Current evidence supports a negative impact of obesity-associated metabolic dysfunction in several cancers. However, the evidence is still controversial regarding high-grade serous ovarian cancer (HGSOC). In this study, we demonstrated that body composition, particularly the presence of high visceral adiposity (with or without sarcopenia) estimated by aCT scan, is associated with worse survival in HGSOC. As a molecular proxy to CT-scan-based assessment of nutritional status and to identify putative biomarkers of metabolic disorders, we evaluated the expression levels of a set of 425 obesity- and lipid-metabolism-disorder-related genes across 273 tumor samples. We identified two obesity- and lipid-metabolism-related clusters with marked differences in survival and that were associated with molecular features predictive of immune checkpoint blocker response. Finally, we assessed the impact of nutritional/pharmacologic interventions affecting body composition/lipid metabolism on patient survival. We observed that the reduction of visceral adiposity, the increase of muscle mass, and the use of metformin/statins improve survival.Although obesity-associated metabolic disorders have a negative impact on various cancers, such evidence remains controversial for ovarian cancer. Here, we aimed to evaluate the impact of body composition (BC) and metabolism disorders on outcomes in high-grade serous ovarian cancer (HGSOC). Methods: We analyzed clinical/genomic data from two cohorts (PUC n = 123/TCGA-OV n = 415). BC was estimated using the measurement of adiposity/muscle mass by a CT scan. A list of 425 genes linked to obesity/lipid metabolism was used to cluster patients using non-negative matrix factorization. Differential expression, gene set enrichment analyses, and Ecotyper were performed. Survival curves and Cox-regression models were also built-up. Results: We identified four BC types and two clusters that, unlike BMI, effectively correlate with survival. High adiposity and sarcopenia were associated with worse outcomes. We also found that recovery of a normal BC and drug interventions to correct metabolism disorders had a positive impact on outcomes. Additionally, we showed that immune-cell-depleted microenvironments predominate in HGSOC, which was more evident among the BC types and the obesity/lipid metabolism cluster with worse prognosis. Conclusions: We have demonstrated the relevance of BC and metabolism disorders as determinants of outcomes in HGSOC. We have shone a spotlight on the relevance of incorporating corrective measures addressing these disorders to obtain better results. Despite recent advances in understanding the biology of high-grade serous ovarian cancer (HGSOC) and the addition of new therapeutic avenues based on molecular targets  polymerase inhibitors (PARPi), or immune checkpoint blockers [ICB]), survival remains very poor ,2. CurreAfter primary treatment is completed, patient surveillance includes management of the side effects of therapy and timely identification of predictors of recurrence and susceptibility to future salvage schemes in those places where such options exist. This approach justifies the search for mutations of somatic/germinal genetic defects (homologous recombination deficiency [HRD] or BRCA1/2 mutations) that predict sensitivity to novel targeted therapies [Recently, in addition to chronological age and baseline performance status, modifiable factors such as obesity and certain co-morbidities  have become more relevant in terms of defining the course of the disease, therapeutic options, and the chance of response in several cancers . DespiteHere, we evaluate whether changes in the status of different predictive factors, measured at diagnosis and after completion of first-line treatment, impact on progression-free (PFS) and overall survival (OS) in HGSOC. Among them, we included (i) reduction of tumor burden , (ii) type of response , (iii) CA-125 levels, and (iv) inflammatory status  ,11,12,13This study was IRB-approved . Electronic records from patients with confirmed epithelial ovarian cancer who were treated at our institution (PUC) between 2004 and 2017 were reviewed. Only stage III and IV HGSOC with at least six months of follow-up were included in the study. All of the cases included in the survival analyses had information on histological confirmation of diagnosis, age, adequate staging, intent of surgery, chemotherapy schemes, and tumor burden/residual disease after planned treatment completion. For those cases undergoing interval debulking surgery (IDS), we registered if the surgery was carried out in timely fashion as planned (after the third or fourth cycle of chemotherapy). All of the cases with ‘incomplete or unclear’ information on these relevant factors were excluded. To guarantee maturity of the survival analyses, we collected data until at least 50% of the patients had experienced death by any cause. Date and cause of death were verified by accessing the Chilean Civil Registry database. The final dataset included the following clinical variables: age, height (cm), weight (kg), body mass index (BMI), FIGO stage, type of surgery , debulking level , type of response , type and number of chemotherapy schemes, the absolute neutrophile count (ANC) to absolute lymphocyte count (ALC) ratio (ANC:ALC), absolute platelet count per ml, the calculated systemic immune-inflammatory index (platelets x ANC/ALC), lactate dehydrogenase (LDH), albumin, and cholesterol levels.2-transformed FPKM-UQ were used to perform further transcriptome-based analyses related to the impact of body composition on progression-free (PFS) and overall survival (OS).In addition, we downloaded publicly available raw counts and upper-quartile normalized fragments per kilo base per million mapped (FPKM-UQ) RNA-seq expression and clinical data from Genomic Data Commons through the TCGA-biolinks R package . As with2-FPKM levels of these 415 genes [https://bioinformatics.mdanderson.org/estimate/ accessed on 10 December 2022) were compiled from different sources [For the RNA-seq analyses, a curated list of 425 genes linked to obesity and lipid metabolism disorders was aggregated from different sources ,16. Unsu15 genes . The cop15 genes ) to calc15 genes ), ShinyG15 genes ), and Ec15 genes ). Availa sources ,24,25,262, VAT/SCAT ≥ 0.4, or VAT/TAT ≥ 0.285 [L4-L5 + 2.5177) [L-S) at a cut-off value of −20 [For clinical analyses, CT scans were downloaded from either our institutional server or the Cancer Imaging Archive . To assess nutritional status and body composition, the visceral adipose tissue (VAT), subcutaneous adipose tissue (SCAT), the psoas muscles, and the vertebral body areas were measured using a single-slice CT scan located between the 4th and 5th lumbar vertebrae. Next, VAT/SCAT ratio, VAT/TAT ratio (VAT/VAT + SCAT), and Psoas to L4 vertebral body index (PLVI) were calculated to estimate visceral obesity and central sarcopenia . Viscera 2.5177) . Liver se of −20 . The imaCo-morbidities such as hypertension, diabetes mellitus, heart failure (CHF), chronic obstructive pulmonary disease, and non-independent functional status were registered to calculate the modified frailty index 5 (mFI-5) ,32,33. At-test, Chi-squared test, Mann–Whitney U-test, or logistic regression. A p-value < 0.05 was considered statistically significant. Survival curves were generated by using the Kaplan–Meier method and analyzed by Wilcoxon and log-rank tests. Univariate Cox proportional hazard regression analyses were carried out for all of the variables. Multivariate Cox proportional hazard regression models were carried out using a stepwise method to assess the effects of different risk factors on survival. To select variables to be included in the modeling, we used a stopping rule or selection criteria based on the p-value of 0.2 [p-value found [JMP16.2.0  was used for the data analyses. Group comparisons and continuous or categorical variable association analyses were performed as required by the Student’s e of 0.2 . We alsoue found ,36,37.A total of 123 and 415 stage III and IV HGSOCs from our institution (PUC) and the TCGA-OV cohort were included in the present study, respectively. Twenty-two cases in the PUC cohort and 102 cases in the TCGA-OV cohorts were excluded for lacking relevant clinical information  or excessive and unjustified delay in accomplishing planned treatment (for the PUC cohort). As shown in Patient BMI remained unchanged in our cohort after six months, by the end of the first-line treatment A. The prBy combining the VAT or WBAT with PLVI estimates, we were able to establish four types of body composition (BC) in our patients. As shown in p < 0.001) independent of BMI status in both cohorts  to establish a formula maybe applicable to other cohorts. Thus, we confirmed a cut-off point already validated by others for the VAT/SCAT (0.4) and we established a cut-off value for the PLVI based on the lowest quartile of the sample, 0.45. The aggregated set included 201 cases. Both the VAT/SCAT and PLVI cut-off points were associated with lower overall survival in univariate analysis A,B. In aAs a molecular proxy to CT-scan-based assessment of nutritional status and to identify putative biomarkers of metabolic disorders, we evaluated the expression levels of a set of in-house precompiled 425 obesity- and lipid-metabolism-disorder-related genes  across 2We compared established patterns and scores that correlate with prognosis and ICB response between the obesity/lipid metabolism type I and obesity/lipid metabolism type II clusters, namely: stemness index, microenvironmental subtype, tumor mutation burden (TMB), and ESTIMATE score ,38,39,40Next, we used the EcoTyper framework that enables the identification of cellular states and communities (cancer ecotypes or CE) from bulk RNA-seq data. Different cell types interact with each other in different functional states (cell states or S) and together condition the relationship between the tumor and host in a specific ecosystem .The obesity and lipid metabolism type II cluster (the cluster with the best prognosis) displayed a significantly higher proportion of CE9 and CE10 ecotypes. We also observed differences in the status of CD8, NK, plasmatic, B, dendritic, and monocyte/macrophage cells but not in CD4 or polymorphonuclear cells .Given that the clusters were identified by gene sets related to obesity and lipid metabolism disorders and not related to sarcopenia, we decided to evaluate the impact of the association of this condition on survival already established by the clusters. We found that the presence of sarcopenia (low PLVI) made the differences in overall survival between clusters more evident, particularly for the cluster with the worst prognosis. More importantly, this association constituted a prognostic variable independent of the patient’s age, achieving complete removal of the disease and obtaining a complete response , we found differences, with the CE1 ecotype predominating in the BC type with the worst prognosis  affecting body composition and lipid metabolism on patient survival. In the PUC cohort, we observed that the improvement in VAT/SCAT and PLVI parameters was associated with reduction of visceral adiposity and recovery of muscle mass at 12 months of follow-up and improved progression-free (PFS) and overall survival (OS) A,B. ThusIn the present study, we confirmed that most women with advanced HGSOC (67.6%) achieve a complete response (in 37.6% of them associated with complete tumor debulking achieved either primarily or after receiving neoadjuvant chemotherapy). In addition to a complete response, other factors associated with survival are the age at diagnosis, the baseline frailty of the patient (related to performance status) and, as we have shown here, the normalization of parameters related to inflammatory  and immune  responses at the end of first-line treatment. Enhancing the concept that ´transitioning from an inflamed to a non-inflamed patient’ besides achieving complete response constitutes a relevant factor to improve long-term outcomes ,41.2 does not correlate with the risk of chronic diseases and death from all causes during aging [Regarding nutritional status and metabolic disorders, we show that BMI does not change significantly after treatment in our cohort and does not correlate well with changes observed in cholesterol or albumin levels. It also does not adequately correlate with the presence of liver steatosis, whole body adiposity (high WBAT), or visceral adiposity (high VAT or VAT/SCAT) or with the presence of central sarcopenia (low PLVI) measured by a CT scan. More importantly, being a normal weight, overweight, or obese based on the BMI parameter does not constitute a predictor of progression-free or overall survival. This contrasts with the significant impact on survival of four body composition (BC) types identified using the combination of VAT or VAT/SCAT or WBAT with PLVI. Our findings confirm the coined concept of the ‘obesity paradox’ where having a BMI ≥ 30 kg/mng aging ,43. Of nng aging . Thus, ang aging ,45,46. Wng aging . This isng aging ,49.Another finding of our research is related to the expression profiles of genes related to obesity and lipid metabolism and their impact on the composition and cell states of the tumor microenvironment (TME) and on patient survival. Here, we showed that the differential expression of these genes within the tumor defines two clusters with distinct survival outcomes. These clusters also differ in the biological processes and signaling pathways that characterize them. However, more importantly, the ecosystems present in them also differ and determine different cellular interactions, particularly among those linked to the anti-tumor immune response. In particular, the cluster named here as obesity/lipid metabolism type I, in addition to exhibiting the worst prognosis, has characteristics that reflect a TME where the anti-tumor immune response is at least deficient, if not permissive. Regardless of the bioinformatic tool or score used, this cluster is characterized by a lower proportion of the immunoreactive molecular subtype, a lower relative proportion of TME defined as immune-enriched, and a higher proportion of TME defined as immunodepleted, particularly of lymphocytes. A more in-depth analysis using EcoTyper revealed that those ecosystems defined as immunoreactive, pro-inflammatory, and more sensitive to immunotherapy (C9 and C10) represent less than 20% of the ecosystems identified in this cluster . When anFinally, we must mention that regardless of the BC type, the carcinoma ecotype that predominates in advanced HGSOC is CE1. CE1-high tumors are described as lymphocyte deficient . Along wAmong the strengths of our study is having used two comparable cohorts (with an adequate size), homogeneous in characteristics (only HGSOC in the advanced stage), and different ways of evaluating the role of body composition and metabolic disorders in survival in HGSOC, along with sufficient follow-up data. One of the limitations of our study is the retrospective nature of our series, where nutritional interventions and the use of statins were not a routine indication and continuity or adherence to them over time was not evaluated. Our discoveries warrant a prospective study where the duration, treatment schemes, adherence to recommendations, and other interacting therapies are evaluated in addition to the primary survival outcomes.Based on our findings, we believe it is relevant to propose the abandonment of BMI as an element of judgment to define or limit therapies at the time of diagnosis or recurrence. Incorporating more precise parameters, measurable by a CT scan, that enable a better definition of body composition can allow more effective interventions and eventually positively condition the response to therapeutic alternatives in this cancer, including immunotherapy.More importantly, frontline therapeutic actions should include nutritional interventions  that tend to normalize body composition in the mid-term and correct lipid metabolism disorders  ,55. Thes


Understanding the regulatory role of enhancer–promoter interactions (EPIs) on specific gene expression in cells contributes to the understanding of gene regulation, cell differentiation, etc., and its identification has been a challenging task. On the one hand, using traditional wet experimental methods to identify EPIs often means a lot of human labor and time costs. On the other hand, although the currently proposed computational methods have good recognition effects, they generally require a long training time.https://github.com/20032303092/StackEPI.git.In this study, we studied the EPIs of six human cell lines and designed a cell line-specific EPIs prediction method based on a stacking ensemble learning strategy, which has better prediction performance and faster training speed, called StackEPI. Specifically, by combining different encoding schemes and machine learning methods, our prediction method can extract the cell line-specific effective information of enhancer and promoter gene sequences comprehensively and in many directions, and make accurate recognition of cell line-specific EPIs. Ultimately, the source code to implement StackEPI and experimental data involved in the experiment are available at The comparison results show that our model can deliver better performance on the problem of identifying cell line-specific EPIs and outperform other state-of-the-art models. In addition, our model also has a more efficient computation speed.The online version contains supplementary material available at 10.1186/s12859-022-04821-9. Enhancer–promoter interactions (EPIs) play a crucial role in the transcriptional regulation of genes and in human disease progression and cell differentiation. A promoter is a short DNA segment with a sequence length varying between 100 and 1000 base pairs, located upstream of a specific gene . PromoteIn the past, with the rapid development of high-throughput sequencing technology, wet experimental methods based on chromosome conformation capture (3C)  and its In recent years, many computational methods using machine learning algorithms to identify EPIs have been proposed. From the perspective of data usage, it can be divided into two categories: one is the prediction method based on the gene sequence, and the other is the prediction method based on the epigenome. For example, Whalen et al.  proposedDifferent from the above methods, we review, analyze and compare 6 different feature coding methods and 7 commonly used machine learning algorithms, in which Multi-Layer Perceptron (MLP) and Logistic Regression (LR) algorithms are only used as meta-classifiers. Then we propose a novel stacking ensemble framework to identify cell line-specific EPIs named StackEPI. Figure We performed pairwise combination of 6 feature encodings  and two statistical methods (LightGBM and XGBoost) to construct 8 base classifiers and selected MLP and LR as the candidate algorithms for the meta-classifier. We first input the enhancer and promoter gene sequences into the 8 baseline models and get 8 predicted probability values. Then we feed these predicted probability values as new features to the MLP classifier or LR classifier. Finally, we get a trained ensemble model and test our model in an independent test set. The final prediction effect of the integrative model on each cell line is shown in Table To verify the effectiveness and superiority of our prediction method, we compare our method with two state-of-the-art methods, namely EPIVAN and EPI-DLMH. To be fair, first we chose the same benchmark dataset of 6 cell lines and used the identical training set and independent test set. Second, each comparison method follows the basic training procedure referred to in the literature for model training. Finally, all our experiments are conducted on the same machine . We give the prediction tables of AUROC, AUPR, and F1-score in Tables Table In addition, in terms of model training time, we further compared the training time of the existing model with that of our proposed method. We found that our approach has unparalleled advantages over other methods. In our experiments, we use the laptop's 3070 GPU to train all models. Table In this study, we propose a method called StackEPI, which combines multiple feature encodings and machine learning algorithms, adopts a stacking ensemble strategy and performs EPI prediction only through enhancer and promoter gene sequences. First, we use 6 feature encodings and 5 machine learning algorithms to construct 30 baseline models based on the benchmark dataset and then obtain a 30-dimensional feature vector generated from all baseline models and composed of predicted probability values. The new vector is input to the meta-classifier to get the final prediction as the ultimate result. In an ensemble manner, our ensemble model synthesizes the advantages of baseline models, which is shown in that our ensemble model has better scores on AUROC, AUPR, and F1-score than the single baseline model. Compared with the most advanced prediction methods, our method has breakneck training speed and better AUROC performance. And theoretically, the input of our approach does not have strict requirements on the length of enhancer and promoter gene sequences. Therefore, it may become a useful tool for fast and accurate prediction of cell line-specific EPIs.http://genome.compbio.cs.cmu.edu/~sss1/SP-EID/all_sequence_data.h5. The details of the dataset are given in Table Our experimental data came from TargetFinder and used the same EPI benchmark dataset as EPIVAN. The dataset was provided initially by SPEID, and the complete data can be downloaded from We tried 6 feature encodings commonly used in biological research and analyzed their ability to identify EPIs under different classifiers. The pros and cons of feature encoding schemes are generally positively correlated with model performance –34. The https://knindex.pufengdu.org. In this study, we used the standardized physicochemical properties of k-tuple nucleotides provided by Zhang. In brief, each feature encoding is described as follows.At present, the physicochemical properties of k-tuple nucleotides are also commonly used in computational prediction of bioinformatics, such as type III secreted effectors , type VICKSNAP is the composition of K-spaced nucleic acid pairs and is widely used in various studies in the bioinformatics field [cs field , 41, 42.Kmer describes the occurrence frequency of k-tuple nucleotide composition from the given sequence. NAC, DNC, TNC, and TeNC will produce 4-, 16-, 64- and 256-dimensional feature vectors. After splicing, the gene sequence will be encoded into 340-dimensional feature vectors.DPCP We used the following 21 dinucleotide physicochemical properties for the computation of feature encoding DPCP, including Base stacking, Protein induced deformability, B-DNA twist; A-philicity, Propeller twist, Duplex stability (free energy), Duplex stability (disrupt energy), DNA denaturation, Bending stiffness, Protein DNA twist, Stabilizing energy of Z-DNA, Aida_BA_transition, Breslauer_dG, Breslauer_dH, Breslauer_dS, Electron_interaction, Hartman_trans_free_energy, Helix-Coil_transition, Ivanov_BA_transition, Lisser_BZ_transition and Polar_interaction. The calculation of this feature vector can be expressed as:TPCP In this study, we considered 10 trinucleotide physicochemical properties: Bendability-DNAse, Bendability-consensus, Trinucleotide GC Content, Nucleosome positioning, Consensus_roll, Dnase I, Dnase I-Rigid, MW-Daltons, Nucleosome and Nucleosome-Rigid. The calculation method of TPCP is similar to DPCP, and its feature vector representation is as follows:EIIP Nair et al. calculated the energy of delocalized electrons in nucleotides as electron–ion interaction pseudopotential, namely EIIP. The EIIP values corresponding to the four bases are {A: 0.1260, C: 0.1340, G: 0.0806, T: 0.1335}, respectively. We considered the EIIP encoding of trinucleotides, calculated as follows:PseKNC is the pseudo k-tuple nucleotide composition widely adopted in researchers' methods due to its powerful information extraction ability for DNA and RNA sequences, such as iTerm-PseKNC [K-tuple nucleotides m-PseKNC . PseKNC Type II PseKNC encoding finally produces a  method, which includes two important parts: cascade forest structure and multi-grained scanning. The model structure is presented in Fig. https://github.com/LAMDA-NJU/Deep-Forest and used the CascadeForestClassifier classifier for it this study.SVM is a traditional supervised machine learning method based on the structural risk minimization (SRM) [https://github.com/Xtra-Computing/thundersvm.on (SRM)  frameworon (SRM) . Commonlon (SRM)  that canLightGBM is another efficient implementation of gradient boosting decision tree (GBDT) [https://github.com/microsoft/Light-GBM/tree/master/python-package.e (GBDT)  algorithe (GBDT) . LightGBXGBoost is a scalable machine learning system implemented by Chen and Guestrin in 2016 based on the gradient boosting framework. First of all, in principle, XGBoost introduces regularization to prevent overfitting of the trained model and uses an approximate algorithm to solve the problem that the greedy algorithm cannot make data read into memory for calculation when the amount of data is too large. Secondly, XGBoost relies on weighted quantile thumbnails to process weighted data to propose candidate split points and then applies the sparsity aware algorithm to deal with sparse data and missing values. Finally, XGBoost adopts fast structure design, out-of-core computing, and cache-aware learning in system design. Therefore, XGBoost has the advantages of supporting distributed and parallel computing and fast, massive data processing. The python version of the XGBoost package is delivered by https://github.com/dmlc/xgboost/tree/master/python-package.RF is one of the most widely used maximum likelihood algorithms. The core idea of RF is to integrate multiple independent and unrelated decision trees in the form of bagging integration and introduce randomness (bootstrap sample method and random selection of feature subsets) to prevent the risk of model overfitting and improve anti-noise ability. The following is the construction process of the decision tree:For the training set with Assuming that the number of features is Each tree grows as much as possible without pruning.MLP is an artificial neural network that structurally includes input, hidden, and output layers, uses backpropagation to update weight computationally and is used to solve nonlinear problems. It has been widely used in problems in different fields.LR is a maximum likelihood algorithm for solving classification problems, and it has been successfully applied to many classification tasks in bioinformatics [ormatics , 50.Our study proposes a stacking ensemble learning framework, StackEPI, which can precisely identify cell-specific EPIs from only genetic sequence data. An overview of StackEPI is illustrated in Fig. The original unbalanced dataset Augment the training set Train the model on the balanced training set Evaluate the model on the independent test set As mentioned above, our dataset is an unbalanced dataset with a positive-to-negative sample ratio of 1:20. In supervised learning, the classification model always tends to focus on the majority class and reduce the prediction accuracy of the minority class. Therefore, we perform data augmentation operations on the training set and test the performance of our model on the independent test set. For a fair comparison with EPIVAN and EPI-DLMH, the specific training process of the ensemble model for any given cell line is described as follows:To make the ensemble model achieve better performance, we use grid search to tune the essential parameters of the base classifiers and meta-classifiers of the ensemble model. If only a one-time grid search adjusts all parameters for some classifiers that need to be tuned with many parameters, it will result in a considerable time cost. Therefore, we group the parameters of such classifiers and then sequentially adjust them layer by layer. The optimized machine learning parameters and search ranges are given in Additional file This study uses AUROC, AUPR, and F1-score to evaluate our method. The receiver operating characteristic curve reflects the relationship between sensitivity and specificity at different thresholds. The precision-recall curve reflects the tradeoff between the model's accuracy for identifying positive examples and the model's ability to cover positive examples. The closer the values of AUROC, AUPR, and F1-score are to 1, the better the model's performance.Additional file 1. Parameters and the value range of parameter adjustment.


Attention-deficit/hyperactivity disorder (ADHD) is a highly prevalent, inheritable, and heterogeneous neurodevelopmental disorder. Children with a family history of ADHD are at elevated risk of having ADHD and persisting its symptoms into adulthood. The objective of this study was to investigate the influence of having or not having positive family risk factor in the neuroanatomy of the brain in children with ADHD. Cortical thickness-, surface area-, and volume-based measures were extracted and compared in a total of 606 participants, including 132, 165, and 309 in groups of familial ADHD (ADHD-F), non-familial ADHD (ADHD-NF), and typically developed children, respectively. Compared to controls, ADHD probands showed significantly reduced gray matter surface area in the left cuneus. Among the ADHD subgroups, ADHD-F showed significantly increased gray matter volume in the right thalamus and significantly thinner cortical thickness in the right pars orbitalis. Among ADHD-F, an increased volume of the right thalamus was significantly correlated with a reduced DSM-oriented t-score for ADHD problems. The findings of this study may suggest that a positive family history of ADHD is associated with the structural abnormalities in the thalamus and inferior frontal gyrus; these anatomical abnormalities may significantly contribute to the emergence of ADHD symptoms.  Attention-deficit/hyperactivity disorder (ADHD) is one of the most prevalent and persistent neurodevelopmental disorders that affects approximately 5% of children worldwide ,2. ThereExisting structural neuroimaging studies in ADHD have shown widely inconsistent results. Several studies reported a lower cortical thickness in the pars opercularis, fusiform, parahippocampal, precentral gyri, temporal pole, and cingulate cortex ,19,20, aWhile there is a large quantity of literature on neural correlation of ADHD symptoms in samples of children/adults with ADHD compared to healthy control groups, there are notably less neuroanatomical imaging studies focused on the familial risk factor that may represent biologically distinct subgroups of ADHD ,38,39. SIn the present study, we proposed to investigate the neuroanatomical substrates and their clinical/behavioral linkages associated with ADHD-F vs. ADHD-NF in a large sample of three independent groups of children . On the basis of findings from our existing neuroimaging and clinical studies ,50,51,52All subjects were obtained from the baseline data in the Adolescent Brain Cognitive Development (ABCD) Study Release 4.0, a large study that took part in a major collaboration between multiple sites across the United States. All participants in the ABCD study were recruited from schools with diverse demographic factors, with the initial recruitment resulting in 11,875 children. All the parents provided written informed consent and the informed assent was obtained from children. The data were downloaded from the National Institute of Mental Health Data Archive. Participants were included in this study if they were in age ranged 9–10 and had Intelligence Quotient (IQ) higher than 80.  was used to assess the ADHD symptomatology in the ABCD study ,58. In tThe short version of the Edinburgh handedness inventory was used to assess a handedness score of “right”, “both”, or “left” based on 4 questions about preferred hand using for writing, throwing, using a spoon, and using a toothbrush . Due to 3; flip angle = 8°; repetition time/echo time for Siemens, Philips, and GE scanners were 2500/2.88, 6.31/2.9, and 2500/2, respectively; field of view = 256 × 256 mm2 for Siemens and GE scanners and 256 × 240 mm2 for Philips scanner. The ABCD neuroimaging data acquisition protocols and parameters have been previously published in detail . Brieflyhttps://surfer.nmr.mgh.harvard.edu (accessed on 13 July 2021) [All participants who had T1-weighted structural images with moderate to severe motion or visible artifacts were excluded from further analyses after careful visual inspection. Three-dimensional cortical reconstruction and segmentation for thickness and area estimations using T1-weighted data were performed utilizing Freesurfer version 7.2.0 (ly 2021) ,67. IndiAfter applying brain masks, each voxel was categorized as white matter or non-white matter based on intensity and estimated bias field. Next, in the white surface formation stage, Freesurfer refined generated surfaces for hemispheres by following intensity gradients between white matter and gray matter. Then, at the pial surface formation, Freesurfer nudged white surface to follow the intensity gradients between gray matter and cerebrospinal fluid. The cortical thickness in each of 68 bilateral cortical regions was then estimated based on the distance between white and pial surfaces at each location. The border surface between gray matter and white matter was inflated into a sphere, and then surface-based labeling was used to segment both left and right hemispheres into different sub-regions in each of 68 bilateral cortical regions. Thirty-seven subcortical structures/nuclei regions’ volumetric segmentations were performed at several stages including affine registration, initial volumetric labeling, inhomogeneities correction, volumetric alignment to the MNI305 atlas, and final segmentation.t-tests were used for continuous variables (displayed as the mean and standard deviation). Chi-square tests were used for categorical variables (displayed as number and frequencies).SPSS  was used to compare clinical, neurocognitive, and demographic measures of the participants between groups of controls and ADHD probands, as well as between the two ADHD subgroups (ADHD-F and ADHD-NF). Independent sample p-values and control potential false positive results of these group comparisons. Additionally, full model analyses were conducted  was used to compare the structural T1 neuroimaging measures including regional cortical thickness and surface area of 68 cortical regions and volume of 37 subcortical structures/nuclei between the groups of TDC and ADHD probands and further between the subgroups of ADHD-F and ADHD-NF. Age and sex were used as covariates; we also added estimated total intracranial volume (ETIV) as covariate for volume and surface area because they scale with size of the head . Bonferrshown in  to invesp-values and control potential false positive results of these comparisons.Pearson correlation was performed to analyze whether the neuroanatomical brain measures that showed between-group differences were correlated with the clinical symptom measures (the raw scores for inattentive and hyperactivity–impulsivity symptoms score collected from KSADS-5 and DSM-oriented t-scores collected from CBCL). Again, Bonferroni correction method was used to adjust The final sample used in the main analysis comprised a total of 606 participants including 309, 165, and 132 in groups of TDC, ADHD-NF, and ADHD-F, respectively. Participants’ demographic information and corresponding group statistical comparisons are shown in p = 0.038) and significantly increased surface areas in the right middle temporal . There were no significant findings for cortical thickness or volume between groups of TDC and ADHD ; significantly increased surface area in the left inferior temporal  and left middle temporal  lobes; significantly greater volume in the right thalamus   .2 = 0.039, p = 0.023)  . No signHere, we present a highly powered study of cortical and subcortical structural brain differences between the subgroups of familial and non-familial of ADHD probands in children, as well as between ADHD and TDC groups. Compared to controls, the ADHD group had a significantly reduced surface area of the cuneus in the left hemisphere. The cuneus is one of the critical regions in processing visual information and has been suggested to be part of the default mode network ,72,73. SWithin the probands, our findings showed significantly increased gray matter volume of the right thalamus in the ADHD-F group compared to the ADHD-NF group. Furthermore, the increased volume of the right thalamus was significantly correlated with the reduced t-score of ADHD problems from DSM-oriented CBCL scales. Thalamus is one of the critical components of the Cortico-Striato-Thalamo-Cortical (CSTC) loops which subserve attention and cognitive processing ,85,86. IFurthermore, within the ADHD probands, we found that children with ADHD-F had significantly thinner cortical thicknesses in the right pars orbitalis compared to children with ADHD-NF. Right pars orbitalis, as a part of the right inferior frontal gyrus (rIFG), plays a key role in behavioral responses inhibition ,95. It hThe present study has a few limitations. One limitation of the study is that the ABCD study only included the ADHD assessment for one of the parents. To minimize the confounding effects in the ADHD-NF group, we further excluded subjects that had any potentially ADHD-related family histories based on the collected family history questions. Another limitation is that the parametric statistical models used in this study have a limited capacity to investigate multiple variables and their interactions simultaneously. Future studies using machine learning methods may effectively extract the high-dimensional information and translate the complex neuroimaging patterns into clinical relevance. Finally, the cross-sectional study design of this study only focused on understanding the anatomical alterations and their interactions with the ADHD symptoms in children with ADHD-F vs. ADHD-NF at 9–10 years of age. Future research with a longitudinal study design will facilitate the understanding of how these common and distinct neuroanatomical features in children with ADHD-F vs. ADHD-NF grow along with age and whether the ADHD-F-specific childhood neural markers  in affected individuals have an important predictive value for future clinical outcomes associated with the persistence/remission of ADHD symptoms.In conclusion, the current study found that relative to group-matched controls, school-aged children with both ADHD-F and ADHD-NF had common gray matter structural abnormalities associated with the cuneus and temporal lobe. Whereas, within the ADHD probands, children with ADHD-F showed a significantly increased volume of the right thalamus and decreased cortical thickness of the right IFG when compared to ADHD-NF; the volume of the right thalamus showed significant negative correlation with the severity of ADHD symptoms  in children with ADHD-F, but not in those with ADHD-NF. These findings resulted from analyses of a big sample of children from community-based recruitment. Therefore, findings reported in our study are statistically much more powerful than those reported in studies with small sample sizes. Meanwhile, this is the first study in the field investigating neuroanatomical substrates of ADHD-F vs. ADHD-NF in two large independent subgroups of ADHD in children. The results suggest that ADHD-F may represent a biologically more homogeneous subgroup of ADHD. These findings have the potential in guiding future studies with longitudinal designs to understand the developmental trajectories of these ADHD-F-specific neural markers and their roles in clinical outcomes associated with the persistence/remission of ADHD symptoms in affected individuals.


While patient-derived iPSC-CMs provide a unique opportunity for disease modeling with isogenic cells, the challenge is that these cells still demonstrate distinct properties which make it functionally less akin to adult cardiomyocytes. In response to this challenge, numerous innovations in differentiation and modification of hiPSC-CMs and culture techniques have been developed. Here, we provide a focused commentary on hiPSC-CMs for use in disease modeling, the progress made in generating electrically and metabolically mature hiPSC-CMs and enabling investigative platforms. The solutions are bringing us closer to the promise of modeling heart disease using human cells  Before the discovery of human pluripotent stem cells (hPSCs), including human embryonic stem cells (hESCs) first isolated by Dr. James Thomson (UW–Madison)  and humaUsing hiPSCs has enabled the investigation of patient-specific conditions in a lab environment . MethodsTBX5Clover2/NKX2-5TagRFP and hES3-TBX5TdTomato/W/NKX2-5eGFP/W double reporter line, respectively, for isolation of FHF- and SHF-derived hPSC-CMs  and posterior (pSHF), contribute to different CM subtypes, in which aSHF gives rise to RV and outflow track (OFT) and pSHF contributes to atria  primarily targets on the left ventricle (LV), while Brugada syndrome (BrS) and arrhythmogenic right ventricular cardiomyopathy (ARVC) mainly affect right ventricle (RV), and atrial fibrillation impacts on atria. Therefore, the generation of chamber-specific hPSC-CM subtypes is needed to model specific heart disease and to develop novel therapies and precision medicine. The hPSC-CM differentiation protocols that have been used in many labs primarily generated majority of ventricular CMs , 8, 9. HhPSC-CMs , 21. Howto atria –25.To realize the full potential of using hPSC-CMs for disease modeling, drug screening, precision medicine, and cardiac regeneration largely rely on our ability to differentiate them to the ideal, specific, and closely akin to adult CMs. Since significant advancement and great progress have been achieved in the past 10 years, we have hope to make it even closer to the goal in the next 10 years.to and IKur) rather than delay rectifier channels Ikr or Iks as in human  and calsequestrin (CASQ2) genes that encode intracellular proteins involved in intracellular Ca2+ regulation , RYR2 (CPVT and CRDS), and hERG1 (LQTS and SQTS), which highlights the need for functional characterization combined with deep clinical phenotype assessment. Other non-ion channel-inherited arrhythmogenic syndromes can involve genes encoding structural and contractile elements in the heart, including arrhythmogenic cardiomyopathy (ACM) (includes ARVC) and laminopathy, and can first present with arrhythmia prior to structural remodeling likely related to the interplay of ion channel regulation with these proteins  in iPSC-CMs which results in control of cellular pacing and AP restitution, in agreement with most mammalian models , pushing the cells to more mature excitation–contraction coupling  is being developed to allow an integrated, mechanistic risk assessment with strong evidence to inform regulatory decision-making and efficiency for drug discovery  in patients  channel . Despiteiscovery . Shortcoed drugs –66. Furted drugs . CiPA iseristics . The utieristics .Understanding iPSC-CM metabolism is key to generating a more mature cell for the study of human disease. One such approach is long-term culture of iPSC-CMs. In a pivotal study, Ebert et al. demonstrated that long-term culture (>200 days) results in divergent control of mitochondrial metabolism, regulated by PKA and proteasome-dependent signaling events . AdditioDeveloped mitochondria are central to cardiac metabolism, as they progress from small, fragmented mitochondria to large organelles that can produce enough ATP to sustain the contractile function of the heart . Mitochovia the pentose phosphate pathway  levels that are involved in cardiomyocyte differentiation and maturation , 85. TimiPSC-CMs , 88. AddiPSC-CMs . FurtheriPSC-CMs . It appeiPSC-CMs .Various techniques have been developed to culture iPSC-CMs in multidimensional constructs to achieve organizational and structure–function relationships that mimic native heart cells.in vivo dynamics of cardiac tissues  range. These conditions even with metabolic enhancements can result in less mature phenotypes in terms of structure, function, and gene expression response with limited ability to reflect  tissues . In cont tissues , 94 have tissues  with imp tissues . Using pUnique engineered substrates have also advanced 2D culture for iPSC-CMs. Micropatterns offer a mechanism to create a platform for cell connectivity and native sarcomere morphology . We and In the native heart, cardiomyocytes make up only part of the heterocellular composition. With the recent capability to create iPSC-cardiac fibroblasts , we haveBeyond 2D culturing and planting platform advances, innovative 3D constructs have distinct advantages of cellular and structural heterogeneity to more closely mimic native cardiac tissue. Some examples include engineered heart tissue (EHT) cardioids, 3D bioprinting, biometric scaffolds such as biowires, bioreactors, and organ-on-a-chip microphysiological system , 101. Hi2+, sarcomere lengths, and membrane potential will help the functionality of 3D platform , Karen Thompson Medhi Professorship to WC, Office of the Vice Chancellor for Research and Graduate Education to WC, and the Gary and Marie Weiner Professor in Cardiovascular Medicine Research to LE.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.


HIV self-testing has been touted as the game-changer in HIV testing uptake with high HIV testing rates reported in studies conducted in low and middle-income countries, a majority of which were conducted in sub-Saharan Africa . In thisAbubakari et al. used community-based interventions, enhanced with mobile platforms and digital technology, as opportunities to increase HIV self-testing and linkage to HIV care among MSM in Ghana. Abubakari et al. worked with community-based organization partners to implement three interventions that successfully engaged and retained MSM which provided an opportunity for linkage to HIV self-testing and medical care. d'Elbee estimated the cost of integrating HIV self-testing into 23 civil society organization (CSO)-led models for key populations in Senegal, Cote d'Ivoire, and Mali and found that providing HIV self-test kits to KPs through CSOs was not only cost-effective but had varying levels of cost-effectiveness. The team found that the cost of reaching female sex workers with HIV self-testing services was much lower than that for reaching MSM and PWUDs . Okoboi et al. found that the cost per new HIV-positive MSM identified ($325 vs. 914) and the cost per HIV transmission averted  through HIV self-testing was much lower than the cost per new HIV-positive MSM identified and new HIV transmissions averted through conventional HIV testing services. The team concluded that HIVST was not only cost-effective but also identified more undiagnosed HIV infections than standard-of-care HIV testing. Kra et al. described the adaptations that the HIV self-testing teams used to navigate the challenges posed by the COVID-19 pandemic including the use of social networks by MSM peer educators to maintain contact with their peers, promote HIV prevention and testing, and organize face-to-face or small group meetings, as needed. These adaptations were essential for the continued provision of HIV self-testing services during the COVID-19 lockdown in Mali, Senegal, and Cote d'Ivoire.Evidence shows that HIV testing uptake among key populations (KPs) remains sub-optimal despite the fact that these populations contribute up to 51% of new HIV infections in sub-Saharan Africa . Four stMantell et al. and Kelvin et al. found that truck drivers preferred blood-based HIV testing over oral-based HIV self-testing. In the study by Kelvin et al. 305 truck drivers were randomized to receive oral HIV self-test kits or stand-of-care HIV testing and followed up for 6 months. At the end of the follow-up period, HIV testing uptake was similar [56.3% in the intervention arm and in the standard-of-care arm (55.6%)], with those who did not test for HIV in both arms citing reasons related to lack of time to test for HIV, low HIV risk behavior, fear of knowing their HIV status and recent HIV testing. When asked to choose between blood-based and oral HIV self-testing, 69.4% preferred blood-based HIV testing. Similar results were reported by Sithole et al. who found that of the men who were given the option to choose between oral or blood-based HIV self-test kits, 62%  preferred to use the blood-based kits while 38%  selected to use the oral fluid kits; suggesting a growing interest in blood-based HIV self-test kits. This interest is usually driven by beliefs, particularly among men, that since HIV is found in blood, then, blood-based HIV self-testing strategies could yield the most realistic results  were reported as used by their recipients. Forty-two per cent of the testers were first-time testers. However, linkage to HIV care remained low with only 9% of the 11 HIV-positive individuals identified were linked to HIV care. McGowan et al. explored PrEP naïve and PrEP-experienced adolescent girls and young women's (AGYW) willingness to engage in a peer-delivered HIV self-testing and referral model for PrEP initiation in Kiambu County, Kenya. Study findings show that PrEP-experienced AGYW were willing to initiate discussions about HIV self-testing and PrEP use among their peers, to deliver HIV self-test kits to them, and to refer them to appropriate HIV prevention, care and treatment services, based on their HIV status. PrEP-naïve AGYW were also willing to receive and use HIV self-test kits delivered to them by their peers and to link to appropriate HIV prevention, care and treatment services based on their HIV test results. Muchedzi et al. distributed 11,983 kits between 2018 and 2020 in Zimbabwe; of these, 99.5%  were used and results were returned to the health care workers. Of the returned HIVST results, 22.3%  were reactive and, of these, 2,610 (98.2%) results were confirmed HIV positive by a trained health care worker using the national testing algorithm. The highest positivity rate was reported among users aged 35–49 years . The prevalence of HIV in the study population was nearly twice as high as the prevalence reported among adults 15 years or older in the 2020 Zimbabwe Population-based HIV Impact Assessment survey (22.3 vs. 12.9%) . However, as Ky-Zerbo et al. reported, stakeholders doubted potential HIVST users' autonomy regarding their ability to use HIVST kits correctly; to ensure quality secondary distribution; to accept a reactive test result; and to use confirmation testing and care services. Similar sentiments have been reported in other settings where HIVST interventions are being introduced for the first time , future interventions will need to enhance HIV disclosure as part of HIV self-testing promotional strategies, especially in populations with traditionally low HIV disclosure rates. As Hamilton et al. have argued, probably there is no one intervention strategy that will work universally to increase HIV testing uptake and linkage to appropriate HIV prevention or care and treatment services. Multiple interventions will be needed to reach men and other unreachable populations with HIV self-testing services including peer-to-peer distribution, use of community health counselors, use of trained lay distributors selected by the community, and integration of HIV self-testing services into other HIV services. Also, multiple interventions will be needed to enhance linkage to HIV care including home-based ART initiation, use of phone reminders, and community-based ART initiation.Evidence from two papers, published as part of this Research Topic, shows that policymakers and other key stakeholders consider HIV self-testing to be an opportunity to reduce stigma; preserve anonymity and confidentiality; reach key populations that do not access HIV testing rst time –7 and suJKBM wrote the initial draft. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.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.


Rhipicephalus microplus ticks.Ticks are blood-feeding ecto-parasites that have a cosmopolitan distribution in tropical and subtropical regions of the world. Ticks cause economic losses in the form of reduced blood, meat and dairy products, as well as pathogen transmission. Different acaricides such as organochlorines, organophosphates, formamidines (e.g. amitraz), synthetic pyrethroids, macrocyclic lactones, fipronil, and fluazuron are currently used sequentially or simultaneously to control tick infestations. Most acaricide treatments now face increasingly high chances of failure, due to the resistance selection in different tick populations against these drugs. Acaricide resistance in ticks can be developed in different ways, including amino acid substitutions that result in morphological changes in the acaricide target, metabolic detoxification, and reduced acaricide entry through the outer layer of the tick body. The current literature brings a plethora of information regarding the use of different acaricides for tick control, resistance selection, analysis of mutations in target sites, and resistance mitigation. Alternatives such as synergistic use of different acaricides, plant-derived phytochemicals, fungi as biological control agents, and anti-tick vaccines have been recommended to avoid and mitigate acaricide resistance. The purpose of this review was to summarize and discuss different acaricides applied for tick control, their mechanisms of action and resistance selection, genetic polymorphisms in their target molecules, as well as the approaches used for diagnosis and mitigation of acaricide resistance, specifically in  Ticks are hematophagous ecto-parasites with global geographical distribution, especially in tropical and subtropical regions, where they parasitize terrestrial and semi-terrestrial vertebrates . Ticks aRhipicephalus microplus was reported for the first time in Australia , amitraz, synthetic pyrethroids (SPs), macrocyclic lactones (MLs), and fipronil are commonly used acaricides that act on the tick central nervous system through various mechanisms, affecting gamma-aminobutyric acid (GABA)-gated chloride channels, inhibitors of acetylcholine-esterase (AChEs), octopamine tyramine receptors, voltage-gated sodium channels, glutamate gated chloride channels (Glu-Cl), and GABA-receptors, respectively . DiffereRhipicephalus annulatus, R. microplus, and Rhipicephalus australis undergo rapid selection in response to selective pressure and higher concentration of acaricides  in a population of ticks selected as a result of the population’s interaction with an acaricide”. The number of ticks that remain alive after exposure to the specific applied concentration of acaricides increases significantly . The mutThree main forms of acaricide resistance are generally known Figure 1R. microplus as early as 1960 in Australia , developed in 1962, uses acaricide-treated filter paper packets to incubate the tick larvae . This teid ticks .R. annulatus and Haemaphysalis bispinosa engorged female ticks , and compounds related to hexa-chlorocyclohexane such as lindane . Organocle ticks . Resistacumented . Organoccumented . This licumented .R. microplus genes targeted by or associated with specific acaricides have been identified. Mutations related to OCs use in R. microplus were different from the observed for certain insects, however they were located in the same second transmembrane protein domain of the GABA-gated channel (TM-2). The effect of this mutation was unknown, but the modified TM-2 may alter the channel properties, inhibiting the entry of chloride ions into the nerve  have been recorded (R. microplus is attributed to increased esterase enzyme activity (R. microplus (R. microplus (strain Tuxpan) were experimentally obtained after the application of OP over seven successive generations , necessary for nervous system functions in  for OPs Figure 3 for OPs . In the  for OPs .R. microplus, resulting in the overexpression of esterases in larvae as well as in adult R. microplus, and consequent resistance to OPs  gene, and metabolic detoxification . Organope to OPs . Even the to OPs . Post-tre to OPs .R. microplus ticks: BmAChE-1 . Synthetic pyrethroids comprised about 17% of the total acaricides used at the beginning of the 21st century  and cause membrane depolarization. The closure of voltage-gated sodium channels for just a few milliseconds after channel activation (fast inactivation) is responsible for the action potential’s falling phase . The mecType-I SPs cause the repetitive release of sodium ions in response to even a single stimulus, while SPs type-II cause membrane depolarization by interfering with membrane action potential. Type-II pyrethroids also delay the inactivation of voltage-gated sodium channels for longer periods compared with type-I SPs .R. microplus, with polymorphisms being identified in two of the four domains (domain-II and domain-III) as shown in Synthetic pyrethroids resistance is linked to mutations in voltage-gated sodium channel genes of R. microplus . Different studies  were also found to be resistant to fipronil , fipronil acts by blocking GABA-receptors , and inhR. microplus ticks  and third transmembrane (TM-3) domain of the GABA-gated chloride channel gene in fipronil resistant us ticks   than any other used acaricides against elopment . In a laus ticks .R. sanguineus nymphs, structures that play essential roles in tick survival  and SPs (cypermethrin)  and fipronil has been reported, which affects the GABA-gated chloride channels of many insects . The seliculatus . It has methrin) . Cross-rus ticks . Cross-re target . Rhipicearicides .Strategies must be developed to minimize the selection of acaricide resistance in ticks. Different acaricides have different modes of action, thus alternate use may help in the reduction of resistance selection. Rotation of acaricides such as pyrethroids with coumaphos (OPs) has been used, and rotation of amitraz with other groups of acaricides may delay a further spread of amitraz resistance . The altDermacentor reticulatus , and the CNPq and CAPES – Brazil for providing research facilities. This work was supported by JSPS KAKENHI Grant Number JP22H02522.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.


O-glycans, sialylation, fucosylation, and complex branched structures have been found to be the most common structural patterns in malignant tumors. In recent years, immunochemical methods, lectin recognition-based methods, mass spectrometry (MS)-related methods, and fluorescence imaging-based in situ methods have greatly promoted the discovery and application potentials of glycomic and glycoprotein biomarkers in various cancers. In particular, MS-based proteomics has significantly facilitated the comprehensive research of extracellular glycoproteins, increasing our understanding of their critical roles in regulating cellular activities. Predictive, preventive and personalized medicine  is an effective approach of early prediction, prevention and personalized treatment for different patients, and it is known as the new direction of medical development in the 21st century and represents the ultimate goal and highest stage of medical development. Glycosylation has been revealed to have new diagnostic, prognostic, and even therapeutic potentials. The purpose of glycosylation analysis and utilization of biology is to make a fundamental change in health care and medical practice, so as to lead medical research and practice into a new era of 3P medicine.Glycosylation is one of the most important post-translational modifications (PTMs) in a protein, and is the most abundant and diverse biopolymer in nature. Glycans are involved in multiple biological processes of cancer initiation and progression, including cell-cell interactions, cell-extracellular matrix interactions, tumor invasion and metastasis, tumor angiogenesis, and immune regulation. As an important biomarker, tumor-associated glycosylation changes have been extensively studied. This article reviews recent advances in glycosylation-based biomarker research, which is useful for cancer diagnosis and prognostic assessment. Truncated  N-linked and O-linked glycosylations according to the modification site. N-glycosylation covalently modifies N-acetylglucosamine (GlcNAc) to the nitrogen atom on the side chain of asparagine (Asn). O-glycosylation covalently modifies N-acetylgalactosamine  to the oxygen atom of a serine (Ser) or threonine (Thr) residue  are chemical modifications of proteins during or after translation , 2, whic residue , 9. Glyc residue . Glycosy residue . GlycosyO-GlcNAcylation, occur in the endoplasmic reticulum (ER) and the lumen of the Golgi apparatus middle 3 core, with the addition or removal of other monosaccharides  Gltability . In additability  and promtability . These hO-linked glycan , T , and Sia-Tn   . Truncat gastric , pancrea gastric , ovarian gastric , bladder gastric , and col gastric . STn thaN-glycans on neural cell adhesion molecule 1 (NCAM1)  . The abn (NCAM1) , breast  (NCAM1) , and neu (NCAM1) , and is  (NCAM1) . The inc (NCAM1) –42. Gang (NCAM1) , lung ca (NCAM1) , cervica (NCAM1) , and breFucosylation plays an important role in tumor pathology, including regulation of signaling pathways and tumor metastasis . Aberranin situ analysis methods. These studies not only focus on the analysis of glycosylation, but also reveal the regulatory mechanism of glycosylation.Since tumor-associated glycosylation alterations are a distinct feature of cancer diagnosis and prognosis, a great deal of effort has been devoted to identify, label, and characterize glycosylation in recent years. Four types of glycosylation research tools have been developed, including immunochemical methods, lectin recognition-based methods, mass spectrometry (MS)-related methods, and fluorescence imaging-based Immunochemical methods mainly include enzyme-linked immunosorbent assay (ELISA) and western blotting (WB). ELISA uses specific antibodies to specifically recognize and quantify glycans/glycosylated proteins of interest . ELISA iin situ tracking of glycan signatures on surfaces; for example, Huang evaluated and compared the glycan structures among 64 cell lines, 37 tissues, and primary colon tumor tissues with 19 fluorescently conjugated lectins  to pre-separate complex mixtures of glycans (N-glycans are released from glycopeptides by peptide-N-glycosidase F (PNGase F) digestion  technology, was first developed by Wollscheid et al. in 2009 for large-scale analysis of surface glycoproteins , possibly due to the large number of sample processing steps resulting in severe loss of samples. Cell surface protein isolation protocols were also optimized to increase cell surface protein coverage. With a special pipette tip, the new workflow is suitable for very small numbers of cells, 10 times less than traditional CSC methods. A total of ~600 cell surface-associated proteins were identified from 1105 cells alone  was compared, and the results showed that GalNAz labelled more cell surface protein glycosylation sites than GlcNAz or ManNAz . Not onlifferent . A compregulated . Furtheregulated . Brieflyegulated , 106. Hoproteins . GlycoprN- and O-glycans, respectively. SEEL in combination with MS identified 37% more sialylated proteins than metabolic labeling methiod. This SEEL study compared the levels of sialylated proteins in undifferentiated vs. differentiated human erythroleukemia cells (HEL), and found that differentiated cells had more N-linked sialylated proteins , a membrane-impermeable crosslinker, was first used to covalently crosslink surface glycoproteins and their interacting proteins. For the extraction of glycoproteins, a strategy similar to the CSC technique was adopted. Galactose oxidase was used to oxidize glycans on surface glycoproteins, which were then enriched for surface glycoproteins and their interactors by hydrazine chemistry, followed by quantitative proteomics. As a result, it identified more than 300 proteins interacting with surface glycoproteins, and the glycoprotein interaction network was constructed  labeling identified these interacting glycoproteins with high sensitivity. At the same time, the glycoprotein interacting with galectin-3 was further verified by in vitro experiments such as WB  MS analysis does not require antibodies that are expensive, cumbersome to obtain, and limited in types. Moreover, the performance of antibodies varies to affect the test results. (ii) MS does not require prior knowledge of the type of protein to be studied. WB, immunoprecipitatioin (IP), and other methods commonly used in traditional biology require pre-judgment of the proteins to be analyzed. MS is a common technique in analysis of proteins. Without prior knowledge of proteins, MS enables large-scale marker screening in complex biological samples. (iii) The advent of tagging methods, such as iTRAQ and TMT labeling, can efficiently perform deep quantification of biomarkers, which has an essential feature in comparison of multiple samples. (iv) Perhaps, the most significant advantage of MS-based methods is able to obtain the detailed structural information on glycan structures and to identify glycosylation sites, which might directly impact cell function. In recent years, the combination of chemical biology and MS has made an indelible contribution to the research of glycosylation at different specificity levels, which deepens one’s understanding of the synergistic regulation of cellular protein activities. We believe that it is the focus and direction of future research Table 2.in vitro research of glycoglycan structure and function, they cannot provide in situ real-time qualitative or quantitative information on glycoglycans on cells, especially the spatial distribution information. Moreover, the complex cleavage and separation process prior to MS analysis might lead to unpredictable loss of glycan information  was conjugated to an azide-modified metabolic sugar precursor, where specific sites of the azide sugar are blocked from being taken up by cells . When careaction . Howeverhemistry . This stc manner . Such stc manner . To avoic manner . Furtherc manner .O-GalNAc as an internal control  is the main method to analyze protein-specific glycans , 124. Me control .The FRET strategy needs to use two different fluorophores that can undergo fluorescence resonance energy transfer when used for protein-specific glycoform imaging. It is very difficult to obtain two pairs of fluorescent donor-acceptor pairs that do not interfere with each other at the same time, so it is difficult for this strategy to simultaneously image multiple glycans in a specific protein. This problem was addressed by designing a hierarchical coding strategy. DNA sequences were used to encode proteins and different classes of glycans. Decoding process started with the addition of a “time code” that exposed the “protein code”. Exposed “protein code” was hybridized with a hairpin that was covalently modified on the sugar, opening the hairpin and exposing the “monosaccharide code”. The clever introduction of “time encoding” in this strategy strictly distinguishes the encoding and decoding process, and the decoding event can be started at any time . A similLCM strategy was proposed to remodel Gal/GalNAcon-specific proteins. Galactose oxidase activity was inhibited in the presence of potassium ferrocyanide, and galactose oxidase activity was “on” when potassium ferricyanide was added. This switch enabled the specific oxidation of Gal/GalNAc at the end of MUC1 on the cell surface. Bioorthogonal labeling was then performed for the purpose of localized glycan analysis . DNA tecIn summary, it clelarly demonstrates that the current protein-specific glycoform imaging strategies need to use a “gating” guarantee mechanism. For example, the FRET strategy relies on distance as the “gating”, the LCM strategy uses ions as the “gating”, and the HieCo strategy designs DNA as the “gating”. These strategies are cleverly designed, but also suffer from disadvantages such as limited generalizability (LCM) and complex design (HieCo). Therefore, a fundamental shift in detection mode is also required in the future to achieve highly sensitive protein-specific glycoform imaging with a simplified “gated-free” design Table 2.As previously discussed, tumor-associated glycans/glycoproteins have been widely used as biomarkers for clinical diagnosis and prognostic assessment of patients . The abnin vitro and in vivo antitumor activity. As a common tumor biomarker, GD2 antibody was also used in the treatment of neuroblastoma (in vivo.(i) tumor suppressive effects of glycan-binding molecules –138. Sinblastoma . It was in vivo was first explored, and the antibody-drug conjugate was further applied to Tn-positive tumor cells in vitro, showing effective cytotoxicity, and the cytotoxicity was positively correlated with the expression level of Tn in the tumor. Antibody-drug conjugates also showed potent antitumor activity in vivo. The study demonstrated for the first time the effectiveness of Tn antibodies as antibody-drug conjugates. Further, Tn antibody was used to provide targeting effect for the drug-encapsulated nanocapsules, the encapsulation efficiency of nanocapsules for antitumor drugs reached 99.9% and the biotin-avidin method was used to attach antibodies to nanocapsules  Targeting function of glycan-binding molecules –149. Glycapsules . In vitrc target . NK cellc target . A novelc target . In addic target . The MUCc target . Bispecic target . Janus lc target . Boronicc target . MonocloAltered glycosylation is a key change in tumors, and aberrant glycosylation is a component of tumor growth, survival, metastasis, and immune evasion . TargetiImmune checkpoint inhibitors, such as anti-PD1 and anti-CTLA-4 blocking antibodies, were widely used in tumor therapy and improved long-term survival of patients , 154. Hoin situ analysis methods have been developed. Immunochemical methods have high sensitivity and specificity, but are overly antibody-dependent and prone to false positives. Lectin microarray is a fast, sensitive, high-throughput glycan analysis technique suitable for studies with large sample sizes. Immunochemical methods, lectin recognition-based methods, and fluorescence imaging-based in situ analysis methods all require prior knowledge of the protein being analyzed, which is disadvantageous for 3P medicine PPPM , the Shandong First Medical University Talent Introduction Funds (to XZ), Shandong First Medical University High-level Scientific Research Achievement Cultivation Funding Program (to XZ), the Shandong Provincial Natural Science Foundation (ZR202103020356 or ZR2021MH156 to XZ), and the Academic Promotion Program of Shandong First Medical University (2019ZL002).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.


Olfactory dysfunction is one of the biomarkers for Alzheimer’s disease (AD) diagnosis and progression. Deficits with odor identification and discrimination are common symptoms of pre-clinical AD, preceding severe memory disorder observed in advanced stages. As a result, understanding mechanisms of olfactory impairment is a major focus in both human studies and animal models of AD. Pretangle tau, a precursor to tau tangles, is first observed in the locus coeruleus (LC). In a recent animal model, LC pretangle tau leads to LC fiber degeneration in the piriform cortex (PC), a cortical area associated with olfactory dysfunction in both human AD and rodent models. Here, we review the role of LC-sourced NE in modulation of PC activity and suggest mechanisms by which pretangle tau-mediated LC dysfunction may impact olfactory processing in preclinical stage of AD. Understanding mechanisms of early olfactory impairment in AD may provide a critical window for detection and intervention of disease progression. Olfactory dysfunction is one of the earliest signs of Alzheimer’s Disease (AD), appearing long before clinical memory symptoms, at which point the disease has already progressed to late stages when intervention is difficult . LongituImpairment at all levels of the olfactory circuit have been implicated in AD development, from olfactory epithelium, nerve, olfactory bulb to olfactory cortex . Import Recently, we presented novel results from a pretangle tau rat model and a new view that dysfunction of norepinephrine (NE) regulation in the PC may underlie preclinical stage olfactory deficiency . In the In mammals, the PC is central to olfactory information processing and is situated in the ventrolateral forebrain. It receives afferent input from the OB, rendering it as a critical site for odor discrimination, contextualization, and associative memory formation . Locatedex vivo slices can produce an N-methyl-D-aspartate receptor (NMDAR) dependent long-term potentiation (LTP) in pyramidal neurons, which is suggested to be essential for olfactory-based learning  within the pyramidal cell layer (II)  . NE signo slices . Computao slices .in vivo studies support the role of LC-NE in a diverse range of olfactory behaviors. Fitting with improving the signal-to-noise ratio, blocking α and β adrenergic receptors within the OB impaired similar odor discrimination in awake-behaving animals , the pretangle tau pathology progressed faster. At 5–6 months post-infusion, LC cells started to reduce, correlating with a deficiency in a simpler version of the odor learning -N-ethyl-bromo-benzylamine (DSP4) also weakened olfactory discrimination ability and short-term memory in the APP/PS1 model . LTCCs are downstream of β-adrenergic receptors, via a Gormation . Recentlormation . Moreoveormation  and coulormation . The incormation . LTCCs cormation . A decreormation . On the ormation , may cauormation . Such dyormation  and may Neurodegeneration of the LC and dysregulated noradrenergic function correlate with AD progression and olfactory dysfunction during preclinical stages. Further exploration of cellular and molecular interactions within these regions may help to characterize preclinical symptoms during this critical window. Future perspectives will require an understanding of AD pathology on multi-regional (OB and PC) extracellular NE, noradrenergic receptor expression, and should involve the interaction of both Aβ and tau pathologies on aging individuals.Preserving LC neuronal health and function could be key to prevent or reverse AD. Promoting phasic LC firing patterns and reducing stress-inducing high tonic activity appear to be beneficial in animal models . Life exVR and QY contributed equally to the conceptualization, wrote the manuscript, and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.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.


To investigate the different impacts on clinical outcomes between regular recall and non-regular recall among incident peritoneal dialysis (PD) patients.A two-center cohort of 216 new PD patients from 1January 2013, to 31 December 2014, was studied. Informative clinical data were collected from baseline until two years after PD initiation, including demographics, laboratory and PD-related parameters, PD-related peritonitis rates, and frequency of hospitalization. Regular in-person recall (RPR) was defined as having a one-month interval and non-regular in-person recall (NRPR) as an interval ranging from more than one month to less than three months.p = .049). PD-related peritonitis rate was higher in the NRPR vs. RPR cohorts . PD-related hospitalization frequency was also higher in the NRPR cohort  over two years. Kt/V values in the NRPR cohort gradually decreased over two years and were at lower levels than in the RPR cohort.Percentage of patients with peritonitis was significantly higher among patients in the NRPR group than among those in the RPR group (27.7% vs. 16.5%, New PD patients with NRPR showed higher rates of PD-related peritonitis and hospitalization frequency than patients with RPR. The incidence of end-stage renal disease is increasing annually, and it is emerging as a critical health issue. Peritoneal dialysis (PD) is one of the two main modalities for renal replacement therapy in end-stage renal disease and is used by approximately 11% of the dialysis population worldwide . In PD sPrior investigations on noncompliance to treatment have commonly focused on the hemodialysis population. Noncompliance in PD patients has been less reported. Studies on compliance of PD patients are based on three aspects of treatment: noncompliance with dialysis procedures/exchanges, medications, and diet/fluid restrictions . The majAlthough some variables were identified as affecting noncompliance, there was almost no consensus among the observed association studies . In one Study design was a retrospective medical information review over two-year study period. New patients diagnosed with end-stage renal disease who initiated PD therapy in an outpatient clinic and no prior hemodialysis were selected for analysis. Patients were recruited from two hospital-facilitated PD centers, Tianjin First Center Hospital in China and Kaohsiung Chang Gung Memorial Hospital in Taiwan, from 1 January 2013, to 31 December 2014. All participants were followed up in the outpatient clinic of their respective hospitals for two years until PD termination, transfer to hemodialysis, receiving kidney transplantation or death.Analytic information included demographics, laboratory, and PD-related parameters and clinical outcomes. Laboratory data and the parameters of standard peritoneal equilibration tests (PET) were generally collected at 1, 12, and 24 months after PD initiation. Data on PD-related parameters were collected from PD home records and PET, including urine output, dialysis adequacy, residual renal function estimated by creatinine clearance, and glucose exposure amount in PD solution dwell. All hemograms and biochemical samples were measured using commercial kits and an autoanalyzer. Albumin levels were measured using the bromocresol green method. Clinical outcomes included drop outs, PD-related peritonitis, and hospitalization. PD-related hospitalizations were defined as PD-related peritonitis, catheter-related infection, catheter displacement, inadequate PD, and fluid overload. PD dropout was defined by PD termination caused by death, ultrafiltration failure, failed PD-related peritonitis treatment, or hemodialysis shifting. Technique failure was defined by PD termination caused by ultrafiltration failure, treatment failure for PD-related peritonitis, or hemodialysis shifting.Supplemental Table S1).The content of follow-up care pathway in PD patients is followed by International Society of PD (ISPD) guidelines  in studyIn practice, PD patients were recalled to PD units at one-month intervals in study hospitals. RPR in an outpatient clinic was defined as a visit involving face-to-face interviews between nephrologists/PD nurses and patients at one-month intervals and NRPR as visits at intervals ranging from more than one month to less than three months. Non-adherence to outpatient recall was defined when patients were unwilling to abide by the rule of one-month interval recall.t-test, chi-squared test, binomial probability test, or Fisher’s exact test . The changes of baseline laboratory measurements within the two groups were estimated using one-way repeated measures ANOVA . The association between the PET-related parameters and visiting frequency in the study period was analyzed using an independent two-sample t-test to determine the difference between the groups , median (interquartile range), and frequency (percentage)  and 2. Des ANOVA . Differee groups . The valectively . The valn groups .t-test (Supplementary Table S2). PD mode changes in patients with incremental PD were estimated by using McNemar’s test (Supplementary Table S3).The clinical outcomes in participants were summarized and presented as frequency (percentage) . The 95% confidence interval (CI) and p-value were used to determine statistical significance. A p value less than 0.05 was considered statistically significant. All statistical analyses were conducted using STATable TA version 11.1 .Cox proportional hazard regression with time-varying covariates were used to derive adjusted hazard ratios (HR) for peritonitis . Comparin = 28), non-adherence for personal preference (n = 22), distance from the hospital (n = 17), disease episodes on follow-up days (n = 13), and financial reasons for traffic transportation (n = 3). Among social features, a higher percentage of smoking was identified in patients with NRPR than in those with RPR (p = .001). Other features including age, sex, body mass index, education, status of employment, and caregiver/marital status were not significantly different between the two groups. Patients in the NRPR group showed a higher percentage of comorbidities with hypertension, coronary artery disease, and cerebral stroke compared with those in the RPR group. Among clinical outcomes, percentage of patients with PD-related peritonitis was significantly higher in the NRPR group than in patients in the RPR group , whereas percentage of drop outs and all-cause hospitalization was not higher in the NRPR group. PD-related peritonitis rate was higher in the NRPR group than the RPR group . Frequency of PD-related hospitalizations was significantly higher in the NRPR group than in the RPR group , whereas not significant in all-cause hospitalizations over two years  and Online Resource 3 , respectively. There were no significant differences in rates of drop out, PD-related peritonitis, and hospitalization based on stratification of missed times of recall in outpatient clinic over the study period.A total of 216 new patients underwent PD were identified for analysis, with 133 patients in the RPR and 83 patients in the NRPR group. Visit interval was one month in the RPR group, and mean visit interval was 1.45 ± 0.35 months  in the NRPR group. The mean age was 53.9 years in the RPR group and 50.3 years in the NRPR group. Sex showed equal distribution in the two groups. The causes of NRPR in the outpatient clinic were personal events on follow-up days (wo years . Detailep < .001). For blood urea nitrogen measurements, the NRPR cohort showed temporary changes in blood levels, whereas the RPR cohort showed a decreasing tendency in blood levels (p = .124). Serum creatinine levels showed significant increases in both groups over two years (p < .001). Similar changes were also observed in serum calcium and phosphate levels in the RPR group (p < .001).Serial laboratory data over two years and comparison between RPR group and NRPR group are shown in p < 0001) group with tendency through two years. The detailed information in PET are shown in Online Resource 4 . Glucose exposure showed an increasing trend over two years in patients in the NRPR group, whereas patients in the RPR group maintained a relatively constant exposure amount. Dialysis adequacy index (Kt/V) showed a decreasing trend in the NRPR group but not in the RPR group. Daily urine output and total weekly creatinine clearance showed significantly decreasing trends in both cohorts, as shown in Regarding PD modes, more patients in the NRPR group used automated PD at baseline than those in the RPR , male sex , primary diabetic kidney disease , and time-varying serum albumin levels  were associated with a reduced risk for peritonitis. In contrast, age  and time-varying hemoglobin levels  were associated with an increased risk for peritonitis, as shown in In analyzing the risk for peritonitis over two years by fully adjusted model, RPR .Clinical outcomes in study patients are summarized in Supplementary Tables S6–S8). Results showed non-significant differences in demographic parameters between smokers and nonsmokers except alcohol drinking and hypertension comorbidity. Smokers demonstrated significantly higher proportion in alcohol drinking and hypertension comorbidity compared to nonsmokers. The comparative results between smokers and nonsmokers in laboratory measurement and PET-related parameters demonstrated that both subgroups had different distribution in blood hemoglobin (Hb), urea nitrogen (BUN), calcium (Ca), sodium (Na), intact parathyroid hormone (iPTH), Kt/V , and weekly creatinine clearance (Ccr) at different time point, while the alteration of these parameters between both subgroups over time did not show significant difference.The comparison results between smokers and nonsmokers were summarized in Online Resource 6–8 (The results of comparison between RPR and NRPR in our study revealed higher percentages of hypertension, coronary artery disease, cerebral stroke, and smoking in the NRPR cohort. This observation implied that physical disability owing to comorbidities in PD patients could interrupt their regular schedule for follow-up in outpatient clinic. To compensate for this inconvenience, a remote monitoring system is likely to promote health management ,17. DemoV demonstrated a gradual decrease over two years in NRPR subjects. Therefore, inadequate dialysis may not have contributed to the relatively constant serum phosphate concentration over two years in the NRPR subjects. Overall, PD patients with NRPR showed lower concentrations in serum albumin, urea nitrogen, and calcium in our study. This result implied that subjects in the NRPR group were not receiving adequate nourishment. Participants in NRPR group also demonstrated lower serum creatinine levels in 12-month and 24-month period compared to these levels in the participants in RPR group (We examined the changes in laboratory parameters in the present study. In the NRPR cohort, serum albumin showed relatively lower concentrations over two years. Serum calcium and creatinine concentrations showed a significant increase over two years, whereas hemoglobin and serum phosphate concentrations did not present significant changes over two years. Our findings disagreed slightly with those of previous studies. According to a cross-sectional self-report questionnaire study, PD patients with lower adherence rates exhibited higher serum creatinine and phosphate concentrations and lower serum albumin and hemoglobin concentrations compared with those with higher adherence rates . Serum pPR group . SimilarPR group . These rV values over two years. Moreover, the amount of daily glucose exposure showed an increased trend in this cohort. We also observed that more NRPR patients used automated PD. This is one of possibilities for the discrepancy between these two trends. Another possible reason was the uncontrolled fluid status in patients in the NRPR group and forward to exchange with a higher glucose-based PD solution. Nevertheless, this assumption needs to be validated by further study. However, the PD population in our study in both the NRPR and the RPR groups achieved target adequacy index in PD based on clinical practice guidelines [In past decades, noncompliant PD patients were commonly reported in skipped or missed PD solution exchanges, which resulted in inadequate dialysis ,6. In thidelines . In otheV values and renal creatinine clearance rate. However, we did not measure the missed exchange rate in our patients. Therefore, the definitive causes of inadequate dialysis cannot be ascertained in our study. Similarly, the exact reasons for nearly equal distribution of all-cause hospitalization frequency and drop out percentage between the two cohorts also were obscure in our study.Noncompliance in PD therapy often increased the incidence of PD-related peritonitis and hospitalization ,6,20,24.The present study was subject to several limitations. First, the definition of non-adherence to outpatient follow-upprotocol was arbitrary. Thus, the beneficial effect of a more rigid schedule in an outpatient clinic may be neglected. Further, a cause–effect relationship might not be clearly delineated in our study. Secondly, participants were recruited from two hospitals. Therefore, the influence from hospital-specific effects cannot be eliminated. For example, manpower in individual PD unit, varied routine laboratory measurements, relationship between medical personnel and patients, local cultural characteristics, and reimbursement policy. Thirdly, several aspects of non-adherence to PD therapeutic measures were not examined in our study, e.g., psychological burden, cognitive skills, informative supporting system for patients, and detailed information for PD exchanges. Although staff assumed these circumstances would not associate with non-adherence in PD patients, patients often felt well in the short-run. Therefore, we assumed that these circumstances may not play a greater role in this two-year observational study. Finally, we cannot refute the possibility of protocol deviation in PD home care procedures in our participants. In addition, invasive procedure and infectious diseases could increase the risk of PD-related peritonitis. We reviewed again the assumed causes of PD-related peritonitis in the participants. Majority are related to ignore hygiene procedure in PD solution exchanges. Rare cases were considered the possibility linking to invasive procedures or indolent infection. Accordingly, we suppose that aforementioned events would not influence our final results. A few strengths were identified in the present study. First, this is the first report to examine the influence of PD outcomes using non-adherence outpatient follow-up protocol among incident PD patients. Secondly, our study evaluated the trajectory of laboratory and PD-related parameters over two years. The data provided a real-world scenario of patients after PD initiation. Given the findings, medical staff have gained insight for the necessity of outpatient follow-up in new PD patients. Thirdly, our study demonstrated that smoking and comorbidities associated the motivation to adhere to outpatient follow-up protocols in PD patients. It is not certain if change in lifestyle behavior and social-support provision would enhance adherence. The ultimate challenge will be to develop a suitable intervention to overcome these barriers. Finally, telemedicine has been reported an alternative to ensure compliance in the medical care especially in the COVID pandemic period. The benefit also include reducing the risk for exposure to infectious organisms. Our study may elicit an emerging medicine to apply in the PD patients with high frequency NRPR.Non-adherence to regular outpatient follow-up could impact clinical outcomes in new PD patients. Smoking and comorbidities contributed to barriers for adherence to regular outpatient follow-up. Consequently, rates of PD-related peritonitis and hospitalization were higher among patients with non-adherence to regular outpatient follow-up. It is noteworthy that an effective strategy and monitoring procedure for recall adherence is essential for outcome improvement in PD patients.Click here for additional data file.