bluuebunny/crossref_metadata_2025 huggingface dataset:
AN AUTOPHAGY PROTEIN ATG16L1 PREVENTS NECROPTOSIS BY REGULATING INTERFERON SIGNALING IN THE INTESTINAL EPITHELIUM
Authors: Matsuzawa Yu , Cadwell Ken
Published: January 2021

Introduction

ATG16L1, an essential component of autophagy, is one of the susceptibility genes of Crohn’s disease. We previously showed mice with decreased expression of Atg16L1 and Crohn’s disease patients homozygous for ATG16L1T300A risk alleles develop Paneth cell abnormalities in the intestinal epithelium. Moreover, the abnormalities were dependent on murine norovirus (MNV) infection in mice. Most recently, we reported mice with deletion of ATG16L1 in the intestinal epithelial cells (Atg16L1ΔIEC) are more susceptible to disease following intestinal injury than wild type mice when they are infected with MNV. Also, we revealed intestinal organoids derived from Atg16L1ΔIEC mouse are susceptible to TNFa-induced necroptosis. However, how ATG16L1 protects IECs against TNFa, and whether the susceptibility is preserved in human IECs harboring ATG16L1T300A risk alleles remains unclear.

Aims

In this study, we first examined the mechanism by which ATG16L1-deficiency renders IECs susceptible to necroptosis. Next, we investigated whether a specific ISG might be involved in the increased susceptibility. Finally, we tested whether human intestinal organoids harboring ATG16L1T300A risk alleles are susceptible to TNFa-induced necroptosis.

Methods & Results

We performed RNA-Seq analysis using small intestinal organoids derived from Atg16L1ΔIEC and control mouse, and found 49 genes were upregulated in naïve Atg16L1ΔIEC organoids (Figure 1). Interestingly, most of them were the genes known as interferon-stimulated genes (ISGs) representing a type I IFN (IFN-I) signature. Importantly, we found Atg16L1ΔIEC organoids pretreated with JAK-STAT inhibitor Ruxolitinib displayed better viability and decreased level of necroptosis-related proteins when they were stimulated with TNFa. Moreover, we found protein kinase R (PKR) was increased in naïve Atg16L1ΔIEC organoids, and inhibition of PKR significantly protected Atg16L1ΔIEC organoids from TNFa. Finally, we generated human intestinal organoids using biopsy specimens mostly collected from Crohn’s disease patients. We compared the viability between individuals with 2 copies vs 0 or 1 copy of ATG16L1T300A risk allele, and found organoids derived from ATG16L1T300A homozygous individuals exhibited heightened susceptibility to TNFa (Figure 2 A and B). Moreover, we tested the effect of Ruxolitinib as well as multiple necroptosis inhibitors on human organoids, and found all the inhibitors successfully protected ATG16L1T300A homozygous organoids from TNFa-induced cell death.

Conclusion

Our data indicate ATG16L1 prevents necroptosis by regulating interferon signaling in both mouse and human IECs, and suggest inhibition of JAK-STAT as well as necroptosis pathway could be promising therapeutic options for Crohn’s disease patients with ATG16L1T300A risk alleles.
FK506 BINDING PROTEIN 8 (FKBP8) INTERACTIONS WITH MYOSIN LIGHT CHAIN KINASE 1 ARE REQUIRED FOR TNF-INDUCED TIGHT JUNCTION BARRIER LOSS
Authors: Zuo Li , Cao Feng , Kuo Wei-Ting , Turner Jerrold
Published: January 2021

Background

Tumor necrosis factor (TNF) regulates intestinal epithelial tight junction permeability by activating myosin light chain kinase 1 (MLCK1) expression and enzymatic activity. MLCK1 recruitment to the apical perijunctional actomyosin ring (PAMR) is, however, required for barrier regulation; Divertin, a small molecule drug that blocks this recruitment, prevents barrier loss and attenuates both acute and chronic experimental diarrheal disease. We therefore hypothesized that MLCK1 recruitment to the PAMR requires interactions with as yet unidentified chaperone protein(s).

Objective

To identify binding partners and define the mechanisms by which they activate MLCK1 recruitment to the PAMR.

Results

We performed a yeast-2-hybrid (Y2H) screen using the MLCK1 domains required for PAMR recruitment as bait. FKBP8, which encodes a peptidyl-prolyl cis-trans isomerase (PPI), was recovered, and direct binding to the MLCK1 domains (Kd=~5mM) was confirmed using microscale thermophoresis (MST). This binding interaction required the FK506-binding PPI domain and was specifically inhibited by FK506 (tacrolimus). Immunofluorescent microscopy demonstrated partial colocalization of MLCK1 and FKBP8 within intestinal epithelial monolayers; TNF caused both to concentrate around the PAMR. To further characterize this interaction, we performed proximity ligation assays (PLA) and found that TNF increased interaction between MLCK1 and FKBP8 > 2-fold. FK506 prevented TNF-induced increases in PLA signal. FK506 was also able to reverse TNF-induced increases in myosin light chain (MLC) phosphorylation and tight junction permeability. In Caco-2 monolayers, FKBP8 knockout blocked TNF-induced MLCK1 recruitment, MLC phosphorylation, and tight junction barrier loss; all of which were restored by FKBP8 re-expression. In mice, MLC phosphorylation and intestinal barrier loss triggered by acute, anti-CD3-induced, T cell activation were blocked by luminal FK506. Importantly, this local FK506 treatment did not prevent anti-CD3-induced increases in mucosal TNF, IL-1b, and IFNg. Immunostains of biopsies from IBD patients documented increased PAMR MLC phosphorylation, MLCK1 recruitment, FKBP8 recruitment, and MLCK1-FKBP8 PLA signal relative to control subjects.

Conclusions

FKBP8 is a chaperone protein required for TNF-induced MLCK1 recruitment and barrier loss. This requires direct interaction between MLCK1 and the PPI domain of FKBP8. FK506 binding to the PPI domain displaces MLCK1 thereby preventing recruitment to the PAMR and barrier loss. These activities are separate from the immunosuppressive effects of FK506. We speculate that molecular blockade of the FKBP8-MLCK1 interaction may be a novel approach to barrier restoration and therapy of diseases associated with intestinal barrier dysfunction.

Support

NIH (DK068271, DK061931) and the NNSF of China (81800464, 82070548).
METHYLTRANSFERASE SMYD5 EXAGGERATES INFLAMMATORY BOWEL DISEASE BY REGULATING PPAR-γ COACTIVATOR 1-α STABILITY
Authors: Hou Yuning , Sun Xiaonan , Gheinani Pooneh , Guan Xiaoqing , Sharma Shaligram , Zhou Yu , Jin Chengliu , Yang Zhe , Naren Anjaparavand , Yin Jun , Denning Timothy , Gewirtz Andrew , Xie Zhonglin , Li Chunying
Published: January 2021

Background and Aims

The expression and role of methyltransferase SET and MYND domain-containing protein 5 (SMYD5) in inflammatory bowel diseases (IBD) is completely unknown. Here, we investigated the role and the underlying mechanism of epithelial SMYD5 in IBD pathogenesis and progression.

Methods

The expression and subcellular localization of SMYD5 and peroxisome proliferator-activated receptor gamma coactivator-1α (PGC-1α) were examined by Western blot analysis, immunofluorescence staining, and immunohistochemistry in intestinal epithelial cells (IECs) and in colon tissues from human IBD patients and mice with experimental colitis. Mice with Smyd5 conditional knockout in IECs and littermate controls were subjected to DSS-induced experimental colitis and the disease severity and inflammation were assessed. SMYD5-regulated mitochondrial biogenesis was examined by RT-qPCR and transmission electron microscopy and mitochondrial oxygen consumption rate was measured in a Seahorse Analyzer system. The interaction between SMYD5 and PGC-1α was determined by co-immunoprecipitation assay. PGC-1α degradation and turnover (half-life) were analyzed by cycloheximide chase assay. SMYD5-mediated PGC-1α methylation was measured via in vitro methylation followed by mass spectrometry to identify the specific lysine residues that were methylated.

Results

Up-regulated SMYD5 and down-regulated PGC-1α were observed in IECs from IBD patients and mice with experimental colitis. However, Smyd5 depletion in IECs protected mice from DSS-induced colitis. SMYD5 was critically involved in regulating mitochondrial biology such as mitochondrial biogenesis, respiration, and apoptosis. Mechanistically, SMYD5 regulated mitochondrial functions in a PGC-1α dependent manner. Further, SMYD5 mediated lysine methylation of PGC-1α and facilitated its ubiquitination and proteasomal degradation.

Conclusion

SMYD5 attenuates mitochondrial functions in IECs and promotes IBD progression by enhancing the proteasome-mediated degradation of PGC-1α protein in a methylation-dependent manner. Strategies to decrease SMYD5 expression and/or increase PGC-1α expression in IECs might be a promising therapeutic approach to treat patients with IBD.
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

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

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

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

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

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 >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< 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

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

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

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<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.
POLYSUBSTANCE USE IN INFLAMMATORY BOWEL DISEASE
Authors: Bogale Kaleb , Vrana Kent , Raup-Konsavage Wesley , Walter Vonn , Stuart August , Dalessio Shannon , Koltun Walter , Bernasko Nana , Tinsley Andrew , Williams Emmanuelle , Clarke Kofi , Coates Matthew
Published: January 2021

Background

Polysubstance Use (PSU), the use of two or more substances of abuse, has been associated with increased risk for development of psychiatric conditions and early death compared to both monosubstance use or no substance use. The vast majority of clinical research on prescription or recreational drug consumption in patients with inflammatory bowel disease (IBD), including both Crohn’s disease (CD) and ulcerative colitis (UC), has focused on use of individual substances. We evaluated the incidence and impact of PSU in IBD and assessed potential predisposing factors in this setting.

Methods

We performed a retrospective analysis using a consented IBD natural history registry from a single tertiary care referral center between 1/1/2015–8/31/2019. Demographics, endoscopic severity (using Mayo endoscopy sub-score for UC and Simple Endoscopic Score for CD), totals and sub-scores of surveys (Harvey-Bradshaw Index, Simple Clinical Colitis Activity Index, Hospital Anxiety and Depression Scale, Short IBD Questionnaire) assessing for symptoms (abdominal pain, fatigue, anxiety/depression, gas, diarrhea, rectal bleeding, and fecal urgency), substance use (tobacco, alcohol, marijuana, cocaine, methamphetamine, heroin, opiates, or benzodiazepine), and antidepressant or anxiolytic medication were abstracted. PSU was defined as concurrent use of two or more non-prescription drugs or substances, and healthcare resource utilization was defined as any IBD-related imaging, emergency room visit, hospitalization, or surgery over the prior 12 months. We computed descriptive statistics and performed contingency table analyses in order to identify associations between PSU and a variety of demographic and clinical characteristics. Multivariable logistic regression models were fit incorporating the clinical factors described above.

Results

315 consecutively enrolled IBD patients (166f:149m; 214 CD and 101 UC) were included. Sixty-six patients (21.0%) were polysubstance users. Of these patients, 40.9% had moderate to severe disease activity, 47.0% had extra-intestinal manifestations (EIM), and 36.4% demonstrated an anxious or depressed state. EIM and antidepressant or anxiolytic use were positively associated with PSU on bivariate analysis (Table 1) and multivariable analysis (Table 2).

Conclusions

PSU is common in IBD, including both CD and UC. Interestingly, disease activity, IBD therapy type, and IBD-related symptoms were not associated with PSU. EIM and antidepressant or anxiolytic use were the only statistically significant predictors of PSU among patients with IBD in the multivariable logistic regression models. Our study represents the first evaluation of PSU within IBD and reinforces the importance of appropriate substance use screening among IBD patients, particularly among those with EIM and antidepressant or anxiolytic use.
THE EVALUATION OF SERUM SEROTONIN LEVEL AMONG PATIENTS WITH CROHN’S DISEASE AND ITS IMPACT ON QUALITY OF LIFE
Authors: Sochal Marcin , Bialasiewicz Piotr , Gabryelska Agata , Talar-Wojnarowska Renata , Fichna Jakub , Szmyd Bartosz , Malecka-Panas Ewa
Published: January 2021

Background and aims

Serotonin affects intestinal physiology, mood, as well as circadian rhythm. Moreover, serotonin has proinflammatory function. Therefore, the aim of this study was to investigate the role of serotonin in clinical severity of Crohn’s Disease (CD) and its effect on pain and sleep quality.

Methods

Fifty-nine CD patients (34 in exacerbation and 25 in remission according to the Harvey-Bradshaw Index-HBI) and 25 health control individuals(HC) were recruited. Sleep quality was assessed by the Pittsburgh Sleep Quality Index (PSQI) and subjective severity of pain by the Visual Analog Scale (VAS). Seventeen patients were treated with anti-TNF-α induction therapy for 14 weeks.

Results

Serotonin level was higher in CD (145.12ng/mL, IQR:98.14–179.25) compared to HC (87.52ng/mL, IQR:70.04–129.39; p=0.002) and in exacerbation of CD (157.66ng/mL, IQR:111.94–197.64) compared to remission (122.33ng/mL, IQR:83.28–163.67; p=0.029). Serotonin level with cut-off point of 92.45 ng/mL is useful for distinguishing participants with CD from HC (sensitivity: 78%, specificity: 60%, positive predictive value: 82%). Positive correlation between serotonin and HBI (r=0.279, p=0.032) and severity of diarrhoea (r=0.260, p=0.047) were found. Serotonin does not correlate with PSQI (r=0.152, p=0.168), but correlates with presence of sleep fragmentation for example by getting up to use the bathroom (joined 5b-5j PSQI questions; r=0.270, p=0.039). Correlations between serotonin and VAS were also obtained (r=0.220, p=0.045). Moreover, serotonin level significantly decreased after anti-TNF-α therapy (192.35ng/mL, IQR:150.36–225.56 vs. 121.11ng/mL, IQR:91.28–188.87; p=0.006). The study was funded by National Science Centre, Poland (#2018/31/N/NZ5/03715).

Conclusions

Serotonin level correlates with the severity of CD and decreases after anti-TNF-α therapy. It is associated with sleep fragmentation, which may be caused by diarrhea.
USTEKINUMAB FOR CHRONIC GRANULOMATOUS DISEASE-ASSOCIATED INFLAMMATORY BOWEL DISEASE
Authors: Bhattacharya Sumona , Marciano Beatriz , Malech Harry , Holland Steven , De Ravin Suk See , Zerbe Christa , Heller Theo
Published: January 2021

Introduction

Chronic granulomatous disease (CGD) is a rare immunodeficiency caused by mutations in the NADPH oxidase complex. Dysregulated immune function may cause inflammatory bowel disease (IBD). Patients with CGD-associated IBD may not respond to or may develop serious infections as a result of traditional IBD therapies such as vedolizumab and infliximab. Ustekinumab is approved for use in Crohn’s disease and ulcerative colitis however there is scarce data on its efficacy and safety in CGD.

Aims

To evaluate the efficacy and safety of ustekinumab for CGD-associated IBD.

Methods

A retrospective chart review was conducted on CGD patients followed at a single center who had consented to participate in a natural history study. Clinical, laboratory, and endoscopic data were extracted in those that had received ustekinumab for IBD.

Results

Eight patients were found. Four were male and four were female. Five were white, one was Asian, one was black, and one was mixed race. Median age at diagnosis of CGD was 3 years (IQR 8) and of IBD was 15.5 years (IQR 20). Median age at initiation of ustekinumab was 27.5 years (IQR 14) and median duration on ustekinumab was 10 months (IQR 7). Six had colonic disease, two had ileocolonic disease, and six had perianal disease. Six failed other biologics (n=5 for vedolizumab, n=1 for infliximab, n=1 for adalimumab). Six patients symptomatically improved whereas two had no improvement. Changes in hemoglobin and C-reactive protein were equivocal. Three patients had improved endoscopic findings, two had unimproved findings, and three patients lacked this data. Overall, four patients achieved clinical remission. However, none of the five patients with endoscopic reevaluation achieved endoscopic remission. Three patients discontinued therapy due to lack of response: two required surgery and one underwent stem cell transplant. Fungal pneumonia (n=2), otitis media (n=1), oral herpes simplex virus 1 (n=1), and viral gastroenteritis (n=1) were reported. One infusion reaction occurred.

Discussion

In our cohort of eight patients with CGD-associated IBD receiving ustekinumab, results were mixed with four patients experiencing some degree of clinical or endoscopic improvement including four who achieved clinical remission. Multiple CGD-related variables may account for the mixed laboratory findings. Four of the five patients with endoscopic reevaluation had pre-existing strictures that would be unlikely to reverse with medical therapy alone. Of these, two had otherwise resolved endoscopic inflammation. Only two patients had no endoscopic improvement. Two serious infections occurred however CGD confers increased infectious susceptibility and no infections lead to discontinuation of therapy. Given these promising results, further formalized study of ustekinumab in CGD-associated IBD is needed.
X-ray-free protocol for pectus deformities based on magnetic resonance imaging and a low-cost portable three-dimensional scanning device: a preliminary study
Authors: Guillot Marc-Samir , Rouchaud Aymeric , Mounayer Charbel , Tricard Jérémy , Belgacem Alexis , Auditeau Emilie , Omraam Olivier , Fourcade Laurent , Ballouhey Quentin
Published: January 2021

OBJECTIVES

To compare a standard protocol using chest computed tomography (CT) to a non-irradiant protocol involving a low-cost portable 3D scanner and magnetic resonance imaging (MRI) for all pectus deformities based on the Haller index (HI).

METHODS

From April 2019 to March 2020, all children treated for pectus excavatum or carinatum at our institution were evaluated by chest CT, 3D scanning (iPad with Structure Sensor and Captevia—Rodin4D) and MRI. The main objectives were to compare the HI determined by CT or MRI to a derived index evaluated with 3D scanning, the external Haller index (EHI). The secondary objectives were to assess the inter-rater variability and the concordance between CT and MRI for the HI and the correction index.

RESULTS

Eleven patients were evaluated. We identified a strong correlation between the HI with MRI and the EHI (Pearson correlation coefficient = 0.900; P < 0.001), with a strong concordance between a radiologist and a non-radiologist using intra-class correlation for the HI with MRI (intra-class correlation coefficient = 0.995; [0.983; 0.999]) and the EHI (intra-class correlation coefficient = 0.978; [0.823; 0.995]). We also identified a marked correlation between the HI with CT and the EHI (Pearson coefficient = 0.855; P = 0.002), with a strong inter-rater concordance (intra-class correlation coefficient = 0.975; [0.901; 0.993]), a reliable concordance between CT and MRI for the HI and the correction index (Pearson coefficient = 0.886; P = 0.033).

CONCLUSIONS

Non-irradiant pectus deformity assessment is possible in clinical practice, replacing CT with MRI and 3D scanning as a possible readily-accessible monitoring tool.
An aorto-oesophageal fistula treated with total arch repair combined with oesophageal repair
Authors: Wang Chaojie , Lei Liming , Zhang Wenqian , Fan Xiaoping
Published: January 2021

Aorto-oesophageal fistula is a rare but fatal disease. Open surgical repair or endotube repair of aorta and oesophagus save patients from circulatory deterioration, major haematemesis and severe infection. Here we present a rare case of an aorto-oesophageal fistula in the right hemi-arch caused by a fish bone treated with an alternative surgical strategy. Total aortic arch repair using the frozen elephant trunk procedure combined with oesophageal repair were performed. No aortic fistula, oesophageal fistula or artificial graft infection occurred in the follow-up.
Stability results on radial porous media and Hele-Shaw flows with variable viscosity between two moving interfaces
Authors: Gin Craig , Daripa Prabir
Published: January 2021

We perform a linear stability analysis of three-layer radial porous media and Hele-Shaw flows with variable viscosity in the middle layer. A nonlinear change of variables results in an eigenvalue problem that has time-dependent coefficients and eigenvalue-dependent boundary conditions. We study this eigenvalue problem and find upper bounds on the spectrum. We also give a characterization of the eigenvalues and prescribe a measure for which the eigenfunctions form an orthonormal basis of the corresponding $L^2$ space. This allows for any initial perturbation of the interfaces and viscosity profile to be easily expanded in terms of the eigenfunctions by using the inner product of the $L^2$ space, thus providing an efficient method for simulating the growth of the perturbations via the linear theory. The limit as the viscosity gradient goes to zero is compared with previous results on multi-layer radial flows. We then numerically compute the eigenvalues and obtain, among other results, optimal profiles within certain classes of functions.
French Abstracts
Authors: Melissa Baysari , Bethany Van Dort , Mirela Prgomet , Wu Yi Zheng , Magda Raban , Luciano Dalla-Pozza , Cheryl Mccullagh , Johanna Westbrook
Published: January 2021

Problème de qualité: les trajets de soin des patients doivent être organisés en fonction des besoins de ces derniers.

Objectif: selon le principe du compromis entre efficacité et rigueur (ETTO, efficiency-thoroughness trade-off), les personnes et les organisations sont souvent obligées de faire un compromis entre être efficace et être rigoureux, car il est difficile d’être à la fois efficace et rigoureux. Cette étude visait à comparer, avant la mise en place de la gestion électronique des médicaments, comment un système de gestion électronique des médicaments pourrait influencer l’efficacité et la rigueur aux expériences post-mise en place de la gestion électronique des médicaments avec le système de gestion électronique et l’ETTO.

Plan: étude qualitative par entretien.

Contexte: centre d’oncologie pédiatrique dans un grand hôpital universitaire tertiaire de pédiatrie à Sydney, Australie.

Participants: 44 entretiens semi-directifs avec des médecins, infirmiers/ères et pharmacien/nes six mois avant et deux ans après la mise en place d’un système de gestion électronique des médicaments.

Résultats: avant la mise en place de la gestion électronique des médicaments, le personnel a identifié les domaines de travail pour lesquels était attendue une amélioration à la fois de l’efficacité et de la rigueur grâce à la gestion électronique des médicaments. Ces domaines incluaient l’accessibilité du dossier de médicaments ainsi que l’organisation et la lisibilité des informations sur les médicaments. Après la mise en œuvre de la gestion électronique des médicaments, le personnel a rapporté des améliorations dans ces domaines. Cependant, le système de gestion électronique des médicaments a été perçu comme apportant des bénéfices en termes de rigueur (sécurité) aux dépens de l’efficacité (temps). Les mesures visant à améliorer la sécurité dans le système de gestion électronique des médicaments ont mis en place des processus chronophages, comme des procédures de double vérification des médicaments.

Conclusions: globalement, le personnel était conscient de l’interaction concurrentielle entre la rigueur et l’efficacité et a rapporté que l’introduction d’un système de gestion électronique des médicaments a imposé des processus en faveur de l’amélioration de la rigueur, aux dépens de l’efficacité.
Processing index of barley grain and dietary undigested neutral detergent fiber concentration affected chewing behavior, ruminal pH, and total tract nutrient digestibility of heifers fed a high-grain diet
Authors: Ran Tao , Saleem Atef M , Beauchemin Karen A , Penner Gregory B , Yang Wenzhu
Published: January 2021

The objective of this study was to investigate the effects of processing index (PI) of barley grain and dietary undigested neutral detergent fiber (uNDF) concentration on dry matter (DM) intake, chewing activity, ruminal pH and fermentation characteristics, total tract digestibility, gastrointestinal barrier function, and blood metabolites of finishing beef heifers. The PI was measured as the density after processing expressed as a percentage of the density before processing, and a smaller PI equates to a more extensively processed. Six ruminally cannulated heifers (average body weight, 715 ± 29 kg) were used in a 6 × 6 Latin square design with three PI (65%, 75%, and 85%) × 2 uNDF concentration (low and high; 4.6% vs. 5.6% of DM) factorial arrangement. The heifers were fed ad libitum a total mixed ration consisting of 10% barley silage (low uNDF), or 5% silage and 5% straw (high uNDF), 87% dry-rolled barley grain, and 3% mineral and vitamin supplements. Interactions (P < 0.01) of PI × uNDF were observed for DM intake, ruminating and total chewing time, and DM digestibility in the total digestive tract. Intake of DM, organic matter (OM), starch, and crude protein (CP) did not differ (P > 0.14) between low and high uNDF diets, but intakes of NDF and acid detergent fiber were greater (P = 0.01) for high uNDF diets regardless of barley PI. Heifers fed high uNDF diets had longer (P = 0.05) eating times (min/d or min/kg DM) and tended (P = 0.10) to have longer total chewing times (min/kg DM) than those fed low uNDF diets. Additionally, heifers sorted (P = 0.01) against long particles (>19 mm) for high uNDF diets but not for low uNDF diets. Altering PI of barley grain did not affect (P > 0.12) total volatile fatty acid (VFA) concentration, molar percentages of individual VFA, or duration of ruminal pH < 5.8 and <5.6. Total VFA concentration was less (P = 0.01), acetate percentage was greater (P = 0.01), and duration of ruminal pH < 5.8 and <5.6 was less (P = 0.05) for high compared with low uNDF diets. Digestibility of DM, OM, and CP was greater (P = 0.02) for low vs. high uNDF diets with PI of 65% and 75%, with no difference between low and high uNDF diets at PI of 85%. Blood metabolites and gastrointestinal tract barrier function were not affected (P ≥ 0.10) by the treatments. These results suggest that increasing dietary uNDF concentration is an effective strategy to improve ruminal pH status in finishing cattle, regardless of the extent of grain processing, whereas manipulating the extent of barley processing did not reduce the risk of ruminal acidosis.