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BackgroundDisruption of central circadian rhythms likely mediated by changes in microbiota and a decrease in gut-derived metabolites like short chain fatty acids (SCFAs) negatively impacts colonic barrier homeostasis. We aimed to explore the effects of isolated peripheral colonic circadian disruption on the colonic barrier in a mouse model of colitis and explore the mechanisms, including intestinal microbiota community structure and function.MethodsColon epithelial cell circadian rhythms were conditionally genetically disrupted in mice: TS4Cre-BMAL1lox (cBMAL1KO) with TS4Cre as control animals. Colitis was induced through 5 days of 2% dextran sulfate sodium (DSS). Disease activity index and intestinal barrier were assessed, as were fecal microbiota and metabolites.ResultsColitis symptoms were worse in mice with peripheral circadian disruption (cBMAL1KO). Specifically, the disease activity index and intestinal permeability were significantly higher in circadian-disrupted mice compared with control animals (TS4Cre) (P < .05). The worsening of colitis appears to be mediated, in part, through JAK (Janus kinase)-mediated STAT3 (signal transducer and activator of transcription 3), which was significantly elevated in circadian-disrupted (cBMAL1KO) mice treated with DSS (P < .05). Circadian-disrupted (cBMAL1KO) mice also had decreased SCFA metabolite concentrations and decreased relative abundances of SCFA-producing bacteria in their stool when compared with control animals (TS4Cre).ConclusionsDisruption of intestinal circadian rhythms in colonic epithelial cells promoted more severe colitis, increased inflammatory mediators (STAT3 [signal transducer and activator of transcription 3]), and decreased gut microbiota–derived SCFAs compared with DSS alone. Further investigation elucidating the molecular mechanisms behind these findings could provide novel circadian directed targets and strategies in the treatment of inflammatory bowel disease.

Posttraumatic stress disorder (PTSD) is a psychiatric disorder, resulting from exposure to traumatic events. Current recommended first-line interventions for the treatment of PTSD include evidence-based psychotherapies, such as cognitive processing therapy (CPT). Psychotherapies are effective for reducing PTSD symptoms, but approximately two-thirds of veterans continue to meet diagnostic criteria for PTSD after treatment, suggesting there is an incomplete understanding of what factors sustain PTSD. The intestine can influence the brain and this study evaluated intestinal readouts in subjects with PTSD. Serum samples from controls without PTSD (

Background Ulcerative Colitis (UC) is a chronic, inflammatory disease, characterized by symptomatic periods (flare) interspersed with asymptomatic periods (remission). Evidence suggests that psychological stress can trigger flare. Studies have shown that mindfulness interventions (MI) reduce stress, foster more adaptive coping, and improve quality of life, but have been minimally used for UC patients. The objective of this study was to determine whether participation in an MI results in improvements in UC disease course and inflammatory cascades, mindfulness, perceived stress, and other psychological outcomes in inactive UC patients with limited or no exposure to past MI. Methods Participants were randomized to an 8-week MI or control group. Biological and psychological assessments were performed at baseline, post 8-week course, and at 6- and 12-months. Results Forty-three participants enrolled. The MI increased the state of mindfulness and mindfulness skills, decreased perceived stress and stress response in patients with inactive UC. The MI intervention significantly decreased the incidence of flare over 12 months (P < .05). None of the UC patients in the MI flared during 12 months, while 5 of 23 (22%) control group participants flared during the same period. Conclusions MIs could be considered as adjuvant treatment for a subset of UC patients with high perceived stress and low state of mindfulness. The trial was registered at clinicaltrials.gov as NCT01491997.

Background Circadian rhythms orchestrate multiple biological processes with 24-hour oscillations that regulate diverse bodily functions. The central clock is in the suprachiasmatic nucleus and orchestrates peripheral circadian clocks in every organ system including the gastrointestinal tract (GIT). Previous work has shown that circadian misalignment in Ulcerative Colitis (UC) is associated with increased colonic permeability, and colonic organoids recapitulate the host circadian phenotype. The goals of this study were to examine whether circadian misalignment would disrupt the colonic barrier integrity in UC. Methods 5 subjects, 2 UC and 3 Healthy Controls (HC), were recruited into the study. All UC subjects were inactive (partial Mayo Score ≤ 1) and stable medications with no flares for the last 3 months. For two weeks prior to entry into the circadian lab all subjects were on a prescribed regular sleep schedule. Subjects were then kept in the circadian lab for 6 days with strict control of their light/dark cycle (Figure 1). All subjects then underwent two unprepped flexible sigmoidoscopies at: (1) baseline with circadian alignment and (2) after three days of simulated night shifts with circadian misalignment. Colonic organoids generated created from the colon tissue biopsies grown in Matrigel and barrier integrity was assessed in apical-out colonoids by FITC-Dextran (4 kDa) Net Fluorescence was estimated by ImageJ. Results Colonic organoids at baseline had significantly decreased ZO-1 in UC versus HC, Figure 2. At baseline there was an increase in UC colonic organoid barrier integrity as measured by net fluorescence (density/area). In UC vs HC barrier integrity was 855 ± 203.8 vs 333.1 ± 52.9, p Conclusion Circadian misalignment by alternation of light:dark cycles (simulated night shift) causes misalignment of host central clock and impacts circadian regulated biological process like tight junction proteins which decreases colonic barrier integrity. This study highlights the importance of studying environmental factors that impact circadian timing in UC including night shift work which may be a significant risk factor for a disease flare. Further in vivo and in vitro studies are ongoing to determine the mechanisms through which circadian misalignment impacts colonic barrier integrity in UC (NCT05180279).

Background & Aims Alcohol intake with circadian rhythm disruption (CRD) increases colon cancer risk. We hypothesized that eating during or around physiologic rest time, a common habit in modern society, causes CRD and investigated the mechanisms by which it promotes alcohol-associated colon carcinogenesis. Methods The effect of feeding time on CRD was assessed using B6 mice expressing a fusion protein of PERIOD2 and LUCIFERASE (PER2::LUC) were used to model colon polyposis and to assess the effects of feeding schedules, alcohol consumption, and prebiotic treatment on microbiota composition, short-chain fatty acid levels, colon inflammation, and cancer risk. The relationship between butyrate signaling and a proinflammatory profile was assessed by inactivating the butyrate receptor GPR109A. Results Eating at rest (wrong-time eating [WTE]) shifted the phase of the colon rhythm in PER2::LUC mice. In TS4Cre × APClox468 mice, a combination of WTE and alcohol exposure (WTE + alcohol) decreased the levels of short-chain fatty acid–producing bacteria and of butyrate, reduced colonic densities of regulatory T cells, induced a proinflammatory profile characterized by hyperpermeability and an increased mucosal T-helper cell 17/regulatory T cell ratio, and promoted colorectal cancer. Prebiotic treatment improved the mucosal inflammatory profile and attenuated inflammation and cancer. WTE + alcohol–induced polyposis was associated with increased signal transducer and activator of transcription 3 expression. Decreased butyrate signaling activated the epithelial signal transducer and activator of transcription 3 in vitro. The relationship between butyrate signaling and a proinflammatory profile was confirmed in human colorectal cancers using The Cancer Genome Atlas. Conclusions Abnormal timing of food intake caused CRD and interacts with alcohol consumption to promote colon carcinogenesis by inducing a protumorigenic inflammatory profile driven by changes in the colon microbiota and butyrate signaling. Accession number of repository for microbiota sequence data: raw FASTQ data were deposited in the NCBI Sequence Read Archive under project PRJNA523141., Graphical abstract

Circadian rhythms regulate much of gastrointestinal physiology including cell proliferation, motility, digestion, absorption, and electrolyte balance. Disruption of circadian rhythms can have adverse consequences including the promotion of and/or exacerbation of a wide variety of gastrointestinal disorders and diseases. Areas covered: In this review, we evaluate some of the many gastrointestinal functions that are regulated by circadian rhythms and how dysregulation of these functions may contribute to disease. This review also discusses some common gastrointestinal disorders that are known to be influenced by circadian rhythms as well as speculation about the mechanisms by which circadian rhythm disruption promotes dysfunction and disease pathogenesis. We discuss how knowledge of circadian rhythms and the advent of chrono-nutrition, chrono-pharmacology, and chrono-therapeutics might influence clinical practice. Expert opinion: As our knowledge of circadian biology increases, it may be possible to incorporate strategies that take advantage of circadian rhythms and chronotherapy to prevent and/or treat disease.

Highlights • Patients with COVID-19 show a prominent Th1 and Th17 cytokine profile. • COVID-19 induces the expression of TGF-β. • TGF-β can be used as a predictive factor of disease severity in patients with COVID-19., Cytokine dysregulation is the proposed mechanism for Coronavirus disease 2019 (COVID-19).The aim of this study was to evaluate the serum levels of interferon (IFN)-γ, interleukin (IL)-5, IL-8, Il-9, IL-17, TGF-β and IFN-γ in patients infected with SARS-CoV-2. The study was conducted between 63 adult patients with COVID‐19 and compared with 33 age and gender-matched healthy subjects as controls. The age range in both groups was 50 to 70 years. The patients were classified into mild group (33 patients) and severe group (30 patients). Serum samples were collected from all participants and tested for the cytokine levels by ELISA (enzyme-linked immunosorbent assay) method. Statistical analysis was performed using the one-way ANOVA. The mean serum levels of IFN-γ, TGF-β, IL-17 and IL-8 in the COVID‐19 patients were significantly higher than those observed in the control group. A comparison of between the mild and severe groups showed significant differences in TGF-β levels. The mean concentration of serum IL-5 and IL-9 in patients with COVID‐19 did not differ from those in the control group. Systemic IL-17 levels correlated positively and significantly with TGF-β in patients with COVID‐19. Th1 (IFN-γ), Treg (TGF-β), and Th17 (IL-17) cytokines concentration were increased in COVID-19 patients. Interferon-γ and IL-17 are involved in inducing and mediating proinflammatory responses. Our data suggest that TGF-β can be used as a predictive factor of disease severity in patients with COVID-19.

Recent studies suggest that circadian rhythms regulate intestinal barrier integrity, but it is not clear whether there are daily variations in barrier integrity. This study investigated daily variations in intestinal barrier integrity, including whether there are differences in alcohol-induced intestinal barrier dysfunction after an alcohol binge at different times of day and whether this is associated with concurrent liver injury. C57BL6/J male mice were fed a standard chow diet, an alcohol-containing liquid diet, or an alcohol control diet for 4 wk. During week 5 (i.e., on days 43–45), mice received three once-daily gavages of alcohol (6 g/kg) or the control (phosphate-buffered saline) at the same time each day. Immediately after the binge on the second day, intestinal permeability was assessed. Four hours after the third and final binge, mice were euthanized and tissue samples collected. The results demonstrated diet-specific and outcome-specific effects of time, alcohol, and/or time by alcohol interaction. Specifically, the alcohol binge robustly influenced markers of intestinal barrier integrity, and liver markers were robustly influenced by time of day. Only intestinal permeability (i.e., sucralose) demonstrated a significant effect of time and also showed a binge by time interaction, suggesting that the time of the alcohol binge influences colonic permeability. NEW & NOTEWORTHY This study investigated daily variations in intestinal barrier integrity, including whether there are differences in alcohol-induced intestinal barrier dysfunction after an alcohol binge at different times of day and whether this is associated with concurrent liver injury. We conclude that 1) alcohol binge significantly impacted markers of intestinal permeability, 2) time of day significantly affected liver outcomes, and 3) the time of day influenced colonic permeability.

Approximately one-third of individuals in the United States experience unsatisfactory bowel habits, and dietary intake, especially one low in fiber, could be partly responsible. We hypothesized that intake of a fermentable fiber (starch-entrapped microspheres, SM) that has a delayed, slow fermentation profile in vitro would improve bowel habit while exhibiting prebiotic capacity in those with self-described unsatisfactory bowel habits, all with minimal adverse effects. A total of 43 healthy volunteers completed a 3-month, double-blind, parallel-arm randomized clinical trial to assess the ability of a daily dose (9 or 12 g) of SM vs psyllium (12 g) to improve bowel habit, including stool consistency and frequency, and modify gut milieu through changes in stool microbiota and short-chain fatty acids while remaining tolerable through minimal gastrointestinal symptoms. All outcomes were compared before and after fiber treatment. Stool frequency significantly improved (P=.0003) in all groups after 3 months, but stool consistency improved only in both SM groups compared with psyllium. In addition, all groups self-reported a similar improvement in overall bowel habit with fiber intake. Both SM and psyllium resulted in minimal changes in microbiota composition and short-chain fatty acid concentrations. The present study suggests that supplementation with a delayed and slow-fermenting fiber in vitro may improve bowel habit in those with constipation, but further investigation is warranted to determine capacity to alter microbiota and fermentation profiles in humans. This trial was registered at ClinicalTrials.gov as NCT01210625.

Background Cholecystectomy (CCY) is one of the most frequently performed abdominal surgeries. However, the impact of CCY in clinical settings with altered gastrointestinal physiology and anatomy, such as Crohn’s disease (CD), has not been fully characterized. We sought to investigate clinical outcomes, disease severity, and quality of life of CD patients after CCY. Methods We utilized a prospective, longitudinal registry of consented CD patients followed at a tertiary center. Crohn’s disease patients that had or had not undergone CCY formed the 2 study groups. The absence or presence of gallbladder was confirmed with abdominal CT scans obtained during routine care. Multiyear clinical, biochemical, and histologic data were collected and analyzed. Results Among 834 CD patients, 151 (18%) had undergone CCY. History of CCY was associated with higher disease activity (median Harvey-Bradshaw index; P Conclusions CCY in CD patients was associated with multiple markers of disease activity and worse quality of life during multiyear follow up. This data suggests that CCY in CD patients may adversely impact the long-term clinical course.

SummaryBackground Microscopic colitis (MC) is a chronic bowel disorder characterised by watery diarrhoea. Nonsteroidal anti-inflammatory drugs (NSAIDs), proton pump inhibitors (PPIs), selective serotonin reuptake inhibitors (SSRIs) and statins have been associated with MC. However, underlying mechanisms remain unclear. Aim To study the association between exposure to these drugs and MC, with attention to time of exposure, duration, dosage and combined exposure, and to test hypotheses on underlying pharmacological mechanisms. Methods A case–control study was conducted using the British Clinical Practice Research Datalink. MC cases (1992–2013) were matched to MC-naive controls on age, sex and GP practice. Drug exposure was stratified according to time of exposure, duration of exposure or dosage. Conditional logistic regression analysis was applied to calculate adjusted odds ratios (AORs). Results In total, 1211 cases with MC were matched to 6041 controls. Mean age was 63.4 years, with 73.2% being female. Current use of NSAIDs (AOR 1.86, 95% CI 1.39–2.49), PPIs (AOR 3.37, 95% CI 2.77–4.09) or SSRIs (AOR 2.03, 95% CI 1.58–2.61) was associated with MC compared to never or past use. Continuous use for 4–12 months further increased the risk of MC. Strongest associations (fivefold increased risk) were observed for concomitant use of PPIs and NSAIDs. Statins were not associated with MC. Conclusions Current exposure to NSAIDs, PPIs or SSRIs and prolonged use for 4–12 months increased the risk of MC. Concomitant use of NSAIDs and PPIs showed the highest risk of MC. Acid suppression related dysbiosis may contribute to the PPI effect, which may be exacerbated by NSAID-related side-effects.

OBJECTIVES The aim of the present study was to examine the changes in bacteria in hospitalized preterm infants during the first month of life. METHODS Rectal swabs were collected daily from 12 preterm infants. DNA was extracted from swabs from day of birth and weekly thereafter. Bacterial taxa were identified with next generation sequencing using universal bacterial primers targeted at the 16S ribosomal DNA on a 454 Roche titanium platform. Sequences were clustered into operational taxonomic units, and taxonomy was assigned against the Greengenes databank using Quantitative Insights Into Microbial Ecology version 1.4. Quantitative polymerase chain reaction was used to determine the abundance of Bifidobacterium spp. Functional assessment of the microbiome was performed with Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt). RESULTS Average birth weight and gestational age were 1055 g and 28 weeks, respectively. There were 6 to 35 different bacterial families identified in the day-of-birth samples, unrelated to the mode of delivery. Richness decreased through hospitalization (week 1, 16.9 ± 7.7 vs weeks 3-5, 10.7 ± 3.4, P

Chronic heavy alcohol use is known to cause gut leakiness and alcoholic liver disease (ALD), but only 30% of heavy drinkers develop increased intestinal permeability and ALD. The hypothesis of this study was that disruption of circadian rhythms is a potential risk factor in actively drinking alcoholics for gut leakiness and endotoxemia. We studied 20 subjects with alcohol use disorder (AD) and 17 healthy controls (HC, 6 day workers, 11 night workers). Subjects wore a wrist actiwatch for 7 days and underwent a 24-h dim light phase assessment and urine collection for intestinal permeability. The AD group had significantly less total sleep time and increased fragmentation of sleep ( P < 0.05). AD also had significantly lower plasma melatonin levels compared with the HC [mean area under the curve (AUC) 322.78 ± 228.21 vs. 568.75 ± 304.26 pg/ml, P = 0.03]. In the AD group, AUC of melatonin was inversely correlated with small bowel and colonic intestinal permeability (lactulose-to-mannitol ratio, r = −0.39, P = 0.03; urinary sucralose, r = −0.47, P = 0.01). Cosinor analysis of lipopolysaccharide-binding protein (marker of endotoxemia) and lipopolysaccharide every 4 h for 24 h in HC and AD subjects had a midline estimating statistic of rhythm of 5,026.15 ± 409.56 vs. 6,818.02 ± 628.78 ng/ml ( P < 0.01) and 0.09 ± 0.03 vs. 0.15 ± 0.19 EU/ml ( P < 0.05), respectively. We found plasma melatonin was significantly lower in the AD group, and lower melatonin levels correlated with increased intestinal permeability and a marker of endotoxemia. Our study suggests the suppression of melatonin in AD may promote gut leakiness and endotoxemia.

BACKGROUND/AIMS Polyethylene glycol (PEG)-3350, approved by Food and Drug Administration (FDA) only for constipation, combined with 1.9 L of sports drink (SD) (GatoradeR) and bisacodyl (B) is commonly used in outpatient practice for bowel preparation due to cited patient satisfaction and tolerability of this specific regimen. We aim to compare PEG-3350 (MiralaxR) with PEG-AA-based (MoviPrepR) in terms of efficacy, patient satisfaction, and the effects of these two regimen on serum electrolytes. MATERIALS AND METHODS This study is a prospective, single-blinded, block randomized trial comparing single-dose PEG-3350+SD+B to split-dose 2-L PEG-AA in the outpatient endoscopy unit in patients undergoing colonoscopy. Basic metabolic profiles were checked on the day of randomization and on the day of procedure. Patients completed a survey on the day of procedure. Bowel preparation quality was assessed using the Boston Bowel Preparation Scale (BBPS) by two endoscopists and a nurse present during the procedure. RESULTS We randomized 150 patients (74 PEG-3350+SD+B and 76 PEG-AA). The PEG-AA group had significantly higher BBPS scores in the right colon by Endoscopist 1, Nurse, and Endoscopist 2 (p 0.005

Both acute and chronic consumption of alcohol (ethanol) have been reported to cause significant gastrointestinal (GI) symptoms. Alcohol-induced GI dysmotihty should not be a surprising finding since it is already known that ethanol can profoundly affect the function of both muscles and nerves in various organs. This chapter discusses the effects of ethanol on motor functions of esophagus, stomach, small and large intestine, and finally gallbladder and biliary tree. It also discusses the clinical picture, and then the in vivo and in vitro effects of both acute and chronic ethanol on motility of the organ. GI symptoms related to esophageal disorders such as postprandial chest pain and heartburn are believed to occur after an acute alcohol binge. The other major esophageal symptom that is sometimes experienced by alcoholics is difficulty in swallowing (dysphagia). Symptoms such as nausea, vomiting, epigastric pain, and postprandial fullness, which are typically related to gastric abnormalities, are common in alcoholics.

ObjectiveRecent evidence suggesting an important role of gut-derived inflammation in brain disorders has opened up new directions to explore the possible role of the gut-brain axis in neurodegenerative diseases. Given the prominence of dysbiosis and colonic dysfunction in patients with Parkinson’s disease (PD), we propose that toll-like receptor 4 (TLR4)-mediated intestinal dysfunction could contribute to intestinal and central inflammation in PD-related neurodegeneration.DesignTo test this hypothesis we performed studies in both human tissue and a murine model of PD. Inflammation, immune activation and microbiota composition were measured in colonic samples from subjects with PD and healthy controls subjects and rotenone or vehicle-treated mice. To further assess the role of the TLR4 signalling in PD-induced neuroinflammation, we used TLR4-knockout (KO) mice in conjunction with oral rotenone administration to model PD.ResultsPatients with PD have intestinal barrier disruption, enhanced markers of microbial translocation and higher pro-inflammatory gene profiles in the colonic biopsy samples compared with controls. In this regard, we found increased expression of the bacterial endotoxin-specific ligand TLR4, CD3+ T cells, cytokine expression in colonic biopsies, dysbiosis characterised by a decrease abundance of SCFA-producing colonic bacteria in subjects with PD. Rotenone treatment in TLR4-KO mice revealed less intestinal inflammation, intestinal and motor dysfunction, neuroinflammation and neurodegeneration, relative to rotenone-treated wild-type animals despite the presence of dysbiotic microbiota in TLR4-KO mice.ConclusionTaken together, these studies suggest that TLR4-mediated inflammation plays an important role in intestinal and/or brain inflammation, which may be one of the key factors leading to neurodegeneration in PD.

Diagnosis and management of inflammatory bowel disease (IBD) requires repeat diagnostic imaging for monitoring of disease activity. Recent evidence has suggested that patients with IBD are at increased risk of radiation exposure from repeat imaging. The aim of this article was to highlight risks associated with increasing radiation exposure and identify alternatives to minimize exposure. The increasing use of computed tomography (CT) in both Crohn's disease and ulcerative colitis has brought additional benefits to guiding management through non-invasive measures. However, the massive increase in use of CT scans poses a risk of exposing patients with IBD to high levels of diagnostic medical radiation. High levels of diagnostic medical radiation are associated with an increased risk of malignancy in several studies. Numerous studies have identified particular risk factors in IBD associated with high levels of diagnostic medical radiation which are also associated with a more severe disease course. Imaging techniques such as magnetic resonance enterography, ultrasound, small bowel follow through, and capsule endoscopy are alternatives to CT scans as they do not utilize radiation. Gastroenterologists managing patients with IBD, particularly Crohn's disease, should be aware of the increased risk of high cumulative doses of radiation exposure, particularly from CT scanning. Alternative forms of imaging should be carefully considered when evaluating patients, in particularly those with identifiable risk factors for an aggressive disease course.

We have shown that alcohol increases Caco-2 intestinal epithelial cell monolayer permeability in vitro by inducing the expression of redox-sensitive circadian clock proteins CLOCK and PER2 and that these proteins are necessary for alcohol-induced hyperpermeability. We hypothesized that alcohol metabolism by intestinal Cytochrome P450 isoform 2E1 (CYP2E1) could alter circadian gene expression ( Clock and Per2), resulting in alcohol-induced hyperpermeability. In vitro Caco-2 intestinal epithelial cells were exposed to alcohol, and CYP2E1 protein, activity, and mRNA were measured. CYP2E1 expression was knocked down via siRNA and alcohol-induced hyperpermeability, and CLOCK and PER2 protein expression were measured. Caco-2 cells were also treated with alcohol or H2O2 with or without N-acetylcysteine (NAC) anti-oxidant, and CLOCK and PER2 proteins were measured at 4 or 2 h. In vivo Cyp2e1 protein and mRNA were also measured in colon tissue from alcohol-fed mice. Alcohol increased CYP2E1 protein by 93% and enzyme activity by 69% in intestinal cells in vitro. Alcohol feeding also increased mouse colonic Cyp2e1 protein by 73%. mRNA levels of Cyp2e1 were not changed by alcohol in vitro or in mouse intestine. siRNA knockdown of CYP2E1 in Caco-2 cells prevented alcohol-induced hyperpermeability and induction of CLOCK and PER2 proteins. Alcohol-induced and H2O2-induced increases in intestinal cell CLOCK and PER2 were significantly inhibited by treatment with NAC. We concluded that our data support a novel role for intestinal CYP2E1 in alcohol-induced intestinal hyperpermeability via a mechanism involving CYP2E1-dependent induction of oxidative stress and upregulation of circadian clock proteins CLOCK and PER2.

Several studies indicate the importance of colonic microbiota in metabolic and inflammatory disorders and importance of diet on microbiota composition. The effects of alcohol, one of the prominent components of diet, on colonic bacterial composition is largely unknown. Mounting evidence suggests that gut-derived bacterial endotoxins are cofactors for alcohol-induced tissue injury and organ failure like alcoholic liver disease (ALD) that only occur in a subset of alcoholics. We hypothesized that chronic alcohol consumption results in alterations of the gut microbiome in a subgroup of alcoholics, and this may be responsible for the observed inflammatory state and endotoxemia in alcoholics. Thus we interrogated the mucosa-associated colonic microbiome in 48 alcoholics with and without ALD as well as 18 healthy subjects. Colonic biopsy samples from subjects were analyzed for microbiota composition using length heterogeneity PCR fingerprinting and multitag pyrosequencing. A subgroup of alcoholics have an altered colonic microbiome (dysbiosis). The alcoholics with dysbiosis had lower median abundances of Bacteroidetes and higher ones of Proteobacteria. The observed alterations appear to correlate with high levels of serum endotoxin in a subset of the samples. Network topology analysis indicated that alcohol use is correlated with decreased connectivity of the microbial network, and this alteration is seen even after an extended period of sobriety. We show that the colonic mucosa-associated bacterial microbiome is altered in a subset of alcoholics. The altered microbiota composition is persistent and correlates with endotoxemia in a subgroup of alcoholics.

Diarrhea is one of the common symptoms that significantly affects quality of life in patients with inflammatory bowel disease (IBD). The clinical manifestation of diarrhea is mainly dependant on the type of IBD and the location, extent and severity of intestinal inflammation. Understanding the pathophysiologic mechanisms of diarrhea in patients with IBD will be beneficial to developing effective treatments for IBD-associated diarrhea. In recent years, modern molecular techniques have been used intensively to dissect the role of the intestinal microbiota, epithelial barrier and the host immune system in the mechanisms of IBD-induced diarrhea. These studies have significantly advanced our knowledge of the mechanisms of IBD-induced diarrhea. In this article, we focus on the new and critical molecular insights into the contributions of the intestinal microbiota, epithelial tight junctions, proinflammatory cytokines and microRNA as potential mechanisms underlying to IBD-induced diarrhea.

The diagnosis of Parkinson's disease rests on motor signs of advanced central dopamine deficiency. There is an urgent need for disease biomarkers. Clinicopathological evidence suggests that α-synuclein aggregation, the pathological signature of Parkinson's disease, can be detected in gastrointestinal tract neurons in Parkinson's disease. We studied whether we could demonstrate α-synuclein pathology in specimens from unprepped flexible sigmoidoscopy of the distal sigmoid colon in early subjects with Parkinson's disease. We also looked for 3-nitrotyrosine, a marker of oxidative stress. Ten subjects with early Parkinson's disease not treated with dopaminergic agents (7 men; median age, 58.5 years; median disease duration, 1.5 years) underwent unprepped flexible sigmoidoscopy with biopsy of the distal sigmoid colon. Immunohistochemistry studies for α-synuclein and 3-nitrotyrosine were performed on biopsy specimens and control specimens from a tissue repository (23 healthy subjects and 23 subjects with inflammatory bowel disease). Nine of 10 Parkinson's disease samples were adequate for study. All showed staining for α-synuclein in nerve fibers in colonic submucosa. No control sample showed this pattern. A few showed light α-synuclein staining in round cells. 3-Nitrotyrosine staining was seen in 87% of Parkinson's disease cases but was not specific for Parkinson's disease. This study suggests a pattern of α-synuclein staining in Parkinson's disease that was distinct from healthy subjects and those with inflammatory bowel disease. The absence of this pattern in subjects with inflammatory bowel disease suggests it is not a sequel of inflammation or oxidative stress. 3-Nitrotyrosine immunostaining was common in all groups studied, suggesting oxidative stress in the colonic submucosa.

Ulcerative colitis (UC) is a chronic inflammatory bowel disease (IBD) associated with multiple colonic and extraintestinal complications, the most severe being the development of colorectal cancer (CRC). Compared to the general population, there is an increased risk of CRC associated with UC. Although the pathogenesis of CRC in UC is unknown, most studies have linked it to long-standing inflammation as well as other risk factors such as duration of disease, extent of inflammation, family history of CRC, and coexisting conditions such as primary sclerosing cholangitis (PSC). UC is a life-long disease for which patients enter a vigilant screening program which includes surveillance colonoscopy to promote early detection of CRC yet some controversies exist regarding the cost effectiveness of surveillance colonoscopy and improving survival. Newer modalities such as chromoendoscopy, narrow band imaging, high definition colonoscopy, and confocal microscopy have aided in developing a more targeted approach for early detection of dysplasia in surveillance colonoscopy. This review focuses on the role of chronic colonic inflammation and dysplasia in development of UC-associated CRC and current methods of screening, detection, chemoprevention, and treatment of UC-associated CRC.

Background: Alcohol consumption is a potential trigger for inflammatory bowel disease (IBD) flare because of alcohol-induced oxidative stress and its deleterious effects on gut barrier function. Additionally, we have recently shown that alcohol consumption is associated with more symptoms in IBD. However, it is not known whether moderate daily alcohol consumption can modify IBD disease activity. To test what effects alcohol may have on patients with IBD, we evaluated the effect of moderate daily red wine for 1 week on two factors associated with recurrent IBD disease activity: intestinal permeability and stool calprotectin. Methods: To assess the effects of moderate daily alcohol consumption on intestinal permeability and inflammation, we recruited 21 patients: 8 with inactive ulcerative colitis (UC), 6 with inactive Crohn’s disease (CD), and 7 healthy controls. All participants with IBD completed a validated questionnaire on disease activity (Crohn’s disease activity index or ulcerative colitis clinical activity index), to confirm they had inactive disease. All subjects then underwent a baseline assessment that included a blood draw, urine collection after sugar challenge, and stool collection. Subjects then consumed 1–3 glasses of red wine a day for 1 week (approx. 0.4 g EtOH/kg), and repeated the three measures. Results: No subjects flared during the study. Moderate alcohol consumption did not significantly change either clinical disease activity scores or C-reactive protein. In contrast to healthy subjects, daily consumption of red wine significantly (1) decreased stool calprotectin in IBD subjects from baseline (p = 0.001) and (2) increased intestinal permeability as measured by urinary lactulose/mannitol excretion (marker of small bowel permeability) in CD (p = 0.028) or urinary sucralose secretion (marker of large bowel permeability) in UC (p = 0.012). Conclusions: One week of moderate consumption of red wine in inactive IBD was associated with a significant decrease in stool calprotectin and a significant increase in intestinal permeability. Our data suggests that patients with inactive IBD who drink red wine daily may be at an increased long-term risk for disease relapse.

Alcohol and non-steroidal anti-inflammatory drugs (NSAIDS) are noxious agents that can disrupt the integrity of the gastroduodenal mucosal and damage the epithelial barrier, and lead to increased gastroduodenal permeability. Moreover, it is not uncommon that patients are exposed to these two barrier stressors at the same time. It is thus important to know how simultaneous exposure affects the gastroduodenal barrier, and acquiring that knowledge was the goal of this study. We used a method that has been widely used for the assessment of injury to the gastroduodenal barrier induced by these noxious agents – measurement of gastroduodenal permeability as indicated by urinary excretion of ingested sucrose. We used gas chromatography to measure the amount of sucrose excreted in the urine over the 5–12 h following ingestion of a bolus of sucrose. The 148 participants in the study included 92 alcoholics and 56 healthy controls. All study subjects had a baseline permeability test. To determine whether addition of a second noxious agent, in addition to chronic alcohol, further decreases gastroduodenal barrier integrity, a subset of 118 study subjects participated in another permeability test in which they were exposed to aspirin. For this test, participants ingested 1300 mg aspirin twice, 12 hours and 1 hour before the final permeability test. The baseline permeability test showed that alcoholics have significantly higher gastroduodenal permeability than controls. Aspirin caused a significant within group absolute increase in gastroduodenal permeability in both alcoholics and controls (+7.72%, p=0.003 and +2.25%, p = 0.011, respectively) but the magnitude of these increases were not significantly different from each other. Baseline permeability did vary by gender, self-reported illegal drug use, and employment type. The extent of the permeability increase after aspirin ingestion varied with illegal drug use and recruitment site (a surrogate marker of socioeconomic status). Our data show that alcoholics have greater gastroduodenal permeability than healthy controls. This difference was independent of the duration of any preceding period of sobriety, gender, smoking history, or illicit drug abuse. The injurious effects of alcohol on the gastroduodenal epithelial barrier are long lasting, persisting even after 7 days of sobriety. Although, acute aspirin and chronic alcohol each increase intestinal permeability in alcoholics, their effects appear to be additive rather than synergistic.

The 'hygiene hypothesis' suggests that a reduction in the microbial exposure due to improved health measures has contributed to an immunological imbalance in the intestine and increased the incidence of autoimmune diseases such as inflammatory bowel diseases (IBD). Accordingly, we investigated associations between oral hygiene practices and IBD.We developed and administered a multiple choice questionnaire to evaluate oral hygiene and dental care practices of 137 subjects (83 with IBD and 54 healthy controls).Of the 83 IBD cases, 31% had ulcerative colitis and 69% had Crohn's disease. For subjects with IBD, the frequency of brushing at disease onset was significantly higher than in controls (P=0.005). Also, the frequency of use of dental floss and breath freshener at disease onset was significantly higher in IBD patients (P=0.005 and0.001, respectively). Also, patients with IBD more frequently visited their dentist at disease onset (P0.001) and continued to visit their dentist more often (P0.001). IBD cases had a higher frequency of dental complications such as tooth caries (P=0.007), oral ulcers (P=0.04) and dry mouth (P=0.001).These findings suggest that oral hygiene practices may cause alterations in the flora of the oral mucosa, which causes imbalance in the gut microbiome (dysbiosis), and thereby contributes to the pathogenesis of IBD. Conversely, the increased frequency of dental problems in IBD patients might be due, at least in part, to alterations in oral flora or to their disease.

Background: Clinical and animal data indicate that gut-derived endotoxin and other luminal bacterial products are necessary cofactors for development of alcoholic liver disease (ALD). Although gut leakiness is clearly an important cause of endotoxemia in ALD, it cannot fully explain endotoxemia in all ALD subjects and thus other factors may be involved. One possible factor is a change in gut microbiota composition (dysbiosis). Thus, the aim of our study was to interrogate the gut bacterial microbiota in alcohol-fed rats to see if chronic alcohol consumption affects gut bacteria composition. Method: Male Sprague–Dawley rats were given either alcohol or dextrose intragastrically by gavage twice daily for up to 10 weeks. A subgroup of rats was also given either a probiotic (lactobacillus GG) or a prebiotic (oats) by gavage. Ileal and colonic mucosal-attached microbiota composition were interrogated by Length Heterogeneity PCR (LH-PCR) fingerprinting. Results: Bacterial microbiota composition in alcohol-fed rats is not different from dextrose-fed rats at weeks 4 and 6. Mucosa-associated microbiota composition in the colon is altered at 10 weeks of daily alcohol gavage. Both LGG and oats prevented alcohol-induced dysbiosis up to 10 weeks of alcohol treatment. Conclusion: Daily alcohol consumption for 10 weeks alters colonic mucosa-associated bacterial microbiota composition in rats. Our data showed, for the first time, that daily alcohol consumption can affect colonic microbiome composition and suggest that dysbiosis may be an important mechanism of alcohol-induced endotoxemia. Further studies are needed to determine how dysbiotic microbiota contributes to development of ALD and whether therapeutic interventions targeted towards dysbiotic microbiota can prevent complications of alcoholism like ALD.

The benefits of dietary fiber on inflammatory bowel disease may be related to the fermentative production of butyrate in the colon, which appears to decrease the inflammatory response. The benefits of dietary fiber against colon cancer may be related to both fermentative and non-fermentative processes, although poorly fermentable fibers appear more influential. Dietary fiber fermentation profiles are important in determining optimal fibers for colonic health, and may be a function of structure, processing conditions, and other food components. A greater understanding of the relationships between fermentation rate and dietary fiber structure would allow for development of dietary fibers for optimum colonic health.

The term non-alcoholic fatty liver disease (NAFLD) has been coined to represent the spectrum of histological changes, from pure non-alcoholic fatty liver (steatosis) to non-alcoholic steatohepatitis (NASH), fibrosis and finally cirrhosis (1, 2) Recent epidemiological data suggest a prevalence of 3–6% for NAFLD in the general population, making NAFLD the most common liver disease in the USA (3, 4). The single most important predictor in the natural history of progression from benign simple steatosis to life-threatening cirrhosis is the presence of inflammation and fibrosis (5). Associated factors such as age and diabetes play a significant but secondary role in NAFLD. Steatosis, the most common form of NAFLD, appears to be a benign process resulting from abnormal lipid metabolism. In contrast, NASH is far less common than steatosis and does not appear to be benign in most patients; not infrequently, it progresses to cirrhosis and liver failure (5). Because only 15–25% of patients with steatosis progress to NASH (6), it is highly likely that a cofactor is required to cause inflammation and liver cell necrosis in a setting of abnormal hepatic lipid homoeostasis and steatosis. This so-called two-hit hypothesis postulates that simple steatosis is the liver’s manifestation of the metabolic syndrome, but it requires a second hit (insult) for progression to NASH (7). One proposed second-hit mechanism is gut-derived endotoxin (8). This is based on two sets of findings. (1) Gut-derived endotoxin is likely a prerequisite cofactor in the development of alcoholic steatohepatitis and cirrhosis in alcoholics (9, 10), fatty liver being extremely common in alcoholics, but steatohepatitis and cirrhosis occur in only 15–25% of alcoholics. This pattern is similar to the pattern for NAFLD. (2) More direct evidence for the role of gut-derived endotoxin in the pathogenesis of NASH comes from animal studies. Animal studies have shown that systemic endotoxaemia leads to steatohepatitis in genetically obese rats, via release of tumour necrosis factor-α (11). In addition, Brun et al. (12) showed that genetically obese mice display enhanced intestinal permeability leading to increased portal endotoxaemia, which makes hepatic stellate cells more sensitive to bacterial endotoxin. Possible mechanisms for endotoxaemia in patients with NASH include (a) bacterial overgrowth and (b) disrupted intestinal barrier integrity (leaky gut) that results in increased absorption of endotoxin (12–14). Indeed, we recently demonstrated increased intestinal permeability (leaky gut) in alcoholics with liver disease (15), a disorder with pathophysiological and histological features similar to NAFLD and NASH. It remains to be determined, however, whether leaky gut is also present in patients with NASH. Accordingly, we hypothesized that patients with NASH have either frank gut leakiness or easily triggered susceptibility to gut leakiness (intermittent gut leakiness), either of which results in increased passage of gut-derived bacterial products such as endotoxin into the portal circulation, where it initiates hepatic necroinflammatory cascades and results in liver cell injury. The aim of our study was to determine whether patients with NASH have leaky guts or susceptibility to gut leakiness.

Background Mast cells have a primary role in atopy. Mast cells may play a unique role in a subgroup of patients with irritable bowel syndrome (IBS). This observation suggests a link between atopic disorders and IBS. Objective To determine whether there is an association between atopic disorders and IBS. Methods We undertook a prospective study using structured questionnaires. We administered questionnaires to 125 consecutive patients seen in the following clinics from July 1 through October 31, 2001: allergy/immunology (AI) (n = 39), gastroenterology (n = 36), and general medicine (n = 50). The survey included questions detailing gastrointestinal and allergic symptoms. Diagnosis of IBS was based on Rome II criteria. Diagnosis of atopy was based on clinical parameters. Results The AI clinic reported a significantly ( P = .015) higher rate of IBS than the general medicine clinic. The IBS incidence reported in the AI clinic was similar to that reported in the gastroenterology clinic. The likelihood of IBS was significantly higher in patients with seasonal allergic rhinitis (2.67 times; 95% confidence interval [CI], 1.10-6.49; P = .03), patients with allergic eczema (3.85 times; 95% CI, 1.72-8.60; P = .001), and patients with depression (2.56 times; 95% CI, 1.05-6.14; P = .04). Patients reporting atopic symptoms (seasonal allergic rhinitis, allergic eczema, and asthma) were 3.20 times (95% CI, 1.20-8.50) ( P = .02) more likely to fulfill the criteria for IBS. Conclusions Adults with atopic symptoms report a high incidence of IBS, suggesting a link between atopy and IBS. We proposed a subgroup of patients with IBS (atopic IBS) who have typical IBS symptoms in association with atopic manifestations. Identifying atopic vs nonatopic IBS may help in identifying the underlying pathophysiologic mechanisms and therapeutic options.

Alcohol-induced intestinal hyperpermeability (AIHP) is a known risk factor for alcoholic liver disease (ALD), but only 20–30% of heavy alcoholics develop AIHP and ALD. The hypothesis of this study is that circadian misalignment would promote AIHP. We studied two groups of healthy subjects on a stable work schedule for 3 mo [day workers (DW) and night workers (NW)]. Subjects underwent two circadian phase assessments with sugar challenge to access intestinal permeability between which they drank 0.5 g/kg alcohol daily for 7 days. Sleep architecture by actigraphy did not differ at baseline or after alcohol between either group. After alcohol, the dim light melatonin onset (DLMO) in the DW group did not change significantly, but in the NW group there was a significant 2-h phase delay. Both the NW and DW groups had no change in small bowel permeability with alcohol, but only in the NW group was there an increase in colonic and whole gut permeability. A lower area under the curve of melatonin inversely correlated with increased colonic permeability. Alcohol also altered peripheral clock gene amplitude of peripheral blood mononuclear cells in CLOCK, BMAL, PER1, CRY1, and CRY2 in both groups, and inflammatory markers lipopolysaccharide-binding protein, LPS, and IL-6 had an elevated mesor at baseline in NW vs. DW and became arrhythmic with alcohol consumption. Together, our data suggest that central circadian misalignment is a previously unappreciated risk factor for AIHP and that night workers may be at increased risk for developing liver injury with alcohol consumption.

Diverticulosis and its complications are important healthcare problems in the USA and throughout the Western world. While mechanisms as to how diverticulosis occurs have partially been explored, few studies examined the relationship between colonic gases such as methane and diverticulosis in humans. This study aimed to demonstrate a significant relationship between methanogenic Archaea and development of diverticulosis. Subjects who consecutively underwent hydrogen breath test at Rush University Medical Center between 2003 and 2010 were identified retrospectively through a database. Medical records were reviewed for presence of a colonoscopy report. Two hundred and sixty-four subjects were identified who had both a breath methane level measurement and a colonoscopy result. Additional demographic and clinical data were obtained with chart review. Mean breath methane levels were higher in subjects with diverticulosis compared to those without diverticulosis (7.89 vs. 4.94 ppm, p = 0.04). Methane producers (defined as those with baseline fasting breath methane level >5 ppm) were more frequent among subjects with diverticulosis compared to those without diverticulosis (50.9 vs. 34 %, p = 0.0025). When adjusted for confounders, breath methane levels and age were the two independent predictors of diverticulosis on colonoscopy with logistic regression modeling. Methanogenesis is associated with the presence of diverticulosis. Further studies are needed to confirm our findings and prospectively evaluate a possible etiological role of methanogenesis and methanogenic archaea in diverticulosis.

Background and Aim: The deleterious effects of stress in inflammatory bowel disease (IBD) have been attributed to activation of the brain–gut axis (BGA) and its end effectors, mast cells (MC). We previously showed that cold pressor stress test (CPT) results in increased activation and degranulation (but not increased proliferation) of mucosal MC, mitochondrial damage to epithelial cells and mucosal protein oxidation in both healthy controls and IBD patients. These changes are more marked in IBD patients. This increased activation of MC in IBD could be due to (i) greater activation of the BGA or (ii) inherited or acquired abnormalities in mucosal MC. In the current study we investigated the latter possibility. Methods: To assess the effects of stress on mucosal MC in patients with IBD, seven controls and 15 subjects with inactive IBD underwent 5 consecutive days of CPT to activate the BGA. Endoscopic mucosal biopsies of the distal sigmoid colon were taken during unprepared sigmoidoscopy before the first CPT and after the last CPT, and formalin-fixed samples were stained for both MC granules (MCg) and for the c-kit receptor, which is present on MC membranes (MCm). Mast cell degranulation was assessed using electron microscopy. Results: Mast cell granule staining suggested that IBD subjects do not have a significantly different number of MC compared with controls, either before or after stress. Mast cell membrane staining, in contrast, suggested that MC c-kit immunostaining was significantly reduced – at both baseline (P = 0.01) and post stress (P = 0.04) samples – in IBD patients compared to controls. MC c-kit immunostaining was independent of stress-induced MC degranulation. There was no significant change in MC number as a result of the stress intervention using either staining method in both groups. Conclusion: These data support our previous report that the size of the mucosal MC population in patients with inactive IBD is not altered by disease or by stress, yet MC in IBD are different (fewer c-kit receptors) and respond differently (greater activation) than MC in control subjects. It remains to be seen whether this abnormality is an inherited or acquired one and to identify its role and mechanism in tissue injury in the pathogenesis of IBD.

Background & Aims: Pouch inflammation after surgery for ulcerative colitis can significantly alter quality of life and thus ideally should be prevented. Dysbiosis or altered microflora is suspected to be the key pathogenic factor for pouchitis. However, dysbiosis in pouchitis has not been characterized carefully because of a lack of available sensitive microbiological technology suitable for in vivo studies in human beings. Thus, the aims of our study were as follows: (1) to show the use of the length heterogeneity polymerase chain reaction (LH-PCR) technique for studying microflora in human beings, and (2) to use the technique to characterize the microfloral patterns in the ileal pouch of patients with pouchitis. Methods: Microfloral patterns initially were assessed using a 16S ribosomal RNA technique (LH–PCR) to determine the qualitative changes in the luminal and mucosal intestinal flora. We subsequently cloned and sequenced the LH-PCR amplification products from the community 16S ribosomal RNA found in patients with pouchitis and in control pouch to identify the microbial species involved in pouchitis. Results: We have shown unique microfloral patterns in pouchitis. Through cloning and sequencing of the LH-PCR amplicons, we have shown the persistence of Fusobacter and Enteric species associated with the disease state and the absence of specific bacteria such as Streptococcus species in the inflamed pouch. Conclusions: We have shown that the LH-PCR technique is suitable for studying microflora in human beings. By using this technique and the clone sequences, we have shown dysbiosis in the microbial biofilm adherent to the mucosa in pouchitis. Our data provide direct evidence of the role of bacteria in the pathogenesis of pouchitis.