bluuebunny/crossref_metadata_2025_split huggingface dataset:
P023 Prevalence of malnutrition, risk of malnutrition and quality of life among patients with Inflammatory Bowel Disease
Authors: Korwel K M , Hjortswang H , Eberhardson M , Pihl Lesnovska K 
Published: January 2022

Abstract

Background
The relapsing and remitting course of inflammatory bowel disease (IBD) with inflammation and gut symptoms may result in an elevated risk of malnutrition and impaired quality of life. Patients with pro-longed active IBD are at the highest risk of developing impaired nutritional status. The aim of this study was to determine the prevalence of malnutrition and risk of malnutrition in outpatients with inflammatory bowel disease. Furthermore, to study their body composition, identify factors associated with malnutrition and investigate the association between malnutrition and quality of life.

Methods
A total of 328 IBD outpatients, (154 [47 %] with Crohn’s disease (CD) and 174 [53%] with ulcerative colitis (UC)) at Linköping University hospital were included in this population-based cross-sectional study. Patients underwent malnutrition assessment using the recently published Global Leadership Initiative on Malnutrition (GLIM) criteria. Data about body mass index, weight loss and dietary problems were collected and used to evaluate risk of malnutrition. The Short Health Scale (SHS) was used to measure quality of life.

Results
Malnutrition was detected in 33 patients (10,1 %) and 162 patients (55,0 %) had an increased risk of malnutrition. Malnutrition was associated with lower quality of life in the entire patient cohort and there was no difference between diagnoses. The prevalence of malnutrition was significantly higher among women (73% vs 27 %). Malnourished patients and those at risk of impaired nutritional status were more likely to have had active disease during the preceding 12 months compared to patients without malnutrition or at risk of malnutrition (27 % vs 12% [p=0.015] and 15 % vs 7% [p=0.037] respectively). Surprisingly, a higher calprotectin value was detected among patients without risk of malnutrition when compared to those who were at risk (median 46 [IQR 17 – 106] vs 59 [IQR 25 – 288] respectively, p=0,009). Women with IBD had significantly lower fat-free mass index (FFMI) compared to men. Patients with CD had reduced quality of life compared to patients with UC.

Conclusion
The prevalence of malnutrition in IBD-outpatients was 10% and more than half of the participants were at risk of malnutrition. Malnutrition was associated with poor quality of life. Malnutrition screening might be important in this population as ongoing inflammation and active disease seem to be apparent even among patients without clear nutrition difficulties or weight problems, but might influence nutritional status and quality of life negatively in the long term. Patients with IBD should therefore be screened for malnutrition and risk of malnutrition on a regular basis.
P024 CD40 and key CD40/CD154 pathway members are significantly upregulated in inflamed lesional IBD biopsies as shown by whole transcriptome AmpliSeq gene expression profiling and corresponding immunohistochemistry
Authors: Raulf F , Delucis-Bronn C , Angevaare E , Caesar N , Marcantonio A , Ceci M , Regnier C , Bonal C , Siegel R , Niess J H , Wieczorek G , Hrúz P 
Published: January 2022

Abstract

Background
Inflammatory bowel diseases (IBDs) are chronic inflammatory disorders with growing evidence of a critical role of the CD40/CD40L pathway (Shenhaji et al Front Immunol 6:529). To confirm and extend these findings, we profiled colonic biopsies from IBD patients with active ulcerative colitis (UC) and Crohn’s disease (CD) as well as non-IBD controls (HV).

Methods
Mucosal inflamed and non-inflamed biopsies of IBD patients and of healthy control subjects were immediately immersed in RNAlater (for RNA) or formaldehyde (for histology). RNA was extracted by RNeasy (Qiagen) with DNase digestion, quantified by UV spectrophotometry, and quality-controlled by Bioanalyzer (Agilent). 10 ng total RNA was subjected to ultrahigh-multiplexed RT-PCR using the AmpliSeq Transcriptome Human Gene Expression kit for 20,802 genes (Thermo Fisher). Barcoded samples were multiplexed and amplicons sequenced on an Ion GeneStudio S5 (Thermo Fisher). The resulting Excel matrix of normalized reads [RPM = Reads Per Million mapped reads; equivalent to TPM, Transcripts Per Million] was analyzed by Qlucore Omics Explorer 3.7 (Qlucore) and GraphPad Prism 9 (GraphPad Software). For histology, 2 μm sections of FFPE colonic biopsies were subjected to standard hematoxylin and eosin stain, and to automated immunohistochemistry (IHC; Ventana Discovery XT & Leica BOND RX) using standard cell-type specific antibodies and anti-CD40 Ab213205 (Abcam) and anti-CD40L AF617 (R&D Systems).

Results
161 colonic biopsies were analyzed from 36 CD and 26 UC patients (from inflamed and non-inflamed mucosa per subject), and 14 HV. Unbiased gene expression analysis by principal component analysis and hierarchical clustering showed upregulation of known inflammation markers and inflammasome components. In inflamed mucosa mRNA levels of CD40 (figure), CD154, CD44, CD86, KYNU, FCRL4, and others revealed a significant consistent upregulation of the CD40/CD154 pathway. Representative corresponding IHC stainings confirmed the upregulation of CD40 and CD154 at the protein level showing strong expression of CD40 in lesional biopsies (inflamed areas and epithelial cells), moderate expression in non-lesional and low in HV biopsies (no expression in epithelial cells), while higher expression of CD154 was observed in lesional and non-lesional biopsies compared to HV.

Conclusion
Consistent upregulation and activation of the CD40/CD154 axis was observed in inflamed mucosa of patients with IBD. Together with other (pre-)clinical evidence pointing to the importance of CD40 signaling, manipulation of this pathway might be a potential therapeutic target in IBD.
P025 A review of the newly implemented Inflammatory Bowel Disease dietetic service
Authors: Norman E 
Published: January 2022

Abstract

Background
Nutrition plays an important role in the onset and later the management of inflammatory bowel disease (IBD). Patients are becoming progressively interested in non-pharmacological methods of managing IBD and the dietary advice around this. It is reported that only 7% of services have enough dietitians to meet the IBD Standards recommendation. A year after the implementation of a new dietetic post at Whipps Cross Hospital, it was decided to review the effectiveness of this post for patients with a survey.

Methods
The inclusion criteria were patients with a diagnosis of IBD, reviewed at least twice by the IBD dietitian and patients who are under the IBD team at Whipps Cross Hospital. Patients were telephoned and asked if they consented to providing their email address for the anonymous survey link to be sent to. The survey was emailed to 40 patients. Six questions were asked on the survey, the survey provider was Survey Monkey.

Results
The final cohort included a 53% response rate to the survey. Age range was 18–64 years old. 95% of patients felt their consultation time was long enough. 95% of patients thought they had sufficient dietetic contact. 95% of patients felt the dietitian involved them in decision making. 67% of patients thought nutrition input improved their health/quality of life, with 29% being unsure. 100% thought having access to an IBD dietitian was useful. 71% of patients rated their dietetic care as excellent and 29% rated it as good.

Conclusion
The response from the majority of IBD patients was that nutrition input was helpful; with all patients finding the care provided by the dietitian was either good or excellent. All patients found it supportive to have dietetic access as part of a multidisciplinary approach to their IBD care. These findings support the best practice guidelines for having a dietitian post as part of the IBD team.
P028 Inhibition of stromal glycolysis by targeting PFKFB3 decreases experimental colitis
Authors: Zhou Z , Plug L G , de Jonge-Muller E S M , Abou Elmagd A , van der Meulen-de Jong A E , Barnhoorn M C , Hawinkels L J A C 
Published: January 2022

Abstract

Background
Studies in fibrotic diseases revealed that glycolysis is the preferred energy source for fibroblasts. 6-phosphofructo-2-kinase/fructose-2, 6-bisphosphatase 3 (PFKFB3) has the highest kinase activity to shunt glucose toward glycolysis. Therefore inhibition of PFKFB3 has been proposed as a potential target for several cancers and inflammatory diseases. However, the metabolic status of fibroblasts in patients with inflammatory bowel disease (IBD) and the role of PFKFB3 are currently unknown.

Methods
Single-sample gene set enrichment analysis (ssGSEA) of GSE16879 was performed to evaluate metabolic changes in IBD. Seahorse real-time cell metabolic analysis was performed to explore the metabolic activity of fibroblasts. Next the expression of PFKFB3 in primary patient derived intestinal fibroblast was determined by quantitative PCR (qPCR) and western blot under normal and inflammatory conditions. Proliferation and migration of fibroblasts were measured using colony formation and wound healing assays. In order to evaluate the effect of the inhibition of PFKFB3 in vivo, PFK15, a specific inhibitor of PFKFB3, was intraperitoneal injected in mice with dextran sodium sulfate (DSS)-induced colitis and in the T-cell transfer model for colitis. Next to clinical parameters, the abundance of α-smooth muscle actin (α-SMA) expressing fibroblasts, immune cells (CD45) and endothelial cells (CD105) was determined by immunohistochemistry.

Results
The ssGSEA analysis revealed glycolysis was significantly higher in IBD patients, compared to healthy controls. Consistently, the expression of PFKFB3 was also elevated in a cohort of inflamed intestinal tissues from IBD patients compared to non-inflamed sites from the same patient or healthy controls. On the cellular levels, this analysis showed that PFKB3 expression was higher in IBD-derived stromal cells compared to healthy or non-inflamed stromal cells. In vitro PFKFB3 expression in fibroblasts was increased after the stimulation with pro-inflammatory cytokines like TNF-α and a mix of cytokines often upregulated in IBD patients: interleukin (IL)-17A, oncostatin M (OSM) and IL-1β. As for the metabolic changes, inflamed fibroblasts had a higher extracellular acidification rate and a lower oxygen consumption rate, which could be reverted by inhibition of PFKFB3 using PFK15. Furthermore, PFK15 suppressed the proliferation and migration of fibroblasts. The in vivo experiments showed that PFK15 reduced the severity of the colitis, accompanied by a reduction of the total amount of immune cells (CD45), activated fibroblasts (α-SMA) and angiogenesis (CD105).

Conclusion
Increased PFKFB3 expression seems to contribute the inflammation and the pathological function of fibroblasts in IBD.
P038 Targeting the hepcidin-ferroportin axis with Nrf2 agonists to treat iron deficiency anaemia in Inflammatory Bowel Disease
Authors: Martin J , Aksan A , Steinhilber D , Ulrich-Rückert S , Stein J M 
Published: January 2022

Abstract

Background
Anaemia of inflammation is a common extraintestinal manifestation of IBD, impacting patient quality of life and hospitalisation rates. High circulating levels of proinflammatory cytokines induce hepatic production of the iron-regulatory peptide hormone hepcidin. By binding to the iron exporter ferroportin, hepcidin induces its internalisation and degradation, restricting the bioavailability and absorption of iron. This study aimed to test the effect of dimethyl fumarate (DMF) and sulforaphane (SFN), known activators of the transcription factor Nrf2, on the hepcidin-ferroportin (hep-fer) axis.

Methods
HepG2 cells were cultivated under standard conditions and treated with DMF or SFN for 16h. For reporter gene assays, plasmids were transfected by lipofection; luciferase activity was measured luminometrically. Quantitative real-time PCR was performed to determine mRNA expression. Proteins were detected by Western blot or LC-MS/MS. For gene knockdown, HepG2 cells were transfected with targeting or nontargeting siRNA duplexes. Graph Pad Prism v.7.00 and IBM SPSS 25 were used for statistical analysis.

Results
Both DMF and SFN reduced basic hepcidin expression and counteracted cytokine-induced hepcidin expression at promoter, mRNA and protein levels in HepG2 cells, independent of Nrf2 activation. Furthermore, DMF caused upregulation of ferroportin and ferritin at mRNA level, dependent on induction of Nrf2 and its downstream target haem oxygenase-1 (HO-1).

Conclusion
The Nrf2 agonists DMF and SFN might ameliorate anaemia of inflammation by disrupting the hep-fer axis by Nrf2-dependent and -independent mechanisms, and may be a promising therapeutic option for anaemia of inflammation for patients with IBD.
P051 IFNγ-macrophages could mediate EMT in Crohn’s disease through the WNT pathway
Authors: Edo M M , Macias-Ceja D C , Bauset C , Lis-Lopez L , Coll S , Seco-Cervera M , Cosin-Roger J , Calatayud S , Barrachina M D , Ortiz-Masiá D 
Published: January 2022

Abstract

Background
Macrophages contribute to fibrosis by releasing different mediators and the pattern of secretion may vary depending on the surrounding environment. We previously described that the mRNA expression of IFNγ was significantly higher in intestinal samples from CD patients.

Methods
The aim of the present study is to analyze the role of IFNγ-treated macrophages in epithelial mesenchymal transition (EMT) through the WNT pathway. The mRNA and protein expression of IFNγ in surgical resections from Crohn′s disease. U937 were differentiated to macrophages and then treated with IFNγ (10 ng/ml) for 4 days, the mRNA expression of WNT2b, WNT6 and TGFβ were determined by RT-PCR and protein. IFNγ-U937 were coculture with HT29 cells for 3 days and the expression of EMT markers, βCATENIN and WNT2b in HT29 cells were analyzed by WB. In some cases, HT29 cells were treated with the inhibitor of the WNT-pathway, XAV939 (1 μM), or were transfected with vectors-targeting human FZD4 (miFzd4). Results are expressed as mean±SEM. Statistical analysis was performed by ANOVA + Newman-Keuls or unpaired t-test.

Results
The mRNA and protein expression of IFNγ were significantly higher in intestinal samples from B2 CD patients (11.4±1.6 fold induction and 70,0 ± 2,3 pg/mg, respectively) and B3 CD patients (14.2±1.8 fold induction and 80,4 ± 6,8 pg/mg, respectively) than in controls (1,0±0,1 fold induction and 18,9 ± 0,3 pg/mg, respectively). U937 cells treated with IFNγ increased significantly the protein expression of WNT2b (1,3 ± 0,07 N=13* vs vehicle) but not the expression of WNT6 or TGFβ. IFNγ-U937 co-cultured with HT29 increased significantly the protein expression of EMT markers, βCATENIN and WNT2b in HT29 cells (VIMENTIN: 2,7 ± 0,3 N=6* vs vehicle-HT29; SNAIL1: 1,5 ± 0,1 N=5* vs vehicle-HT29; βCATENIN: 1,4 ± 0,09 N=5* vs vehicle-HT29; WNT2b: 2,0 ± 0,3 N=8* vs vehicle-HT29). Treatment HT29 cells with XAV939 (VIMENTIN: 1,4 ± 0,4 N=6 vs vehicle-HT29 XAV; SNAIL1: 1,0 ± 0,1 N=5 vs vehicle-HT29 XAV; βCATENIN: 0,8 ± 0,1 N=5 vs vehicle-HT29 XAV) or with miFzd4 (VIMENTIN: 0,3 ± 0,07 N=12* vs IFNγ-HT29 mock; SNAIL1: 0,5 ± 0,06 N=12* vs IFNγ-HT29 mock; βCATENIN: 0,6 ± 0,04 N=12* vs IFNγ-HT29 mock; FZD4: 0,5 ± 0,04 N=12* vs IFNγ-HT29 mock) partially reversed the increases produced by IFNγ-U937 in HT29 cells.

Conclusion
IFNγ-macrophages may stimulate the expression of WNT ligands in an autocrine and paracrine manner. The expression of WNT ligands at the epithelial level could favor mesenchymal epithelial transition through WNT2b ligand and FZD4 receptor.
P055 Factors from human mesenteric adipose tissue control intestinal stem cell differentiation and proliferation in vitro
Authors: Ziegler J , Gerbeth L , Brunkhorst M F , Glauben R , Weidinger C , Siegmund B 
Published: January 2022

Abstract

Background
In Crohn’s disease, hyperplastic mesenteric adipose tissue wrapping around inflamed small intestinal segments is a pathognomonic finding. Factors secreted by this so-called creeping fat can influence immune cells and affect disease severity in animal models. However, data on the interactions of adipose tissue and the intestinal epithelium are still sparse and derive almost exclusively from cell lines and animal models. Therefore, we employed primary human intestinal organoid culture and primary human adipose tissue explants to elucidate the role of adipose tissue-derived factors on the epithelium.

Methods
Creeping fat and unaffected mesenteric adipose tissue from a Crohn’s disease patient was used to produce adipose tissue-conditioned medium (ATcM). ATcM or control medium was added to growth medium (contains Wnt3a) and differentiation medium (Wnt3a removal) during primary human ileum-derived organoid culture. We analysed organoid morphology, proliferation, and phenotype by light microscopy, live cell imaging, resazurin viability assay, and reverse transcription quantitative polymerase chain reaction (RT-qPCR).

Results
In the presence of ATcM, differentiation of organoids was accompanied by a subtle and only transient delay in the development of morphologic signs of differentiation, namely organoid budding and darkness. In line, RT-qPCR at day 4 showed high expression of the differentiation markers MUC2 and ALPI in ATcM as well as in control conditions. In stem cell-enhancing growth medium, however, ATcM from both unaffected mesenteric adipose tissue and creeping fat induced a profound alteration of organoid morphology, perfectly reflecting so-called spheroids. Additionally, this ATcM-treated organoids were larger than the control, which corresponded with increased proliferation as measured by resazurin assay. This stem-like phenotype was further confirmed by a reduced expression of the differentiation markers MUC2 and ALPI in ATcM-treated organoids compared to the control.

Conclusion
Overall, our data suggest that factors from human mesenteric adipose tissue have profound effects on intestinal epithelial stem cells, while differentiated cells appear less affected. In the stem cell niche, they promote stem cell proliferation and inhibit differentiation. In Crohn’s disease, factors released from creeping fat could thus play a significant role in the pathophysiology, e.g. by enhancing epithelial regeneration via a promotion of stem cell proliferation.
P059 Diet controls segmented filamentous bacteria in driving Crohn’s disease-like inflammation in TNFdeltaARE mice
Authors: Metwaly A , Calasan J , Waldschmitt N , Khaloian S , Häcker D , Ahmed M , Butto L F , Hammoudi N , Le Bourhis L , Mayorgas A , Siebert K , Basic M , Zeissig S , Schwerd T , Allez M , Panes J , Salas A , Bleich A , Cominelli F , Haller D 
Published: January 2022

Abstract

Background
Crohn’s disease (CD)-like inflammation in TNFdeltaARE mice (ARE) mice is triggered by dysbiotic gut microbial communities. Similar to the therapeutic effect of exclusive enteral nutrition in CD, dietary intervention using semi-synthetic experimental diet prevents disease development in ARE mice. The aim of this study is to identify the causal microbial cues responsible for CD-like inflammation and to dissect the protective role of diet in ARE mice.

Methods
Bacterial communities in ARE and wildtype (WT) mice were profiled by 16S rRNA gene sequencing, FISH and qPCR. Germfree WT and ARE mice were colonized with segmented filamentous bacteria (SFB) from dysbiotic ARE mice. Disease activity, cytokine expression, mucosal immune cell infiltrates as well as Paneth and goblet cells phenotypes were assessed in murine ileal and colonic tissue. To assess the influence of diet on disease development, ARE mice were fed either chow, purified semi-synthetic experimental diet (SED) or fiber-rich diet. Impact of SFB on ileo-colonic inflammation was tested in additional mouse models (XIAP-/- and IL10 -/-). Mucosal biopsies from adult and pediatric IBD patients (N=407) were screened for the presence of human SFB using SFB-specific PCR primers.

Results
Under specific pathogen-free (SPF) conditions, ARE mice developed CD-like ileal inflammation which correlated strongly with increasing abundance of SFB. Mono-colonization of germfree ARE mice with SFB resulted in severe enterocolitis. Parallel to high tissue levels of TNF and IL-17, SPF-mediated inflammation was associated with neutrophil infiltration and the expansion of IFNγ expressing Th1 cells in the mucosa. Loss of Paneth and goblet cell function allowed SFB to penetrate mucus layers towards the epithelium. SED eradicated SFB and completely abolished inflammation in ARE mice. In contrast, fiber-rich diet partially restored SFB colonization and led to increased levels of inflammatory cytokines. SFB mono-colonization of GF XIAP-/- and IL10 -/- failed to induce inflammation, emphasizing host specificity to the mucosa-adherent SFB. Presence of SFB was confirmed in 12 ileal and 11 colonic mucosal biopsies from IBD patients with active or inactive disease.

Conclusion
We identified a novel pathogenic role of SFB in driving severe CD-like ileo-colonic inflammation characterized by loss of Paneth and goblet cell functions in ARE mice. Detection of SFB in mucosal biopsies of patients with IBD opens new perspectives about therapeutic strategies targeting SFB-mediated processes. Purified diet antagonized SFB colonization and prevented disease development in ARE mice in contrast to fiber-rich diet, clearly demonstrating the important role of diet in modulating a novel IBD-relevant pathobiont.