Introduction

Metabolic Dysfunction–Associated Steatotic Liver Disease (MASLD) is currently the leading cause of chronic liver disease worldwide, affecting more than 65% of patients with Type 2 Diabetes Mellitus (T2DM). The aim of our study was to determine the prevalence of MASLD among T2DM patients, with or without coexisting significant fibrosis, to assess its association with diabetic complications, and to identify the factors linked to MASLD in our population.

Aims & Methods

We conducted a retrospective, descriptive, and analytical study including T2DM patients admitted to the Endocrinology Department in 2019. MASLD diagnosis was based on the presence of hepatic steatosis detected by abdominal ultrasound or a Hepatic Steatosis Index (HSI) >36. Fibrosis was assessed using the Fibrosis-4 (FIB-4) index (with a threshold of ≥1.3 or <1.3). Chronic Kidney Disease (CKD) associated with T2DM was defined by an estimated glomerular filtration rate (eGFR) <60 ml/min/1.73 m² and/or dipstick-positive proteinuria (>1+). Diabetic Peripheral Neuropathic Pain (DPNP) was assessed using the Douleur Neuropathique 4 (DN4) questionnaire, and Diabetic Retinopathy was diagnosed via dilated eye examination.

Results

Among the 202 patients included, the median age was 57 years (±12 years) with a marked female predominance (sex ratio = 2.6). Regarding comorbidities, 58.4% of patients had hypertension and 34.2% had dyslipidemia. The median Body Mass Index (BMI) was 27 kg/m² (±5.62), with overweight or obesity present in 61.4% of the cohort. The median duration of diabetes was 10 years (±7.8 years). Impaired renal function (eGFR <60 ml/min/1.73 m²) was found in 9.9% of patients, and proteinuria in 6.9%. Diabetic neuropathy (DN4 score ≥4) was present in 7.9% of patients, while diabetic retinopathy was diagnosed in 37.6% of cases.
The prevalence of MASLD in our cohort was 73.3%. The median FIB-4 score was 0.91 (±0.63), with 25.7% of patients presenting a FIB-4 >1.3. Statistical analysis identified several factors significantly associated with the presence of MASLD: female sex (p=0.004), dyslipidemia (p=0.004), BMI (p<0.001), alanine aminotransferase (ALT) levels (p<0.001), low-density lipoprotein cholesterol (LDL-C) levels (p=0.006), and triglyceride levels (p=0.013). In particular, higher BMI was strongly associated with MASLD (Odds Ratio [OR] = 1.970; 95% Confidence Interval [CI]: 1.580–2.460; p<0.001), as were elevated ALT levels (OR = 1.115; 95% CI: 1.053–1.18; p<0.001).
However, after adjustment, LDL-C levels (p=0.163), triglycerides (p=0.402), dyslipidemia (p=0.329), and sex (p=0.360) were no longer significantly associated with MASLD. Finally, a significantly higher proportion of CKD was observed in patients with MASLD (22.3%) compared to those without MASLD (9.2%, p=0.036), although this association was not maintained in the subgroup with advanced fibrosis (FIB-4 >1.3).

Conclusion

Our study confirmed the high prevalence of MASLD among patients with T2DM. Early screening is therefore essential, particularly in female patients and in those presenting with chronic kidney disease or other associated cardiometabolic risk factors such as obesity and dyslipidemia.

References

Younossi ZM, Golabi P, Price JK, Owrangi S, Gundu-Rao N, Satchi R, Paik JM. The Global Epidemiology of Nonalcoholic Fatty Liver Disease and Nonalcoholic Steatohepatitis Among Patients With Type 2 Diabetes. Clin Gastroenterol Hepatol. 2024 Oct;22(10):1999-2010.e8. doi: 10.1016/j.cgh.2024.03.006. Epub 2024 Mar 21. PMID: 38521116.

Introduction

Pain in chronic pancreatitis (CP) varies between patients in pattern, severity, and psychological impact. CP pain assessment has been focused on intensity, missing the complexity. To address the importance of multidimensional pain evaluations, we aimed to identify pain phenotypes based on multidimensional pain assessments and examine their association with clinical factors.

Aims & Methods

We studied 248 patients with painful CP from Asia, Europe, and the USA. A cluster analysis was performed on patients with painful CP, including five pain dimensions from the COMPAT-SF questionnaire (severity, fluctuation, provocative factors, spreading pain, and qualitative descriptors). The resulting clusters were compared based on demographics and clinical data, including patient-reported outcomes and quantitative sensory testing.

Results

		Cluster 1	Cluster 2	Cluster 3	p-value​
Questionnaires	Quality of life	66.3 (22.5)	64.8 (27.7)	51.1 (24.6)	<0.001
	Catastrophizing	10.7 (11.2)	19.1 (12.4)	24.3 (13.0)	<0.001
	Anxiety	5.3 (4.1)	6.2 (4.2)	8.7 (4.5)	<0.001
	Depression	4.3 (4.3)	5.5 (4.6)	7.3 (4.7)	<0.001
	Sleep	5.0 (3.5)	5.9 (3.8)	7.5 (4.5)	<0.001
QST	PPDT sum, kPa	2284 (997)	1811.5 (574.6)	1640.1 (600.4)	<0.001
	PPTT sum, kPa	2909.6 (1247.3)	2227.1 (691.3)	2184.9 (903.2)	<0.001
	Temporal summation, abdomen	1.7 (1.8)	1.9 (1.5)	1.4 (1.8)	0.178
	CPM (%)	2.3 (18.3)	5.1 (23.7)	7.2 (17.7)	0.135

Three distinct pain phenotypes were identified in the cluster analysis. Cluster 1 (n=73) included patients with low-severity pain; Cluster 2 (n=80) was characterized by severe, fluctuating pain; and Cluster 3 (n=95) comprised patients reporting constant, severe pain. Patients in Clusters 1 and 2 reported low scores across the remaining dimensions, whereas Cluster 3 consistently scored higher. Quality of life and sleep were best in Cluster 1, declined in Cluster 2, and were lowest in Cluster 3 (both p<0.001). Scores for anxiety, depression, and catastrophizing increased progressively across clusters (all p<0.001). Sensory testing revealed a pattern of increasing hyperalgesia, with Cluster 3 showing the most pronounced responses (p<0.001).

Conclusion

The COMPAT-SF questionnaire identifies three pain phenotypes in CP. Greater pain severity and persistence correlate with increased hyperalgesia, higher psychological distress, and worse overall well-being.

Introduction

Intestinal barrier integrity is emerging as a key therapeutic target in inflammatory bowel disease (IBD). Advanced endoscopic imaging techniques, including ultra-high magnification endocytoscopy (ECS) and probe-based confocal laser endomicroscopy (pCLE), enable real-time, in vivo structural and functional intestinal barrier assessment. Raman Spectroscopy (RS) can add a molecular dimension through vibrational fingerprinting of barrier-associated metabolites in biological samples.¹

Aims & Methods

This study explores the potential of RS integration with advanced endoscopy to characterise the intestinal barriers in IBD, targeting on tryptophan-derived metabolites and carbohydrates. A total of 46 IBD patients [25 Crohn’s disease (CD) and 21 ulcerative colitis (UC)] and 7 healthy controls (HC) undergoing endoscopic assessment or surveillance were prospectively enrolled from two academic centres. ECS or pCLE were used to evaluate epithelial and vascular barrier integrity. Barrier healing was defined as ECS score ≤ 1 or pCLE score ≤ 4, with preserved villi/crypt architecture, normal vasculature, and also absence of fluorescein leakage for pCLE. Advanced endoscopy-guided biopsies were collected from representative bowel segments. Mucosal healing (MH) was assessed by experienced pathologists using Nancy’s Histological Index ≤1. Epithelial (Claudin-2 and ZO-1) and vascular (PV-1 and CD-31) barrier markers were analysed through confocal microscopy, and their expression was automatically quantified using QuPath. Micro-RS was applied to tissue samples and matched sera to measure key tryptophan metabolites (kynurenine, indolepropionic acid, and melatonin) and carbohydrates.^2,3 Six-month major adverse outcomes (MAOs) were recorded. Group differences in Raman spectra were analysed with the Mann-Whitney test; correlations were assessed via Pearson’s test.

Results

Among patients undergoing ECS, barrier healing was observed in 10/22 (45%) colonic and 6/9 (67%) ileal cases. pCLE-based assessment revealed healing features in 5/16 (31%) colonic and 2/15 (13%) ileal evaluations. Raman spectral analysis showed a significant (p<0.05) reduction in tryptophan metabolites and carbohydrate-associated peaks in IBD tissues and sera compared to HC, especially in inflamed patients. When stratified by advanced endoscopy-identified barrier integrity, tryptophan metabolites -known barrier modulators-, and carbohydrates -likely reflecting mucin barrier layer- were markedly reduced (p<0.05) in patients with impaired barrier function. Notably, tryptophan derivatives positively correlated with Claudin-2 and ZO-1 in UC (r>0.4), and with PV-1 in CD (r>0.5). Similarly, carbohydrates correlated positively with Claudin-2 in UC (r=0.4) and negatively with CD-31 in CD (r=-0.4). Lower levels of indolepropionic acid and carbohydrates correlated with worse short-term MAOs (r<-0.4).

Conclusion

The integration of RS with advanced endoscopic images offers a novel, multi-layered approach to assess intestinal barrier integrity in IBD. RS can identify novel non-invasive biomarkers of mucosal and barrier healing, with potential real time applications for monitoring disease activity and predicting therapeutic outcome. Tryptophan metabolites may serve as a predictive biomarker of barrier healing paving the way toward personalised, barrier-targeted therapies.

References

1 Smith SCL, Banbury C, Zardo D, et al. PLoS One 2021; 16: e0252210.
2 Michaudel C, Danne C, Agus A, et al. Gut 2023; 72: 1296–307.
3 Arras W, Breugelmans T, Oosterlinck B, et al. J Crohns Colitis 2025; 19.

Introduction

Microscopic colitis (MC) is an inflammatory bowel disease (IBD) that causes chronic non-bloody watery diarrhea and abdominal pain that significantly reduces the quality of life¹. MC can be subdivided into collagenous colitis (CC) and lymphocytic colitis (LC), which are clinically indistinguishable but differ histologically². The activation of immune responses and the alteration of the intestinal epithelial barrier via water channel aquaporin downregulation and crypt hyperproliferation are characteristic features of MC³, often shared by other gastrointestinal disorders, such as “classical” IBD (Crohn’s disease and ulcerative colitis, UC), or irritable bowel syndrome (IBS). Enteric glial cells (EGC) are key regulators of gut homeostasis and epithelial permeability, that respond to inflammation by entering a reactive state, potentially driving epithelial dysfunction. Whether EGC promote resolution or perpetuate inflammation remains unclear.⁴

Aims & Methods

MC is poorly understood and EGC role in MC is still unknown. Therefore, our aim is to investigate the involvement of EGC – due to their neuroimmune functions and involvement in intestinal dysmotility – in patients with MC. In addition, mast cells will be investigated since altered interactions with EGC have been demonstrated during IBS⁵.
Paraffin-embedded colonic biopsies from patients with active CC, LC, UC, IBS-diarrhea (D) and healthy controls (HC) were collected (n=13-26 per cohort). In addition, a subset of CC patients was also sampled during successful treatment with glucocorticosteroid 9mg budesonide (n=13). The amount of EGC was quantified using glial fibrillary acidic protein (GFAP) and S100 calcium-binding protein B (S100β). Mast cells were quantified using tryptase.

Results

Immunofluorescence analysis of EGC revealed an increased amount of EGC^GFAP+ in CC compared to HC (p<0.001). Of note, CC EGC^GFAP+ levels were comparable to those shown by biopsies from patients suffering from IBS-D and UC. Among the 13 CC patients with paired samples before and under budesonide treatment, we observed a trend toward reduced EGC^GFAP+ levels during treatment (p = 0.08). In contrast, LC patients displayed similar EGC^GFAP+ levels as to HC, but lower than those observed in biopsies from patients with CC, IBS-D, and UC (p<0.001). On the other hand, EGC^S100β+ were increased only in active CC (p<0.0001). Yet, UC patients displayed the highest level of EGC^S100β+ (p<0.0001). Mast cells analysis showed an increased counts for tryptase in CC before and under treatment compared to HC (p<0.01).

Conclusion

Our findings indicate that EGC and mast cells are involved in CC pathogenesis due to an increased reactivity of EGC and mast cells in the colonic mucosa. CC patients exhibit signs of EGC^GFAP+ levels comparable to those observed in UC and IBS-D. Oppositely, LC seems to be independent of neuroinflammation as EGC and mast cells presence was comparable to HC. These preliminary findings support the theory that CC and LC are distinct disease entities. Future work will include RNA sequencing of EGC-related genes, counts of eosinophils as well as in situ analysis of mast cells and eosinophils degranulation, and assessment of budesonide effects on EGC in vitro.

References

1. Nyhlin N, Wickbom A, Montgomery SM, et al. Long-term prognosis of clinical symptoms and health-related quality of life in microscopic colitis: a case-control study. Aliment Pharmacol Ther 2014; 39: 963-972. 20140311. DOI: 10.1111/apt.12685.
2. Carrasco A, Esteve M, Salas A, et al. Immunological Differences between Lymphocytic and Collagenous Colitis. J Crohns Colitis 2016; 10: 1055-1066. 20160229. DOI: 10.1093/ecco-jcc/jjw058.
3. Escudero-Hernandez C, Munch A, Ostvik AE, et al. The Water Channel Aquaporin 8 is a Critical Regulator of Intestinal Fluid Homeostasis in Collagenous Colitis. J Crohns Colitis 2020; 14: 962-973. DOI: 10.1093/ecco-jcc/jjaa020.
4. Gonzales J and Gulbransen BD. The Physiology of Enteric Glia. Annu Rev Physiol 2024 20241115. DOI: 10.1146/annurev-physiol-022724-105016.
5. Meira de-Faria F, Casado-Bedmar M, Marten Lindqvist C, et al. Altered interaction between enteric glial cells and mast cells in the colon of women with irritable bowel syndrome. Neurogastroenterol Motil 2021; 33: e14130. 20210402. DOI: 10.1111/nmo.14130.

Introduction

Dysregulated activation of myeloid cells, including monocytes and macrophages, contributes to intestinal inflammation in Inflammatory Bowel Disease (IBD) through excessive release of pro-inflammatory cytokines. Microbial-derived short-chain fatty acids (SCFAs) modulate the inflammatory and metabolic nature of these cells through activation of a wide-range of cellular signalling pathways. Several of these pathways—including targeting Free-fatty acid receptors (FFA1, FFA2, FFA3) and other metabolic pathways (Glycolysis and Oxidative phosphorylation) have exhibited potential by influencing the polarisation from inflammatory to regulatory macrophages. In this study, we aim to unravel these mechanisms and explore the interplay between myeloid cells and SCFAs in IBD.

Aims & Methods

We first investigated the immunomodulatory effects of SCFA on THP-1 and monocyte-derived macrophage (MDM) models of inflammation. These cells were polarised toward an M1-like phenotype (LPS+ IFN-γ) in the presence of SCFAs or pharmacological agonist of FFA3 (AR420626). We analysed how these treatments impacted cytokine secretion and markers of polarisation using ELISA and RT-qPCR. To gain insight with a disease context, CD14+ monocytes from the peripheral blood of IBD patients (n=6), were exposed to M1 polarising stimuli (LPS+IFN-γ), in the presence of SCFAs or AR420626. To explore potential disease-specific responses, monocytes from actively inflamed IBD patients (n=7) and healthy controls (HC) (n=3) were cultured in presence of butyrate (1mM) in the absence of inflammatory stimulus. In both instances, RT-qPCR and ELISA were conducted to assess secretion of pro-inflammatory cytokines and activation of metabolic pathways.

Results

In THP-1 and MDM models, treatment with butyrate (4 mM) markedly suppressed IL-6 secretion, an effect that was mimicked by the FFA3 agonist AR420626 (50 µM). These responses extended to IBD patient-derived monocytes, where strong induction of IL-6 and TNF-α were repressed by both butyrate and propionate (vs M1; p< 0.001). FFA3, which plays an important role in mediating responses to SCFAs, was also upregulated in M1 polarised patient monocytes (vs Unt; p< 0.05). However, this elevated expression was abrogated in monocytes that were exposed to butyrate (1mM), giving insight toward how butyrate can regulate SCFA sensing mechanisms under inflammatory conditions. Interestingly, this pro-inflammatory monocyte phenotype was accompanied by raised expression of the glycolysis markers HK2 and GLUT1 —which, again, were repressed by butyrate (vs Unt; p< 0.05). In untreated monocytes derived from IBD patients, we observed an elevated inflammatory state relative to HCs, suggesting prior disease-associated activation. While these cells displayed heightened baseline expression of HK2 than HC monocytes, this was attenuated by exposure to butyrate (vs Unt; p< 0.05), and cells were rescued from this hyper-inflammatory phenotype.

Conclusion

Taken together, these results highlight that butyrate helps to counteract inflammation in IBD monocytes —possibly stemming from an influence on glycolysis. This work demonstrates that monocyte metabolic activity and SCFA-sensing pathways are dysregulated under inflammatory conditions. These findings provide new insight into the altered immune-metabolic state of monocytes in IBD and highlight the important role of microbial-derived SCFAs to restoring immunometabolic balance.

Introduction

There is evidence linking inflammatory bowel disease (IBD) and cardiovascular diseases. However, the roles of histologic and clinical activity of IBD in the development of arrhythmias remain unknown.

Aims & Methods

In this nationwide cohort study in Sweden, we included IBD patients with histologic data (n = 61,383; diagnosed 1969–2017; classified as histologic inflammation vs. remission based on the Swedish version of the Systematized Nomenclature of Medicine codes) and clinical activity data (n = 96,154; diagnosed 1969–2020; classified as clinically active vs. quiescent IBD based on IBD-related hospitalization, surgery, and treatment escalation) and followed them through December 31, 2021. The primary outcome was incident arrhythmias (including atrial fibrillation/flutter, bradyarrhythmias, other supraventricular arrhythmias, and ventricular arrhythmias/cardiac arrest) within 2 years after documented histologic inflammation (vs. remission) or clinically active IBD (vs. quiescent IBD). Poisson regression models estimated incidence rates (IRs) of arrhythmias, while Cox proportional hazards models estimated adjusted hazard ratios (aHRs) and 95% confidence intervals (CIs). We adjusted for age at IBD diagnosis, sex, calendar year at index date, county of residence, educational attainment, country of birth, number of health care visits, and history of CVD-related comorbidities in the model.

Results

Histologic inflammation was associated with an increased risk of developing any arrhythmias compared to histologic remission (IR: 62.0 vs. 47.2 per 10,000 person-years; aHR = 1.35, 95%CI: 1.18–1.54), corresponding to one additional event of arrhythmia per 338 IBD patients over 2 years after histologic inflammation. We found an excess risk for atrial fibrillation/flutter (aHR = 1.38, 95%CI: 1.18–1.62) and ventricular arrhythmias/cardiac arrest (aHR = 1.73, 95%CI: 1.22–2.44), but not for bradyarrhythmias or other supraventricular arrhythmias. Compared to clinically quiescent IBD, active IBD was also related to a higher risk of arrhythmias (IR: 67.4 vs. 108.4; aHR = 1.77, 95%CI: 1.66–1.88), affecting all subtypes except bradyarrhythmias. In patients with clinically quiescent IBD, histologic inflammation was also associated with an increased risk of arrhythmias (aHR = 1.16, 95%CI: 1.00–1.36, P=0.054).

Conclusion

Histologic and clinical activity were associated with a slightly increased risk of arrhythmias in patients with IBD.

Disclosure

All authors have completed the ICMJE uniform disclosure form and declare:
OO has been PI on projects at Karolinska Institutet financed by grants from Janssen, Pfizer, AbbVie, Takeda, Galapagos/Alfasigma, Ferring, and Bristol Myer Squibb. JH served as speaker or advisory board member for AbbVie, BMS, Celltrion, Eli Lilly, Ferring, Galapagos, Gilead, Index Pharma, Janssen, Medtronic, Merck, MSD, Novartis, Pfizer, Sandoz, Shire, Takeda, Thermo Fisher Scientific, Tillotts Pharma, and Vifor Pharma and received grant support from Takeda, Janssen, and MSD. FE has served as an advisory board member for Boehringer Ingelheim. JFL has coordinated a study for the Swedish IBD Quality Register (SWIBREG), which received funding from Janssen Corporation. JFL has also received financial support from MSD to develop a paper reviewing national healthcare registers in China. JFL has a research collaboration on celiac disease with Takeda and has a research collaboration on fibrosis in IBD with MSD. The other authors report no disclosures relevant to the manuscript.