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Article ; Online: Measuring Myeloperoxidase Activity as a Marker of Inflammation in Gut Tissue Samples of Mice and Rat.

Hanning, Nikita / De Man, Joris G / De Winter, Benedicte Y

2023 Volume 13, Issue 13, Page(s) e4758

Abstract: Myeloperoxidase (MPO) is an enzyme contained in lysosomal azurophilic granules of neutrophils. MPO activity has been shown to correlate with the number of neutrophils in histological sections of the gastrointestinal tract and is therefore accepted as a ... ...

Abstract	Myeloperoxidase (MPO) is an enzyme contained in lysosomal azurophilic granules of neutrophils. MPO activity has been shown to correlate with the number of neutrophils in histological sections of the gastrointestinal tract and is therefore accepted as a biomarker of neutrophil invasion in the gut. This protocol describes an easy, cost-effective kinetic colorimetric assay to quantify myeloperoxidase activity in intestinal tissue samples. It is explained using tissue collected in mice but can also be used for other laboratory animals. In a first step, tissue specimens are homogenized using a phosphate buffer containing 0.5% hexadecyltrimethylammonium bromide (HTAB), which extracts MPO from neutrophils. The obtained supernatant is added to a reagent solution containing o-dianisidine dihydrochloride, which is a peroxidase substrate. Finally, the change in absorption is measured via spectrophotometry and converted to a standardized unit of enzyme activity. The assay is illustrated and compared to a commercially available enzyme-linked immunoassay (ELISA), demonstrating that MPO activity does not necessarily correlate with MPO protein expression in tissue samples. Key features Optimized for use in mice and rats but can also be used for samples of other species. Measures enzymatic activity instead of mRNA or protein expression. Requires a spectrophotometer. Can be performed in duplo using 10 mg of (dry-blotted) gut tissue or more. Graphical overview.
Language	English
Publishing date	2023-07-05
Publishing country	United States
Document type	Journal Article
ZDB-ID	2833269-6
ISSN	2331-8325 ; 2331-8325
ISSN (online)	2331-8325
ISSN	2331-8325
DOI	10.21769/BioProtoc.4758
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Article ; Online: Single-day and multi-day exposure to orogastric gavages does not affect intestinal barrier function in mice.

Hanning, Nikita / Verboven, Rosanne / De Man, Joris G / Ceuleers, Hannah / De Schepper, Heiko U / Smet, Annemieke / De Winter, Benedicte Y

American journal of physiology. Gastrointestinal and liver physiology

2023 Volume 324, Issue 4, Page(s) G281–G294

Abstract: Animals involved in common laboratory procedures experience minor levels of stress. The direct effect of limited amounts of stress on gastrointestinal function has not been reported yet. Therefore, this study aimed to assess the effect of single-day and ... ...

Abstract	Animals involved in common laboratory procedures experience minor levels of stress. The direct effect of limited amounts of stress on gastrointestinal function has not been reported yet. Therefore, this study aimed to assess the effect of single-day and multi-day orogastric gavages on gut physiology in mice. To this end, 12-wk-old female C57Bl6/J mice were randomized to receive treatment with sterile water (200 µL) delivered by orogastric gavages twice daily for a total of 1 or 10 day(s). Control animals did not receive any treatment. Subsequently, gastrointestinal function was assessed by measuring fecal pellet production. Furthermore, ex vivo intestinal barrier and secretory function of the distal colon, proximal colon, and terminal ileum were quantified in Ussing chambers. In mice, single-day gavages did neither influence corticosterone levels nor gastrointestinal function. In mice exposed to multi-day gavages, corticosterone levels were slightly but significantly increased compared with controls after 10 days of treatment. Gastrointestinal motor function was altered, as evidenced by increased fecal pellet counts and a small increase in fecal water content. However, exposure to repeated gavages did not lead to detectable alterations in gastrointestinal barrier function as quantified by the paracellular flux of the probe 4 kDa FITC-dextran as well as transepithelial resistance measurements. Thus, the administration of drugs via single-day or multi-day orogastric gavages leads to no or minor stress in mice, respectively. In both cases, it does not hamper the study of the intestinal barrier function and therefore remains a valuable administration route in preclinical pharmacological research.
MeSH term(s)	Animals ; Female ; Mice ; Colon ; Corticosterone/pharmacology ; Gastrointestinal Tract ; Ileum ; Intestinal Mucosa ; Permeability
Chemical Substances	Corticosterone (W980KJ009P)
Language	English
Publishing date	2023-02-07
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	603840-2
ISSN	1522-1547 ; 0193-1857
ISSN (online)	1522-1547
ISSN	0193-1857
DOI	10.1152/ajpgi.00203.2022
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Article ; Online: IL-22-Activated

Breugelmans, Tom / Arras, Wout / Oosterlinck, Baptiste / Jauregui-Amezaga, Aranzazu / Somers, Michaël / Cuypers, Bart / Laukens, Kris / De Man, Joris G / De Schepper, Heiko U / De Winter, Benedicte Y / Smet, Annemieke

Cells

2023 Volume 12, Issue 9

Abstract: Overexpression of the transmembrane mucin MUC13, as seen in inflammatory bowel diseases (IBD), could potentially impact barrier function. This study aimed to explore how inflammation-induced MUC13 disrupts epithelial barrier integrity by affecting ... ...

Abstract	Overexpression of the transmembrane mucin MUC13, as seen in inflammatory bowel diseases (IBD), could potentially impact barrier function. This study aimed to explore how inflammation-induced MUC13 disrupts epithelial barrier integrity by affecting junctional protein expression in IBD, thereby also considering the involvement of MUC1. RNA sequencing and permeability assays were performed using LS513 cells transfected with
MeSH term(s)	Animals ; Mice ; Colitis/chemically induced ; Colitis/metabolism ; Inflammation/metabolism ; Inflammatory Bowel Diseases/metabolism ; Intestinal Mucosa/metabolism ; Mucins/metabolism ; rho-Associated Kinases/metabolism ; Interleukin-22
Chemical Substances	Mucins ; rho-Associated Kinases (EC 2.7.11.1) ; Rock2 protein, mouse (EC 2.7.11.1) ; Ly64 protein, mouse
Language	English
Publishing date	2023-04-23
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2661518-6
ISSN	2073-4409 ; 2073-4409
ISSN (online)	2073-4409
ISSN	2073-4409
DOI	10.3390/cells12091224
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Article ; Online: Review, Performance Comparison, and Validation of Models Predicting Type 2 Diabetes Remission After Bariatric Surgery in a Western European Population.

Plaeke, Philip / Beunis, Anthony / Ruppert, Martin / De Man, Joris G / De Winter, Benedicte Y / Hubens, Guy

Obesity surgery

2021 Volume 31, Issue 4, Page(s) 1549–1560

Abstract: Purpose: The majority of patients with type 2 diabetes (T2DM) achieve remission after bariatric surgery. Several models are available to preoperatively predict T2DM remission. This study compares the performance of these models in a Western population ... ...

Abstract	Purpose: The majority of patients with type 2 diabetes (T2DM) achieve remission after bariatric surgery. Several models are available to preoperatively predict T2DM remission. This study compares the performance of these models in a Western population one year after surgery and explores their predictive value in comparison to a model specifically designed for our study population. Materials and methods: Prediction models were retrieved using a literature search. Patients were retrospectively selected from a database of the Antwerp University Hospital. Performance of the models was assessed by determining the area under the receiver operating characteristic curve (AUROC), the accuracy, and the goodness of fit, and by comparing them to a custom-made logistic model. Results: The probability of T2DM remission was calculated using 11 predictive scoring models and 8 regression models in a cohort of 250 patients. Complete T2DM remission occurred in 64.0% of patients. The IMS score (AUROC = 0.912; accuracy = 83.6%), DiaBetter score (0.907; 82.0%), and Ad-DiaRem score (0.903; 82.8%) best predicted T2DM remission and closely approached the performance of the custom-constructed model (0.917; 84.0%). The model by Ioffe et al. (0.630; 69.2%), Umemura et al. (0.692; 71.4%), and the ABCD score (0.757; 72.8%) were the least accurate. Conclusion: Most T2DM remission models reliably predicted one-year T2DM remission, with limited inter-model differences. The accuracy of most models approached that of the custom-constructed model, indicating a high predictive capability and performance in our patient cohort. To date, most models are only validated to estimate T2DM remission one year after surgery and they do not predict long-term remission.
MeSH term(s)	Bariatric Surgery ; Diabetes Mellitus, Type 2/surgery ; Humans ; Obesity, Morbid/surgery ; Remission Induction ; Retrospective Studies ; Treatment Outcome
Language	English
Publishing date	2021-01-04
Publishing country	United States
Document type	Journal Article
ZDB-ID	1070827-3
ISSN	1708-0428 ; 0960-8923
ISSN (online)	1708-0428
ISSN	0960-8923
DOI	10.1007/s11695-020-05157-0
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Article ; Online: Mucin-microbiome signatures shape the tumor microenvironment in gastric cancer.

Oosterlinck, Baptiste / Ceuleers, Hannah / Arras, Wout / De Man, Joris G / Geboes, Karen / De Schepper, Heiko / Peeters, Marc / Lebeer, Sarah / Skieceviciene, Jurgita / Hold, Georgina L / Kupcinskas, Juozas / Link, Alexander / De Winter, Benedicte Y / Smet, Annemieke

Microbiome

2023 Volume 11, Issue 1, Page(s) 86

Abstract: Background and aims: We aimed to identify mucin-microbiome signatures shaping the tumor microenvironment in gastric adenocarcinomas and clinical outcomes.: Methods: We performed high-throughput profiling of the mucin phenotypes present in 108 gastric ...

Abstract	Background and aims: We aimed to identify mucin-microbiome signatures shaping the tumor microenvironment in gastric adenocarcinomas and clinical outcomes. Methods: We performed high-throughput profiling of the mucin phenotypes present in 108 gastric adenocarcinomas and 20 functional dyspepsia cases using validated mucin-based RT-qPCRs with subsequent immunohistochemistry validation and correlated the data with clinical outcome parameters. The gastric microbiota was assessed by 16S rRNA gene sequencing, taxonomy, and community composition determined, microbial networks analyzed, and the metagenome inferred in association with mucin phenotypes and expression. Results: Gastric adenocarcinomas with an intestinal mucin environment or high-level MUC13 expression are associated with poor survival. On the contrary, gastric MUC5AC or MUC6 abundance was associated with a more favorable outcome. The oral taxa Neisseria, Prevotella, and Veillonella had centralities in tumors with intestinal and mixed phenotypes and were associated with MUC13 overexpression, highlighting their role as potential drivers in MUC13 signaling in GC. Furthermore, dense bacterial networks were observed in intestinal and mixed mucin phenotype tumors whereas the lowest community complexity was shown in null mucin phenotype tumors due to higher Helicobacter abundance resulting in a more decreased diversity. Enrichment of oral or intestinal microbes was mucin phenotype dependent. More specifically, intestinal mucin phenotype tumors favored the establishment of pro-inflammatory oral taxa forming strong co-occurrence networks. Conclusions: Our results emphasize key roles for mucins in gastric cancer prognosis and shaping microbial networks in the tumor microenvironment. Specifically, the enriched oral taxa associated with aberrant MUC13 expression can be potential biomarkers in predicting disease outcomes. Video Abstract.
MeSH term(s)	Humans ; Stomach Neoplasms/genetics ; Stomach Neoplasms/metabolism ; Stomach Neoplasms/pathology ; Mucin-2/genetics ; Tumor Microenvironment ; RNA, Ribosomal, 16S/genetics ; Mucin-6/genetics ; Adenocarcinoma ; Microbiota ; Phenotype
Chemical Substances	Mucin-2 ; RNA, Ribosomal, 16S ; Mucin-6
Language	English
Publishing date	2023-04-21
Publishing country	England
Document type	Video-Audio Media ; Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2697425-3
ISSN	2049-2618 ; 2049-2618
ISSN (online)	2049-2618
ISSN	2049-2618
DOI	10.1186/s40168-023-01534-w
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Article: Volatile organic compound profiling as a potential biomarker in irritable bowel syndrome: A feasibility study.

Van Malderen, Kathleen / Hanning, Nikita / Lambrechts, Helen / Haverhals, Tine / Van Marcke, Silke / Ceuleers, Hannah / De Man, Joris G / De Winter, Benedicte Y / Lamote, Kevin / De Schepper, Heiko U

Frontiers in medicine

2022 Volume 9, Page(s) 960000

Abstract: Background: Irritable bowel syndrome (IBS) is a chronic gastrointestinal disorder for which no diagnostic tools are currently available. Patients are diagnosed using the Rome IV criteria and subtyped into a diarrhea, constipation, or mixed phenotype ... ...

Abstract	Background: Irritable bowel syndrome (IBS) is a chronic gastrointestinal disorder for which no diagnostic tools are currently available. Patients are diagnosed using the Rome IV criteria and subtyped into a diarrhea, constipation, or mixed phenotype based on their dominant stool pattern. A recent development in the biomarker area is the analysis of volatile organic compounds (VOCs). The aim of this study was to evaluate the potential of VOCs as diagnostic and phenotypic biomarkers for IBS in breath and fecal samples. Materials and methods: Breath and fecal samples from IBS patients and healthy asymptomatic controls (HC) were analyzed with multicapillary column/ion mobility spectrometry (MCC/IMS) and classification models were created based upon VOCs and clinical characteristics. Discussion: Irritable bowel syndrome patients were differentiated from HC by means of volatile profiling in both breath and fecal samples with area under the curve (AUCs) of respectively 0.62 and 0.80. Patient subtypes could also be differentiated from each other with AUCs ranging between 0.65 and 0.78. Furthermore, VOC models could differentiate IBS patients based on clinical characteristics like psychological comorbidities and microbiota-influencing therapies. Conclusion: This study is the first to demonstrate the use of VOC profiling with the help of MCC/IMS to differentiate IBS patients. Furthermore, the importance of clinical characteristics beside the dominant stool pattern in the differentiation of IBS patients was emphasized.
Language	English
Publishing date	2022-08-04
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2775999-4
ISSN	2296-858X
ISSN	2296-858X
DOI	10.3389/fmed.2022.960000
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Article ; Online: Volatomics in inflammatory bowel disease and irritable bowel syndrome.

Van Malderen, Kathleen / De Winter, Benedicte Y / De Man, Joris G / De Schepper, Heiko U / Lamote, Kevin

EBioMedicine

2020 Volume 54, Page(s) 102725

Abstract: Volatile organic compounds (VOCs) are produced by the human metabolism, inflammation and gut microbiota and form the basis of innovative volatomics research. VOCs detected through breath and faecal analysis hence serve as attractive, non-invasive ... ...

Abstract	Volatile organic compounds (VOCs) are produced by the human metabolism, inflammation and gut microbiota and form the basis of innovative volatomics research. VOCs detected through breath and faecal analysis hence serve as attractive, non-invasive biomarkers for diagnosing and monitoring irritable bowel syndrome (IBS) and inflammatory bowel disease (IBD). This review describes the clinical applicability of volatomics in discriminating between IBS, IBD and healthy volunteers with acceptable accuracy in breath (70%-100%) and faecal (58%-85%) samples. Promising compounds are propan-1-ol for diagnosing and monitoring of IBD patients, and 1-methyl-4-propan-2-ylcyclohexa-1,4-diene as biomarker for IBS diagnosis. However, these VOCs often seem to be related to inflammation and probably will need to be used in conjunction with other clinical evidence. Furthermore, three interventional studies underlined the potential of VOCs in predicting treatment outcome and patient follow-up. This shows great promise for future use of VOCs as non-invasive breath and faecal biomarkers in personalised medicine. However, properly designed studies that correlate VOCs to IBD/IBS pathogenesis, while taking microbial influences into account, are still key before clinical implementation can be expected.
MeSH term(s)	Biomarkers/analysis ; Biomarkers/metabolism ; Breath Tests/methods ; Feces/chemistry ; Humans ; Irritable Bowel Syndrome/diagnosis ; Irritable Bowel Syndrome/metabolism ; Metabolomics/methods ; Volatile Organic Compounds/analysis ; Volatile Organic Compounds/metabolism
Chemical Substances	Biomarkers ; Volatile Organic Compounds
Language	English
Publishing date	2020-04-21
Publishing country	Netherlands
Document type	Journal Article ; Review
ZDB-ID	2851331-9
ISSN	2352-3964
ISSN (online)	2352-3964
DOI	10.1016/j.ebiom.2020.102725
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Article: The Effect of Serine Protease Inhibitors on Visceral Pain in Different Rodent Models With an Intestinal Insult.

Ceuleers, Hannah / Hanning, Nikita / De Bruyn, Michelle / De Man, Joris G / De Schepper, Heiko U / Li, Qian / Liu, Liansheng / Abrams, Steven / Smet, Annemieke / Joossens, Jurgen / Augustyns, Koen / De Meester, Ingrid / Pasricha, Pankaj J / De Winter, Benedicte Y

Frontiers in pharmacology

2022 Volume 13, Page(s) 765744

Abstract: Background: ...

Abstract	Background:
Language	English
Publishing date	2022-06-02
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2587355-6
ISSN	1663-9812
ISSN	1663-9812
DOI	10.3389/fphar.2022.765744
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Article: Intestinal barrier dysfunction in irritable bowel syndrome: a systematic review.

Hanning, Nikita / Edwinson, Adam L / Ceuleers, Hannah / Peters, Stephanie A / De Man, Joris G / Hassett, Leslie C / De Winter, Benedicte Y / Grover, Madhusudan

Therapeutic advances in gastroenterology

2021 Volume 14, Page(s) 1756284821993586

Abstract: Background and aim: Irritable bowel syndrome (IBS) is a complex and heterogeneous disorder. Sensory, motor and barrier dysfunctions are the key physiological endophenotypes of IBS. Our aim is to review studies evaluating barrier dysfunction in adults ... ...

Abstract	Background and aim: Irritable bowel syndrome (IBS) is a complex and heterogeneous disorder. Sensory, motor and barrier dysfunctions are the key physiological endophenotypes of IBS. Our aim is to review studies evaluating barrier dysfunction in adults and children with IBS, as well as to link those changes with IBS symptomatology and quality of life. Methods: A comprehensive and systematic review of multiple databases was performed up to March 2020 to identify studies comparing intestinal permeability in IBS patients with healthy controls. Both Results: We identified 66 studies, of which 27 used intestinal probes to quantify barrier function. The prevalence of barrier dysfunction differed between PI-IBS (17-50%), IBS-D (37-62%) and IBS-C (4-25%). At a group level, permeability was increased compared with healthy controls in IBS-D (9/13 studies) and PI-IBS (4/4 studies), but only a minority of IBS-C (2/7 studies) and not in the only IBS-M study. All four studies in children with IBS demonstrated loss of barrier function. A heterogeneous set of tight junction genes were found to be altered in small and large intestines of adults with IBS, but these have not been evaluated in children. Positive associations were identified between barrier dysfunction and bowel disturbances (6/9 studies), abdominal pain (9/13 studies), overall symptom severity (1/6 studies), depression and anxiety (1/1 study) and quality of life (1/4 studies). Fecal slurry or supernatants of IBS patients were found to induce barrier disruption in animal models (5/6 studies). Conclusions: Barrier dysfunction is present in a significant proportion of adult and all pediatric IBS studies, especially in the IBS-D and PI-IBS subtype. The majority of studies indicated a positive association between loss of barrier function and symptoms such as abdominal pain and changes in the bowel function.
Language	English
Publishing date	2021-02-24
Publishing country	England
Document type	Journal Article ; Review
ZDB-ID	2440710-0
ISSN	1756-2848 ; 1756-283X
ISSN (online)	1756-2848
ISSN	1756-283X
DOI	10.1177/1756284821993586
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Article: The Effect of a Novel Serine Protease Inhibitor on Inflammation and Intestinal Permeability in a Murine Colitis Transfer Model.

Van Spaendonk, Hanne / Ceuleers, Hannah / Smet, Annemieke / Berg, Maya / Joossens, Jurgen / Van der Veken, Pieter / Francque, Sven M / Lambeir, Anne-Marie / De Man, Joris G / De Meester, Ingrid / Augustyns, Koen / De Winter, Benedicte Y

Frontiers in pharmacology

2021 Volume 12, Page(s) 682065

Abstract: Background: ...

Abstract	Background:
Language	English
Publishing date	2021-06-24
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2587355-6
ISSN	1663-9812
ISSN	1663-9812
DOI	10.3389/fphar.2021.682065
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