Fecal bacteria and short-chain fatty acids in irritable bowel syndrome: Relations to subtype.

BACKGROUND: The relationship between gut microbiota and irritable bowel syndrome (IBS) subtype is unclear. We aimed to explore whether differences in fecal bacteria composition and short-chain fatty acid (SCFA) levels were associated with subtypes and symptoms of IBS. METHODS: All participants delivered fecal samples and self-reports on IBS Symptom Severity Score (IBS-SSS), Bristol Stool Scale (BSS), and Gastrointestinal Symptom Rating Scale (GSRS). Fecal bacteria composition was assessed by the GA-map® Dysbiosis Test based on 16S rRNA sequences of bacterial species/groups. SCFAs were analyzed by vacuum distillation followed by gas chromatography. KEY RESULTS: Sixty patients with IBS were included (mean age 38 years, 46 [77%] females): Twenty-one patients were classified as IBS-D (diarrhea), 31 IBS-M (mixed diarrhea and constipation), and eight IBS-C (constipation). Forty-two healthy controls (HCs) (mean age 35 years, 27 [64%] females) were included. Patients had a significantly higher relative frequency of dysbiosis, lower levels of Actinobacteria, and higher levels of Bacilli than HCs. Eight bacterial markers were significantly different across IBS subgroups and HCs, and 13 bacterial markers were weakly correlated with IBS symptoms. Clostridia and Veillonella spp. had a weak negative correlation with constipation scores (GSRS) and a weak positive correlation with loose stools (BSS). Diarrhea scores (GSRS) and looser stool (BSS) were weakly correlated with levels of total SCFAs, acetic and butyric acid. Levels of total SCFAs and acetic acid were weakly correlated with symptom severity (IBS-SSS). CONCLUSIONS & INFERENCES: Patients with IBS had a different fecal bacteria composition compared to HCs, and alterations of SCFAs may contribute to the subtype.

Nutritional safety and status following a 12-week strict low FODMAP diet in patients with irritable bowel syndrome.

BACKGROUND: A low FODMAP diet (LFD) is an established dietary treatment for patients with irritable bowel syndrome (IBS). However, knowledge on the extended effects of the restriction phase regarding nutrient intake, symptom severity, and quality of life (QoL) is sparse. Therefore, our objectives were to evaluate the safety of a dietitian-led 12-week strict LFD on measures of blood biochemistry, nutritional status, symptom severity, and QoL. METHODS: In this open-label dietitian-led 12-week strict LFD intervention for IBS patients with predominantly diarrhea or mixed stool pattern (IBS-D/-M), we collected data on diet intake (3-day dietary record), overnight fasting routine blood samples, body weight, IBS symptoms (IBS Severity Scoring System (IBS-SSS)), and IBS-related QoL (IBS-QoL) at baseline and after 12 weeks. KEY RESULTS: Thirty-six participants completed the 12-week follow-up (mean age: 37 years, 67% women, IBS-SSS: 242 (101)). All blood parameters measured were within established reference values at both time points. We found no change in intake of macro- or micronutrients, but several micronutrients were below the recommendations both before and after 12 weeks. BMI slightly decreased, primarily driven by participants with BMI >25 (p < 0.005). QoL improved among most subdomains (p ≤ 0.002), except food avoidance and social reaction. CONCLUSION: An extended dietitian-guided LFD (12 weeks) is not inferior to the participants' baseline diet, since no clinically meaningful changes in nutritionally related blood samples and no changes in macro- or micronutrient intake were observed. However, the intake of several nutrients was below the recommendations at both time points indicating low diet quality.

Cognitive function in patients with irritable bowel syndrome: impairment is common and only weakly correlated with depression/anxiety and severity of gastrointestinal symptoms.

OBJECTIVE: To investigate cognitive function in patients with irritable bowel syndrome (IBS) and its relation to anxiety/depression and severity of gastrointestinal (GI) symptoms. METHODS: Patients with IBS (n = 65) and healthy controls (HCs, n = 37) performed the ten subtests of the Repeatable Battery for the Assessment of Neuropsychological Status (RBANS). Age-normed index scores of five cognitive domains (Immediate memory, Visuospatial function, Language function, Attention, Recall) and a total (Fullscale) score were derived from the performance. Emotional function was assessed using the Hospital Anxiety and Depression Scale (HADS), and the IBS Symptom Scoring System (IBS-SSS) was used to define the severity of GI symptoms. RESULTS: Patients with IBS reported significantly higher scores than the HC group on symptom measures of anxiety and depression, and significantly lower scores on the Immediate memory, Recall, and Fullscale RBANS indexes. Approximately 30% of the IBS patients obtained index scores at least one standard deviation below the population mean, and more than 50% scored above the screening threshold for an anxiety disorder. The severity of GI symptoms was significantly correlated with the severity level of anxiety symptoms (p=.006), but neither the severity level of emotional nor GI symptoms was significantly correlated with the RBANS index scores in the IBS group. CONCLUSION: Cognitive and emotional function were more severely affected in patients with IBS than in HCs. The weak correlation between the two functional areas suggests that both should be assessed as part of a clinical examination of patients with IBS.

Cognitive and emotional function should be assessed in patients with IBS.Cognitive impairment was less closely related to symptoms of anxiety/depression and severity of GI symptoms than expected.An independent contribution of both emotional symptoms and cognitive function should be considered when developing treatment programs for patients with IBS.

A psychological symptom based machine learning model for clinical evaluation of irritable bowel syndrome.

Background: Irritable bowel syndrome (IBS) is a chronic functional gastrointestinal disorder characterized by recurrent abdominal pain associated with alterations  in stool form and/or stool frequency. Co-morbidities such as anxiety, depression, fatigue, and insomnia are frequently reported by patients suffering from IBS. Identification of these symptoms should thus be an integral part of an IBS assessment.      However, an optimal tool to screen for core psychological symptoms in IBS is still  missing. Here, we aim to develop a psychological symptom based machine learning model to efficiently help clinicians to identify patients suffering from IBS. Methods: We developed a machine learning workflow to select the most significant psychological features associated with IBS in a dataset including 49 patients with IBS and 35 healthy controls. These features were used to train three different types of machine learning models: logistic regression, decision trees and support vector machine classifiers; which were validated on a holdout validation dataset and an unseen test set. The performance of these models was compared in terms of balanced accuracy scores. Results: A logistic regression model including a combination of symptom features associated with anxiety and fatigue resulted in a balanced accuracy score of 0.93 (0.81-1.0) on unseen test data and outperformed the other comparable models. The same model correctly identified all patients with IBS in a test set (recall score 1) and misclassified one non-IBS subject (precision score 0.91). A complementary post-hoc leave-one-out cross validation analysis including the same symptom features showed similar, but slightly inferior results (balanced accuracy 0.84, recall 0.88, precision 0.86). Conclusions: Inclusion of machine learning based psychological evaluation can complement and improve existing clinical procedure for diagnosis of IBS.

Assessment of Self-Reported Executive Function in Patients with Irritable Bowel Syndrome Using a Machine-Learning Framework.

Introduction: Irritable bowel syndrome (IBS) is characterized as a disorder of the gut-brain interaction (DGBI). Here, we explored the presence of problems related to executive function (EF) in patients with IBS and tested the relative importance of cognitive features involved in EF. Methods: A total of 44 patients with IBS and 22 healthy controls (HCs) completed the Behavior Rating Inventory of Executive Function (BRIEF-A), used to identify nine EF features. The PyCaret 3.0 machine-learning library in Python was used to explore the data, generate a robust model to classify patients with IBS versus HCs and identify the relative importance of the EF features in this model. The robustness of the model was evaluated by training the model on a subset of data and testing it on the unseen, hold-out dataset. Results: The explorative analysis showed that patients with IBS reported significantly more severe EF problems than the HC group on measures of working memory function, initiation, cognitive flexibility and emotional control. Impairment at a level in need of clinical attention was found in up to 40% on some of these scales. When the nine EF features were used as input to a collection of different binary classifiers, the Extreme Gradient Boosting algorithm (XGBoost) showed superior performance. The working memory subscale was consistently selected with the strongest importance in this model, followed by planning and emotional control. The goodness of the machine-learning model was confirmed in an unseen dataset by correctly classifying 85% of the IBS patients. Conclusions: The results showed the presence of EF-related problems in patients with IBS, with a substantial impact of problems related to working memory function. These results suggest that EF should be part of an assessment procedure when a patient presents other symptoms of IBS and that working memory function should be considered a target when treating patients with the disorder. Further studies should include measures of EF as part of the symptom cluster characterizing patients with IBS and other DGBIs.

Gut bless you: The microbiota-gut-brain axis in irritable bowel syndrome.

Irritable bowel syndrome (IBS) is a common clinical label for medically unexplained gastrointestinal symptoms, recently described as a disturbance of the microbiota-gut-brain axis. Despite decades of research, the pathophysiology of this highly heterogeneous disorder remains elusive. However, a dramatic change in the understanding of the underlying pathophysiological mechanisms surfaced when the importance of gut microbiota protruded the scientific picture. Are we getting any closer to understanding IBS' etiology, or are we drowning in unspecific, conflicting data because we possess limited tools to unravel the cluster of secrets our gut microbiota is concealing? In this comprehensive review we are discussing some of the major important features of IBS and their interaction with gut microbiota, clinical microbiota-altering treatment such as the low FODMAP diet and fecal microbiota transplantation, neuroimaging and methods in microbiota analyses, and current and future challenges with big data analysis in IBS.

Gastrointestinal transit and contractility in diabetic constipation: A wireless motility capsule study on diabetes patients and healthy controls.

BACKGROUND: Diabetic constipation is traditionally attributed to slow colonic transit, despite limited evidence. More than half of patients find treatment unsatisfactory. To improve treatment, there is a need for better diagnostic understanding of the condition. OBJECTIVE: In this wireless motility capsule study, we aimed to investigate gastrointestinal transit and contractility in diabetes patients with and without constipation, and in healthy controls. METHODS: We prospectively included type 1 or type 2 diabetes patients with gastrointestinal symptoms. Based on the Gastrointestinal Symptom Rating Scale we distinguished into two groups: with constipation and without constipation. Non-diabetic controls were asymptomatic. All were examined with wireless motility capsule, determining transit times and contractility parameters. RESULTS: 57 patients (42 women, 46 with type 1 diabetes) and 26 healthy controls (14 women) were included. We found no difference in transit times between diabetes patients with and without constipation. Compared to healthy controls (35:55, h:min), whole-gut transit was slower in both diabetes patients with constipation (66:15, p = 0.03) and without constipation (71:16, p < 0.001). Small bowel motility index correlated rs  = -0.32 (p = 0.01) with constipation symptoms. CONCLUSIONS: Diabetes patients with constipation had similar transit times as those without constipation. Both groups had slower whole-gut transit than healthy controls. Constipation was associated with reduced small bowel, but not colonic contractility. Our results imply that other mechanisms than slow colonic transit may be more important in the pathogenesis of diabetic constipation.

Study protocol of the Bergen brain-gut-microbiota-axis study: A prospective case-report characterization and dietary intervention study to evaluate the effects of microbiota alterations on cognition and anatomical and functional brain connectivity in patients with irritable bowel syndrome.

INTRODUCTION: Irritable bowel syndrome (IBS) is a common clinical label for medically unexplained gastrointestinal (GI) symptoms, recently described as a disturbance of the brain-gut-microbiota (BGM) axis. To gain a better understanding of the mechanisms underlying the poorly understood etiology of IBS, we have designed a multifaceted study that aim to stratify the complex interaction and dysfunction between the brain, the gut, and the microbiota in patients with IBS. METHODS: Deep phenotyping data from patients with IBS (n = 100) and healthy age- (between 18 and 65) and gender-matched controls (n = 40) will be collected between May 2019 and December 2021. Psychometric tests, questionnaires, human biological tissue/samples (blood, faeces, saliva, and GI biopsies from antrum, duodenum, and sigmoid colon), assessment of gastric accommodation and emptying using transabdominal ultrasound, vagal activity, and functional and structural magnetic resonance imaging (MRI) of the brain, are included in the investigation of each participant. A subgroup of 60 patients with IBS-D will be further included in a 12-week low FODMAP dietary intervention-study to determine short and long-term effects of diet on GI symptoms, microbiota composition and functions, molecular GI signatures, cognitive, emotional and social functions, and structural and functional brain signatures. Deep machine learning, prediction tools, and big data analyses will be used for multivariate analyses allowing disease stratification and diagnostic biomarker detection. DISCUSSION: To our knowledge, this is the first study to employ unsupervised machine learning techniques and incorporate systems-based interactions between the central and the peripheral components of the brain-gut-microbiota axis at the levels of the multiomics, microbiota profiles, and brain connectome of a cohort of 100 patients with IBS and matched controls; study long-term safety and efficacy of the low-FODMAP diet on changes in nutritional status, gut microbiota composition, and metabolites; and to investigate changes in the brain and gut connectome after 12 weeks strict low-FODMAP-diet in patients with IBS. However, there are also limitations to the study. As a restrictive diet, the low-FODMAP diet carries risks of nutritional inadequacy and may foster disordered eating patterns. Strict FODMAP restriction induces a potentially unfavourable gut microbiota, although the health effects are unknown. TRIAL REGISTRATION NUMBER: NCT04296552 (ClinicalTrials.gov).

Association between gastrointestinal complaints and psychopathology in patients with anorexia nervosa.

OBJECTIVE: Gastrointestinal (GI) symptoms appear frequently in patients with anorexia nervosa (AN), but the associations between psychopathological, GI, and eating disorder (ED) symptoms remain unclear. This study aimed to determine the relationships of GI complaints with psychopathological measures, ED symptoms, and body mass index (BMI) in patients with AN. METHOD: Thirty outpatients with AN aged >16 years were included. Psychopathological measures (Symptom Checklist-90-Revised, Beck Depression Inventory-II, and Beck Anxiety Inventory), ED symptoms (Eating Disorder Examination Questionnaire), ED-associated impairment (Clinical Impairment Assessment Questionnaire), GI complaints (Irritable Bowel Syndrome Severity Scoring System [IBS-SSS]), and BMI were assessed prior to starting treatment, and correlation and multiple regression analyses were applied to data from 19 patients. RESULTS: IBS-symptoms were significantly correlated only with ED symptoms (r = 0.583, p = .009) and somatization (r = 0.666, p = .002). Multiple regression analysis revealed that somatization significantly predicted worse IBS symptoms (beta = 0.5, p = .04), while ED symptoms did not. DISCUSSION: Higher IBS-SSS scores were associated with higher severities of other somatic complaints. GI complaints and somatization should be addressed in treatments for AN in order to prevent these factors impeding the establishment of healthy eating patterns. CLINICAL TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT02745067.

MicroRNA-34a Acutely Regulates Synaptic Efficacy in the Adult Dentate Gyrus In Vivo.

Activity-dependent synaptic plasticity involves rapid regulation of neuronal protein synthesis on a time-scale of minutes. miRNA function in synaptic plasticity and memory formation has been elucidated by stable experimental manipulation of miRNA expression and activity using transgenic approaches and viral vectors. However, the impact of rapid miRNA modulation on synaptic efficacy is unknown. Here, we examined the effect of acute (12 min), intrahippocampal infusion of a miR-34a antagonist (antimiR) on medial perforant path-evoked synaptic transmission in the dentate gyrus of adult anesthetised rats. AntimiR-34a infusion acutely depressed medial perforant path-evoked field excitatory post-synaptic potentials (fEPSPs). The fEPSP decrease was detected within 9 min of infusion, lasted for hours, and was associated with knockdown of antimiR-34a levels. AntimiR-34a-induced synaptic depression was sequence-specific; no changes were elicited by infusion of scrambled or mismatch control. The rapid modulation suggests that a target, or set of targets, is regulated by miR-34a. Western blot analysis of dentate gyrus lysates revealed enhanced expression of Arc, a known miR-34a target, and four novel predicted targets (Ctip2, PKI-1α, TCF4 and Ube2g1). Remarkably, antimiR-34a had no effect when infused during the maintenance phase of long-term potentiation. We conclude that miR-34a regulates basal synaptic efficacy in the adult dentate gyrus in vivo. To our knowledge, these in vivo findings are the first to demonstrate acute (< 9 min) regulation of synaptic efficacy in the adult brain by a miRNA.

The RNA-Binding Protein hnRNP K Mediates the Effect of BDNF on Dendritic mRNA Metabolism and Regulates Synaptic NMDA Receptors in Hippocampal Neurons.

Brain-derived neurotrophic factor (BDNF) is an important mediator of long-term synaptic potentiation (LTP) in the hippocampus. The local effects of BDNF depend on the activation of translation activity, which requires the delivery of transcripts to the synapse. In this work, we found that neuronal activity regulates the dendritic localization of the RNA-binding protein heterogeneous nuclear ribonucleoprotein K (hnRNP K) in cultured rat hippocampal neurons by stimulating BDNF-Trk signaling. Microarray experiments identified a large number of transcripts that are coimmunoprecipitated with hnRNP K, and about 60% of these transcripts are dissociated from the protein upon stimulation of rat hippocampal neurons with BDNF. In vivo studies also showed a role for TrkB signaling in the dissociation of transcripts from hnRNP K upon high-frequency stimulation (HFS) of medial perforant path-granule cell synapses of male rat dentate gyrus (DG). Furthermore, treatment of rat hippocampal synaptoneurosomes with BDNF decreased the coimmunoprecipitation of hnRNP K with mRNAs coding for glutamate receptor subunits, Ca2+- and calmodulin-dependent protein kinase IIß (CaMKIIß) and BDNF. Downregulation of hnRNP K impaired the BDNF-induced enhancement of NMDA receptor (NMDAR)-mediated mEPSC, and similar results were obtained upon inhibition of protein synthesis with cycloheximide. The results demonstrate that BDNF regulates specific populations of hnRNP-associated mRNAs in neuronal dendrites and suggests an important role of hnRNP K in BDNF-dependent forms of synaptic plasticity.

NMDA receptor-dependent regulation of miRNA expression and association with Argonaute during LTP in vivo.

microRNAs (miRNAs) are major regulators of protein synthesis in the brain. A major goal is to identify changes in miRNA expression underlying protein synthesis-dependent forms of synaptic plasticity such as long-term potentiation (LTP). Previous analyses focused on changes in miRNA levels in total lysate samples. Here, we asked whether changes in total miRNA accurately reflect changes in the amount of miRNA bound to Argonaute protein within the miRNA-induced silencing complex (miRISC). Ago2 immunoprecipitation was used to isolate RISC-associated miRNAs following high-frequency stimulation (HFS)-induced LTP in the dentate gyrus of anesthetized rats. Using locked-nucleic acid-based PCR cards for high-throughput screening and independent validation by quantitative TaqMan RT-PCR, we identified differential regulation of Ago2-associated and total miRNA expression. The ratio of Ago2/total miRNA expression was regulated bidirectionally in a miRNA-specific manner and was largely dependent on N-methyl-D-aspartate receptor (NMDA) activation during LTP induction. The present results identify miRNA association with Ago2 as a potential control point in activity-dependent synaptic plasticity in the adult brain. Finally, novel computational analysis for targets of the Ago2-associated miRNAs identifies 21 pathways that are enriched and differentially targeted by the miRNAs including axon guidance, mTOR, MAPK, Ras, and LTP.

MicroRNA regulation of the synaptic plasticity-related gene Arc.

Expression of activity-regulated cytoskeleton associated protein (Arc) is crucial for diverse types of experience-dependent synaptic plasticity and long-term memory in mammals. However, the mechanisms governing Arc-specific translation are little understood. Here, we asked whether Arc translation is regulated by microRNAs. Bioinformatic analysis predicted numerous candidate miRNA binding sites within the Arc 3'-untranslated region (UTR). Transfection of the corresponding microRNAs in human embryonic kidney cells inhibited expression of an Arc 3'UTR luciferase reporter from between 10 to 70% across 16 microRNAs tested. Point mutation and deletion of the microRNA-binding seed-region for miR-34a, miR-326, and miR-19a partially or fully rescued reporter expression. In addition, expression of specific microRNA pairs synergistically modulated Arc reporter expression. In primary rat hippocampal neuronal cultures, ectopic expression of miR-34a, miR-193a, or miR-326, downregulated endogenous Arc protein expression in response to BDNF treatment. Conversely, treatment of neurons with cell-penetrating, peptide nucleic acid (PNA) inhibitors of miR-326 enhanced Arc mRNA expression. BDNF dramatically upregulated neuronal expression of Arc mRNA and miR-132, a known BDNF-induced miRNA, without affecting expression of Arc-targeting miRNAs. Developmentally, miR-132 was upregulated at day 10 in vitro whereas Arc-targeting miRNAs were downregulated. In the adult brain, LTP induction in the dentate gyrus triggered massive upregulation of Arc and upregulation of miR-132 without affecting levels of mature Arc-targeting miRNAs. Turning to examine miRNA localization, qPCR analysis of dentate gyrus synaptoneurosome and total lysates fractions demonstrated synaptic enrichment relative to small nucleolar RNA. In conclusion, we find that Arc is regulated by multiple miRNAs and modulated by specific miRNA pairs in vitro. Furthermore, we show that, in contrast to miR-132, steady state levels of Arc-targeting miRNAs do not change in response to activity-dependent expression of Arc in hippocampal neurons in vitro or during LTP in vivo.