bluuebunny/crossref_metadata_2025_split huggingface dataset:
14 COLONIC EPITHELIAL CELL-SPECIFIC AFTIPHILIN KNOCKDOWN REGULATES EPITHELIAL BARRIER FUNCTION THROUGH MYOSIN LIGHT CHAIN-ASSOCIATED ACTIN ORGANIZATION IN VITRO AND INTESTINAL LENGTH IN VIVO
Authors: Man Law Ivy Ka , Rankin Carl , Pothoulakis Charalabos 
Published: January 2020

Abstract

Background and Aims
Colonic epithelial integrity is often compromised during colonic inflammation and Inflammatory Bowel Disease. Aftiphilin (AFTPH) is a downstream target of microRNA-133a and its expression is reduced in colonic tissues of wild type mice from experimental colitis models and colonic biopsies from patients with ulcerative colitis. We have previously shown that AFTPH is involved in regulating intestinal epithelial barrier function and actin organization in human colonic epithelial cells in vitro (DDW 2016). On the other hand, our results suggested that global aftiphilin knock-out is embryonic lethal in mouse models (DDW 2019). Here, we further examined the role of AFTPH in regulating actin organization in vitro and characterize the colonic epithelial cell-specific aftiphilin knock-out mice.

Methods
Human colonic epithelial NCM460 cells were transfected with si-RNA against AFTPH to achieve transient AFTPH gene-silencing. Stable AFTPH knock-down clones were generated by transducing Caco2-BBE cells with recombinant lentivirus carrying sh-AFTPH or control sh-RNA. To create intestinal epithelial cell-specific aftiphilin knock-out mice, Aftph flox/flox mice were cross-bred with B6.Cg-Tg(Vil1-cre)997Gum/J mice, which express Villin-driven Cre recombinase (Vil-Cre), to generate intestinal epithelial cell-specific aftiphilin knock-out mice (Aftph Vil-/Vil-). Protein expression of F- and G-actin and p70S6K were detected using Western blot. Tissues from various organs were collected with Aftph Vil-/Vil- and its wildtype counterparts at 12 weeks.

Results
Results from western blot analysis showed that F-/G-actin ratio in AFTPH gene-silenced NCM460 cells were 0.6±0.17 fold, when compared to the treatment control. In addition, AFTPH gene-silencing in human colonic epithelial cells activated p70S6K, a kinase that is involved in actin organization, when compared to treatment control (1.2±0.15 vs. 2.0±0.15, p=0.0354). Furthermore, transepithelial electric resistance (TER) of Caco2-BBE cells deficient in AFTPH is significantly lower than that of control cells (0.5±0.07 fold). Lastly, in vivo intestinal epithelial cell-specific Aftph knock-out increased the length of small intestine, when compared to that of wild type mice (30.7±0.33 vs. 34.8±0.97, p=0.02), while the tissue weight of spleen to body weight was reduced (0.30±0.011 vs. 0.26±0.006, p=0.0169).

Summary and Conclusions
Our results indicate that AFTPH directly regulates epithelial barrier function and actin organization through mediating F-/G-actin ratio in human colonic epithelial cells, possibly through p70S6K. Importantly, intestinal epithelial cell-specific knock-out in vivo increased intestinal length and reduced size of the spleen. Our results suggested that AFTPH is crucial in regulating colonic epithelial barrier function in vitro and intestinal homeostasis.
20 ELUCIDATING THE ROLE OF FUSOBACTERIUM NUCLEATUM IN INTESTINAL INFLAMMATION
Authors: Engevik Melinda , Danhof Heather , Britton Robert , Versalovic James 
Published: January 2020

Abstract

Background
While the direct cause of inflammatory bowel disease (IBD) is unknown, the gut microbiota is speculated to play a key role. The complexity of the microbiome has made it difficult to pinpoint whether bacterial species are specifically associated with IBD exacerbations; although select microbes have emerged as compelling candidates. Several groups have identified increased abundance in Fusobacterium in IBD patients. Fusobacterium nucleatum drives inflammation in the oral cavity, but few studies have examined the potential for F. nucleatum to promote intestinal inflammation. We hypothesize that F. nucleatum secretes Outer Membrane Vesicles (OMVs) that activate epithelial Toll-like receptor 4 (TLR4) to drive inflammation.

Methods & Results
Given the prevalence of F. nucleatum in IBD specimens, we sought to determine if this pathobiont could promote pro-inflammatory responses in human epithelial cultures. Using fluorescently tagged F. nucleatum, we demonstrate that F. nucleatum subspecies polymorphum adheres to the mucus layer of human colonic HT29-MTX cells. Application of F. nucleatum metabolites to HT29-MTX cells resulted in upregulation of pro-inflammatory cytokines IL-8 and TNF by qPCR and ELISA. Purified OMVs from F. nucleatum alone were able to stimulate IL-8 and TNF production. This demonstrates the robust response of colonic epithelial cells to F. nucleatum. Additionally, we used human jejunum and colon enteroid monolayers treated with F. nucleatum metabolites in an anti-oxidant free enteroid media and found that F. nucleatum secreted products promoted TNF secretion by ELISA. Using the Luminex Magpix Platform we further queried the enteroid system to assess which pathways were activated. Enteroid monolayers treated with F. nucleatum metabolites exhibited increased phosphorylated ERK and CREB, downstream effectors of TLRs. We next sought to address whether F. nucleatum alone could elicit pro-inflammatory responses in a mouse model. Mice harboring a human microbiota, or humanized mice, were treated for 5 days with a cocktail of antibiotics and treated with F. nucleatum (108 CFU) by oral gavage; a regimen designed to mimic IBD patient treatment. Compared to control mice that received antibiotics and PBS vehicle control, mice treated with F. nucleatum exhibited disruption of the colonic architecture, with increased immune infiltrate and depleted mucus layer, resulting in a closer proximity of luminal contents to the epithelium. Analysis of mucosal gene expression revealed increased levels of epithelial TNF and KC (mouse homolog of IL-8), in addition to immune cell derived IL-6 in F. nucleatum-treated mice compared for controls.

Conclusions
These data provide evidence that F. nucleatum is capable of driving a pro-inflammatory signaling cascade in vitro and in vivo and F. nucleatum may represent a specific target for drug therapy.
P077 A NEW TREATMENT APPROACH FOR INFLAMMATORY BOWEL DISEASE: INTRACOLONIC BIFIDOBACTERIUM AND XYLOGLUCAN APPLICATION
Authors: Bozkurt Hüseyin 
Published: January 2020

Abstract

Background
Inflammatory bowel disease (IBD) pathogenesis includes the altered gut microbiota, environmental factors, human immune responses and genetic. Reduced bifidobacteria level is associated with IBD. Xyloglucan is a plant based prebiotic oligosaccharide. Bifidobacteria level is increased in the presence of xyloglucan.
In this article we aim to share the results of our cases; Ulcerative colitis (UC) patients treated by intracolonic single administration of Bifidobacterium animalis subsp. lactis and Xyloglucan combination.

Methods
Ten UC patients were evaluated; before and after intracolonic single administration of Bifidobacterium animalis subsp. lactis and Xyloglucan combination with colonoscopic laboratory and clinical examination.

Results
Age, sex, diagnosis, disease location, previous medications are summarized in Table 1. All the patients had active ulcerative colitis disease before the administration. The Mayo Score was used to assess the severity of UC. 2 cases had extensive colitis and 8 patients had left-sided colitis. After 6 weeks of the administration mucosal healing and resolution of colonic symptoms were seen. These results are summarized in Table 2. Of the 10 cases, 7 were undertaken 5-ASA +Azathiopurine and three were undertaken vedolizumab treatment. Intracolonic single Bifidobacterium animalis subsp. lactis and xyloglucan administration was found effective in the mucosal healing and resolution of colonic symptoms in ulcerative colitis patients.

Conclusions
Herein we reported the importance of Bifidobacterium and xylooligosaccharide combination in IBD. Colonoscopic single Bifidobacterium animalis subsp. lactis and xyloglucan administration is a new method that has no side effect and easy to apply for treatment of IBD.This application might provide enhancement of non-stimulatory status and higher biodiversity in colonic mucosa so mucosal healing may be improved rapidly. However, it would be necessary to develop diagnostic strategies in order to discriminate which patients would benefit from this strategy.
P167 ADULT CROHN’S DISEASE TREATED WITH THE SPECIFIC CARBOHYDRATE DIET (SCD): A CASE REPORT USING OBJECTIVE MARKERS OF RESPONSE
Authors: Arjomand Ali 
Published: January 2020

Abstract
The specific carbohydrate diet (SCD) has been shown to resolve symptoms in pediatric IBD patients, inducing clinical remission and normalizing or significantly reducing inflammatory markers. Demonstration of similar effects, including changes to objective disease markers, are lacking in the adult IBD patient. Presented here are clinical, biomarker, microbiome, imaging and histologic observations after successful implementation of the SCD in an adult with moderate-to-severe Crohn’s disease. HISTORY: A 50-year-old male patient, diagnosed with Crohn’s disease in 2001, implemented the SCD in 2017 according to the published protocol. Prior to diet intervention, medical management consisted of mesalamine (1 g orally 4x/day for 6 mo.); infliximab (300 mg IV/8 wks for 5 yrs), natalizumab (300 mg IV/4 wks for 6 mo.), adalimumab (40 mg sc/14 d for 7 yrs; every 7 d for 1 yr) and vedolizumab (300 mg IV/8 wks for 1 yr). Corticosteroids were administered with adalimumab and vedolizumab over a continuous span of 8 yrs (prednisone 15–20 mg/d or budesonide 9 mg/d). Two resections of the terminal ileum were performed in 2003 and 2015. History included annual ER or hospital admissions and severe malnutrition (albumin 1 g/dL). TPN was provided in 2008 and 2015. DIETARY INTERVENTION: The patient was advised to switch to ustekinumab after LOR to vedolizumab and return of active inflammation. The patient refused further biologic treatment and proceeded with the SCD intervention. Prednisone (40 mg/d) was initiated during diet transition. The patient adhered to a strict SCD protocol, eliminating sugar, grains, starch and all forms of processed foods while adding home-fermented yogurt.
Results: By day 10, the patient reported loss of abdominal and lower back pain along with a 5 lb weight drop. Cooked, non-starchy vegetables, nut flours and other foods were then introduced according to the SCD protocol and as tolerated. At month 3, the patient reported 1–2 formed BM/d, increased energy and mobility, loss of joint pain and a stable weight. The patient initiated yoga 3x/wk. Advanced SCD foods continued to be introduced one at a time. Stool was well-formed and of large mass. Prednisone was tapered to 20 mg/d at month 3 and 0 mg/d by month 8. At month 12, MRE and colonoscopy showed resolution of inflammation and normal mucus in the terminal ileum assessed by biopsy. At month 24, FCP and CRP remained in the normal range. Microbiome analysis of fecal samples showed poor diversity at all times with minor phyla-level changes at month 12. International travel resulted in genus-level increase in diversity. The patient has completed 3 years with the SCD and plans to maintain the diet as part of a lifestyle change. Prospective studies are needed in adult IBD patients, including moderate-to-severe cases, to assess the long-term efficacy of the SCD using objective markers of response.
P163 DYSREGULATED MICROBIAL METABOLISM IN ULCERATIVE COLITIS INCREASES THE RISK OF CLOSTRIDIODES DIFFICILE INFECTION
Authors: Kitamoto Hiroko , Higgins Peter , Young Vincent , Kamada Nobuhiko 
Published: January 2020

Abstract
Clostridioides difficile infection (CDI) is recognized as a major clinical complication in patients with ulcerative colitis (UC). However, the mechanism associated with increased susceptibility to C. difficile in UC patients remains poorly understood. Given the evidence that the gut microbiota plays a critical role in the prevention of CDI and that gut microbiota is perturbed in UC patients, we hypothesized that UC-associated gut dysbiosis contributes to the increased susceptibility of patients to CDI. To address this hypothesis, we employed the human microbiota-associated (HMA) mouse model. Germ-free mice were colonized with the gut microbiotas isolated from either UC patients or healthy control (HC) subjects. Utilizing this model, we have found that UC-HMA mice are susceptible to C. difficile, while HC-HMA mice are completely protected. An isogenic mutant C. difficile strain, which lacks the succinate utilization operon, exhibited impaired colonization to UC-HMA mice, indicating that succinate is a key metabolite that promotes the colonization of C. difficile in UC-HMA mice. Restoration of healthy microbiotas by fecal microbiota transplantation (FMT) reduced luminal concentration of succinate in UC-HMA mice, thereby rendering the mice protective against CDI. The abundance of succinate-consuming bacteria likely regulates the availability of luminal succinate. We found that host mucus/epithelial N-glycosylation plays a crucial role in the growth of succinate-consuming bacteria in the gut. The expression of N-glycosylation–related enzymes were reduced in UC patients, which may lead to the gut dysbiosis with decreased succinate-consuming bacteria and increased luminal succinate. Taken together, impaired host N-glycosylation in UC induces gut dysbiosis and subsequent dysregulation of gut microbial metabolism. The impaired microbial metabolic activities - reduced succinate utilization - increases host susceptibility to C. difficile.
P166 MICROBIOME AND EPIGENETIC PREDICTORS OF RESPONSE TO BIOLOGIC THERAPY IN PATIENTS WITH ULCERATIVE COLITIS
Authors: Shah Rajesh , Shen Lanlan 
Published: January 2020

Abstract

Introduction
Current ulcerative colitis (UC) treatments have variable efficacy and may take several weeks to assess improvement. Emerging data suggest the intestinal microbiota may serve as a biomarker and mechanistically may influence immune system activity through epigenetic regulation of host gene expression. The aim of this pilot project was to examine whether the colonic mucosal microbiota and mucosal DNA methylation patterns are associated with a response to treatment in UC.

Methods
We conducted a retrospective cross-sectional study of patients with active UC. Fresh frozen colon biopsy samples were obtained from the Texas Medical Center IBD Tissue Bank. Patients were included if they had a colonoscopy performed to assess disease activity and follow up through 14 weeks to assess response. Disease activity was defined using the Partial Mayo Score and response was defined as a 2 point or greater decrease in the score after 14 weeks of follow up. 16s rRNA gene sequencing and DNA methylation pyrosequencing were used to define the microbiome and quantify DNA methylation. Comparisons were performed of alpha and beta diversity, taxonomy and DNA methylation between responders and non-responders at 14 weeks. Additionally, a random forest machine learning model was developed to identify predictors of response.

Results
We identified 16 patients with tissue samples from the cecum/ascending, transverse and rectum/sigmoid available for analysis. After excluding patients with limited rectal disease, recent antibiotic use and inadequate follow up, 4 patients (2 per group) were included and thus provided 12 tissue samples for analysis. The mean age of responders was 38 and non-responders was 24. 50% of patients were male and all were Caucasian. Responders had a numerically lower alpha diversity, though this did not reach statistical significance (p=0.18). Responders and non-responders separated in unweighted (p=0.001) and weighted (p=0.001) beta diversity analysis. Responders had a greater abundance of Firmicutes and Bacteroidetes and lower abundance of Proteobacteria compared to non-responders. This corresponded to responders having a greater abundance of genera for Bifidobacterium, Faecalibacterium and Roseburia. Additionally, we saw increased methylation in the P16, HOXC5 and B4GALNT1 genes of responders compared to non-responders. Finally, in a random forest machine learning model, predictors of response included left sided disease extent and OTUs for the genus Roseburia.

Discussion
Patients with UC and response to therapy had a significantly different pre-treatment microbiome and methylation of genes related to intestinal barrier function, including B4GALNT1. Larger studies will be needed to validate these findings, but these results suggest the microbiome and DNA methylation changes may be effective biomarkers of response to therapy and warrant further study.
11 ENTERAL NUTRITION THERAPY IS ASSOCIATED WITH FEWER READMISSIONS AND DEATHS AMONG MALNOURISHED INPATIENTS WITH INFLAMMATORY BOWEL DISEASE
Authors: LeBrett Wendi , Sauk Jenny , Limketkai Berkeley 
Published: January 2020

Abstract

Background
Malnutrition is a common complication observed in hospitalized patients with inflammatory bowel disease (IBD). Enteral nutrition therapy can be used to support the nutritional needs of inpatients with IBD. However, evidence on the impact of inpatient enteral nutrition on clinical outcomes is equivocal. This study assesses post-hospitalization outcomes associated with enteral nutrition therapy amongst inpatients with IBD in a large nationwide database.

Methods
We conducted a retrospective propensity score-matched study among IBD inpatients diagnosed with protein-calorie malnutrition using the Nationwide Readmissions Database from 2010–2015. ICD9 codes associated with each admission were used to identify patients who received enteral nutrition. Using propensity score matching, patients who received inpatient enteral nutrition were matched with patients who did not receive enteral nutrition based on the following variables: age, sex, elective admission, patient income, teaching hospital, and hospital urban or rural locality. Primary endpoints included 30-day readmissions, 90-day readmissions, 30-day mortality and 90-day mortality.

Results
Among the 1,588 IBD patients (822 Crohn’s disease, 755 ulcerative colitis, 11 unclassified IBD) with protein-calorie malnutrition, patients who received enteral nutrition (n=794) had fewer 30-day readmissions (OR 0.73; 95% CI 0.55 – 0.96) and 90-day readmissions (OR 0.77; 95% CI 0.61 – 0.97). None of the patients (0%) in the enteral nutrition group died on a subsequent admission within 30 days of discharge, compared to 6 patients (0.8%) in the control group (p=0.027). Inpatient mortality within 90 days of discharge did not differ significantly between the two groups (0.8%, enteral nutrition vs. 1.6%, control; p=0.086).

Discussion
Enteral nutrition therapy among IBD inpatients with malnutrition was associated with lower odds of readmission and 30-day mortality, but not 90-day mortality. The findings of our study support the use of enteral nutrition in IBD inpatients and motivate the need for prospective studies assessing the impact of enteral nutritional support in IBD inpatients.
P027 TRANSITION OF CARE OF ACTIVE DUTY PATIENTS WITH IBD TO THE VETERANS ADMINISTRATION HEALTHCARE SYSTEM: TOOLKIT FOR SUCCESS
Authors: Patel Anish , Keefer Laurie 
Published: January 2020

Abstract
The shift from being an active duty military patient with inflammatory bowel disease (IBD) seen in a military treatment facility (MTF) to being a veteran receiving chronic illness care within the Veterans Administration (VA) is a time of increased vulnerability and risk for loss of continuity of care, and poor disease outcomes. A transitional care program must prioritize the psychosocial growth, self-efficacy and disease-specific knowledge of active military patients who are transitioning to VA care as well as enhance collaborative management between military and VA providers. The Department of Defense (DoD) has established the Transition Assistance Program (TAP) to assist military members to civilian life, but limit preparedness in skills to navigate/understand medical care. There is currently no available data on transition readiness from active duty military medical care to VA medical care and our objective is to determine the feasibility and acceptability of a self-management intervention.
Methods: We prospectively measured readiness with the use of the IBD Self-Efficacy Scale (IBD-SES) and Transition Readiness Assessment Questionnaire (TRAQ). All enrolled patients (50 active duty servicemembers) were in disease remission and underwent TAP and 50% were further randomized to undergo TRAQ-based educational and behavioral interventions via mobile applications and clinical consults.
Results: Servicemembers who underwent further TRAQ-based interventions compared to those only undergoing TAP demonstrated significant improvement in medication/appointment management, tracking/managing health issues, and talking with providers (p&lt;0.001). Furthermore, these servicemembers also demonstrated significant improvement in self-efficacy and self-management which was maintained up to 3 months post military discharge (p&lt;0.001). Servicemembers had an average VA primary care wait time of 3.5 months and for Gastroenterology specialist of 5.4 months, resulting in &gt;60% of servicemembers having to discontinue therapy due to lapse in care (50% rate of flares) and unnecessary therapy switch.
Conclusion: This is the first and only study to assess transitioning from active duty service to VA medical care for a chronic illness. We demonstrated improved and sustained self-efficacy outcomes with a transition program for IBD along with potential limitations in the VA transition. We have an opportunity to influence the conversation of appropriate DoD transition of medical care and to develop a model of care with wider applicability.