bluuebunny/crossref_metadata_2025_split huggingface dataset:
OP28 Gene expression (GE) values in a phase 2 trial of mirikizumab in ulcerative colitis (UC) correlate better with histopathology (HP) than endoscopy (EN) and Mayo scores
Authors: Steere B , GOTTLIEB K , Schmitz J , Higgs R , Jia B , Milch C , Tuttle J , Krishnan V , Belin R , Reinisch W 
Published: January 2020

Abstract

Background
HP is considered integral to the definition of mucosal healing of UC. The regulated transcription of genes determines cell identity and function, and, consequentially, HP and EN appearance. Here we correlate GE in a UC clinical trial with EN and HP indices, and the modified Mayo score (MMS).

Methods
249 patients. with moderately-to-severely active UC were randomised to placebo or different doses of mirikizumab, a p19-directed IL-23 antibody (NCT02589665). Gene expression in baseline biopsy samples from the most affected area in the colon was evaluated using Affymetrix HTA2.0 exon-format microarray with a typical Bioconductor workflow (DOI: 10.1186/gb-2004-5-10-r801). Genes were represented by their largest groups of highly correlated exon groups. Expression values were correlated with Robart‘s Histopathology Index (RHI), Geboes score (GS), Ulcerative Colitis Endoscopic Index of Severity (UCEIS), and modified Mayo score (MMS) at baseline. A threshold of false-discovery rate (FDR)-adjusted p-value ≤0.05 was applied to the significance of the r-values. Subsets of genes of interest were subjected to pathway enrichment using MetaCore (Clarivate Analytics). Simulation analysis (SIM) was performed to estimate any bias in correlation estimates caused by the number of levels used for different clinical scores.

Results
The plot (Figure 1) shows more genes with expression values that correlate significantly (q &lt; 0.05) with RHI (4647), than UCEIS (1232), or MMS (869) (refer to legend). There are 498 genes that intersect all scores, representing predominantly the pathways/networks in Table 1. The RHI covers 86 % (1064/1232) of the genes that correlate with UCEIS. The genes unique to the UCEIS (168) do not significantly map to a specific pathway/network. Highest absolute r-values were RHI 0.56, GS 0.45, UCEIS 0.4 and MMS 0.3. The corresponding lowest q values expressed in −Log10 are 15, 9, 4, and 2.5. SIM shows only a minimal bias of &lt;0.03 units (Pearson’s and Spearman’s r).

Conclusion
In our analysis, RHI tracked baseline gene expression values best, followed by another HP index (GS), followed by EN (UCEIS) and finally MMS. Judging by the orders-of-magnitude greater significance of correlation with gene expression values, the histology indices are particularly information-rich. Regulated gene expression, fundamental to a phenotypic appearance in HP, seems to best mirrored by the RHI, followed by GS. The genes that track with all indices predominantly modulate cell adhesion and leukocyte trafficking. The genes that are unique to the UCEIS, compared with the RHI, do not significantly belong to any recognisable pathway or network. Further evaluation of what current EN scores contribute relative to HP disease activity assessment in UC may be useful.
P026 The impact of different dietary factors in the etiopathogenesis of inflammatory bowel disease: a retrospective case–control study
Authors: Preda C M , Manuc T , Chifulescu A E , Istratescu D , Louis E , Diculescu M , Cristian T , Gheorghe L S , Manuc M 
Published: January 2020

Abstract

Background
Inflammatory bowel disease development has been associated with several environmental factors, among which diet can play a key role, probably due to a westernised lifestyle. However, its involvement in the pathogenesis of IBD is difficult to be demonstrated. The aim of this study was to analyse dietary composition in a Romanian and Belgian population with IBD.

Methods
We conducted an observational retrospective comparative study using two European cohorts (Romanian and Belgian). The IBD group included 76 Romanian and 53 Belgian patients with an IBD diagnosis while the control group included 56 healthy persons (35 Romanians and 21 Belgians). All subjects were interviewed and asked to fill in a questionnaire regarding diet.

Results
The baseline characteristics of the patients included in the two cohorts were described in the table. Figures 1 and 2 illustrated the consumption of different foods in Romanian, respectively, Belgian IBD patients before the onset of the disease vs. healthy controls (variables were compared with Fisher’s exact test). In the entire IBD cohort (Romanian+Belgian), we found a significantly increased consumption of sweets (OR 3.36[95% CI 1.6,7]), processed and high-fat meat (OR 2.5[95% CI1.4, 4.7], fried food (OR 9.5[3.8, 23.6]), salt (OR 2.8[1.5, 5.3]), ice cream (OR 3.25[1.1, 9.8]), mayonnaise (OR 3.49[1.1, 10.3]), margarine (OR 5.63[1.64, 19.33]), chips/nachos/other snacks (OR 2.3[0.97, 5.73]), compared with the healthy control group. The intake of seeds, nuts (OR 0.26[0.14,0.52]) and the yoghurt consumption (OR 0.44[0.23, 0.83]) was lower in the IBD group compared with the control group.

Conclusion
In Romanian and Belgian IBD patients, a westernised diet with increased consumption of sweets, processed food, high-fat meat, fried food, salt, margarine, snacks, ice cream and mayonnaise seems to be a risk factor for IBD. Food intake of seeds, nuts and yogurt may be a protective factor.
P035 NUDT15 variance increases DNA-incorporated thiopurine metabolites and lymphocyte apoptosis in patients with inflammatory bowel disease
Authors: Kiyohara H , Toyonaga T , Kuronuma S , Ueno A , Okabayashi S , Ozaki R , Takeuchi O , Coulthard S A , Redfern C P F , Terai H , Tanaka Y , Nakano M , Hibi T , Kobayashi T 
Published: January 2020

Abstract

Background
A novel thiopurine metabolizing enzyme, nucleotide diphosphate-linked moiety X-type motif 15 (NUDT15) was associated with drug-induced leukopenia in patients with non-synonymous genetic polymorphisms. Thiopurine-induced leukopenia in Japanese patients with genetic variance in NUDT15 (c.415C&gt;T) appears to be independent of the 6-thioguanine nucleotide concentration in red blood cells. However, detailed molecular mechanism how NUDT15 variance causes thiopurine-induced leukopenia remains unclear and NUDT15-associated subcellular thiopurine metabolism has not been investigated in patients with inflammatory bowel diseases (IBD).

Methods
DNA-incorporated deoxythioguanosine (dTG) was measured in the peripheral blood mononuclear cells (PBMCs) of Japanese patients with IBD under thiopurine treatment. Association of a single-nucleotide polymorphism for NUDT15 (c.415C&gt;T) with dTG in PBMCs (dTGPBMC) was examined. Peripheral blood T lymphocytes were cultured in vitro with 6-thioguanine (6-TG) to examine the Impact of NUDT15 genotypes on incorporation into DNA, cell proliferation and apoptosis.

Results
NUDT15 variants had significantly higher dTGPBMC per thiopurine dosage than non-variants (homozygous variants (TT) vs. heterozygous variants (CT) vs. non-variants (CC), 4418.0 vs. 663.0 vs. 295.3 dTG mol/106 moles dA per mg/kg/day of 6-MP (Figure A)). dTGPBMC and peripheral lymphocyte counts showed a negative correlation (r = −0.30, p = 0.015) (Figure B). Peripheral blood lymphocytes from patients with NUDT15 variance showed a higher DNA-incorporated dTG associated with increased apoptosis (increase of Annexin V+ PI+ CD4+ lymphocytes; TT vs. CT vs. CC, 158.5 % vs. 80.1 % vs. 57.9 % (p = 0.0427)) (Figure C) and decreased proliferation (decrease of proliferative CD4+ lymphocytes, TT vs. CT vs. CC, 49.0 % vs. 25.0 % vs. 19.1 % (p = 0.0098)) (Figure D) when cultured with 6-thioguanine in vitro.

Conclusion
DNA-incorporated dTG affected by NUDT15 genotypes induces T lymphocyte apoptosis in patients with IBD.
P053 3-Oxo-C12:2, a quorum sensing molecule from the human gut, exerts anti-inflammatory effects through a bitter taste receptor
Authors: Coquant G , Aguanno D , Peyrottes A , Brot L , Thenet S , Grill J P , De Sordi L , Seksik P 
Published: January 2020

Abstract

Background
Acyl-homoserine lactones (AHLs) are quorum sensing molecules involved in bacterial communication network and can also have an impact on host’s cells. We recently showed the presence of AHLs in the gut ecosystem and identified one that has never been described: 3-oxo-C12:2. This molecule was decreased in faecal samples of inflammatory bowel disease (IBD) patients, especially during flare, and its absence was correlated to dysbiosis. 3-Oxo-C12:2 is structurally close to an AHL well described and synthesised by P. aeruginosa, 3-oxo-C12. We intent to describe 3-oxo-C12:2 effects on gut inflammation and to identify which signalling pathways are involved. Given its analogous structure to 3-oxo-C12, we hypothesised that 3-oxo-C12:2 can interact with the same cellular partners, in particular a bitter taste receptor, called T2R138, which is a GPCR expressed by immune and epithelial gut cells.

Methods
To test our hypothesis, we used murine macrophages cell line RAW264.7, stimulated by interferon-γ (IFN-γ; 20 U/ml) and lipopolysaccharide (LPS; 10 ng/ml). Inflammatory response was monitored by measuring cytokine secretion (TFNα, IL-6 and IL-10) via ELISA. Protein expression was assessed by Western blot. Probenecid, a known allosteric inhibitor for T2R138, was used to study T2R138 role in AHL signalling. Cytotoxicity was checked by measuring LDH release.

Results
In steady state, challenged by 3-oxo-C12:2, RAW264.7 macrophages showed no change in cytokine expression (TNFα, IL-6 and IL-10). After LPS/IFN-γ activation, we observed a decrease in the ratio of secreted TNFα/IL-10 when cells are exposed to 3-oxo-C12:2, in a dose-dependent manner: 15 μM (−30%, p &lt; 0.05), 25 μM (−50%, p &lt; 0.001) and 50 μM (−65%, p &lt; 0.0001). This reflects an anti-inflammatory effect, without increasing cytotoxicity. Those immune-modulatory effects were lost in presence of Probenecid. Moreover, the amount of T2R138 protein was not changed in the presence of Probenecid and/or 3-oxo-C12:2. Therefore, the loss of anti-inflammatory effects with the inhibitor was not due to a decrease of the receptor expression in our experimental conditions.

Conclusion
3-Oxo-C12:2 exerts a dose-dependent anti-inflammatory effect on murine immune cells. This response is partly mediated by the bitter taste receptor T2R138. This receptor is a potential target of our AHL. We are currently studying via a multiplex assay the cytokine response after exposing cells to 3-oxo-C12:2. Studying the signalling between the receptor and the anti-inflammatory response would allow us to understand better the inter-kingdom dialogue between microbiota and the host and to what extend AHLs are involved.
We are grateful for the FRM for the financial support (ECO201806006843)
P056 Kefir (Lactobacillus kefiranofaciens) promotes autophagy and protects the intestinal barrier against DSS-induced colitis
Authors: Da Silva K N , Cordeiro T S , Fávero A G , Bogsan C S , Sinigaglia R D C , Miszputen S J , Ambrogini–Júnior O , Paiotti A P R 
Published: January 2020

Abstract

Background
Inflammatory bowel disease occurs when luminal antigens from microbiota stimulate the immune system of an intestinal barrier through defects in the tight junctions lead to an exacerbated response in the genetically predisposed individual. It is well known that the consumption of probiotics improves intestinal health. Specifically, the Lactobacillus sp. strains play a key role in the epithelial intestinal barrier, promote mucin secretion, better performance of tight junctions and prevent apoptosis of epithelial cells. This study aimed to evaluate the role of Kefir (Lactobacillus kefiranofaciens) on inflammation of dextran sodium sulphate (DSS)-induced colitis.

Methods
Forty male Wistar rats were randomised into five groups: Sham; 5% DSS-induced colitis; control Kefir; 5% DSS-induced colitis treated with Kefir; 5% DSS-induced colitis treated with Kefir + 5-ASA (125mg/kg/day) (Ethics Committee on the Use of Animals number 3627290119). The lactic culture of Kefir (Danisco Biolacta, Olstyn, Poland) was administered ad libitum per 6 days. The preparation of the matrix was carried out as follows: whole milk powder was reconstituted to 10% with distilled water and was incubated at 85°C for 15 min using a mechanical shaker. Subsequently, 20 mg freeze-dried Kefir culture was added to 100 ml of the treated milk and fermentation were carried out at 23°C for 16 h. When the desired pH of 4.6 was reached, the fermentation was stopped by cooling the flasks in ice bath and storing them under refrigeration at 4°C until utilisation. The rats were euthanised on day 9. Colonic samples were processed for transmission electron microscope (TEM) and conventional microscopy. Samples were analysed in a TEM EX II 1200 (JEOL, Japan) coupled to a digital camera system GATAN Orius (USA). Histopathological evaluation was performed by a pathologist under a light microscope. Parameters such as inflammation extent, regeneration and crypt damage were graded according to Dieleman et al.3 Statistical analysis was performed by one-way analysis of variance (ANOVA), followed by Tukey’s test using Graph Pad Prism (version 6.0).

Results
Kefir treatment reduces the severity of DSS-induced colitis evidenced by decreased abdominal discomfort and rectal bleeding. Histologically, there was a significant decreased of colonic damage and preservation of goblet cells (p &lt; 0.0001) compared with DSS-colitis control group (Figure 1). The effect of Kefir was also verified by TEM analysis at the ultrastructural level, demonstrating the tight junctions preserved, decreased of reticulum edema and increased of autophagosomes (Figure 2).

Conclusion
Kefir promotes autophagy and protects the intestinal barrier against damage caused by DSS.
P073 Compartmentalised human gut immunity: studies of innate and adaptive lymphocyte distribution in the epithelium, lamina propria and GALT of healthy gut mucosa by flow cytometry
Authors: Carrasco Garcia A , Rao A , Kokkinou E , Haapaniemi S , Lindforss U , Winberg M E , Casado-Bedmar M , Fenton T , Jørgensen P , Nordenvall C , Agace W , Keita Å V , Söderholm J D , Mjösberg J 
Published: January 2020

Abstract

Background
The human gut mucosal immune system is compartmentalised in distinct and specialised immune niches. The epithelium and the lamina propria have been proposed as effector sites, while gut-associated lymphoid tissues (GALTs) constitute inductive immune niches. The major mucosal GALTs are the Peyer’s patches in the ileum and the colonic isolated lymphoid follicles (ILFs), scattered in the submucosa of the colon. The majority of studies of human gut immune function in health and disease have analysed unfractionated mucosal tissue samples. Hence, in contrast to mice, little is known about compartmentalised immune cell specialisation in the human gut. The aim of this study was to use novel dissection methods to analyse separate human gut immune niches.

Methods
Macroscopically healthy margins from colorectal cancer colectomies were obtained at a minimum distance of 10 cm from the tumour border. After faeces, mucus, fat and muscle removal, Peyer’s patches were identified and dissected using a stereomicroscope (based on Keita et al., Lab Invest, 2006). Colonic mucosa and submucosa (containing ILFs) fractions were mechanically separated by forceps (based on the method developed by Fenton et al., Immunity, under revision). Isolation of epithelial and lamina propria fractions from the mucosal compartment was performed by calcium chelation (DTT and EDTA) and enzymatic digestion (Collagenase II and DNAse), respectively. Cell suspensions from each fraction were analysed by flow cytometry (BD LSR-Fortessa and BD FACSymphony).

Results
As expected, mucosal GALTs were characterised by an enrichment of germinal centre B cells (CD19+CD20+CD38+), lymphoid tissue-like innate lymphoid cells (Lin−CD127+CD117+Nrp1+) and a higher CD4+/CD8+ T-cell ratio vs. mucosa, whereas the mucosal fraction was enriched for plasma cells (CD19+CD20−CD38high) and distinguished by a decreased CD4+/CD8+ T-cell ratio as compared with the GALT in both ileum and colon. CD19+/CD3+ ratios were only higher in Peyer’s patches but not in colonic submucosa enriched with ILFs, possibly due to the smaller size of the B-cell follicles in the latter. The intraepithelial compartment lacked B cells and contained more γδ-T cells as compared with the GALT and lamina propria.

Conclusion
We have used novel dissection methods in human intestinal tissues that reveal a compartmentalised immune cell specialisation that is in line with what has previously been described in mice. The method will allow for future deeper analysis of the human gut immune niches in health and disease, such as in inflammatory bowel disease.
P181 Triage straight to test (tSTT) improves early diagnosis of inflammatory bowel disease (IBD)
Authors: Jora S 
Published: January 2020

Abstract

Background
It is well known that there is a significant delay in diagnosing IBD. On average it takes 4–9 months to diagnose Crohn’s disease (CD) and 4 months for ulcerative colitis (UC)1. Barts Health NHS Trust in the UK is one of the pioneer hospitals in establishing Triage Straight to Test (tSTT) for patients with lower gastrointestinal (GI) symptoms. The aim of the programme is to reduce the time from a primary care physician (General practitioner) referral to the secondary care to achieve a definitive diagnosis. This minimises the number of outpatient appointments required and makes an efficient use of medical and surgical resources. We aim to study whether tSTT pathway reduces the time to diagnose IBD.

Methods
Patients with lower GI symptoms were referred to a specialist tSTT triage nurse, where they were allocated into two pathways—routine (18-week wait) and upgraded (2-week wait). A comprehensive telephone assessment, calprotectin levels and clinical information provided by the GP would determine the route and urgency of the investigation. Pathologies such as cancer, IBD and polyps were treated via a predefined management pathway. The patients with a benign disease were discharged and in some cases called for a follow-up appointment.

Results
During July 2013 to November 2019 a total of 2270 patients have been triaged. One hundred and one (4.45%) patients were diagnosed with IBD, where 36 patients (36%) were histologically characterised as CD and the other 65 patients (64%) were diagnosed as UC. The mean age of diagnosis for both CD and UC patients was 40 years (range 16–79). The average time from referral to endoscopy on tSTT pathway was 22.6 days for upgraded referrals and 43.5 days for non-upgraded. There is a significant difference in time to diagnosis between routine and upgraded pathways (22.6 days vs. 43.5 days, p &lt; 0.001).

Conclusion
The above data suggest a significant reduction in time to diagnose IBD through tSTT pathway. Early diagnosis ensures that the patient receives an early customised care and avoids disease-related complications. It also means that the NHS resources can be used wisely by avoiding a delay in getting outpatient consultant appointments.
P194 Risk factors of axial spondyloarthritis among inflammatory bowel disease patients
Authors: JEMMALI C , Laabidi A , Hafi M , Ben Mustapha N , Serghini M , Fekih M , Boubaker J 
Published: January 2020

Abstract

Background
Spondyloarthritis (SpA) is the most common extraintestinal manifestation (EIM) in inflammatory bowel disease (IBD) patients. However, the delay for the diagnosis of SpA in IBD patients is still unacceptably long, and may be the main cause of impairment due to ankylosing spondylitis. The aim of this study was to identify predictors of axial SpA in IBD patients, in order to prevent complications.

Methods
We conducted a retrospective study between 2001 and 2018 including all IBD patients diagnosed with symptomatic axial Spondyloarthritis (SpA) (Group1) and non-SpA patients among IBD (Group2). Diagnosis of SpA was based on ASAS 2009 criteria and confirmed by rheumatologists. Medical data were collected then uni and multivariate statistical analysis (p significant if ≤0.05) were performed.

Results
Seventy-eight patients with mean age 38 ± 11 years and sex ratio H/F = 1.6 were included. There were 58 (74.4%) Crohn’s disease (CD) patients and twenty patients (25.6%) had ulcerative colitis (UC). The average duration of IBD in all patients was 89.7 ± 69.1 months. SpA group included 26 patients (33.3%), six of whom (23%) had a late-stage disease (ankylosis). Most of the SpA patients (53.8%) had their disease before IBD onset with an average delay of 52.5 ± 68 months between SpA diagnosis and IBD symptoms. In univariate analysis, factors associated with SpA in IBD patients were familial history of IBD (p = 0.025) and the type of IBD (39.7% in CD vs. 15% in UC; p = 0.04). Among CD patients’, structuring-type was significantly associated with SpA (p = 0.04). Multivariate analysis showed that familial history of IBD (OR = 0.17; IC95%; 0.03–0.93; p = 0.04) and CD (OR = 3.72; IC 95%; 0.98–14.15; p = 0.05) were independent risk factors of axial SpA during IBD. EIMs other than SpA were more frequent in group 1 than in group 2 such as peripheral arthropathies (19.2% vs. 8.3% respectively; p = 0.02), anterior uveitis (11.5% vs. 0% respectively; p = 0.01) and erythema nodosum (7.7% vs. 0% respectively; p = 0.04).

Conclusion
In conclusion, predictors of symptomatic axial SpA were familial history of IBD and CD with structuring behaviour. Early-stage diagnosis is important to avoid ankylosis, which is a major cause of handicap in younger patients.