bluuebunny/crossref_metadata_2025_split huggingface dataset:
THE TIGHT JUNCTION PROTEIN ZO-1 REGULATES MITOTIC SPINDLE ORIENTATION TO ENABLE EFFICIENT MUCOSAL REPAIR
Authors: Kuo Wei-Ting , Zuo Li , Turner Jerrold 
Published: January 2021

Abstract
The intestinal damage can be caused by physical, infectious, and immune-mediated injury. Rapid repair, which is essential for a return to mucosal homeostasis, depends on rapid epithelial migration, i.e., restitution, as well as epithelial proliferation and differentiation. Barrier restoration is a critical component of this repair process, but the contributions of structural proteins that form the barrier to mucosal repair have not been defined.
Aim: To determine the contributions of the intestinal epithelial tight junction protein zonula occludens-1 (ZO-1) to mucosal repair.
Results: Intestinal epithelial ZO-1 transcript and protein expression are reduced in biopsies from inflammatory bowel disease (IBD) patients relative to healthy controls. To determine if this ZO-1 loss contributes to disease pathogenesis, we created intestinal epithelial ZO-1-deficient Tjp1f/f x villin-Cre+ mice. When stressed, for example, with 2% DSS, ZO-1-deficient mice displayed much greater mucosal damage and weight loss relative to ZO-1-sufficient mice and failed to recover fully, even 4 weeks after DSS discontinuation. To better define the defect, mice were injected with nucleoside analogs (BrdU and EdU) to label proliferating cells 1 and 3 days after DSS discontinuation. When assessed 2 hours after labeling, numbers of labeled nuclei were significantly reduced in ZO-1-deficient mice relative to ZO-1-sufficient mice. More striking, however, was the nuclear fragmentation and staining for cleaved caspase-3 at 24 hours after labeling. In vitro studies of colonoids were used to better define the mechanisms of epithelial loss. Colonoids from ZO-1-deficient mice were smaller, had reduced numbers of buds (crypt domains) and Lgr5+ stem cells, and were nonresponsive to enhanced Wnt signaling induced by the GSK3 inhibitor CHIR99021, relative to ZO-1-sufficient colonoids. Live imaging of mitosis showed misoriented mitotic spindles in ZO-1-deficient colonoids that caused one daughter cell to lose contact with the extracellular matrix. Misoriented mitotic spindles were also present in tissues from DSS-treated ZO-1-deficient, not ZO-1-sufficient, mice. Live imaging of colonoids from mRFP1-ZO-1 transgenic mice detected transient accumulation of ZO-1 at the cleavage furrow, suggesting that ZO-1 interactions at this site required for accurate mitotic spindle orientation.
Conclusion: ZO-1 has a previously unappreciated, noncanonical function in mitotic spindle orientation that is independent of barrier maintenance but central to epithelial proliferation and repair. We postulate that, in the absence of ZO-1, loss of contact with the basement membrane leads to anoikis, i.e., detachment-induced apoptosis, and an abortive proliferative response that compromises repair. We speculate that ZO-1 downregulation in IBD may similarly interfere with mucosal healing.
Support: NIH (DK068271, DK061931)
TARGETING SIDEROPHORES TO REDUCE COLONIZATION OF ADHERENT INVASIVE E. COLI IN CROHN’S DISEASE
Authors: Gerner Romana , Hossain Suzana , Siada Kareem , Sargun Artur , Neumann Wilma , Nolan Elizabeth , Raffatellu Manuela 
Published: January 2021

Abstract
The pathovar adherent-invasive Escherichia coli (AIEC) is a mucosa-associated bacterium that is frequently found in patients with Crohn’s disease (CD), but targeted treatment options are currently lacking. During intestinal inflammation, host-mediated iron limitation is a key strategy to restrict the outgrowth of pathogens. Various bacterial pathogens evade this nutrient limitation by secreting small, iron-chelating molecules known as siderophores. The siderophore enterobactin (Ent) is an important virulence factor for Gram-negative pathogens such as AIEC by promoting their expansion and competition over commensal microbes during colitis. Here, we developed an immunization-based approach to target bacterial siderophores during colitis and to reduce AIEC colonization in mice. We demonstrate that immunization with Ent (conjugated to the mucosal adjuvant cholera toxin subunit B, CTB) potently elicits fecal antibodies against Ent. Immunized animals show reduced AIEC colonization along with milder weight loss, less colonic shortening, and decreased histological severity scores. This phenotype is largely B cell-dependent, as immunized mice lacking mature B lymphocytes do not show protection during AIEC infection. Ent-specific B cells are primarily found in small intestinal Peyer’s patches and can be isolated by a flow cytometry-based sorting approach. This project paves the way to develop monoclonal antibodies against siderophores, which could provide a more targeted and narrow-spectrum strategy to reduce AIEC colonization in patients with CD.
ACTIVATED INTESTINAL MUSCLE CELLS PROMOTE PREADIPOCYTE MIGRATION: A NOVEL MECHANISM OF CREEPING FAT FORMATION IN CROHN’S DISEASE
Authors: Mao Ren , Doyon Genevieve , Gordon Ilyssa , Li Jiannan , Lin Sinan , Wang Jie , Le Thi Hong Nga , Elias Michael , Kurada Satya , Southern Brian , Olman Mitchell , Zhao Shuai , Dejanovic Dina , Chandra Jyotsna , Pranab Mukherjee , West Gail , van Wagoner David , Fiocchi Claudio , Rieder Florian 
Published: January 2021

Abstract

Background and Aims
Creeping fat, the wrapping of mesenteric fat around the bowel wall, is a typical feature of Crohn’s disease, and is associated with stricture formation and bowel obstruction. How creeping fat forms is unknown, and we interrogated potential mechanisms using novel intestinal tissue and cell interaction systems.

Methods
Tissues from normal, ulcerative colitis, non-strictured and strictured Crohn’s disease intestinal specimens were obtained. Fresh and decellularized tissue, mesenteric fat explants, primary human adipocytes, pre-adipocytes, muscularis propria cells, and native extracellular matrix were used in multiple ex vivo and in vitro systems involving cell growth, differentiation and migration, proteomics, and integrin expression.

Results
Crohn’s disease muscularis propria cells produced an extracellular matrix scaffold which is in direct spatial and functional contact with the immediately overlaid creeping fat. The scaffold contained multiple proteins, but only fibronectin production was singularly upregulated by TGF-b1. The muscle cell-derived matrix triggered migration of pre-adipocytes out of mesenteric fat, fibronectin being the dominant factor responsible for their migration. Blockade of α5β1 on the pre-adipocyte surface inhibited their migration out of mesenteric fat and on 3D decellularized intestinal tissue extracellular matrix.

Conclusion
Crohn’s disease creeping fat appears to result from the migration of pre-adipocytes out of mesenteric fat and differentiation into adipocytes in response to an increased production of fibronectin by activated muscularis propria cells. These new mechanistic insights may lead to novel approaches for prevention of creeping fat-associated stricture formation.
DEVELOPMENT OF SMALL MOLECULE EPIGENETIC THERAPEUTICS FOR INFLAMMATORY BOWEL DISEASE
Authors: Li Yi , Uribe Gabriela , Pingyuan Wang , Chen Haiying , Liu Zhiqing , Cong Yingzi , Brasier Allan , Pinchuk Irina , Tian Bing , Zhou Jia 
Published: January 2021

Abstract
Long-term persistence of chronic inflammation in inflammatory bowel disease (IBD) including ulcerative colitis (UC) and Crohn’s disease (CD) is among the major factors contributing to neoplastic transformation and the development of colitis-associated colorectal cancer. There exists a lack of efficient medications for IBD, primarily due to emergence of resistance or side effects. Antibodies against tumor necrosis factor-α (anti-TNFα) are effective therapies in the armamentarium; however, up to 40% of patients become resistant to this therapy. Thus, target-based improved therapy with higher efficacy and enhanced safety profile is urgently needed to fill these gaps.
We have demonstrated that bromodomain-containing protein 4 (BRD4), an epigenetic regulator, is required for stabilization of NFkB binding on the promoters of inflammatory genes, activation of RNA polymerase II, and histone H3 Lys122 acetylation (H3K122ac) to permit high levels of inflammatory gene expressions in sentinel immune cells, epithelial cells, and fibroblasts. Our compelling data using human IBD patient samples suggest that BRD4 activation is coincident with the initiation of colonic inflammation. Inhibition of the BRD4 activation is an attractive target for the development of superior therapeutics for IBD, especially for anti-TNFα-resistant patients.
We have successfully identified proprietary highly potent and specific BRD4 inhibitors (patent WO 2018/112037 A1) with direct binding modes validated by the solved co-complex crystal structures. We observed that our lead compounds exhibit low toxicity both in vitro and in vivo. Therapeutic administration of our lead inhibitors ZL0516 and ZL0590 significantly reduces mucosal inflammation in several animal models of IBD and restores tissue architecture. Our data show that inhibition of BRD4 results in a decrease of key inflammatory cytokines associated with pathological responses in IBD such as TNFα, IL-6, IL-17A, and IL-1β expression in human colonic epithelial cells and peripheral blood mononuclear cells as well as several animal models of IBD colitis.
Collectively, our data suggest that BRD4 activation is critical to the initiation of human colonic inflammation during IBD. BRD4 inhibition disrupts its protein-protein interactions with acetylated histone lysine residues, thereby blocking the pathological activation of the BRD4-NFκB signaling and suppressing colonic inflammation. Thus, BRD4 represents a unique drug target, and novel BRD4 inhibitors ZL0516 and ZL0590 have been identified as promising drug candidates towards the development of small molecule epigenetic therapeutics for the treatment of IBD and its chronic sequelae.
Funding support: Crohn’s & Colitis Foundation Entrepreneurial Investing (EI) Initiative award, Litwin IBD Pioneers Program, and a research fellowship award from the Crohn’s & Colitis Foundation of America.
EVALUATION OF A DIET AND ANTIBIOTICS TARGETING THE MICROBIOTA FOR TREATMENT OF MILD TO MODETARE ACTIVE PEDIARTIC ULCERATIVE COLITIS: AN OPEN LABEL PILOT STUDY
Authors: Sarbagili-Shabat Chen , Albenberg Lindsey , Van Limbergen Johan , Weiner Dror , Yaakov Michal , Levine Arie 
Published: January 2021

Abstract

Background
Newer strategies that target the microbiome may offer an alternative therapeutic approach for Ulcerative Colitis (UC). We developed a novel diet that targets changes in the microbiome and barrier function that have been reported in UC. The goal of the current study was to evaluate the efficacy of two sequential induction of remission strategies that target the microbiota: the novel diet termed the ulcerative colitis diet (UCD) and an antibiotics cocktail combination in dietary non responders.

Methods
This was a prospective, single arm, open label, pilot study in patients aged 8–19, with a pediatric UC activity index (PUCAI) scores &gt;10 and ≤45 on stable maintenance therapy (5ASA or thiopurines). PUCAI score was assessed at week 3 and 6. Patients failing to enter remission or intolerant to dietary therapy could receive an open label 14-day course of Amoxycillin, Metronidazole and Doxycycline (AMD), and had PUCAI scored at day 21. Response was defined a decline in PUCAI ≥ 10 points, remission as PUCAI&lt; 10. The primary endpoint was intention to treat (ITT) remission at week 6 with diet as the sole intervention.

Results
Twenty-three children mean age of 15.1±2.9 years were enrolled. Two patients (1 responder, 1 remission) withdrew by 3 weeks, four required additional therapy by week 3, all were considered failures by ITT. Mean PUCAI decreased at week 3 and 6 from 34.5±9.8 to 21.7±14.9 and 17.6±17.2 respectively (P=0.005, P=0.001) at ITT analysis including all patients. Sixteen out of twenty-three patients (69.6%) responded by week 6. Ten of twenty-three (43.5%) achieved remission by week 6, and nine (39.1%) had clinical remission at week 6. The median fecal calprotectin (FC) level decreased in patients (n=5) who achieved remission from 630 (IQR, 332–1586) μg/g at week 0 to 230 (75–1298) μg/g at week 6. Eight patients received treatment with antibiotics after failing diet, 4/8 (50.0%) subsequently entered remission.

Conclusion
A dietary intervention called the UC Diet appears to be effective for induction of remission in children with mild to moderate UC. Sequential use of diet, followed by antibiotic therapy in dietary non responders, needs further evaluation as a microbiome directed steroid sparing therapy in patient’s refractory to 5ASA and thiopurines.
ADHESIONS ARE NOT ALWAYS THE ENEMY: POUCH VOLVULUS
Authors: Dionigi Beatrice , Lavaryk Olga , Holubar Stefan , Lightner Amy , Hull Tracy 
Published: January 2021

Abstract

Introduction
Proctocolectomy with ileal pouch anal anastomosis (IPAA) was introduced over 40 years ago, prior to the era of laparoscopy. Since then, minimally invasive surgery (MIS) techniques have been applied to pelvic pouch surgery. One advantage of MIS is the reduction in adhesion-related complications. However, the lack of adhesions may result in a different set of complications. Specifically, when the pouch is not adherent in the pelvis, pouch volvulus can occur around the mesenteric axis. The aim of this study was to describe our experience with pelvic pouch volvulus.

Methods
Our prospectively maintained pelvic pouch registry and our enterprise wide electronic medical record were queried for keyword combinations of “pouch volvulus” between 1994 and 2020. Pouch volvulus was defined as torsion of the ileal pouch on its own mesenteric axis or around small bowel while maintaining the proper orientation of the ileo-anal anastomosis. Patients with pouches constructed with twisted mesenteric axis from ill-aligned anastomoses were excluded. Data for these patients was collected from the pouch registry and additional chart review.

Results
We identified 17 patients with pouch volvulus; of these, 11 patients did not meet our selection criteria and were excluded. Of the 6 patients (5 female; median age 25 range 20–33 yr at time of volvulus surgery) with true volvulus, the diagnosis at IPAA was ulcerative colitis (n=5) and Lynch syndrome with a rectal cancer. All pelvic pouches were constructed with MIS techniques, including standard laparoscopy (n=4) and single incision laparoscopic technique (n=2). All 6 patients presented with diffuse abdominal pain and abdominal distention. The average time from pouch construction to pouch volvulus was 2.5 years (range: 5.2 – 97.5 months). Computed tomography with or without rectal contrast was the initial diagnostic test in 4/6 with findings highly suspicious for pouch volvulus. Surgery was performed urgently in 5/6 of patients; all 6 had open surgery. At reoperation, all had minimal adhesions and a gap between the pouch mesentery and the retroperitoneum. Interventions included pouch-pexy (n=3), closure of gap between pouch and sacrum (n=2), and pouch excision and ileostomy (n=1). At a median of 9 months (IQR: 4–97) of follow up, pouch survival was 83%; functional outcomes included mild fecal incontinence (n=1) and paradox requiring intermittent pouch intubation for stool evacuation (n=1).

Conclusions
Pelvic pouches constructed by minimally invasive techniques may be at risk for pouch volvulus due to minimal adhesions. Surgeons should have a high index of suspicion for patients with unexplained abdominal pain, distension, and other obstructive symptoms. Cross-sectional imaging with rectal contrast, may help clarify the diagnosis. Immediate surgical care can allow for pouch salvage.
EXPLORING THE REPRODUCTIVE HEALTH DECISION EXPERIENCES AND PREFERENCES OF WOMEN WITH INFLAMMATORY BOWEL DISEASES
Authors: Picciarelli Zach , Stransky Olivia , Michel Hilary , Kim Sandra , Schwartz Marc , Gray Whitney , Kazmerski Traci 
Published: January 2021

Abstract

Background
Women with inflammatory bowel diseases (IBD), such as Crohn’s disease or ulcerative colitis, have normal fertility and can have healthy pregnancies and children. However, many have fewer children than the general population, with up to a third of women choosing “voluntary childlessness.” Given that IBD diagnoses can elicit reproductive concerns related to contraception choice, pregnancy, and parenthood, this study explored the reproductive health decision-making experiences and preferences of women with pediatric-onset IBD.

Methods
In this qualitative study, we recruited women ages 18–44 years old with a childhood diagnosis of IBD who are followed by gastroenterology at one pediatric and one adult academic medical center in Western Pennsylvania to participate in individual, semi-structured interviews. We conducted interviews between July 2019 and July 2020, exploring experiences and attitudes toward parenthood, pregnancy, contraception, and family planning care provision. Interviews were audio-recorded and transcribed verbatim. Two independent coders performed analyses using a deductive and inductive coding approach and identified dominant themes and representative quotations. We conducted interviews until thematic saturation was reached.

Results
Twenty-one women with IBD participated in interviews (average age 24.7 +/-5.9 years, range 18–43 years; average age of diagnosis 14.1 +/- 2.0 years). Five women had been pregnant, and 16 wanted to have children in the future. We identified 4 key themes: (1) Nulliparous women who do not currently desire pregnancy appear to lack reproductive health knowledge; (2) Women with IBD lack clarity regarding the role IBD plays in contraceptive choice; (3) Related to pregnancy, women are concerned about the heredity of IBD, antepartum disease activity, and the safety of their current medications; (4) Women with IBD typically default to their women’s health provider for reproductive healthcare and counseling, but they expect their gastroenterologist to initiate relevant reproductive health discussions with them and to provide information in the context of their disease.

Conclusions
Women have concerns about the effects of IBD on pregnancy, parenthood, and contraceptive choice; however, many have had limited or no discussion with their gastroenterologist about these topics. They have gaps in their reproductive health knowledge and report receiving conflicting advice from different providers. Obtaining the perspectives of a larger sample of women with IBD and both women’s health and gastroenterology providers may further guide the development of interventions to improve this aspect of comprehensive care.
IMPACT OF TIME-TO-TEST FOR CLOSTRIDIOIDES DIFFICILE INFECTION ON LENGTH OF STAY IN INFLAMMATORY BOWEL DISEASE PATIENTS HOSPITALIZED WITH FLARE
Authors: Verma Abhishek , Varma Sanskriti , Freedberg Daniel , Hudesman David , Chang Shannon , Axelrad Jordan 
Published: January 2021

Abstract

Background
Guidelines recommend testing inflammatory bowel disease (IBD) patients hospitalized with flare for Clostridioides difficile infection (CDI), though little is known about whether a delay in testing for CDI is related to adverse outcomes. We examined the relationship between time-to-C. difficile PCR test order, collection, and result with adverse IBD outcomes.

Methods
We performed a retrospective cohort study of IBD patients hospitalized with flare through the emergency department (ED) between 2013 and 2020 at an urban academic medical center. The time from ED presentation to C. difficile test order (time-to-order), sample collection (time-to-collection), and test result (time-to-result) were collected. Time-to-result was stratified by within 6 hours, 6–24 hours, and 24 hours or longer. The primary outcome was length of stay (LOS). Secondary outcomes were inpatient anti-TNF administration and surgery. We used hemodynamic and laboratory values at presentation to evaluate disease severity as a confounding variable between length of stay and time-dependent variables.

Results
We identified 122 IBD patients hospitalized with flare. There were no significant differences in baseline characteristics among time-to-result groups. Despite a shorter time-to-result, the average LOS in the 6 hours group was 7.3 days, longer compared to the 6–24 hours group (4.3 days, p=0.018) and the 24 hours group (4.2 days, p=0.035; Table 1). There were no differences in inpatient anti-TNF administration (p=0.10) or surgery (p=0.08) among time-to-result groups. The markers of disease severity that correlated with longer LOS were C-reactive protein (CRP) (0.28 days, p=0.003), heart rate (0.478 days, p&lt;0.001), diastolic hypotension (0.228 days, p=0.01), and hypoalbuminemia (0.215 days, p=0.02). Higher CRP correlated with earlier time-to-result (-0.218 hours, p=0.02). Patients with more markers of disease severity had earlier times-to-result (12.8 hours vs. 32.2 hours, p=0.014) and had a longer LOS (7.9 vs. 3.4 days, p=0.007) (Table 2). Patients with more severe disease had an earlier time-to-order (4.48 hours) compared to those with less severe disease (17.4 hours), though this difference did not meet statistical significance (p=0.09; Table 2).

Conclusion
Earlier time-to-result for CDI is associated with longer LOS in IBD patients hospitalized with flare. This inverse relationship is confounded by disease severity at presentation: patients with more severe disease have a shorter time-to-result and a longer LOS. It may be that these patients produce a stool sample more readily; however, the near significance of differences in time-to-order among severity groups suggest a role for provider bias, which must be studied further. Delay in testing was not associated with higher rates of inpatient anti-TNF administration or surgery.