The impact of various meditation interventions on modifying human health outcomes has been demonstrated at both cognitive and cellular levels. Preksha Dhyana (PD) meditation has previously shown effects on cognitive skills, transcriptome, and the DNA methylome in healthy, novice college students. In this study, we performed a cross-platform metabolomic and lipidomic analysis of serum samples collected from 38 healthy, meditation-naïve participants and five age-matched participants who served as controls. Serum samples were collected from both groups, at baseline (pre-meditation) and after an 8-week PD intervention. Analysis was conducted using capillary electrophoresis and reversed-phase liquid chromatography coupled to electrospray ionization–mass spectrometry, and higher serum concentration levels for four circulating metabolites, namely, hypoxanthine, oxoproline, choline, and cystine, were identified along with a series of lysophosphatidylcholine (LPC) and lysophosphatidylethanolamine (LPE) species (LPC 20:0, LPC 18:0, LPC 18:1, LPC 16:0, LPE 18:0, LPE 18:1, and LPE 22:6), following the PD intervention that satisfied a Bonferroni correction (p < 0.05, FDR). Pathway enrichment analysis revealed that these metabolites and lipids are primarily associated with the purine degradation cycle, glycerophospholipid metabolism, and glutathione metabolism, which are critical for maintaining cellular energy balance, preserving membrane integrity, and protecting against oxidative stress. A multi-omics analysis with Data Integration Analysis for Biomarker discovery using Latent approaches for Omics studies (DIABLO) revealed positive and negative correlations between these metabolites/lipids, DNA methylated sites (DMSs), and cognitive outcomes (r > 0.5). Overall, our findings suggest that the wellbeing experienced after PD meditation might be associated with bioactive metabolites and lipids, which may also function as epigenetic modifiers regulating gene expression and biological pathways.Clinical Trial RegistrationClinicalTrials.gov, Identifier NCT03779269.

ObjectivesCeliac disease (CeD) is an autoimmune enteropathy triggered by gluten in genetically predisposed individuals, causing intestinal damage and impaired gut barrier integrity. Current diagnostic methods, such as serology and intestinal biopsy, are costly and invasive. Intestinal fatty acid binding protein (FABP2), exclusively expressed in the cytosol of mature enterocytes and released during injury, offers a potential non-invasive alternative biomarker. This study aimed to evaluate urinary FABP2 levels in pediatric patients undergoing gastrointestinal assessment or CeD follow-up to determine its potential as a non-invasive biomarker of intestinal integrity.MethodsIn this retrospective cross-sectional study, urinary FABP2 was measured in 103 pediatric patients evaluated for intestinal permeability using the lactulose/mannitol test. Based on serology and symptomatology, patients were categorized as controls (n = 65), CeD on gluten-free diet (GFD) > 6 months (n = 26), or active CeD (n = 12). Associations between FABP2 levels, serology, symptoms, and diagnosis were analyzed.ResultsFABP2 levels were significantly elevated in active CeD patients (47.7 pg/mL) compared with controls (9.65 pg/mL) (p < 0.05), but not in CeD patients on GFD (15.0 pg/mL). A receiver operating characteristic curve analysis confirmed the diagnostic potential of FABP2 as a biomarker for celiac disease (area under the curve = 0.7269; cutoff = 23.7 pg/mL; sensitivity = 66.7%, specificity = 76.9%).ConclusionOur findings indicate that urinary FABP2 reflects intestinal epithelial injury and may serve as a non-invasive biomarker for assessing gut integrity and disease activity in CeD. However, larger prospective studies with histological correlation are needed to validate its diagnostic utility.

Common indications to evaluate exocrine pancreatic function in children include chronic diarrhea, steatorrhea, failure to thrive, cystic fibrosis and those with chronic abdominal pain due to chronic pancreatitis where imaging studies are normal [1]. Exocrine Pancreatic Insufficiency (EPI) has a spectrum of severity. In children often remains an underdiagnosed condition, particularly in its mild, partial, and isolated enzyme deficiency forms. The purpose of this review is to help understand the different varieties of EPI including isolated pancreatic enzyme deficiencies as possible causes of malnutrition and growth failure in pediatric patients. Among the indirect diagnostic methods, the fecal elastase-1 (FE-1) testing is the most widely used one. While it has good sensitivity and specificity in severe pancreatic damage, like cystic fibrosis in children, its performance in the diagnosis of mild, partial, and isolated enzyme deficiencies is poor. Direct pancreatic function testing performed during endoscopy (ePFT), has emerged as a more sensitive and specific method for assessing all forms of exocrine pancreatic function. Notably, recent guidelines from the North American Society for Pediatric Gastroenterology, Hepatology, and Nutrition (NASPGHAN) emphasize the importance of ePFT in pediatric patients. Most of the pediatric practitioners taught that the pancreas has only two diseases, cystic fibrosis and pancreatitis. They are missing the fact that pancreas, like other digestive organs, can have different, many times secondary, dysfunctions that influence the growth of children. Most pediatric gastroenterologists still use the fecal elastase-1 (FE-1) test, however, this lacks sufficient specificity and sensitivity [2–5] especially in patients with mild or early pancreatic disease or those with isolated enzyme deficiencies [5]. The most accurate diagnostic modality to explore these conditions is ePFT. In this review we highlighted the critical importance of direct pancreatic function testing. Enhancing clinical awareness and incorporating direct testing methods can ultimately improve outcomes for affected children.

OBJECTIVES:Exocrine pancreatic insufficiency (EPI) results from insufficient pancreatic enzyme and bicarbonate production, impairing digestion and causing malnutrition. Limited studies have been conducted to fully understand the underlying mechanisms contributing to pediatric EPI. The aim of this study was to measure interleukin-8 (IL-8) levels in pancreatic fluid of EPI patients to determine if IL-8 could be an inflammatory mediator in pediatric EPI. METHODS:In this retrospective study, IL-8 protein levels were measured in pancreatic fluid collected from 193 pediatric patients evaluated for EPI. Based on the endoscopic pancreatic function test (ePFT) results, patients were categorized into normal and EPI groups, and statistical analyses explored associations between IL-8, enzyme deficiency, symptoms, and clinical diagnosis. RESULTS:IL-8 levels were elevated in EPI patients (30%), versus normal group (6%) (p < 0.001). Symptoms significantly associated with EPI were constipation, diarrhea and weight loss. However, it is important to note that many of these symptoms overlapped with other symptoms. No significant association was observed between IL-8 and the disease subgroups, except for disaccharidase deficiency. Overall, there was a strong association of IL-8 with pancreatic enzyme deficiency, suggesting its role in EPI pathogenesis. CONCLUSION:This is the first study to measure IL-8 levels in pancreatic fluid of a large cohort of pediatric EPI patients. The elevated IL-8 levels suggest its potential role as an inflammatory mediator in pediatric EPI. Furthermore, these findings open avenues for investigating other inflammatory factors that may contribute to EPI development.

Genetic sucrase-isomaltase deficiency (GSID) is an inherited deficiency in the ability to digest sucrose and potentially starch due to mutations in the sucrase-isomaltase (SI) gene. Congenital sucrase-isomaltase deficiency is historically considered to be a rare condition affecting infants with chronic diarrhea as exposure to dietary sucrose begins. Growing evidence suggests that individuals with SI variants may present later in life, with symptoms overlapping with those of irritable bowel syndrome. The presence of SI genetic variants may, either alone or in combination, affect enzyme activity and lead to symptoms of different severity. As such, a more appropriate term for this inherited condition is GSID, with a recognition of a spectrum of severity and onset of presentation. Currently, disaccharidase assay on duodenal mucosal tissue homogenates is the gold standard in diagnosing SI deficiency. A deficiency in the SI enzyme can be present at birth (genetic) or acquired later, often in association with damage to the enteric brush-border membrane. Other noninvasive diagnostic alternatives such as sucrose breath tests may be useful but require further validation. Management of GSID is based on sucrose and potentially starch restriction tailored to the individual patients' tolerance and symptoms. As this approach may be challenging, additional treatment with commercially available sacrosidase is available. However, some patients may require continued starch restriction. Further research is needed to clarify the true prevalence of SI deficiency, the pathobiology of single SI heterozygous mutations, and to define optimal diagnostic and treatment algorithms in the pediatric population.

BACKGROUND:Recurrent upper endoscopies are essential for monitoring therapy response and disease activity in patients with eosinophilic esophagitis (EoE), leading to increased costs, procedural complications, and anesthesia exposure. The aim of this study was to examine an office-based model using serial sedation-free blind esophageal epithelial brushing (BEEB) to monitor therapy response through eosinophil-derived neurotoxin (EDN) levels and guide therapy plans in pediatric EoE patients. METHODS:EoE patients (≤21 years of age) were enrolled in this prospective study. Subjects were placed on dietary, pharmacologic, or combination therapy with the goal of inducing or maintaining remission. To assess response to sequential interventions, subjects underwent sequential sedation-free BEEBs through nasogastric tubes to measure EDN levels. Based on serial brushings, an individual plan of diet, medications, or a combination of both was created for each subject, and a final endoscopy was then performed to validate the accuracy of the individual plans. RESULTS:Twenty-four subjects completed the study. The average peak eosinophil count in patients with active EoE was 58.1 ± 30.8 eosinophils per high-power field and mean EDN level was 165.2 ± 191.3 μg/mL. A total of 42 BEEBs were completed. Individual therapy plans based on sequential BEEB were accurate in 19 out of the 24 patients (79%) and specifically nine out of 10 patients (90%) treated with elimination diets. CONCLUSION:This study suggests that office-based sedation-free BEEBs can be used to monitor therapy response and disease activity in pediatric EoE patients.

OBJECTIVES:Eosinophil-derived neurotoxin (EDN) is a viable marker of eosinophilic esophagitis (EoE) disease activity. We studied the utility of measuring EDN from esophageal epithelial brushings for diagnosing EoE, focusing on two scenarios: (1) cases of exclusive distal eosinophilia and (2) cases of discrepancy between endoscopy and histology. METHODS:Records of patients who underwent esophagogastroduodenoscopy (EGD) with EDN measured via esophageal brushings at Arnold Palmer Hospital for Children in Orlando, Florida from January 2014 to October 2018 were retrospectively reviewed. Demographics, clinical, endoscopic, and histologic data were collected. RESULTS:We reviewed 231 patient records (66.7% male, mean age 10.3 years, range 1-22 years). EDN values correlated with endoscopic reference score (EREFS) and peak eosinophil count (PEC) (Spearman's rho = 0.756 (p < 0.001) and 0.824 (p < 0.001) respectively). Average PEC, EREFS, and EDN concentrations were higher in patients with active EoE than in controls or patients with EoE in remission (inactive). When grouping patients based on esophageal eosinophilia distribution, EDN mirrored PEC, and EREFS. Patients with exclusive distal eosinophilia had lower EDN concentrations than those with eosinophilia in >1 level of the esophagus (23.8 ± 46.1 mcg/mL vs. 171.3 ± 205.8 mcg/mL respectively, p < 0.001). EDN values were more consistent with EREFS in cases of discrepancies between endoscopic findings and pathology (p < 0.001). CONCLUSION:EDN measured in esophageal brushing samples reflects disease activity objectively and accurately. It also offers significant value in cases of exclusive distal esophageal eosinophilia and when discrepancies exist between endoscopy and histology.

Background and Objective: Aspiration is common in mechanically ventilated patients and may predispose patients to aspiration pneumonia, chemical pneumonitis, and chronic lung damage. Pepsin A is a specific marker of gastric fluid aspiration and is often detected in ventilated pediatric patients. We investigated the effect of oral care and throat suctioning in the detection of pepsin A in tracheal aspirates (TAs) up to 4 hours after these procedures. Methods: Twelve pediatric patients between age 2 weeks to 14 years who underwent intubation for cardiac surgery were enrolled in this study. Six of the 12 patients were consented before their surgery with initial specimen collected at the time of intubation and last one shortly before extubation (intubation duration < 24 hours). The remaining 6 patients were consented after cardiac surgery. All specimens were collected per routine care per respiratory therapy protocol and shortly before extubation (intubation duration > 24 hours). Tracheal fluid aspirates were collected every 4 to 12 hours in the ventilated patients. Enzymatic assay for gastric pepsin A and protein determination were performed. The time of oral care and throat suctioning within 4 hours prior was recorded prospectively. Results: A total of 342 TA specimens were obtained from the 12 intubated pediatric patients during their course of hospitalization; 287 (83.9%) showed detectable total pepsin (pepsin A and C) enzyme activity (> 6 ng/mL) and 176 (51.5%) samples had detectable pepsin A enzyme levels (>6 ng/mL of pepsin A). Only 29 samples of 76 samples (38.2%) had evidence of microaspiration after receiving oral care, while 147 of 266 (55.3%) samples were pepsin A positive when no oral care was provided. Odds ratio is 0.50 (Cl 0.30–0.84), and the number needed to treat is 5.8 (Confidence interval 3.4–22.3). Testing air filters for pepsin was not beneficial. Conclusion: Oral care is a highly effective measure to prevent microaspiration of gastric fluid in ventilated pediatric patients. The number needed to treat (5.8) suggests this is a very effective prevention strategy. Our study suggests that pepsin A is a useful and sensitive biomarker that allows identification of gastric aspiration.

Introduction: The stress and psychological factors affect the human transcriptomic and epigenomic landscapes. Preksha Dhyana meditation (PM) was found to be effective, in novice healthy college student meditators, at the cognitive skills and transcriptomic levels. Recently published data showed that PM induced alterations at the transcriptome level in healthy and novice college students.Methods: To decipher potential mechanisms underlying the PM effect at the cellular level, array-based methylation analyses in peripheral blood were performed at baseline and 8 weeks postintervention in 34 participants.Results: Overall, 470 CpG sites were nominally differentially methylated (p <= 0.05 and change magnitude from >= 3% to <= -3%) between baseline and 8 weeks postintervention with 180 sites hypermethylated and 290 sites hypomethylated. Pathway analysis of the genes linked to the differentially methylated sites revealed the enrichment of several molecular and cellular signaling pathways, especially metabolic and brain function signaling pathways.Conclusions: Besides its beneficial effects on cognitive skills and transcriptome alterations, the current data indicate that PM meditation also affects the DNA methylation profile of novice and healthy college students 8 weeks postintervention.Clinical Trial Registration number: NCT03779269.

The impact of different meditation protocols on human health is explored at the cognitive and cellular levels. Preksha Dhyana meditation has been observed to seemingly affect the cognitive performance, transcriptome, and methylome of healthy and novice participant practitioners. In this study, we performed correlation analyses to investigate the presence of any relationships in the changes in cognitive performance and DNA methylation in a group of college students practicing Preksha Dhyāna (N = 34). Nine factors of cognitive performance were assessed at baseline and 8 weeks postintervention timepoints in the participants. Statistically significant improvements were observed in six of the nine assessments, which were predominantly relating to memory and affect. Using Illumina 850 K microarray technology, 470 differentially methylated sites (DMS) were identified between the two timepoints (baseline and 8 weeks), using a threshold of p-value < 0.05 and methylation levels beyond −3% to 3% at every site. Correlation analysis between the changes in performance on each of the nine assessments and every DMS unveiled statistically significant positive and negative relationships at several of these sites. The identified DMS were in proximity of essential genes involved in signaling and other important metabolic processes. Interestingly, we identified a set of sites that can be considered as biomarkers for Preksha meditation improvements at the genome level.

The exocrine pancreas plays an important role in digestion. Understanding of the physiology and regulation of exocrine function provides insight into disease processes and basis of functional testing. Specifically, exocrine pancreatic insufficiency (EPI) can cause maldigestion and thus a proper assessment of exocrine pancreatic function is important. There are indirect and direct methods for evaluating pancreatic function. Indirect methods are varied and include stool, serum, urine, and breath tests. Fecal elastase is a commonly used indirect test today. Direct methods involve stimulated release of pancreatic fluid that is collected from the duodenum and analyzed for enzyme activity. The most used direct test today is the endoscopic pancreatic function test. Indirect pancreatic function testing is limited in identifying cases of mild to moderate EPI, and as such in these cases, direct testing has higher sensitivity and specificity in diagnosing EPI. This review provides a comprehensive guide to indirect and direct pancreatic function tests as well as an in-depth look at exocrine pancreatic function including anatomy, physiology, and regulatory mechanisms.

Background:The human transcriptome across a variety of cell types and tissues are affected by stress and other psychological factors. Preksha Dhyana meditation (PM) is effective at improving cognitive skills in novice healthy college student meditators after 8 weeks of intervention, but the molecular and cellular mechanisms involved in these improvements are still largely unknown.Methods:In order to decipher potential mechanisms at the cellular level, transcriptomic profiling analyses, from peripheral blood, were performed at baseline and 8 weeks post-intervention in 18-paired participants (RNASeq).Results:At the transcriptomic level, 494 genes were nominally differentially expressed (p-value ≤ 0.05) between baseline and 8 weeks post-intervention. Our data showed that 136 genes were upregulated, while 358 genes were downregulated. These genes were enriched in several cellular pathways including innate and adaptive immunity, cell signaling, and other metabolic processes.Conclusions:Overall, our findings indicate that PM meditation affects gene expression patterns from whole blood in novice healthy college students. Improvements at the cognitive skills were also mirrored with changes at RNA expression profiling.

Background: Functional Abdominal Pain disorders (FAPDs) are a group of heterogeneous gastrointestinal disorders with unclear pathophysiology. In children, FAPDs are more common in the winter months than summer months. The possible influence of school stressors has been proposed. Previously, our group showed differences in bacterial relative abundances and alpha diversity in the gut microbiome and its relationship with stressors in a cross-sectional evaluation of children suffering from FAPDs compared to a healthy control group. We present longitudinal data to assess whether the gut microbiome changes over school terms in the control and FAPDs groups. Methods: The longitudinal study included children with FAPDs (n = 28) and healthy controls (n = 54). Gastrointestinal symptoms, as well as stool microbiome, were assessed in both groups. Stool samples were serially collected from all participants during both the school term and summer vacation. The stool samples were subjected to total genomic extraction, 16S rRNA amplicon sequencing, and bioinformatics analysis. The gut microbiome was compared at school and during vacation. Other metrics, alpha diversity, and beta diversity, were also compared between the two school terms in every group. Results: In the healthy group, there were differences in microbiome composition between school terms and summer vacation. Conversely, we found no differences in the FAPDs group between the two terms. The healthy control group revealed differences (p-value < 0.05) in 55 bacterial species between the school term and vacation. Several of the differentially abundant identified bacteria were involved in short-chain fatty acids production (SCFAs), inflammation reduction, and gut homeostasis. Alpha diversity metrics, such as the Shannon index, were different in the control group and remained unchanged in the FAPDs group. Conclusion: Although preliminary, our findings suggest that the gut microbiome is static in FAPDs. This compares with a more dynamic healthy gut microbiome. Further studies are warranted to corroborate this and understand the interplay between stress, symptoms, and a less diverse and static microbiome. Future studies will also account for different variables such as diet and other patient demographic criteria that were missing in the current study.

The pancreas has both endocrine and exocrine function and plays an important role in digestion and glucose control. Understanding the development of the pancreas, grossly and microscopically, and the genetic factors regulating it provides further insight into clinical problems that arise when these processes fail. Animal models of development are known to have inherent issues when understanding human development. Therefore, in this review, we focus on human studies that have reported gross and microscopic development including acinar-, ductal-, and endocrine cells and the neural network. We review the genes and transcription factors involved in organ formation using data from animal models to bridge current understanding where necessary. We describe the development of exocrine function in the fetus and postnatally. A deeper review of the genes involved in pancreatic formation allows us to describe the development of the different groups (proteases, lipids, and amylase) of enzymes during fetal life and postnatally and describe the genetic defects. We discuss the constellation of gross anatomical, as well as microscopic defects that with genetic mutations lead to pancreatic insufficiency and disease states.

Introduction: Mind body techniques such as meditation improve symptoms in children and adults with IBS. Typical courses, however, are lengthy and difficult to administer. We report our experience with a short course of Preksha Dhyana (PD), a child-friendly focused meditation with yoga. Method: Physicians deliver focused meditation while medical assistants taught yoga. Three sessions were administered biweekly with recommendations for daily practice. Pain severity Likert scores were compared with a treatment as usual (TAU) historical control. Anxiety scores were compared from baseline in the PD group. Results: Thirty PD patients aged 9–17 (20 female) and 52 consecutive TAU group aged 5–17 (33 female) were reviewed. The biweekly sessions had high (71%) completion rates. Utilization rates of PD were similar to TAU despite added sessions. The PD group had an average time of follow-up of 8.9 ± 9.4 vs. 6.0 ± 3.9 months in the TAU group ( p = 0.522). Changes in pain scores from baseline showed improvement in the PD group, 0.67 ± 0.13 vs. TAU 1.39 ± 0.11 ( p = 0.0003). In the PD group, anxiety scores improved significantly from baseline (0.5 vs. 1, P < 0.001). Pain improved in 93% (28/30) and resolved in 47% (14/30). Conclusion: A short course of PD was successfully embedded in a busy pediatric office without additional staffing. The approach proved cost-effective without increasing overall healthcare utilization and showed significant benefits over TAU. Pending RCT confirmation, this offers a cost-effective method to incorporate mind–body techniques into a pediatric office practice.

The gastrointestinal microbiome is established during foetal development and continuously altered throughout the life span. The microbial ecosystem is complex and it plays a vital role in human health, including digestion and immune response. The human gastrointestinal environment is colonized by numerous species of microorganisms with substantial variations. The symbiotic interactions between host metabolism and the resident microflora contribute largely to the maintenance of gut homeostasis and perturbations in the microbial balance (dysbiosis) have been correlated with a wide array of diseases. Various factors have been implicated in microbial dysbiosis including environmental changes and drug effects. This review discusses the microbial composition of the gastrointestinal environment and the factors causing microbial dysbiosis. It highlights the evolution of the microbiota in the human gut with special emphasis on the role of different bacterial species in the gut homeostasis. In addition, it aims to provide critical analysis on the role of microbiota in gut related diseases, especially irritable bowel syndrome (IBS), inflammatory bowel disease (IBD), and Crohns’ disease. A better understanding of the gut ecosystem would enable more targeted interventions in gut microbial composition to prevent or manage these disorders by restoring gut homeostasis.

OBJECTIVES:The aim of the study was to determine prevalence and characterize sucrase-isomaltase (SI) gene variants of congenital sucrase-isomaltase deficiency in non-Hispanic white pediatric and young adult patients with functional gastrointestinal disorders (FGIDs), and abnormal sucrase activity on histologically normal duodenal biopsy. METHODS:Clinical symptoms and disaccharidase activities data were collected for an abnormal (low) sucrase (≤25.8 U, n = 125) activity group, and 2 normal sucrase activity groups with moderate (≥25.8-≤55 U, n = 250) and high (>55 U, n = 250) sucrase activities. SI gene variants were detected by next-generation sequencing of DNA from formalin-fixed paraffin-embedded tissues of these patients. FGIDs symptoms based on Rome IV criteria and subsequent clinical management of abnormal sucrase activity cases with pathogenic SI gene variants were analyzed. RESULTS:Thirteen SI gene variants were found to be significantly higher in abnormal sucrase cases with FGIDs symptoms (36/125, 29%; 71% did not have a pathogenic variant) compared to moderate normal (16/250, 6.4%, P < 0.001) or high normal (5/250, 2.0%, P < 0.001) sucrase groups. Clinical management data were available in 26 of abnormal sucrase cases, and only 10 (38%) were correctly diagnosed and managed by the clinicians. Concomitant lactase deficiency (24%; 23/97) and pan-disaccharidase deficiency (25%; 13/51) were found in the abnormal sucrase group. CONCLUSIONS:Heterozygous and compound heterozygous mutations in the SI gene were more prevalent in cases with abnormal sucrase activity presenting with FGIDs, and normal histopathology. This suggests heterozygous pathogenic variants of congenital sucrase-isomaltase deficiency may present as FGIDs. Concomitant lactase or pan-disaccharidase deficiencies were common in abnormal sucrase cases with SI gene variants.