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1.
Dig Dis Sci ; 65(5): 1423-1428, 2020 05.
Article in English | MEDLINE | ID: mdl-31617129

ABSTRACT

BACKGROUND: Clostridium difficile infection (CDI) is one of the leading causes of health-care-associated infections in the USA. There are limited data available regarding CDI in hospitalized patients with inflammatory bowel disease-related ileal pouch. AIMS: This study aimed to evaluate the demographics, clinical features, risk factors, and admission outcomes among hospitalized patients with CDI-related pouchitis (CDP). METHODS: Retrospective chart review was performed for patients who were admitted to our institute for pouchitis between 2013 and 2016 to identify patients with CDP. Logistic regression analysis was performed to assess the risk factors associated with CDP. RESULTS: A total of 160 subjects with pouchitis had a total of 218 admissions during the study period. Primary admission diagnosis was pouchitis or inflammatory bowel disease flare-up for 202 (93%) admissions. Clostridium difficile was tested at least once for 72 patients, and the diagnosis of CDP was established for 16 (10%) patients. All patients with CDP were symptomatic, 13 (81%) had diarrhea, 8 (50%) had abdominal pain, 7 (44%) had nausea/vomiting, and 2 (13%) had gastrointestinal bleeding. On multivariable analysis, only body mass index > 25 (OR 0.25, 95% CI 0.06-0.94, p = 0.048) was significantly associated with decreased risk of CDP. No patients in CDP cohort were admitted to ICU, died at the hospital, or readmitted in 30 days after the discharge. CONCLUSIONS: In our cohort, obesity was associated with low risk of CDP among hospitalized patients with pouchitis. This finding warrants further validation in prospective studies.


Subject(s)
Anti-Bacterial Agents/adverse effects , Clostridioides difficile , Cross Infection/chemically induced , Enterocolitis, Pseudomembranous/chemically induced , Obesity/complications , Postoperative Complications/chemically induced , Pouchitis/drug therapy , Adult , Aged , Aged, 80 and over , Colonic Pouches/microbiology , Cross Infection/microbiology , Enterocolitis, Pseudomembranous/microbiology , Female , Hospitalization , Humans , Male , Middle Aged , Obesity/microbiology , Obesity/surgery , Postoperative Complications/microbiology , Pouchitis/microbiology , Retrospective Studies , Risk Factors , Young Adult
2.
Colorectal Dis ; 18(5): O154-7, 2016 May.
Article in English | MEDLINE | ID: mdl-26945555

ABSTRACT

AIM: Clostridium difficile infection (CDI) of the ileal pouch following restorative proctocolectomy (RPC) is becoming increasingly recognized. We aimed to understand better (i) the associated risk factors, (ii) treatment practices and (iii) the pouch diversion and failure rate in patients who developed CDI of the pouch after RPC for ulcerative colitis (UC). METHOD: Patients who tested positive for C. difficile of the pouch between 2007 and 2010 were included in the analysis. Data collected included patient demographics, time from RPC to documented CDI, the treatment of CDI and rate of excision of the pouch. RESULTS: Of 2785 patients recorded in the hospital CDI database, 15 had had an RPC with ileal pouch anal anastomosis. The median age was 44 years and the median interval from RPC to first documented episode of CDI was 3 years. Thirteen (81%) patients had had multiple episodes of pouchitis before and after CDI infection, and all were symptomatic at the time of testing for CDI. Within 30 days of the diagnosis of CDI, six (40%) patients were taking immunosuppressive medication, seven (47%) were taking a proton pump inhibitor and 12 (80%) had received antibiotics. Five patients required hospitalization for CDI and four had severe infections characterized by a serum creatinine more than 1.5 times baseline (n = 3) and a white cell count above 15 000 (n = 1). Six patients who underwent endoscopy had severe inflammation of the pouch including the presence of a pseudomembrane in one case. Ten patients were treated with metronidazole alone and five with vancomycin. Two patients had recurrent CDI of the pouch during a median follow-up period of 2.9 years and one had CDI refractory to medical management. This patient required diversion of the pouch with an ileostomy for refractory CDI but no patient required excision of the pouch. CONCLUSION: All 15 patients developing CDI of the pouch were successfully treated with antibiotics and only one required surgery in the form of an ileostomy.


Subject(s)
Clostridioides difficile , Enterocolitis, Pseudomembranous/microbiology , Postoperative Complications/microbiology , Pouchitis/microbiology , Adolescent , Adult , Anal Canal/surgery , Anastomosis, Surgical/adverse effects , Anti-Bacterial Agents/therapeutic use , Colitis, Ulcerative/surgery , Colonic Pouches/adverse effects , Colonic Pouches/microbiology , Enterocolitis, Pseudomembranous/drug therapy , Female , Humans , Male , Middle Aged , Postoperative Complications/drug therapy , Pouchitis/drug therapy , Proctocolectomy, Restorative/adverse effects , Retrospective Studies , Young Adult
3.
Surg Today ; 46(8): 939-49, 2016 Aug.
Article in English | MEDLINE | ID: mdl-26510664

ABSTRACT

PURPOSE: Ulcerative colitis (UC) is a chronic, relapsing, and refractory disorder of the intestine. Total proctocolectomy with ileal pouch anal anastomosis (IPAA) is the preferred and standard surgical procedure for patients' refractory to medical therapy. Pouchitis is one of the most common long-term complications after IPAA. In the present study, the safety and efficacy of Clostridium butyricum MIYAIRI (CBM) as a probiotic were examined. METHODS: A randomized and placebo-controlled study was performed. Seventeen patients were recruited from 2007 to 2013. Nine tablets of MIYA-BM(®) or placebo were orally administered once daily. The cumulative pouchitis-free survival, pouch condition (using the modified pouch disease activity index), and blood parameters were evaluated. A fecal sample analysis was also performed. RESULTS: Subjects were randomly allocated to receive MIYA-BM or placebo (9 and 8 subjects, respectively). One subject in the MIYA-BM group and four subjects in the placebo group developed pouchitis. No side effects occurred in either group. Characteristic intestinal flora was observed in each group. CONCLUSIONS: Our results suggest that probiotic therapy with CBM achieved favorable results with minimal side effects and might be a useful complementary therapy for the prevention of pouchitis in patients with UC who have undergone IPAA.


Subject(s)
Clostridium butyricum , Colitis, Ulcerative/microbiology , Colitis, Ulcerative/surgery , Gastrointestinal Microbiome , Postoperative Complications/prevention & control , Pouchitis/prevention & control , Probiotics/administration & dosage , Administration, Oral , Adult , Anastomosis, Surgical , Colonic Pouches/microbiology , Female , Humans , Male , Middle Aged , Proctocolectomy, Restorative
4.
Am J Gastroenterol ; 109(7): 941-7, 2014 Jul.
Article in English | MEDLINE | ID: mdl-24989088

ABSTRACT

Clostridium difficile (C. difficile) infection (CDI) following total proctocolectomy and ileal pouch-anal anastomosis has been increasingly recognized over the past 5 years. CDI of the ileal pouch has been recognized in ∼10% of symptomatic patients seen at a tertiary referral center for pouch dysfunction. In contrast to colonic CDI in the general population or in patients with inflammatory bowel disease, postoperative antibiotic exposure and the use of immunosuppressive agents or proton pump inhibitors do not appear to be associated with CDI of the pouch. Male gender, recent hospitalization, and presurgery antibiotic use were shown to be risk factors for ileal pouch CDI. The ileal pouch may be susceptible to CDI owing to similarities with the colon at physiological and structural levels. Postcolectomy CDI likely represents a spectrum of disease processes, varying from asymptomatic colonization to severe symptomatic infection. CDI should be considered in any patient with an ileal pouch presenting with a change in "normal" symptom pattern or treatment-refractory disease. Sensitive and specific methods for the detection of CDI are available, and pouchoscopy is a valuable tool in the evaluation of the patient with symptomatic CDI of the pouch. At a referral center for pouch dysfunction, vancomycin is used as the first-line therapy for ileal pouch CDI. Fecal microbiota transplantation may find use in the management of severe or antibiotic refractory CDI-related pouchitis.


Subject(s)
Clostridioides difficile , Clostridium Infections/diagnosis , Clostridium Infections/microbiology , Colonic Pouches/microbiology , Postoperative Complications/diagnosis , Postoperative Complications/microbiology , Anti-Bacterial Agents/therapeutic use , Clostridium Infections/drug therapy , Endoscopy , Humans , Postoperative Complications/drug therapy , Proctocolectomy, Restorative , Risk Factors , Sex Factors , Vancomycin/therapeutic use
5.
Dig Dis Sci ; 59(6): 1262-8, 2014 Jun.
Article in English | MEDLINE | ID: mdl-24504594

ABSTRACT

BACKGROUND: Clostridium difficile infection (CDI) might contribute to a refractory course of pouchitis. However, the association between preoperative CDI and postoperative CDI in ileal pouch patients has not been investigated. AIM: Our study aimed to evaluate whether preoperative CDI had an impact on the occurrence of postoperative CDI in pouch patients. METHODS: Consecutive eligible ileal pouch patients from February 2005 to December 2012 were identified from the Pouchitis Registry at the Cleveland Clinic. Patients in the registry with known status of CDI of the pouch were surveyed with a structured questionnaire regarding preoperative C. difficile test and its treatment. Medical records were also reviewed. Demographics and clinical characteristics and outcomes were evaluated with univariable and multivariable analyses. RESULTS: A total of 102 patients with preoperative C. difficile test were identified for this study and 21 patients (20.6%) tested positive for C. difficile test after colectomy. In logistic regression analysis, male patients were 7.85 (P = 0.003) times more likely to have CDI than women. In addition, preoperative significant comorbidities (P = 0.037) and preoperative use antibiotics for other indications (P = 0.005) were found to be associated with postoperative CDI of the pouch. However, there was no evidence to suggest that the preoperative CDI was associated with the occurrence of postoperative CDI (P = 0.769). CONCLUSIONS: Postoperative CDI occurred frequently in male patients with IPAA. In addition, preoperative comorbidities and antibiotic use were found to be risk factors for CDI of the pouch. However, preoperative CDI did not appear to be associated with an increased risk for postoperative CDI in pouch patients.


Subject(s)
Clostridioides difficile , Clostridium Infections/microbiology , Colonic Pouches/adverse effects , Adult , Anti-Bacterial Agents/therapeutic use , Clostridium Infections/drug therapy , Colonic Pouches/microbiology , Data Collection , Female , Humans , Male , Middle Aged , Postoperative Complications , Risk Factors , Surveys and Questionnaires
6.
Int J Colorectal Dis ; 27(11): 1455-63, 2012 Nov.
Article in English | MEDLINE | ID: mdl-22430887

ABSTRACT

BACKGROUND: Fecal antibodies against bacterial products may directly reflect the interaction between luminal bacteria and mucosal immunity, and assays for these antibodies may be clinically useful in the diagnosis and differential diagnosis of Crohn's disease-like (CDL) condition of the pouch. AIMS: This study aims to evaluate stool and serum anti-Saccharomyces cerevisiae antibodies (ASCA) in normal and diseased pouches, to assess the correlation between ASCA levels and endoscopic disease activity, and to ascertain the diagnostic utility of ASCA for CDL of the pouch. METHODS: One hundred eighty-nine patients with ileal pouches were prospectively enrolled and corresponding serum and pouch aspirate samples were collected. Fecal and serum ASCA levels were measured with enzyme-linked immunosorbent assay in a blinded fashion. Statistical analysis was then conducted using the signed rank test, Spearman correlation coefficients, and analysis of variance. RESULTS: Forty-three patients (22.8 %) had irritable pouch syndrome or normal pouches, 74 (39.2 %) had pouchitis/cuffitis, 52 (27.5 %) had CDL, 9 (4.8 %) had familial adenomatous polyposis, and 11 (5.8 %) had surgical complications of the pouch. Receiver operating characteristic curves to distinguish CDL from other categories of pouch dysfunction had an area under the curve (AUC) of 0.608 for fecal ASCA and an AUC of 0.517 for serum ASCA. Neither fecal nor serum ASCA correlated with endoscopic disease activity scores. There was a significant difference in the mean values of fecal ASCA between inflammatory and fistulizing CDL (0.27 vs. 0.03 ELISA units/ml, P < 0.05). CONCLUSIONS: Fecal ASCA appears to be better than serum ASCA in differentiating CDL from other pouch disorders, although this distinction may be of limited clinical utility.


Subject(s)
Antibodies, Fungal/blood , Colonic Pouches/microbiology , Crohn Disease/diagnosis , Crohn Disease/immunology , Feces/microbiology , Saccharomyces cerevisiae/immunology , Colonic Pouches/immunology , Crohn Disease/blood , Demography , Diagnosis, Differential , Endoscopy , Female , Humans , Male , Middle Aged , Phenotype , ROC Curve
7.
Dig Dis Sci ; 57(11): 2965-74, 2012 Nov.
Article in English | MEDLINE | ID: mdl-22539039

ABSTRACT

BACKGROUND: We previously investigated fecal flora of the pouch after total proctocolectomy using terminal restriction fragment polymorphism analysis. Although the results of the cluster analysis demonstrated clearly that bacterial populations, including an unidentified bacteria generating a 213-bp PCR fragment, moved toward a colon-like community in the pouch, it did not track changes in the individual species of fecal bacteria. AIMS: The aim of the present study was to estimate genome copy number of ten bacterial species, clusters, groups, or subgroups (including the bacteria generating 213-bp fragment in the previous study) in feces samples from pouches at various times following ileostomy closure. METHODS: A total of 117 stool samples were collected from patients with ulcerative colitis after surgery as well as healthy volunteers. We used real-time polymerase chain reaction of the 16S rRNA gene to estimate genome copy numbers for the nine bacterial populations and the bacteria generating 213-bp fragment after identification by DNA sequencing. RESULTS: We demonstrated a time-dependent increase in the number of anaerobic and colon-predominant bacteria (such as Clostridium coccoides, C. leptum, Bacteroides fragilis and Atopobium) present in proctocolectomy patients after stoma closure. In contrast, numbers of ileum-predominant bacterial species (such as Lactobacillus and Enterococcus faecalis) declined. CONCLUSIONS: Our data confirm previous findings that fecal flora in the pouch after total proctocolectomy changes significantly, and further demonstrate that the number and diversity of ileal bacteria decreases while a more colon-like community develops. The present data are essential for the future analysis of pathological conditions in the ileal pouch.


Subject(s)
Colitis, Ulcerative/microbiology , Colitis, Ulcerative/surgery , Colonic Pouches/microbiology , Feces/microbiology , Proctocolectomy, Restorative , Adolescent , Adult , Aged , Case-Control Studies , Colon/microbiology , DNA, Bacterial/analysis , Female , Humans , Ileum/microbiology , Male , Middle Aged , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Statistics, Nonparametric
8.
Cells ; 10(11)2021 11 19.
Article in English | MEDLINE | ID: mdl-34831464

ABSTRACT

The pathogenesis of ulcerative colitis (UC) is unknown, although genetic loci and altered gut microbiota have been implicated. Up to a third of patients with moderate to severe UC require proctocolectomy with ileal pouch ano-anastomosis (IPAA). We aimed to explore the mucosal microbiota of UC patients who underwent IPAA. METHODS: For microbiome analysis, mucosal specimens were collected from 34 IPAA individuals. Endoscopic and histological examinations of IPAA were normal in 21 cases, while pouchitis was in 13 patients. 19 specimens from the healthy control (10 from colonic and 9 from ileum) were also analyzed. Data were analyzed using an ensemble of software packages: QIIME2, coda-lasso, clr-lasso, PICRUSt2, and ALDEx2. RESULTS: IPAA specimens had significantly lower bacterial diversity as compared to normal. The microbial composition of the normal pouch was also decreased also when compared to pouchitis. Faecalibacterium prausnitzii, Gemmiger formicilis, Blautia obeum, Ruminococcus torques, Dorea formicigenerans, and an unknown species from Roseburia were the most uncommon in pouch/pouchitis, while an unknown species from Enterobacteriaceae was over-represented. Propionibacterium acnes and Enterobacteriaceae were the species most abundant in the pouchitis and in the normal pouch, respectively. Predicted metabolic pathways among the IPAA bacterial communities revealed an important role of immunometabolites such as SCFA, butyrate, and amino acids. CONCLUSIONS: Our findings showed specific bacterial signature hallmarks of dysbiosis and could represent bacterial biomarkers in IPAA patients useful to develop novel treatments in the future by modulating the gut microbiota through the administration of probiotic immunometabolites-producing bacterial strains and the addition of specific prebiotics and the faecal microbiota transplantation.


Subject(s)
Colitis, Ulcerative/microbiology , Colonic Pouches/immunology , Colonic Pouches/microbiology , Intestinal Mucosa/microbiology , Metabolome , Microbiota , Adult , Biodiversity , Entropy , Female , Humans , Male , Microbiota/genetics , Middle Aged , Phylogeny , Principal Component Analysis , RNA, Ribosomal, 16S/genetics
9.
Ann Surg ; 252(1): 90-8, 2010 Jul.
Article in English | MEDLINE | ID: mdl-20562611

ABSTRACT

OBJECTIVE: To identify, compare, and contrast the microbiota in patients with and without pouchitis after restorative proctocolectomy (RPC) for ulcerative colitis (UC) and familial adenomatous polyposis (FAP). SUMMARY BACKGROUND DATA: Pouchitis is the most common complication following RPC. An abnormal host-microbial interaction has been implicated. We investigated the pouch microbiota in patients with and without pouchitis undergoing restorative proctocolectomy for UC and FAP. METHODS: Mucosal pouch biopsies, taken from 16 UC (pouchitis 8) and 8 FAP (pouchitis 3) patients were analyzed to the species (or phylotype) level by cloning and sequencing of 3184 full-length bacterial 16S rRNA genes. RESULTS: There was a significant increase in Proteobacteria (P = 0.019) and a significant decrease in Bacteroidetes (P = 0.001) and Faecalibacterium prausnitzii (P = 0.029) in the total UC compared with the total FAP cohort, but only limited differences were found between the UC nonpouchitis and pouchitis groups and the FAP pouchitis and nonpouchitis groups. Bacterial diversity in the FAP nonpouchitis group was significantly greater than in UC nonpouchitis (P = 0.019) and significantly greater in UC nonpouchitis compared with UC pouchitis (P = 0.009). No individual species or phylotype specifically associated with either UC or FAP pouchitis was found. CONCLUSIONS: UC pouch patients have a different, less diverse, gut microbiota than FAP patients. A further reduction in bacterial diversity but no significant dysbiosis occurs in those with pouchitis. The study suggests that a dysbiosis occurs in the ileal pouch of UC RPC patients which predisposes to, but may not directly cause, pouchitis.


Subject(s)
Pouchitis/microbiology , RNA, Ribosomal, 16S/genetics , Adenomatous Polyposis Coli/microbiology , Adenomatous Polyposis Coli/surgery , Adult , Bacteroidetes/genetics , Bacteroidetes/isolation & purification , Base Sequence , Biopsy , Cloning, Molecular , Colitis, Ulcerative/microbiology , Colitis, Ulcerative/surgery , Colonic Pouches/microbiology , DNA, Bacterial/analysis , Humans , Male , Middle Aged , Molecular Sequence Data , Proctocolectomy, Restorative , Proteobacteria/genetics , Proteobacteria/isolation & purification
10.
Aliment Pharmacol Ther ; 52(8): 1323-1340, 2020 10.
Article in English | MEDLINE | ID: mdl-32955120

ABSTRACT

BACKGROUND: There is expanding interest in the role that diet plays in ileoanal pouch function and in the pathogenesis of pouchitis. AIMS: To present a narrative review of published literature regarding the relationship of diet with pouch function and the pathogenesis of pouchitis, and to provide potentially beneficial dietary strategies. METHODS: Current relevant literature was summarised and critically examined. RESULTS: Dietary components influence pouch function via their effect on upper gastrointestinal transit, small bowel water content and the structure and fermentative activity of the pouch microbiota. FODMAPs in fruits and vegetables appear to affect pouch function the most, with intake positively associated with increased stool frequency and reduced consistency. Dietary factors that influence the pathogenesis of pouchitis appear different and, at times, opposite to those better for optimising function. For example, risk of pouchitis appears to be inversely associated with intake of fruits. The food components mechanistically responsible for this observation are not known, but a rich supply of fermentable fibres and micronutrients in such foods might play a beneficial role via modulation of microbial community structure (such as increasing diversity and/or changing microbial communities to favour 'protective' over 'pathogenic' bacteria) and function and/or anti-inflammatory effects. CONCLUSION: Available data are weak but suggest tailoring dietary recommendations according to pouch phenotype/behaviour and pouchitis risk might improve outcomes. More sophisticated dietary strategies that utilise the physiological and pathophysiological effects of dietary components on ileoanal pouches have potential to further improve outcomes. Well designed, adequately powered studies are required.


Subject(s)
Colonic Pouches/physiology , Diet , Pouchitis/etiology , Colonic Pouches/microbiology , Diet/adverse effects , Humans , Microbiota/physiology , Pouchitis/diet therapy , Pouchitis/prevention & control , Risk Factors
11.
Cell Host Microbe ; 27(4): 659-670.e5, 2020 04 08.
Article in English | MEDLINE | ID: mdl-32101703

ABSTRACT

Secondary bile acids (SBAs) are derived from primary bile acids (PBAs) in a process reliant on biosynthetic capabilities possessed by few microbes. To evaluate the role of BAs in intestinal inflammation, we performed metabolomic, microbiome, metagenomic, and transcriptomic profiling of stool from ileal pouches (surgically created resevoirs) in colectomy-treated patients with ulcerative colitis (UC) versus controls (familial adenomatous polyposis [FAP]). We show that relative to FAP, UC pouches have reduced levels of lithocholic acid and deoxycholic acid (normally the most abundant gut SBAs), genes required to convert PBAs to SBAs, and Ruminococcaceae (one of few taxa known to include SBA-producing bacteria). In three murine colitis models, SBA supplementation reduces intestinal inflammation. This anti-inflammatory effect is in part dependent on the TGR5 bile acid receptor. These data suggest that dysbiosis induces SBA deficiency in inflammatory-prone UC patients, which promotes a pro-inflammatory state within the intestine that may be treated by SBA restoration.


Subject(s)
Bile Acids and Salts/metabolism , Colonic Pouches/microbiology , Dysbiosis/complications , Feces/microbiology , Receptors, G-Protein-Coupled/metabolism , Adenomatous Polyposis Coli/microbiology , Animals , Bile Acids and Salts/pharmacology , Colitis/etiology , Colitis/microbiology , Disease Models, Animal , Humans , Inflammation/drug therapy , Inflammation/etiology , Intestines/drug effects , Intestines/pathology , Metagenome , Mice , Microbiota , Receptors, G-Protein-Coupled/drug effects , Ruminococcus/isolation & purification , Transcriptome
12.
Am J Gastroenterol ; 104(4): 1013-23, 2009 Apr.
Article in English | MEDLINE | ID: mdl-19259080

ABSTRACT

Ileal pouch-anal anastomosis is the procedure of choice in the surgical management of refractory ulcerative colitis. Pouchitis affects up to 60% of patients following ileal pouch-anal anastomosis for ulcerative colitis. It overlaps significantly with ulcerative colitis such that improvements in our understanding of one will impact considerably on the other. The symptoms are distressing and impinge significantly on patients' quality of life. Despite 30 years of scientific and clinical investigation, the pathogenesis of pouchitis is unknown; however, recent advances in molecular and cell biology make a synergistic hypothesis possible. This hypothesis links interaction between epithelial metaplasia, changes in luminal bacteria (in particular sulfate-reducing bacteria), and altered mucosal immunity. Specifically, colonic metaplasia supports colonization by sulfate-reducing bacteria that produce hydrogen sulfide. This causes mucosal depletion and subsequent inflammation. Although in most cases antibiotics lead to bacterial clearance and symptom resolution, immunogenetic subpopulations can develop a chronic refractory variant of pouchitis. The aims of this paper are to discuss proposed pathogenic mechanisms and to describe a novel mechanism that combines many hypotheses and explains several aspects of pouchitis. The implications for the management of both pouchitis and ulcerative colitis are discussed.


Subject(s)
Bacteria/growth & development , Intestinal Mucosa/pathology , Pouchitis , Anal Canal/microbiology , Anal Canal/pathology , Anal Canal/surgery , Anastomosis, Surgical/methods , Anti-Bacterial Agents/therapeutic use , Colitis, Ulcerative/surgery , Colonic Pouches/microbiology , Colonic Pouches/pathology , Diagnosis, Differential , Humans , Intestinal Mucosa/microbiology , Metaplasia/pathology , Postoperative Complications , Pouchitis/diagnosis , Pouchitis/drug therapy , Pouchitis/etiology
13.
Dis Colon Rectum ; 52(8): 1492-500, 2009 Aug.
Article in English | MEDLINE | ID: mdl-19617766

ABSTRACT

PURPOSE: Previous studies on dysbiosis and pouchitis using conventional culture techniques have been disappointing because of inherent limitations associated with the technique. This study was designed to use terminal restriction fragment length polymorphism to evaluate patients with and without pouchitis. METHODS: Bacterial microbiota in 20 pouch patients (15 healthy and 5 with inflamed) were studied. DNA was extracted from feces, and polymerase chain reaction was performed using primers (V6-V8 region) that were modified at the 5' end with cyanine dyes. Amplicons were digested with merozoite surface protein-1 enzyme. The restricted fragments were analyzed by capillary electrophoresis, and the electrophenograms were studied. Electrophenograms provide information about operational taxonomic units, which correspond to specific organisms. Principal component analysis was performed to identify dominant and important operational taxonomic units in the 20 patients. Bacterial diversity and counts of these operational taxonomic units were compared in the two groups of patients. RESULTS: Total bacterial diversity in patients with pouchitis was similar to that in patients with healthy pouches (16 (11-20) vs. 12 (9-13), P = 0.279). Using principal component analysis, 29 operational taxonomic units were found to be important. Bacterial counts of seven dominant organisms (operational taxonomic unit 79 (enterococci), 85 (Pantoea), 88 (Enterobacteriaceae), 90 (eubacteria), 91 (Pseudomonas), 146 (clostridia), and 148 (bacilli)) were similar in patients with pouchitis and those with a healthy pouch (P > 0.05). Seventeen (operational taxonomic unit 73 (Leptospira), 93 (Pseudoalteromonas), 96, 100 (Desulfosporosinus), 114, 121, 134, 137, 141 (Microcystis), 159, 174 (Methylobacter), 193 (uncultured proteobacteria), 232, 376, 381, 414, and 465) of the remaining 22 nondominant organisms were seen exclusively in patients with pouchitis. The majority of these organisms were novel. CONCLUSION: Terminal restriction fragment length polymorphism can be used to identify candidate organisms that may be associated with pouchitis.


Subject(s)
Bacteria/genetics , Bacterial Infections/microbiology , Colonic Pouches/microbiology , DNA, Ribosomal/analysis , Polymorphism, Restriction Fragment Length , Pouchitis/microbiology , RNA, Bacterial/analysis , Adult , Bacteria/growth & development , Bacteria/isolation & purification , Colonic Pouches/adverse effects , Colony Count, Microbial , Female , Humans , Intestinal Mucosa/microbiology , Male , Middle Aged , Polymerase Chain Reaction , Risk Assessment
14.
Clin Transl Gastroenterol ; 10(5): 1-7, 2019 05 22.
Article in English | MEDLINE | ID: mdl-31117112

ABSTRACT

INTRODUCTION: Pouchitis is a common complication after ileal pouch-anal anastomosis (IPAA). However, there is a poor correlation between symptoms and endoscopic appearance of the pouch, and many patients can have debilitating symptoms in the absence of overt inflammation. It is unknown whether these clinical symptoms are independently associated with the microbiota. The objective of this work was to examine whether the individual clinical components of the pouch activity scoring systems are associated with specific microbiota. METHODS: Pouch biopsies from 233 patients (50% male, 100% IPAA/ulcerative colitis) post-IPAA were included. Clinical phenotyping was performed, and patients were classified using both clinical and endoscopic components of the Pouch Activity Scale. Scoring for symptoms examined 24-hour stool frequency, urgency, incontinence, and rectal bleeding as described by the Pouchitis Disease Activity Index Score. RESULTS: In the absence of inflammation, an increase in stool frequency reported over 24 hours was associated with a decrease in Bacteroidetes relative abundance, and this was the strongest association found. Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) analysis in inflamed groups showed that an increase in 24-hour stool frequency was associated with an increase in biofilm formation. DISCUSSION: These findings indicate that in patients with IPAA, the composition of mucosa-associated microbiota of the pouch may contribute to clinical symptoms, particularly stool frequency, independent of endoscopic disease activity.


Subject(s)
Colonic Pouches/microbiology , Gastrointestinal Microbiome/immunology , Ileum/microbiology , Intestinal Mucosa/microbiology , Pouchitis/diagnosis , Proctocolectomy, Restorative/adverse effects , Adult , Aged , Biopsy , Colitis, Ulcerative/microbiology , Colitis, Ulcerative/surgery , Colonic Pouches/pathology , Colonoscopy , Female , Follow-Up Studies , Humans , Ileum/diagnostic imaging , Ileum/pathology , Ileum/surgery , Intestinal Mucosa/diagnostic imaging , Intestinal Mucosa/immunology , Intestinal Mucosa/pathology , Male , Middle Aged , Pouchitis/immunology , Pouchitis/microbiology , Severity of Illness Index
15.
Clin Immunol ; 127(3): 270-9, 2008 Jun.
Article in English | MEDLINE | ID: mdl-18499066

ABSTRACT

A patient from the University of North Carolina Hospitals is presented who developed Crohn's disease of the ileal J-pouch following restorative proctocolectomy for ulcerative colitis. Inflammation of the ileal pouch in human inflammatory bowel disease (IBD) represents the best clinical example of the importance of host-enteric microbial interactions, and this case highlights rapid advances in our understanding of the role of the enteric microbiota in the immunopathogenesis of IBD, impacting on clinical care. Successful management of this patient necessitated accurate diagnosis as there are several inflammatory and non-inflammatory conditions of the pouch that present with similar symptoms. Diagnostic measures included serologic assays of response to microbial antigens, including ASCA, anti-OmpC, anti-Cbir1, and pANCA with DNAse sensitivity. Although the serologic detection of selective loss of tolerance to microbial antigens defines clinically important subgroups of inflammatory bowel disease patients, the clinical value of these serodiagnostic tests is a matter of debate. Genome wide screens have also identified NOD2/CARD15, IL23 receptor, and ATG16L1 variants as important in IBD susceptibility and pathogenesis. These genetic associations have also provided new insights into the importance of interaction between the host and microbes in the pathogenesis of IBD, but the precise mechanisms by which these gene variants contribute to disease development remain to be determined. Genetic associations and serological markers will ultimately be used to define important clinical subgroups of disease, predict natural history, and ultimately identify patient populations for early therapeutic intervention.


Subject(s)
Colitis, Ulcerative/immunology , Crohn Disease/immunology , Immunity, Mucosal , Antibodies/blood , Antibodies/immunology , Antigens, Bacterial/immunology , Bacteria/immunology , Bacteria/pathogenicity , Biomarkers/blood , Colitis, Ulcerative/diagnosis , Colitis, Ulcerative/microbiology , Colitis, Ulcerative/surgery , Colonic Pouches/immunology , Colonic Pouches/microbiology , Crohn Disease/diagnosis , Crohn Disease/genetics , Crohn Disease/microbiology , Diagnosis, Differential , Genetic Predisposition to Disease , Humans , Intestinal Obstruction/diagnosis , Pouchitis/genetics , Pouchitis/immunology , Pouchitis/microbiology , Proctocolectomy, Restorative , Risk Factors
17.
Dis Colon Rectum ; 51(5): 561-7, 2008 May.
Article in English | MEDLINE | ID: mdl-18299930

ABSTRACT

PURPOSE: We characterized the expression of sialomucin and sulphomucin in pouches fashioned for familial adenomatous polyposis and ulcerative colitis. We correlated sulphomucin expression with bacterial colonization and mucosal inflammation. METHODS: Ethical approval and informed consent were obtained. Mucosal biopsies from 9 patients with familial adenomatous polyposis and 12 with ulcerative colitis were obtained. Sulphomucin levels were assessed by using the high iron-diamine stain. Mucous gel layer composition was correlated with villous height, crypt depth, and total mucosal thickness. Mucous gel layer composition was correlated with acute and chronic inflammatory infiltrates. Colonization by a panel of seven bacterial species (including sulphate reducing bacteria) was established and correlated with sulphomucin levels. RESULTS: High-iron-diamine positivity (i.e., sulphomucin expression) was greater in ulcerative colitis pouch mucous gel (2.083 +/- 0.5 vs. 0.556 +/- 0.4, P = 0.003). Sulphomucin expression correlated with reduced crypt depth, villous height, and total mucosal thickness. In the ulcerative colitis group, chronic inflammatory infiltrate scores were significantly greater for high-iron-diamine-positive patients. Colonization by sulphate reducing bacteria was increased in high-iron-diamine-positive patients. CONCLUSIONS: Sulphomucin expression is increased in the mucous gel layer of the ulcerative colitis pouch compared with that of the familial adenomatous polyposis pouch. Sulphomucin expression is associated with colonization by sulphate-reducing bacteria and increased chronic inflammation.


Subject(s)
Adenomatous Polyposis Coli/metabolism , Colitis, Ulcerative/metabolism , Colonic Pouches/microbiology , Mucins/metabolism , Adenomatous Polyposis Coli/surgery , Biopsy , Colitis, Ulcerative/surgery , Female , Humans , Male , Statistics, Nonparametric
18.
Dis Colon Rectum ; 51(7): 1113-9, 2008 Jul.
Article in English | MEDLINE | ID: mdl-18483827

ABSTRACT

PURPOSE: This study was designed to identify the mucosa-associated microflora in patients with severe ulcerative colitis before and after restorative proctocolectomy with ileoanal pouch construction in comparison with historic controls. METHODS: Ten patients with a diagnosis of ulcerative colitis were evaluated. Mucus was collected during colonoscopy from all segments of the colon and terminal ileum before surgery, and from the ileal pouch two and eight months after ileostomy closure. The prevalence and mean concentration of the mucosa-associated microflora were compared over time and with historic controls. RESULTS: Veillonella sp was the most prevalent bacterium in patients and controls. Klebsiella sp was significantly more prevalent in the ileum of controls, was not found in patients with ulcerative colitis, and after proctocolectomy returned to values found in controls. Some bacteria such as Enterobacter sp, Staphylococcus sp (coag-), Bacteroides sp (npg), Lactobacillus sp, and Veillonella sp had higher mean concentrations in the ileal pouch of patients after surgery than in controls. CONCLUSION: No bacterium was identified that could be exclusively responsible for the maintenance of the inflammatory process. The mucosa-associated microflora of patients with ulcerative colitis underwent significant changes after proctocolectomy with ileal pouch construction and returned to almost normal values for some bacteria.


Subject(s)
Bacteria/isolation & purification , Colitis, Ulcerative/surgery , Colon/microbiology , Colonic Pouches/microbiology , Intestinal Mucosa/microbiology , Proctocolectomy, Restorative/methods , Rectum/microbiology , Adolescent , Adult , Aged , Aged, 80 and over , Colitis, Ulcerative/microbiology , Colitis, Ulcerative/pathology , Colon/pathology , Colon/surgery , Colonoscopy , Female , Follow-Up Studies , Humans , Intestinal Mucosa/pathology , Male , Middle Aged , Prognosis , Prospective Studies , Rectum/pathology , Rectum/surgery
19.
Aliment Pharmacol Ther ; 47(4): 466-477, 2018 02.
Article in English | MEDLINE | ID: mdl-29205422

ABSTRACT

BACKGROUND: The resident gut microbiota is essential for physiological processes; the disturbance of its balance is linked to intestinal inflammation. The ileoanal pouch is a model for the study of intestinal inflammation, as inflammation of the pouch is common and mostly develops within 12 months following ileostomy closure. This allows the longitudinal study of the microbiota, giving insight into the microbiota changes during transition from a normal to an inflamed pouch. AIM: To explore the literature on the microbiota of the ileoanal pouch in health and disease. METHODS: A systematic computer search of the on-line bibliographic databases MEDLINE and EMBASE was performed between 1966 and February 2017. Randomised controlled trials, cohort studies and observational studies were included. Studies were included if they reported microbiota analysis on faecal samples or tissue from the ileoanal pouch. RESULTS: Twenty-six papers were eligible. Following ileostomy closure, anaerobic bacteria are the abundant species in the ileoanal pouch with presence of a diverse microbiota key to maintaining a healthy ileoanal pouch. Acute pouchitis is associated with an increase in Clostridia species, while chronic pouchitis is associated with an increase in Staphylococcus aureus. In the treatment of pouchitis, a decrease in Clostridia species appears to be associated with treatment response. CONCLUSION: The microbiota plays an important role in both the inflamed and the healthy ileoanal pouch. A direct causal relationship between individual microbiota changes and inflammation has not yet been established, but manipulation of the ileoanal pouch microbiota may be a novel therapeutic avenue to explore.


Subject(s)
Colonic Pouches/microbiology , Gastrointestinal Microbiome/physiology , Health , Pouchitis/microbiology , Adult , Feces/microbiology , Female , Humans , Longitudinal Studies , Male , Pouchitis/etiology
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