Disorders . A ques(3−monthfollowup) – PDA(Baseline)] was used in those fMRI analyses, which investigated the relation between the neurophysiology of alcohol-specific inhibition and drinking outcomes.Assessment of alcohol consumption was assessed at baseline (assessing drinking in the 90 days prior to detoxification entry) and 3-month follow-up (assessing drinking in the 90 days following treatment discharge) with the TLFB interview . From TLALC, 416 GoNEU, 64 NoGoALC, and 64 NoGoNEU trials, leading to a Go/NoGo ratio of 6.5. The trials were presented in a pseudorandomized order with a mean of 7.5 Go-trials  between two NoGo-trials. The task was subdivided into two blocks with a break of 1 min and 1 s. Photographs were displayed for 900 ms with a 100 ms inter-stimulus interval blank screen. The GNG-task was created and response data was logged with the E-Prime 2.0 software .The task used in this study to assess BOLD-responses of alcohol-specific and neutral inhibition was equal to the task in Batschelet et al. . BuildinFunctional and anatomical MRI data acquisition was conducted at the University Hospital of Bern, using a Siemens Magnetom Prisma scanner with 3 Tesla magnetic field strength and a head coil with 64 channels. For functional image acquisition during the above described GNG-task, a multi-band echo planar imaging (EPI) sequence was run . For subsequent image distortion correction, a b0 protocol was run to acquire 4 field map images (2 phase/amplitude each) with the same image geometry as in the EPI-sequence, with TR/TE1/TE2 = 591/4.92/7.38 ms. Anatomical images were obtained using an MP2RAGE sequence . Note that, while the combination of a fast event-related task design with BOLD-fMRI is not optimal, such a combination has—despite the drawbacks and a reduced signal-to-noise-ratio—yielded important insights into the neural basis of inhibition in prior studies , 63.3 FWHM.Task-fMRI images were preprocessed using the routines implemented in SPM12. Initially, the origins of all functional and anatomical images were reoriented to the anterior commissure. The field map images were used to construct the voxel displacement map for unwarping, which was applied after realignment that involves participant's motion correction. Subsequently, slice time correction was run as well as coregistration of the functional images to the anatomical image. A brain tissue segmentation was performed to obtain forward deformation fields for the image normalization procedure, which transformed the images to the MNI standard space. Finally, all functional images were smoothed using a 3D-Gaussian Kernel with 6 mmALC_correct, NoGoNEU_correct, GoALC_correct, GoNEU_correct, NoGoALC_error, NoGoNEU_error, GoALC_error, and GoNEU_error). The six individual motion parameters derived from realignment were entered as regressors of no interest to the design matrix. Finally, the GLM was estimated. For calculation of contrast images, only parameter estimates of regressors corresponding to event types with correct participant responses were included, since this study focused exclusively on successful inhibition. Thus, the following four contrasts were computed: (NoGoALC + NoGoNEU) > (GoALC + GoNEU), NoGoALC > GoALC, NoGoNEU > GoNEU, (NoGoALC > GoALC) > (NoGoNEU > GoNEU).In order to extract functional activation images for each GNG stimulus condition, contrast images at the subject level were generated . For this purpose, the preprocessed images were entered to a general linear model (GLM). In detail, a design matrix was constructed with the predefined GNG task events consisting of stimulus type , response type , and participant's response accuracy . We used the canonical hemodynamic response function (HRF) as a basis function that was convoluted with the onsets of the resulting eight event types . To this end, a whole-brain linear regression was performed with NoGoALC > GoALC) > (NoGoNEU > GoNEU), which isolates alcohol-specific inhibitory activation, was used for exploring whether the neurophysiological signature of alcohol-specific inhibition is predictive for drinking outcomes. More detailed, the individual planned contrast whole brain images (from the pre-training fMRI session) of N = 35 patients with AUD were used in a whole brain linear regression with the drinking outcome (indicated by PDA Δ) as the covariate of interest. This analysis thus used the fMRI data from the pre-training session to predict drinking outcome at 3-months follow-up. For this analysis, only gray matter voxels were used by applying a binary mask derived from the mean over all individuals' MP2RAGE gray matter segmented image. Brain regions yielded by this analysis as reflecting a neural correlate of alcohol-specific inhibition which is predictive of relapse, will be used as regions of interest (ROIs) in the third analysis (see below).Second, a planned contrast  and the three within-subject factors response type (NoGo/Go) × stimulus type (ALC/NEU) × pre-post (pre-training/post-training). A significant 4-way interaction was needed to confirm a training-effect on alcohol-specific inhibition in the potential ROI that is predictive for drinking outcome.Third, to investigate whether the thus functionally defined, relapse-predicting, alcohol-specific inhibitory activation can be altered through training, the MNI-coordinates of potential effects yielded by the second whole-brain analysis served as regions of interest (ROIs). From these ROIs, beta-values were extracted for each subject, for which a complete dataset was obtained. The analytic sample of this third analysis consisted of t-tests run separately for the AUD and HC groups with the aim of contributing to the research on general inhibition in AUD. As general inhibition was not the main focus of this study, these results can be found in the For completeness, a basic analysis used the (NoGoALC + NoGoNEU) > (GoALC + GoNEU) contrast (pre-training images only) in one-sample ALC > GoALC and OCDSimp, linear regression with NoGoNEU > GoNEU and OCDSimp, linear regression with planned contrast and PDA Δ, and one-sample t-tests with NoGo > Go), SPM12 routines were applied with a family-wise error (FWE) correction at critical p-value threshold of 0.05, with a cluster size of 0. The repeated-measures ANOVA involving beta-values of the ROI was computed with SPSS. Anatomical brain and activation illustrations were created using the MRIcroGL software (https://www.nitrc.org/projects/mricrogl/). Finally, statistics necessary to produce illustrations  was used to assess group differences of demographics and clinical scores. All variables except the BDI II score were non-normally distributed. Therefore, to test differences between the two AUD subgroups, non-parametric Kolmogorov-Smirnov tests were run. For differences over all three groups, non-parametric Kruskal-Wallis tests were computed. Behavioral GNG data  were non-normally distributed and therefore analyzed with ANOVA-type non-parametric statistics using the nparLD package in R . This antrations , 4 were p-values can be found in While the three study groups did not differ in age, the HC group showed a higher education level than the AUD group. As expected, the HC group scored lower in BSCL GSI, OCDSimp, and OCDSsum scores than the AUD group. All group means as well as group statistical (1) = 7.25, p = 0.007]. EOCs, the main indicator of inhibitory performance, did however not differ between patients with AUD and HCs. The RTs of EOC were significantly shorter in NEU than ALC , and shorter in HC compared to AUD , although the two-way interaction was not significant . Concerning the number of EOOs the AUD group performed worse than the HC group . Again, the interaction was not significant . RTs in correct Go trials were shorter in HC compared to AUD , with no significant stimulus type × group interaction .Analyzing the GNG behavioral data from the pre-training measurement, both AUD and HC made more EOCs with NEU trials than ALC trials as confirmed by a significant main effect of stimulus type . The two-way  and three-way interaction (stimulus type × pre/post-training × training type) were also significant . Follow-up analyses of these interactions showed that only in the NEU condition, but not in the ALC condition, the two-way interaction (pre/post-training × training type) was significant . Subsequent simple effects analyses indicated that EOCs in the NEU condition only decreased in the control training , and not the Alc-IT training type . When analyzing the two-way interaction of stimulus type and pre/post-training separately for each training group, this interaction was not significant in both groups .The next analysis compared the patients' GNG behavioral data between the two training groups  and both timepoints (pre taining/post training) and stimulus types . This analysis indicated that numbers of EOC were higher in NEU than ALC . In the number of EOO, main effects of pre/post-training , and stimulus type  were found. In particular, the number of EOO was higher pre- compared to post-training, and higher for ALC compared to NEU. Finally, correct response RTs in the Go condition were shorter post than pre training RTs . Means and standard deviations of the analyzed GNG behavioral variables can be found in Such effects were not found in the RTs of EOC , showing higher activation in NoGo compared to Go trials. Moreover, a significant three-way interaction of response type × pre- post-training × training type  indicated that Alc-IT has an effect on inhibition activation in general, but not specifically on alcohol-related inhibition. This interaction can be seen in F = 1.6, p = 0.213, η2 = 0.053], opposing to our hypothesis of a beneficial effect of Alc-IT on alcohol-specific inhibition .This analysis aimed at investigating whether Alc-IT had a beneficial effect on the alcohol-related inhibitory activation in the rIFG ROI identified in the previous analysis. Analyzing fMRI data collected during an alcohol-related GNG task, this study set out to investigate whether neurophysiological correlates of alcohol-specific inhibition (I) vary with craving ; (II) are related to drinking outcomes in AUD  and (III) can be altered through alcohol-specific inhibition training .In a small cluster of the right inferior frontal gyrus (rIFG), the neurophysiological correlates of alcohol-related  inhibition increased with craving, indicating that brain activation during alcohol-related inhibition was higher in those subjects, which experienced higher levels of craving. This finding extends earlier reports of craving being linked to enhanced neurophysiological activation in cingulate areas during alcohol-specific inhibition , 42 to tThe rIFG is part of the cognitive control network , 67 and Again in the rIFG, neurophysiological activation during alcohol-specific inhibition was related to a better outcome, as indicated by a higher percentage of days abstinent at 3-month follow-up. Those patients displaying a higher activation difference in the rIFG for alcohol-related  inhibition reported more days abstinent at 3-months follow-up. The outcome was thus better in those patients, who managed to recruit enhanced neuronal resources during inhibition when the inhibitory system had to oppose cue-induced appetitive processes.As summarized above, the rIFG is closely linked to effective inhibition and has been linked to response suppression in the face of craving and cue-induced reactions , 70. OurTaken together, the present results indicate that higher rIFG activation is necessary to inhibit responses to alcohol-related cues when craving is high. In line with this, as most patients with AUD probably experience situations with high craving when returning to everyday life after residential treatment, enhanced IFG activation during alcohol-specific inhibition predicts a better outcome at 3-months follow-up.post-hoc with no significant results). To be complete, one should mention that the neurophysiological correlate of general  inhibition was differentially affected in the group receiving Alc-IT when compared to control training. As there was however no theoretical justification for a ROI-analysis focusing on this three-way interaction in the rIFG, this finding has to be seen as highly exploratory.A training effect on the neurophysiological correlate of alcohol-specific inhibition in the rIFG could not be observed; rIFG activation was not modulated by patients engaging in alcohol-related inhibition training (Alc-IT) vs. an unspecific inhibition training (control). Thus, we cannot conclude that this neurophysiological correlate, which—according to the analyses reported above—varies with craving and predicts drinking outcome, is affected by the alcohol-specific inhibition training. In that respect, our results differ from reports on another cognitive training intervention, approach bias training, where training effects on brain activity could be observed, albeit in ROI analyses focusing on other brain regions and during different tasks , 73. OnepFWE−corr = 0.030; T = 5.95; kE = 3; x = 48 mm; y = 12 mm; z = 8 mm). Besides the fact that the application of the FWE-correction used here is a common way to minimize type-I errors, one should however take into account two important aspects: (I) Alcohol-specific inhibitory activation, as we attempted to isolate it in this study, is reflected in a highly specific effect. Our analyses thus had to concentrate on those inhibitory sub-processes that vary with the context in which inhibition had to be carried out. On the one hand, such a highly specific effect might be represented on smaller sub-regions. On the other hand, such an effect might also be harder detectable in a noisy signal such as BOLD. One might therefore consider the application of higher field strengths , where a higher signal should be expected and where the higher spatial resolution might also allow a better investigation of subregions of IFG. (II) The spatial extend of the rIFG clusters amounted to 32 and 72 mm3, respectively. With a neuronal density of several thousand per mm3, we should not preclude that activation of such a specific cognitive process is possible within these small regions, unless there exists indication that it is impossible. In addition, it appears unlikely that two different whole-brain analyses yielded exclusively two highly proximal clusters in a region central to inhibition  during NoGo trials). This is inconsistent with the overall pattern yielded by meta-analyses summarizing behavioral studies on inhibitory performance , 9 and wA second aspect concerns the fact that no effect of the Alc-IT training intervention on alcohol-related EOCs could be detected. As studies on behavioral effects of cognitive training interventions are often larger, this might either be due to a limited power or to the fact that Alc-IT really did not affect alcohol-related errors of commission.In summary, the present study corroborated the central role of the rIFG in the inhibitory control network in AUD by indicating that rIFG activation during alcohol-related inhibition is related to craving and drinking outcome. Therefore, as has been proposed before in the context of cocaine addiction , future maria.stein@upd.unibe.ch.The datasets presented in this article are not readily available because no consent for this was collected from patients. Processed data and corresponding processing subroutines can be requested from the corresponding author. Requests to access the datasets should be directed to ClinicalTrials.gov (NCT02968537). The patients/participants provided their written informed consent to participate in this study.The studies involving human participants were reviewed and approved by the Local Ethics Committee (KEK-number: 2016-00988) and registered at the Swiss National Clinical Trials Portal (SNCTP000002043) and MG collected the data, conceptualized and performed data and statistical analyses, and wrote the manuscript. LS and FM designed the study and supervised data collection and data curation. RT and HB collected the data. AF conceptualized and performed data and statistical analyses. SS collected the data and conceptualized data analyses. YM supervised data collection and data curation. MS designed the study, supervised data collection and data curation, conceptualized data and statistical analyses, and wrote the manuscript. All authors contributed to discussion and interpretation of the results, revised, and approved the final version of the submitted manuscript.This study was funded by the Swiss Foundation for Alcohol Research (SSA-Nr. 283) and the Swiss National Science Foundation .The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Health literacy (HL) is considered an important prerequisite for health. HL research often identifies migrants as vulnerable for low HL. However, in-depth data on HL among migrants and its determinants are still scarce, especially in Germany. Therefore, the analysis presents first time data on HL among migrants in Germany, specifically in the domains of health care, disease prevention and health promotion, considering migrant-specific and universal factors, such as social and psychological aspects.Around 1.000 first- and second-generation adult migrants from two of the largest migrant groups in Germany, from Turkey and former Soviet Union (FSU), were interviewed face-to-face in German, Turkish or Russian in late summer 2020. HL was measured using the HLS19-Q47 instrument. Bi- and multivariate statistical analyses were conducted.More than half of the migrants have limited general HL on average. HL in disease prevention and health promotion was lower than in health care. Low social status, financial deprivation, low literacy skills and low self-efficacy were negatively correlated with each HL domain. Social integration, by contrast, goes along with a higher HL. Duration of stay in Germany and country of origin were only partly associated with HL.Our study goes beyond existing studies by analyzing HL in its domains and including the explanatory power of self-efficacy and social integration for migrants’ HL. Moreover, we reveal that migrants can't generally be considered as vulnerable, as oftentimes outlined. There is need for targeted interventions, e.g. enhancing usability of health information, for the abovementioned subgroups regarding differences in domains.• HL among migrants varies across domains and is only partly linked to migrant-specific features but mainly associated with universal aspects.• Measures to increase HL should consider differences in domains and address vulnerable subgroups including targeted interventions, respectively.


Tubulator, enables automated analysis of t-tubules and dyadic proteins alike, in both tissue sections and isolated myocytes. The program measures densities of subcellular structures and proteins in individual cells, quantifies their distribution into transversely and longitudinally oriented elements, and supports detailed co-localization analyses. Importantly, Tubulator provides tools for three-dimensional assessment and rendering of image stacks, extending examinations from the single plane to the whole-myocyte level. To provide insight into the consequences of dyadic organization for synchrony of Ca2+ handling, Tubulator also creates ‘distance maps', by calculating the distance from all cytosolic positions to the nearest t-tubule and/or dyad. In conclusion, this freely accessible program provides detailed automated analysis of the three-dimensional nature of dyadic and t-tubular structures.During cardiac disease, t-tubules and dyads are remodelled and disrupted within cardiomyocytes, thereby reducing cardiac performance. Given the pathological implications of such dyadic remodelling, robust and versatile tools for characterizing these sub-cellular structures are needed. While analysis programs for continuous and regular structures such as rodent ventricular t-tubules are available, at least in two dimensions, these approaches are less appropriate for assessment of more irregular structures, such as dyadic proteins and non-rodent t-tubules. Here, we demonstrate versatile, easy-to-use software that performs such analyses. This software, called This article is part of the theme issue ‘The cardiomyocyte: new revelations on the interplay between architecture and function in growth, health, and disease’. A high density of well-organized t-tubules and dyads facilitate synchronized Ca2+ release across the entire myocyte and efficient triggering of contraction [2+ handling, and hence reduced whole-heart contractility [In cardiomyocytes, invaginations of the sarcolemmal membrane form a complex system, known as the t-tubular network. Although t-tubules are primarily oriented transversely across the cardiomyocyte, a substantial fraction of ‘axial' or ‘longitudinal' tubules may be oriented along the long axis of the cell. Both transverse and longitudinal elements form dyadic junctions with the sarcoplasmic reticulum (SR). This close localization of the dyadic membranes enables the opening of L-type Catraction –5. T-tubmageJ plugin using these methods was previously published [AutoTT that some of these shortcomings were circumvented [in situ preparations and cryosections. In addition, the plugin relies on continuous structures, thereby limiting its use on tissue where t-tubules or dyadic proteins are more sparsely distributed. Another approach has used matched-filter-based algorithms to circumvent the pitfalls of thresholding [First and foremost, assessing dyadic remodelling requires quality imaging of cardiomyocyte structures, acquired with optimized labelling specificity and intensity. Once recorded, dyadic structure has traditionally been quantified in two ways; firstly, by density analyses (e.g. fraction of cell covered by structure of interest) in binarized images, and secondly, by regularity analysis using fast Fourier transformation (FFT). While these approaches are practical and easily implemented, both methods are sensitive to variations in image quality and pixel intensity. A common recurring problem when analysing density is that automated thresholding algorithms are sensitive to the overall t-tubule signal, thus biasing threshold settings towards lower values in cells with fewer t-tubules. By comparison, FFT analyses are, in addition to being susceptible to intensity-driven bias, also sensitive to cell size and require regular structures to yield readily interpretable results. An automated Iublished , but it umvented . Their psholding . HoweverTubulator software employs novel methods to circumvent the most common pitfalls occurring when analysing such data, while enabling quantification of the density, orientation and co-localization of t-tubule membranes and proteins. The current software is more versatile than previously published plugins, as it supports automated analysis of different cell types and multicellular preparations, and because it enables interpretation and reconstruction of the three-dimensional nature of dyadic structure.Here, we developed an automated approach to analyse t-tubule and dyadic structures in images captured by confocal and Airyscan microscopy. The . 2 (a)Animal experiments were approved by the ethics committee at the University of Oslo and performed in accordance with the Norwegian Animal Welfare Act and NIH Guidelines . Wistar rat cardiomyocytes were isolated by retrograde Langendorff perfusion, and either imaged directly or fixated in 4% paraformaldehyde as previously described .Methods for collection of human tissue were approved by the Regional Ethics Committee (project S-05172) in agreement with The Declaration of Helsinki and the Council of Europe Convention on Human Rights and Biomedicine. Left ventricular tissue was obtained from non-diseased donor hearts deemed unsuitable for transplant owing to surgical concerns. Tissue was snap-frozen in liquid nitrogen and, following cryosectioning 10–20 µm), fixed in 4% paraformaldehyde for 30 min Experimental imaging of t-tubules was performed by staining live cells with di-4-ANEPPS (Sigma-Aldrich), di-8-ANEPPS (Sigma-Aldrich), RH237 (ThermoFisher), CellMask (ThermoFisher) or FM1–43FX (ThermoFisher). In fixed cells and tissue sections, t-tubules were stained with wheat germ agglutinin conjugated to Alexa Fluor . Dyadic proteins were labelled with primary antibodies against Caveolin-3  and bridging integrator 1 , and secondary antibodies coupled to Alexa Fluor 488 or 546 (ThermoFisher). Presented images of dyadic proteins and t-tubules were obtained using an LSM800 Airyscan confocal microscope  using a 63× magnification oil immersion objective. To test the versatility of the plugin, analysis of images captured with other confocal microscopes (Zeiss LSM 510 and 710) was also performed. (c)Tubulator was written in Matlab and compiled to a stand-alone application . The software requires installation of Matlab compiler runtime. The Tubulator executable file can then be copied to anywhere on a Windows PC. Technical details and limitations of Tubulator can be found in the electronic supplementary material. (d)Tubulator is provided in the electronic supplementary material. The workflow of the program is demonstrated in figure 1a for a representative, isolated rat ventricular cardiomyocyte immunolabelled for the dyadic proteins Caveolin-3 and Bin-1. Based on rough manual estimation of cell orientation and contours, Tubulator rotates and trims the raw image to fit and centre the cardiomyocyte in the analysis window. The ‘active contour' [Tubulator calculates the fraction of the cell covered by t-tubules or protein of interest.A step-by-step user guide for contour'  function2+ release [Tubulator analyses the fractions of transverse and longitudinal elements by analysing the direction of neighbouring pixels in the skeleton.Dyadic structures are examined in more detail, as the binarized image is skeletonized followed by morphological closing. Based on the resulting skeleton, distances to the skeleton are calculated for each pixel in the cytosol. These distances are then summarized in a ‘distance plot', illustrating the distance from all points in the cytosol to the nearest t-tubule or sarcolemmal membrane; a critical determinant of the synchrony of cellular Ca release . In addiTubulator is the ability to analyse cells in tissue sections directly . The software enables analysis of several individual cells in each tissue section and can, provided that the analysed image is captured as a z-stack, also calculate distances to the nearest t-tubule or membrane in three-dimensional space . Three-dimensional reconstructions of the calculated metrics can subsequently be assembled using other programs  as shown in figure 2b, and explained in more detail in [An important feature of directly a, left. al space a, right.etail in .Figure  (e)Tubulator, an array of synthetic cardiomyocyte phantoms (n = 37 cells) was created in Matlab , with known t-tubule density and quantities of transverse and longitudinal elements. In brief, these synthetics cells were created using an image matrix consisting of 1024 × 1024 pixels, at a resolution of 6.67 pixels μm−1. In each matrix, the cell's surface membrane was simulated with a 900 × 200 pixel rectangle 7 pixels in thickness. Within this rectangle, transverse and longitudinal lines representing t-tubules were drawn. Transverse lines were spaced at intervals of 1.8 ± 0.3 µm, while longitudinal lines had an average distance of 5.6 ± 1.2 between them. To simulate a variable t-tubule density in both orientations, a random number generator was used to determine whether a given line would be drawn. The chance of losing a given line was set by the operator, varying from 0% to 100%. Exact densities of transverse and longitudinal tubules were calculated by counting the number of pixels containing transverse lines inside the cell, divided by the area of the cell. To simulate noise acquired during experimental imaging, a noise map was superimposed on simulated t-tubule images using random Gaussian noise created and filtered with convolution of a circular averaging filter. After analysing the final simulated images with Tubulator, comparison was made with the performance of previously published software [To test the accuracy and analytic power of oms n = 3 cells wa. 3Tubulator was developed to improve upon existing methods for structural analysis of t-tubules and dyadic proteins . Unl. UnlTubuthe cell .Tubulator's adaptive thresholding method ensures binarization unbiased by cell contents , while at the same time eliminating bias owing to variations in pixel intensity . Importantly, estimations of density are made prior to the skeletonization step, meaning that variations in t-tubule diameter are accounted for. The subsequent skeletonization step in the analysis pipeline enables extraction of key features of t-tubule/dyadic organization, allowing delineation of transversely and longitudinally oriented structures . Unlike currently available software, the technique used for these calculations is not based on FFT analysis, which is dependent on structural regularity, but rather determines the directionality of each pixel's neighbours.Tubulator's performance, synthetic cardiomyocyte phantoms were created, with t-tubule densities varying between 0% and 30% (a). Importantly, the images comprised differing compositions of transverse and longitudinal components. Example cells with high and low t-tubule density are shown in figure 3a. We observed that Tubulator accurately estimated t-tubule density, as outputs were strikingly similar to known values in the simulated images (figure 3b). Measurements of the densities and fractions of transverse and longitudinal elements were also found to be robust (figure 3b). To compare Tubulator's capabilities with previously published analysis tools . As expected, these macros were often unable to analyse cells with no transverse elements, or very low overall t-tubule density. Indeed, of the 37 cells analysed, TTorg and AutoTT failed to yield outputs for the six images with the sparsest and least regular t-tubule arrangements. This shortcoming highlights the pitfalls of traditional FFT-based analyses, and the advantage of not assuming the presence of regular structures. Notably, all programs estimated densities and fractions of transverse and longitudinal elements which were significantly correlated with actual values (figure 3b–d). However, Tubulator achieved the strongest associations, and the improved performance of Tubulator was not limited to cells with sparse t-tubules. Thus, the program is well suited for analysis of cardiomyocytes with a wide range of t-tubule or protein density.To benchmark  and 30% a. Imports (TTorg  and Autog [utoTT ), the saz-stacks, distances are additionally calculated in three-dimensional space to yield valuable information about the true three-dimensional nature of myocyte substructure.As the arrangement of dyads in transverse and longitudinal orientations is frequently complex, greater insight is given by the produced distance maps. In addition to producing colour-coded images, average, median, and maximum distance and variability are conveyed to give a nuanced picture of both density and regularity of the analysed structures. For Tubulator is also convenient to use for analysis of co-localization between proteins or structures of interest. The output image file contains both the binarized and skeletonized cell which can be readily employed for subsequent co-localization analyses. In addition, individual skeletons of transverse and longitudinal elements are produced, which provides a scaffold for assessing co-localization within these two orientations. As an example, a rat ventricular cardiomyocyte labelled with Bin-1 and Cav-3 is shown in figure 1a. Although previous reports have indicated that there is robust expression of these proteins along t-tubules [Tubulator showed that Bin-1 and Cav-3 do not fully co-localize. Indeed, large fractions of transversely oriented Bin-1 were present in the absence of Cav-3 staining, and longitudinal co-localization was virtually non-existent (figure 1b).-tubules , co-locaTubulator performs analyses in tissue sections directly (Tubulator enables easy identification of individual cells' localization in a tissue section. This feature is especially useful when, for example, investigating regional differences in cardiac chambers [in situ preparations, where individual cardiomyocytes can readily be selected for analysis [In contrast to previously published software, directly . The filchambers ,17. Anotanalysis ,19.. 4Tubulator provides the possibility to analyse pre-binarized images, or binarization based on mean fluorescence.The main challenge for obtaining robust quantification of t-tubules and dyadic proteins is proper thresholding of staining signals. Here, a custom-made adaptive thresholding technique was developed to allow more accurate binarization. Although, this approach improves upon existing methods, it does not overcome all pitfalls of binarization entirely, and some risk of over- or under-estimating signal density remains. However, we have sought to minimize binarization bias by allowing the user to adjust the adaptive thresholding in the program. Importantly, our method relies on good image quality for optimal outputs and on rare occasions a different thresholding method may be preferred. Thus, Tubulator does not directly assess t-tubule widths, although previous work has shown that t-tubule dimensions may be significantly altered during cardiac disease [Tubulator outputs, by comparing t-tubule densities between two groups before and after skeletonization. For example, we previously have shown that higher t-tubule density observed during heart failure with preserved ejection fraction was attributed to t-tubule dilation, since skeletonization reduced t-tubule signals to control values [ disease . Neverthl values . Alternal values .. 5Tubulator's versatility and novel features for data extraction will aid researchers in interpreting data and generate new hypotheses. The software is freely available and can be accessed by contacting the authors.In summary, we developed software suited for detailed analysis of the three-dimensional organization of t-tubules and dyadic proteins, with easy application to isolated cardiomyocytes or tissue sections. We anticipate that


In the non-paroxysmal AF (non-PAF) patients, the non-O-type blood groups have significantly higher incidences of very late recurrence  than those in the O-type blood group. The multivariate analysis revealed the non-O blood group  and amiodarone  were independent predictors for very late recurrence in the non-PAF patients after catheter ablation, which could be applied as a useful disease marker. This work highlighted the potential link between the ABO blood types and inflammatory activities that contribute to the pathogenic development of AF. The presence of surface antigens on cardiomyocytes or blood cells in patients with different ABO blood types will have an impactful role in risk stratification for AF prognosis after catheter ablation. Further prospective studies are warranted to prove the translational benefits of the ABO blood types for the patients receiving catheter ablation.Chronic inflammation harbors a vulnerable substrate for atrial fibrillation (AF) recurrence after catheter ablation. However, whether the ABO blood types are associated with AF recurrence after catheter ablation is unknown. A total of 2106 AF patients  who underwent catheter ablation were enrolled retrospectively. The patients were separated into two groups according to the ABO blood types, the O-type  and the non-O-type groups  . The clinical characteristics, AF recurrence, and risk predictors were investigated. The non-O type blood group had a higher incidence of diabetes mellitus (11.90 vs. 9.03%,  Atrial fibrillation (AF) is a common global health issue in the aging society. Not only causing poor quality of life, but the AF also increases adverse cardiovascular (CV) events, including death, heart failure (HF), and stroke. The present therapeutic guideline recommended the importance of stroke prevention, rhythm, and rate control are considered the cornerstone of a standardized AF treatment . Early iCatheter ablation reduces the incidence of stroke and all-cause mortality in selected patients compared with AADs . HoweverTo be especially noted in patients with atrial fibrillation (AF), the non-O-type blood groups  were associated with active inflammatory status, as reflected by increased levels of von Willebrand factors (VWF) and inflammatory factors. As cardiac inflammation underlies AF pathogenesis by the induction of atrial electrical and structural remodeling, increases in AF vulnerability or recurrence after catheter ablation in patients with increased cardiac inflammation are observed . The infCatheter ablation is a standard therapy for AF patients to maintain sinus rhythm. The AF recurrence at one year remains approximately 20% of AF patients receiving catheter ablation. Developing biomarkers to predict AF recurrence is crucial, which helps the physician identify the patients at risk for AF recurrence. Therefore, the therapies could be adjusted, in the early stages, and the patient could be followed up more closely. In the present work, we aimed to investigate whether ABO blood groups could be used as a biomarker for the prediction of arrhythmia recurrence in AF patients receiving catheter ablation in the single-center retrospective cohort.2DS2-VASc score, underlying comorbidities, 12-lead electrocardiogram (ECG), AAD, echocardiogram, and AF recurrence were collected from the hospital’s medical system. The risk factors of AF recurrence including patient’s age, HTN, diabetes mellitus (DM), HF, dyslipidemia, and LAD were also investigated. This study protocol was reviewed and approved by the Ethics of Institutional Review Board of the Taipei Veterans General Hospital, Taiwan (IRB: 2021-01-026CC).All symptomatic drug-refractory AF patients who underwent catheter ablation (January 2004 to January 2018) were enrolled at Taipei Veterans General Hospital, Taiwan. The AF diagnosis and documentation were guided by the 9th version of the International Classification of Diseases (ICD) and Clinical Modification (ICD-9-CM) codes. The ICD codes were registered by cardiologists in charge of the patient’s diagnosis and treatment. Any patient who presented with a bed-ridden status, pregnancy, hemodynamic unstable condition, intracardiac thrombus, younger than 18 years old, older than 80 years old were excluded from this investigation. All AF patients were further divided into 2 groups (O-type blood group and non-O-type blood groups). This study included the baseline characteristics of personal history, AF types, blood group, age, gender, CHAThe AF-related symptoms included fatigue, dizziness, palpitations, dyspnea, chest pain, and anxiety. The European Heart Rhythm Association (EHRA) symptom score was applied by the physician to identify and quantify AF-related symptoms for guidance in a symptom-driven treatment approach . The AF-According to the latest AF guideline, paroxysmal AF (PAF) was defined as AF that terminated spontaneously or terminated with intervention within 7 days of onset . PersistAccording to our protocol, symptomatic drug-refractory AF patients discontinued their AADs for >5 half-lives before the procedure. Amiodarone was stopped 2 weeks before the procedure because of the long half-life. Patients all received adequate oral anticoagulant before catheter ablation according to the guideline . During The catheter ablations were performed by an anatomic-based linear ablation approach at the PV antrum. The ablation lesions were achieved through a point-by-point approach using ablation catheters based on the selection of the EAM systems . SuccessIn patients with non-PAF, substrate modification  was performed if the AF did not terminate after PVIs. The endpoint was a prolongation of the cycle length, elimination of CFAEs, or abolishment of local fractionated potentials. Linear ablation included roof or mitral lines in the left atrium (LA) . Linear After catheter ablation, all patients received AADs for at least 6–8 weeks to prevent acute recurrence during the blanking period. Acute recurrence after catheter ablation is defined as the attack of atrial arrhythmias within three months of the blanking period . A blankAll patients received a series of outpatient follow-ups 1–2 weeks, 1 month, and every 3 months after catheter ablation. One year after the catheter ablation, the outpatient follow-ups were changed to every 6 months. All patients received a resting 12-lead ECG during every outpatient follow-up. The 12-lead ECGs were performed by trained cardiology technicians with a 30-s observation on the 12-lead ECG monitor to determine whether the atrial arrhythmias were sustained [Therapeutic strategies that improve long-term rhythm control were recommended and educated, including weight loss, strict control of chronic diseases, and avoiding AF-related triggers. Chronic diseases included HTN, DM, HF, and dyslipidemia. The AF-related triggers included alcohol drinking or smoking . The impp-value of less than 0.05 was considered significant.All analyses were calculated with the IBM SPSS Statistics 22 . Continuous variables were expressed as mean ± standard deviation, and categorical variables as counts (percentages). The two-sample independent T-test was used to analyze continuous variables. Categorical data were compared using a Chi-square test with Yates correction or the Fisher exact test. Logistic regression analysis was used for identifying the independent predictor of very late AF recurrence. A n = 1196, 56.8%) were enrolled. A total of 2106 patients . In PAF patients after catheter ablation, there were no differences in late and very late recurrences between the O-type and the non-O-type blood groups, respectively .2DS2-VASc score, AADs, alcohol consumption, smoking, and echocardiogram parameters showed no difference between the no recurrence and very late recurrence groups  and amiodarone  were independent predictors for very late recurrence in the non-PAF patients after catheter ablation  have higher incidences of very late AF recurrence after catheter ablation. The non-O type blood group was an independent predictor for AF recurrence. This suggests that different ABO blood groups could be applied as a biomarker for risk stratification of AF recurrence in patients after catheter ablation.Accurate blood group typing is critical before a blood transfusion. This information is considered essential for all the patients before catheter ablation of AF because blood transfusion is sometimes unfortunately needed for bleeding during the procedure. However, this test is rarely applied as an alternative application. In the present work, we showed that the ABO blood groups were an independent predictor of very late AF recurrence in the non-PAF patients. This extends the potential application of the ABO blood group from blood transfusion to a clinical biomarker to predict AF recurrence. In addition to the conventional risk factors, including age, gender, chronic diseases, non-pulmonary vein foci, and AF persistence, the non-O-type blood groups  are a simple and effective biomarker with an independent predictive power for AF recurrence ,15. The The non-O-type blood groups  are an independent risk factor for AF recurrence. Several potential clinical applications of ABO blood groups could be considered in the future. First, the risk prediction model, including ABO blood groups, will help doctors set up a treatment plan and make shared decision-making. This includes the explanation of recurrent risk and post-procedure patient care. Moreover, the patients and families will be informed of their relevant risky condition after the procedure. As these patients have a higher risk for AF recurrence, some relevant symptoms related to AF recurrence could be explained thoroughly to avoid any negligence of arrhythmia attack. The AF patients who neglect the presence of an AF recurrence after catheter ablation could potentially suffer from AF-related symptoms and have a poor quality of life. Early detection of AF recurrence is not only associated with improved quality of life but also changes the therapeutic plan. Following a 3-month blanking period, it is reasonable to incorporate an additional 1–3 months of therapy for consolidation. The adjustment of AADs and/or electrical cardioversion can be performed if AF recurrence could be detected as early as possible. The appropriate treatment for these patients might mitigate complications from AF recurrence [Second, the patients after catheter ablation are usually followed regularly after catheter ablation by cardiac monitoring devices. In order to detect AF recurrence in the early stages, the non-O-type blood groups  might need a more frequent and longer duration of arrhythmia monitoring. The typical monitoring duration of the non-invasive recorders ranges from 24 h to months. As some of the attacks are minimally symptomatic, patch monitoring for 1–3 weeks, rather than 24-h Holter recording might be considered if AF recurrence is suspected in the non-O-type blood groups .Third, AF recurrence is related to the reconnection of PVs or the triggers from non-PV areas. In the patients with the non-O-type blood groups , a more intensive induction of arrhythmic triggers might be considered. The PV reconnection could be checked cautiously by PV exit pacing after catheter ablation at the end of the procedure. Although it remains controversial, the adenosine test can be used to unmask dormant conduction after PVI, ensure persistent bidirectional PV conduction block, and possibly prevent future reconnection.Finally, anti-inflammatory therapy might be used for patients with non-O-type blood groups . This is an interesting hypothesis and needs forthcoming solid clinical evidence to prove its feasibility. However, anti-inflammatory therapy is not a new story for AF. For example, colchicine, statins, steroid, or non-steroid anti-inflammatory drugs have been reported to decrease AF recurrence in small cohorts.The United Biobank investigation suggests that the ABO genotypes are strongly associated with common inflammatory and CV diseases. In addition, the ABO blood groups are associated with different levels of VWF and platelet endothelial cell adhesion molecule-1 .In a genome-wide association study that investigated patients with the acute coronary syndrome, genetic variants at the ABO blood group locus were associated with interleukin-10 levels . FurtherThe ABO blood groups are determined by antigen modifications on glycoproteins and glycolipids. The expressions of the ABO blood group antigens are found on the cardiomyocytes of the endocardium . DiffereAmong these inflammatory biomarkers, the VWF level might be the most important factor that links the ABO blood groups and AF recurrence. The human ABO blood group led to different plasma levels of VWF and factor VIII (FVIII), which contributed to coagulopathy and CV diseases ,21. PatiThe present study has several limitations. First, this study was a retrospective design within a single medical center, therefore, it cannot represent the general population. A prospective multicenter randomized controlled trial might be needed to clarify our findings. Second, the catheter ablation procedural variables or ablation strategies that could affect long-term outcomes still needs to be thoroughly investigated. Different strategies between operators and operating systems might be related to procedure outcomes, although the procedure endpoints have been standardized in our center. Third, the influence of drug-drug interaction in underlying chronic diseases needed to be thoroughly investigated. The follow-up of the patients in the present retrospective cohort is not as rigorous as those in the clinical trials. The monitoring devices for post-procedure arrhythmia monitoring, as physicians decided, could change the monitoring durations of arrhythmias. Finally, the biomarkers and molecular mechanisms of the ABO blood group and AF recurrence were not thoroughly inspected in this retrospective study, which demands further investigations. The timeframe to examine the biomarker and the optimal cut-off values was uncertain . The effThe non-O-type blood groups  were independent predictors for very late AF recurrence in the non-PAF patients after catheter ablation. The ABO blood group could be a useful biomarker for risk stratification of AF recurrence. The ABO blood groups in AF patients who underwent catheter ablation could help set up the pre-procedure treatment plan and trigger early awareness of AF recurrence in patients and physicians. This might change post-procedure care from actively detecting AF recurrence at an earlier stage to prompt diagnosis and therapy and hope to improve the long-term AF outcome.


The outcomes of endoscopic ultrasonography-guided drainage (EUSD) in treatment of pancreas fluid collection (PFC) after pancreas surgeries have not been evaluated systematically. The current systematic review and meta-analysis aim to evaluate the outcomes of EUSD in patients with PFC after pancreas surgery and compare it with percutaneous drainage (PCD).PubMed and Web of Science databases were searched for studies reporting outcomes EUSD in treatment of PFC after pancreas surgeries, from their inception until January 2022. Two meta-analyses were performed: (A) a systematic review and single-arm meta-analysis of EUSD  and (B) two-arm meta-analysis comparing the outcomes of EUSD and PCD . Pooled proportion of the outcomes in meta-analysis A as well as odds ratio (OR) and mean difference (MD) in meta-analysis B was calculated to determine the technical and clinical success rates, complications rate, hospital stay, and recurrence rate. ROBINS-I tool was used to assess the risk of bias.The literature search retrieved 610 articles, 25 of which were eligible for inclusion. Included clinical studies comprised reports on 695 patients. Twenty-five studies (477 patients) were included in meta-analysis A and eight studies (356 patients) were included in meta-analysis B. In meta-analysis A, the technical and clinical success rates of EUSD were 94% and 87%, respectively, with post-procedural complications of 14% and recurrence rates of 9%. Meta-analysis B showed comparable technical and clinical success rates as well as complications rates between EUSD and PCD. EUSD showed significantly shorter duration of hospital stay compared to that of patients treated with PCD.EUSD seems to be associated with high technical and clinical success rates, with low rates of procedure-related complications. Although EUSD leads to shorter hospital stay compared to PCD, the certainty of evidence was low in this regard.The online version contains supplementary material available at 10.1007/s00464-022-09137-6. Post-operative complications after pancreatic surgery can be life threatening and lead to major morbidity and mortality . The mosPFCs often extend toward visceral organs and form irregular shapes, which make it difficult to drain the fluid , 10. ForRecent studies have focused on endoscopic ultrasonography-guided drainage (EUSD) of PFC. EUSD has a high clinical efficacy and low morbidity and is considered the best option for managing pancreatic pseudocysts and PFCs –17. SeveThe current systematic review and meta-analysis evaluated the outcomes of EUSD in treatment of patients with PFC after pancreatic surgery and compared them with PCD.This study was reported in accordance with the preferred reporting items for systematic reviews and meta-analyses (PRISMA) 2020 guidelines  and a PRPopulation Patients with post-pancreatectomy PFCIntervention EUSD to treat PFC after pancreatic resectionComparator None in meta-analysis A; PCD in meta-analysis BOutcome Technical success, time to drainage, clinical success, repeated drainage, hospital stay, and relevant perioperative data, including intra- and post-operative complications, as well as data regarding incidence of post-operative complications.Study design All study designs, except editorials and letter to editors.The study question was formulated based on the population, intervention, comparison, outcome, and study design (PICOS) strategy. Studies were included in the study if they met the following criteria:To avoid analyzing the same patients more than once, the studies were thoroughly assessed and double publications and overlapping reports removed.A systematic literature search in Medline (via PubMed) and ISI Web of Science was conducted using the following search terms: “ AND (Collection[tiab] OR Pancreas Collection[tiab] OR Pancreatic Collection[tiab] OR Fistula[tiab] OR Pancreas fistula[tiab] OR Pancreatic fistula[tiab]).” The search was not restricted to a specific study type or year of publication. The last query was performed in January 2022.After screening titles and abstracts in selected electronic databases, the full texts of appropriate studies were evaluated, and their data were extracted by three investigators  independently. Discrepancies among these investigators were resolved through discussions with a senior author (AM). For each study, the following data were extracted: study characteristics, patient characteristics, study quality, and the abovementioned outcome measures.Baseline data, including indication and type of primary pancreas surgery, PFC-related symptoms, and time to drainage, were obtained.Technical success was defined as successful insertion of the stents into the PFC endoscopically, as well as access to and drainage of the contents. Repeated drainage was defined as an unsuccessful initial drainage that needed additional interventions. Clinical success was defined as the resolution of the PFC and its associated symptoms. Procedure-related complications were stent migration, perforation, bleeding, sepsis, infection, and PFC recurrence. The duration of hospital stay after interventions was also recorded.robvis tool was used [Two investigators  assessed bias in non-randomized studies using the ROBINS-I (Risk of Bias in Non-randomized Studies of Interventions) tool . The riswas used .https://gradepro.org).The certainty of evidence was assessed by applying “The Grading of Recommendations Assessment, Development and Evaluation (GRADE)” approach , and a sχ2 and inconsistency (I2) analyses, and the threshold for heterogeneity was a P value lower than 0.05 or an I2 value greater than 50%. Publication bias was assessed using a funnel plot. The R software (version 4.0.1) and Meta package were used for data analysis.For the single-arm meta-analysis, data proportions were analyzed using a random effects model. For the two-arm meta-analysis, dichotomous data were presented as odds ratios (OR) and continuous data as weighted mean differences (MD). Summary effect measures were presented along with their corresponding 95% confidence intervals (CIs). Statistical heterogeneity was assessed using The literature search identified 264 articles, 25 of which met our eligibility criteria. These clinical studies reported 695 cases and were included in this meta-analysis , 22–39 . The artA detailed summary of the risk of bias for non-randomized two-arm studies using ROBINS-I tool is listed in Supplementary Fig. 1. The level assessment for each outcome was varied between moderate and serious risk of bias. Overall risk of bias for technical success, clinical success, post-procedural complications, and PFC recurrence were assessed as serious due to missing data and selection of results domain. Overall risk of bias for hospital stay was reported at moderate. In Table I2 = 0%; Heterogeneity-P = 0.94)  .The one-arm analysis of 24 studies indicated that the clinical success rate in the EUSD group was 87%       Below is the link to the electronic supplementary material.