Introduction

Fecal microbiota transplantation (FMT) is an effective therapy for recurrent Clostridioides difficile infection. A pool of healthy stool donors is essential for any FMT program. However, donor
recruitment is not easy due to screening failures and the commitment requested.

Aims & Methods

We aimed to report data from the first two years of stool donor screening, recruitment, and stool banking from the first public Italian FMT center. We prospectively collected data from donor screening questionnaires, laboratory evaluations, and donations. Donor screening has been performed according to the Italian Health Ministry regulation and the FMT European consensus FMT (1): potential donors, all aged below 50 years old, were initially screened for the presence of concomitant gastrointestinal and systemic diseases or for the anamnestic drugs intake or recent antibiotic intake. The subsequent clinical evaluation aimed at excluding at-risk conditions for infectious diseases, while the microbiological screening evaluated the colonization by potential pathogens. Bacterial counts in terms of aerobes and anaerobes concentration [colony forming unit (CFU)/mL] from aliquots of each frozen fecal material after aerobic samples preparation were assessed after 1 week and subsequently monthly until 18 months from the donation.

Results

Two hundred and 12 patients completed screening questionnaires. Of these, 29 (13.7%) were excluded for concurrent gastrointestinal symptoms (altered bowel habits) or diseases, 50 (23.6%) for concomitant systemic diseases, and 8 (3.8%) for antibiotic intake in the previous 6 months. Finally, of the 120 donors clinically evaluated, 79 (37.3%) were excluded for at-risk conditions for infectious disease (e.g., use of illegal drugs, risky sexual behavior, recent body tattoo, piercing, travel in tropical countries, healthcare workers, etc.). Among the remaining 41 donors microbiologically screened, 19 (9%) were excluded for pathogens or potential pathogens presence (6 pathogenic Escherichia coli, 1 Vancomycin-resistant Enterococci, 2 Helicobacter pylori, 1 Rotavirus, 1 Giardia, and 8 for other reasons). Finally, 22 subjects (10.4%) were suitable for donation. Among the 10 healthy donors actively donating stools (50% females), 6 were excluded for pathogens infections diagnosed during donation screenings after a median of 3 donations (3 pathogenic Escherichia coli, 1 Salmonella enterica, 1 Campylobacter jejuni, 1 Cryptosporidium). Among the remaining 4 subjects, 2 discontinued the donation program due to the commitment requested. Finally, 22 subjects (10.4%) were suitable for donation. Ten healthy donors actively donated stools (50% females), but 6 were excluded for pathogens/potentially pathogenic bacteria diagnosed during donation screenings after a median of 3 donations. Among the remaining 4 subjects, 2 discontinued the donation program due to the commitment requested.
Aliquots evaluation showed no differences in aerobic counts until 18 months, while significant differences were found for anaerobic counts already after 1 month (1.33x10^10 CFU/mL vs 7.6x10^9 CFU/mL at donation and after 1 month, respectively, p=0.028).

Conclusion

Only one in ten potential healthy donors is finally eligible for stool donation. Frozen stool samples showed a significant decrease in anaerobic concentration already after 1 month from storage, compared to fresh material.

References

1. Cammarota G., et al. Gut. 2017 Apr;66(4):569-580.

Disclosure

Lecture grants from EG Pharma, AlfaSigma, Bromatech, Echosens, Ferring, Mayoly Spindler and Schwabe Pharma.

Introduction

Radiation therapy is widely used in the treatment of carcinoma in the abdomen and pelvic cavity, but as a side effect, acute and chronic damage to the gastrointestinal tract may occur. It is well known that the rectum is more prone to radiation damage since it is fixed in the pelvis. Acute symptoms of radiation enteritis are mainly associated with inflammation of the intestinal mucosa and loss of normal mucosa, but changes in gastrointestinal motility due to exposure to high-concentration radiation may also be a cause. Previous studies are demonstrating that patients who received radiation therapy had increased bowel movements, symptoms of urgency, and decreased rectal compliance. But there are only limited studies investigated the pattern of changes in the motility of human rectal smooth muscle following irradiation.

Aims & Methods

This study was conducted to investigate changes in motility and their mechanisms after irradiation in human rectum.
Human rectal tissues were obtained immediately in the operating room from patients undergoing colorectal resection. After removing the mucosal layer of rectum, muscle layer was cut in strips of 5-6 mm in length and 2-3 mm in width. The muscle strips were mounted to an isometric force transducer (Biopac Systems, Inc., Goleta, CA, USA) and suspended in 10 ml organ bath containing aerated (97% O₂ and 3% CO₂) and warmed (36.5 ± 0.5°C) KRB solution. The area under the curve (AUC) for 5 min and tone of spontaneous contractions were analyzed to compared the responses to KCl, ACh (acethylcholine), SNP (sodium nitroprusside), a nitric oxide donor, and ATP (adenosine triphosphate), a purinergic receptor agonist. The mechanical responses were recorded and digitized using Acknowledge software (Biopac Systems, Inc., Goleta, CA, USA). Data were analyzed offline using Clampfit (version 10.2. Molecular devices, San Jose, CA, USA). qPCR was performed for expression of VChAT, NOS1 and PDGFRα to quantify cells mediating cholinergic, nitrergic and purinergic neurotransmission.

Results

KCl and ACh increased the tonic contraction and area under the curve (AUC) in a concentration-dependent manner in both normal and irradiated rectum. The difference in contractile pattern between the two groups was not statistically significant. SNP and ATP decreased AUC and reduced tonic contractions in a concentration-dependent manner in both normal and irradiated rectum. The response to SNP between the two groups was not statistically significant. However, the response to ATP reduced significantly in irradiated rectum compared to normal rectum. In addition, expression of VAChT and NOS1 which are related with cholinergic and nitrergic neurotransmission was not different in both normal and irradiated rectum, however, PDGFRα expression which is related with purinergic neurotransmission was significantly decreased in irradiated rectum compared to normal rectum.

Conclusion

In this study, we analyzed contractile changes in irradiated rectum using neurotransmitters. There were no differences in contractile responses to KCl and ACh and in relaxation response to SNP, however, the relaxation response to ATP was significantly reduced in irradiated rectum. Cholinergic and nitrergic neurotransmission were not changed, however, purinergic neurotransmission was decreased in in irradiated rectum.
These results may explain one of the causes by which increased bowel movements, symptoms of urgency, and decreased rectal compliance in irradiated patients.

Introduction

Irritable bowel syndrome (IBS) is a bothersome condition with long term pain and bowel symptoms and the most common cause of referral to gastroenterologists in the United Kingdom. Despite this, IBS is poorly understood as a clinical entity. Furthermore, IBS has numerous comorbidities but the full spectrum of these and how they relate to IBS has not been studied.

Aims & Methods

Using clustering analysis on the UK Biobank dataset, an adult cohort (age > 40 years), we aimed to define the top co-morbidities associated with IBS and develop a machine learning model to predict IBS. Co-morbidities in patients based on ICD-10 Summary Diagnosis codes 2 years prior to and 2 years following diagnosis with IBS were assessed and clusters defined. These were compared with a control group of non-IBS patients. Several machine learning algorithms (K-Means, Linear Discriminant Analysis, Gaussian Mixture Models) were compared for accuracy.

Results

Amongst 18000 total adults (aged over 40), 8168 patients coded for IBS, 75% of whom were female. The top ten comorbidities (Table 1) found in IBS patients included diverticular disease, hypertension, hernia, haemorrhoids, osteoarthritis, asthma, gastro-oesophageal reflux disease without oesophagitis, history of psychoactive substance abuse, hypercholesterolaemia, and cholelithiasis. K-Means achieved the highest accuracy (83%) for predicting IBS based on the co-morbidity profile.

Conclusion

Most clusters identified contained obesity, hypercholesterolemia, and hypertension, indicating a possible link with metabolic syndrome. Asthma and allergies suggest a possible cohort of IBS patients with atopy. Further research is needed to establish these links. In clinical practice, this could help both in predicting development of IBS, and also identify the cohorts of IBS on which certain therapies are more likely to work.

References

Drossman, D. A., Camilleri, M., Mayer, E. A., & Whitehead, W. E. (2002). AGA technical review on irritable bowel syndrome. Gastroenterology, 123(6), 2108-2131. doi:10.1053/gast.2002.37095
Snyder, N. (2023, 10 Feb 2023). Irritable bowel syndrome. BMJ Best Practice. Retrieved from https://bestpractice.bmj.com/topics/en-gb/122
Staller, K., Olén, O., Söderling, J., Roelstraete, B., Törnblom, H., Khalili, H., . . . Ludvigsson, J. F. (2021). Diagnostic yield of endoscopy in irritable bowel syndrome: A nationwide prevalence study 1987-2016. Eur J Intern Med, 94, 85-92. doi:10.1016/j.ejim.2021.08.001

Introduction

Coeliac disease (CeD) is a chronic immune-mediated enteropathy driven by dietary gluten proteins. Previous studies have indicated an increased risk of cardiovascular disease (CVD) in individuals with CeD [1-3]. However, results are inconsistent and specific guidelines on the management of CVD risk and comorbidities for individuals with CeD do not exist. In this study, we aimed to compare the distribution of traditional CVD risk factors in CeD patients and non-CeD controls from a population-based cohort.

Aims & Methods

We performed a CeD screening of 54,541 serum samples by anti-transglutaminase 2 immunoglobulin A and G serology from in total 56,042 adult participants in the fourth wave of the Norwegian Trøndelag Health Study (HUNT4) [4]. All seropositive participants were invited to upper endoscopy with duodenal biopsies to confirm a CeD diagnosis. Seronegative participants with previously diagnosed CeD were identified by linkage to hospital records and the Norwegian Patient Registry. We calculated SCORE2 and SCORE2-OP as predictors for 10-year risk of first-onset CVD [5, 6]. The groups “no CeD”, previously diagnosed CeD (“known CeD”) and CeD diagnosed in HUNT4 (“new CeD”), as well as “CeD” (known + new cases) were examined in this study. To compare group characteristics, we used the Wilcoxon rank sum test. Associations between groups and SCORE2(-OP) were assessed using ordinal logistic regression analysis.

Results

468 (0.80%) individuals received a CeD diagnosis in HUNT4, while 309 (0.57%) seronegative individuals had known CeD. Individuals with newly diagnosed CeD were younger, better educated, and had lower blood lipids compared to controls. Individuals with CeD smoked significantly less, had lower BMI and waist-hip-ratio, glycated hemoglobin levels, serum creatinine, and hsCRP levels compared to controls without CeD (n=52,438; p<0.05 for all variables mentioned, see table). We found a significantly lower SCORE2 for CeD cases (n=589) compared to controls (n=39,747; p<0.05). Adjustment for alcohol consumption did not affect group characteristics. A significantly lower SCORE2 was observed in CeD cases also after adjustment for level of education (OR .84, 95% CI .72-.98), high sensitivity C-reactive protein (hsCRP) and serum creatinine (OR .85, 95% CI .78-.99), but not for the combination of all three variables (OR .90, 95% CI .76-1.05).

Conclusion

Individuals with CeD appear to have a lower cardiovascular risk based on traditional risk factors like smoking habits, body fat, diabetes, renal function and inflammation compared to control individuals without CeD. In line with that, SCORE2 and SCORE2-OP as predictors for 10-year risk of first-onset CVD were lower in individuals with CeD.

References

1. Bathrellou, E., M.D. Kontogianni, and D.B. Panagiotakos, Celiac disease and non-celiac gluten or wheat sensitivity and health in later life: A review. Maturitas, 2018. 112: p. 29-33.
2. Ludvigsson, J.F., et al., Nationwide cohort study of risk of ischemic heart disease in patients with celiac disease. Circulation, 2011. 123(5): p. 483-90.
3. Figtree, G.A., et al., Mortality in STEMI patients without standard modifiable risk factors: a sex-disaggregated analysis of SWEDEHEART registry data. Lancet, 2021. 397(10279): p. 1085-1094.
4. Asvold, B.O., et al., Cohort Profile Update: The HUNT Study, Norway. Int J Epidemiol, 2023. 52(1): p. e80-e91.
5. SCORE2 working group and ESC Cardiovascular risk collaboration, SCORE2 risk prediction algorithms: new models to estimate 10-year risk of cardiovascular disease in Europe. Eur Heart J, 2021. 42(25): p. 2439-2454.
6. SCORE2-OP working group and ESC Cardiovascular risk collaboration, SCORE2-OP risk prediction algorithms: estimating incident cardiovascular event risk in older persons in four geographical risk regions. European Heart Journal, 2021. 42(25): p. 2455-2467.

Disclosure

None.

Introduction

Mast cells are the primary cells that cause acute and chronic colitis due to irradiation causing radiation-induced colitis (RIC). Herein, we investigated whether pre-treatment with tranilast, mast cell inhibitor, could alleviate RIC.

Aims & Methods

A total of 23 Sprague Dawley (SD) rats were randomly divided into three groups control (n=5, C), irradiation group (n=9, RG), and tranilast pre-treated irradiation group (n=9, TG). Tranilast (100 mg/kg) was administered with meal for 10 d to TG, then Gamma ray (20 gray single dose) was irradiated to the pelvis of RG and TG. Ten weeks after irradiation, the rats were euthanized. Rectal tissue samples were stained and the total inflammation score (TIS) and mast cell count were assessed histopathologically. Expression of MUC2, MUC5AC and matrix metalloproteinase (MMP)-9 were also assessed immunohistochemically

Results

Either TIS or some components of TIS including epithelial atypia, vascular sclerosis and colitis cystica profunda, were significantly higher in RG than in TG (p=0.02, 0.038, 0.025, 0.01, respectively). The median number of infiltrating mast cells was significantly higher in the RG (27.5±18.40)) than in TG (6.0±6.02) (P=0.034). Quantitatively, the number of MMP-9-positive cells was significantly higher in the RG (23.67±19.00) than in the TG (10.25±8.45) (p<0.05).

Conclusion

TIS and MMP9 exhibited strong associations (correlation coefficient r=0.6, p<0.05) Immunohistochemically, mucin-lake of CCP showed MUC2 positivity only but was negative for MUC5AC. Conclusively, tranilast pretreatment in RIC shows an anti-inflammatory effect on the RIC with reducing mast cell infiltration and MMP expression

Introduction

Inflammatory bowel disease (IBD) shares many features with other chronic inflammatory diseases (CIDs), such as rheumatoid arthritis and psoriasis, which are characterized by dysregulated immune responses and systemic manifestations. While IBD is primarily associated with inflammation in the gut, it can also lead to extra-intestinal symptoms, including joint pain and skin lesions, which are commonly observed in these other diseases. Understanding commonalities and differences among these chronic inflammatory conditions is important for developing more effective therapies that can target the underlying immune dysregulation and improve patients' overall quality of life.

Aims & Methods

We aim to identify both the common disease signatures shared between IBD and other CIDs, as well as the disease-specific signatures unique to each condition. A cross-sectional cohort of 650 patients with CIDs, including Crohn's disease, ulcerative colitis, rheumatoid arthritis, psoriatic arthritis, psoriasis, and systemic lupus erythematosus (SLE), were recruited from the University Hospital Kiel. Peripheral blood samples were collected from this cohort, along with samples from 202 healthy controls, and subjected to RNA sequencing and methylation profiling. At the time of sample collection, the patients in the cohort underwent detailed clinical phenotyping including assessments of disease activity and symptoms. Additionally, these patients were followed up for up to 5 years to record any potential future complications that may arise.

Results

We performed differential expression and methylation analysis between each CID diagnosis and the healthy control group to identify shared molecular signatures. Our analysis showed that IBD shared the most differential signatures with rheumatoid arthritis, indicating common pathways in their pathogenesis. Psoriatic diseases also shared a significant proportion of differentially expressed and methylated patterns, while SLE had the least in common with other CIDs, suggesting distinct molecular pathways driving its pathogenesis. Interestingly, we were able to identify core transcriptomic changes that persisted even during inactive disease by separating the analysis by disease activity. These signatures were shared across different CIDs and were correlated with DNA methylation changes, reflecting the molecular mechanism of chronicity. Commonly upregulated genes across all CIDs were enriched in pathways related to inflammatory response, cytokine signaling, neutrophil chemotaxis, and humoral immune response. Disease-specific pathways included interferon-beta production in SLE, several metabolic processes in psoriasis, and complement activation in ulcerative colitis. Disease-specific methylation signatures were mainly located in CpG islands, while methylation signatures in enhancers were shared among most diseases. Our large cohort design enabled us to identify subgroup of patients across CIDs that share molecular characteristics, including a cluster of genes related to interleukin and interferon signaling upregulated in 71 samples belonging to different CIDs.

Conclusion

In conclusion, our study identified common and disease-specific molecular signatures in CIDs. Our findings suggest shared immune dysregulation pathways underlying the pathogenesis of IBD and rheumatoid arthritis, as well as potential commonalities between psoriatic diseases. We show the clear presence of core signatures of disease regardless of disease activity, which could represent insights into actionable targets for causative treatments and better disease control of CIDs.