Marker Of Intestinal Inflammation Research Articles

P0574 Correlations Between Inflammatory Biomarkers and Hematological Parameters in Assessing Disease Activity in IBD

Abstract Background Monitoring ulcerative colitis (UC) and Crohn's disease (CD) involves assessing inflammatory and hematological biomarkers to evaluate disease activity, treatment response, and the impact of inflammation on overall health. The aim of this study was to assess if hematological and inflammatory biomarkers along with clinical parameters correlate with IBD inflammatory activity and severity. Methods The study retrospectively analyzed 100 IBD patients, of which 72 patients with UC and 38 patients with CD. Age, hematological parameters (hemoglobin, iron and ferritin), inflammatory biomarkers (CRP, fibrinogen and fecal calprotectin) and clinical scores such as Truelove and Witts for UC and UCDAI for CD were analyzed. Pearson's correlation coefficient demonstrated relationships between hematological and inflammatory biomarkers, while the Kruskal-Wallis test evaluated associations between these biomarkers and clinical scores. Results The mean age was 44.02 years, with UC patients averaging 43.03 years and CD patients 49.3 years, reflecting a slightly older demographic for CD. Age was negatively correlated with iron (-0.93) and positively with ferritin (0.88), indicating that older individuals have less iron deposits. Hemoglobin levels (mean 11.81 g/dL) were reduced across both diseases and exhibited significant negative correlations with CRP (-0.93) and fecal calprotectin (-0.91). Systemic (CRP) and intestinal (fecal calprotectin) inflammation can diminish hemoglobin levels in IBD patients, causing inflammatory anemia. Iron levels were lowest in CD (61.78 µg/dL) and correlated negatively with CRP (-0.93) and ferritin (-0.99) during active systemic inflammation, while ferritin levels (107.89 ng/mL) were elevated, particularly in active inflammation in CD group. Intestinal inflammation impacts iron status (-0.40), while ferritin and CRP are positively correlated (0.96), showing that systemic inflammation enhances iron storage. Fecal calprotectin emerged as a particularly sensitive marker of intestinal inflammation, showing the highest levels in UC patients (517.18 µg/g) and significant correlations with clinical activity scores (p = 0.02) for both Truelove-Witts and UCDAI. Hemoglobin was the only parameter that showed a weak trend of correlation with clinical scores (p=0.06), without statistical significance. Conclusion Hematological and inflammatory biomarkers along with clinical parameters correlate with IBD inflammatory activity and severity of the inflammatory process. The results highlight the effectiveness of biomarker evaluation in the personalized management of patients, enabling the adaptation, configuration, or establishment of monitoring schedules and therapeutic optimization based on the inflammatory profile.

Associations between dietary patterns and intestinal inflammation among HIV-infected and uninfected adults: A cross-sectional study in Tanzania.

The increased burden of non-communicable diseases (NCDs) is fueled by lifestyle factors including diet. This cross-sectional study explored among Tanzanian adults whether unhealthy dietary patterns are associated with intestinal and systemic inflammation which could increase the risk of NCDs. The study included 574 participants, with both diet and inflammatory markers data. Dietary patterns were derived using principal component analysis and reduced rank regression, revealing three main patterns: vegetable-rich, vegetable-poor, and carbohydrate-dense diets. Fecal myeloperoxidase (MPO) and neopterin (NEO) were markers of intestinal inflammation whereas plasma lipopolysaccharide-binding protein (LBP) and C-reactive protein (CRP) were assessed as markers of systemic inflammation. Ordinal logistic regression was used to assess associations between terciles of dietary patterns and quintiles of the inflammatory markers adjusting for potential confounders. High adherence to a vegetable-poor dietary pattern was associated with elevated MPO (adjusted OR, 1.7 95% CI 1.1, 2.8). NEO tended to be higher in people with high adherence to both vegetable-poor pattern (adjusted OR, 2.6 95% CI 1.0, 6.4) and vegetable-rich pattern (adjusted OR, 2.7, 95% CI 1.1, 6.5). No associations were found between dietary patterns and systemic inflammation markers (LBP and CRP). We found links between dietary vegetable intake and intestinal inflammation but not systemic inflammation. However, the cross-sectional nature of the study limits establishing causality and the sample size for some variables may have been inadequate, emphasizing the need for further studies to understand how dietary habits influence inflammation in this population.

Dietary Patterns and Fibre Intake Are Associated with Disease Activity in Australian Adults with Inflammatory Bowel Disease: An Exploratory Dietary Pattern Analysis.

Few studies have explored the relationship between habitual dietary patterns and disease activity in people with Inflammatory Bowel Disease (IBD). This cross-sectional study explored the association between dietary patterns and clinical and objective markers of inflammation in adults from the Australian IBD Microbiome Study. Dietary patterns were derived using principal component analysis (PCA) of baseline food frequency questionnaire data. Food intake was quantified using 3-day food record data. Associations between dietary intake and both clinical disease activity index (CDAI) and faecal calprotectin (FCP) were analysed. Participants included 412 adults (IBD = 223, Healthy controls (HC) = 189). Both cohorts consumed poor-quality diets with inadequate servings of most food groups compared to Australian reference standards. IBD participants without FCP inflammation had significantly higher fibre intake than those with moderate FCP. In the Crohn's Disease group, high adherence to 'High plant diversity' and 'Meat eaters' dietary patterns were associated with increased CDAI and FCP, respectively. In the combined IBD cohort, high adherence to a 'Vegan-style' dietary pattern was associated with increased FCP. There is a need for dietary modifications among Australian adults, both with and without IBD, to improve dietary fibre intake and adherence to dietary guidelines. Dietary patterns characterised by a high intake of plant foods or meat products were both positively associated with indicators of active IBD. It is possible that some participants with active IBD were modifying their diet to try to manage their disease and reduce symptoms, contributing to the association between healthier dietary patterns and active disease. Further clinical and longitudinal studies are needed to expand upon the findings. This study offers a unique contribution by utilising FCP as an objective marker of intestinal inflammation and applying dietary pattern analysis to investigate the relationship between diet and inflammatory markers.

Assessment of calprotectin levels preceding and succeeding colonoscopy

Background: Faecal calprotectin (FC) is an easily applicable faecal surrogate marker of intestinal inflammation; it has provided a new means for objectively assessing disease activity in patients with various bowel diseases. In this study, the authors assessed levels of FC before colonoscopy preparation, during bowel cleansing and 1 week after colonoscopy. Methods: This cross-sectional study was conducted on 58 individuals referred to Razi Hospital, Rasht, Iran, for colonoscopy between 2021 and 2022. For all individuals, stool sampling was repeated three times before colonoscopy, during bowel preparation and 1 week after colonoscopy. All demographical and clinical data were collected through a questionnaire. FC results, colonoscopy and pathology findings were also recorded. Results: The mean age of the participants was 48.22±11.69 years and approximately 62.1% were female. There was no statistically significant difference between the average levels of FC before colonoscopy preparation, during bowel cleansing and 1 week after colonoscopy (P>0.05). The average amount of FC at different time points was not significantly different in patients with abnormal histopathological results compared to individuals with normal findings (P>0.05). Conclusions: The study findings suggest that bowel cleansing and colonoscopy do not significantly alter FC levels.

The impact of cooling and Moringa supplementation on oxidative stress in serum and milk, including milk cytokines, in heat stressed lactating sows and their litters.

Heat stress (HS) poses a significant challenge to the United States swine industry. Sows and their piglets are particularly vulnerable to HS, as the periparturient phase is characterized by heightened metabolism and increased oxidative stress and inflammation. The study examined the effects of using conductive electronic cooling pads (ECP) and dietary supplementation with 4% Moringa (M) leaf powder on controlling oxidative stress and inflammation caused by HS in sows and their piglets. Forty-eight late gestation sows were assigned to four treatment groups: HS-fed corn-soybean meal (HS + CS), ECP-fed corn-soybean meal (ECP + CS), HS + M, and ECP + M. Blood was collected from sows on gestation (G) day 112, and lactation (L) day 14 and L20, and from piglets (2 males and 2 females) in each litter on postnatal (PN) day 1 and PN20. Colostrum was collected within 2h of birth of the first neonate, and mature milk was collected on L14. Piglet fecal samples were collected on PN14 to measure calprotectin concentration as a marker of intestinal inflammation. Biological antioxidant potential (BAP), derivatives of reactive oxygen metabolites (dROMs). and oxidative stress index (OSi) were measured in blood and milk samples using a Free Radical Elective Evaluator. Milk samples pooled by day of lactation and treatment group were analyzed using cytokine array. Levels of inflammatory cytokines in colostrum were affected by Moringa supplementation and cooling, but not mature milk. Notably, the anti-inflammatory cytokines interleukin (IL)-10 and IL-1ra were 2.14 and 1.57 Log2 higher in the colostrum of HS + M compared to other groups. The OSi of colostrum was higher (P = 0.0002) than mature milk. Level of BAP in sow serum was greater in ECP + CS and HS + M (P = 0.0291) compared to other groups. Moringa had an overall effect of increasing dROMs (P = 0.0035) and levels of OSi were lowest in ECP + CS (P = 0.0296) sow serum. Treatments did not affect piglet serum oxidative index (P > 0.05) or calprotectin levels (P > 0.05). Findings support further studies to investigate the efficacy of using ECP and Moringa supplementation to mitigate inflammation and oxidative stress imposed by heat stress conditions in lactating sows.

Oral Iron-Hydroxide Polymaltose Complex Versus Sucrosomial Iron for Children with Iron Deficiency with or without Anemia: A Clinical Trial with Emphasis on Intestinal Inflammation.

Iron deficiency anemia (IDA) is a major public health problem among children worldwide. Iron deficiency without anemia (IDWA) is at least twice as common as IDA. Some studies propose that oral iron fortification can modify the infant's gut microbiome, leading to intestinal inflammation. To determine whether oral iron therapy can lead to intestinal inflammation in children with IDA or IDWA. Fifty-six patients aged 6 months to 16 years (median age 7.6 years) with IDA or IDWA were randomly assigned to receive either iron (III)-hydroxide polymaltose complex (IPC) 5 mg/kg once daily (maximum dose 100 mg) or sucrosomial iron (SI)1.4 mg/kg once daily (maximum dose 29.4 mg). Safety and efficacy were studied after 30 and 90 days of treatment. In addition, fecal calprotectin as a marker of intestinal inflammation was measured simultaneously and compared to results obtained before therapy. A significant increase in serum ferritin was noted in both groups as the median ferritin level at baseline was 6.7 μg/L in the IPC group and 6.6 μg/L in the SI group, increasing to 15.9 μg/L and 12.1 μg/L respectively, after 90 days of treatment. However, there was no significant change in fecal calprotectin in either group. In addition, no differences in the trend over time were observed between the two groups regarding fecal calprotectin, serum ferritin, and hemoglobin. IPC and SI were equally effective in treating IDA and IDWA. At the recommended doses, oral iron therapy does not seem to induce intestinal inflammation.

Effects of Exposure to Different Types of Metal-Organic Framework Nanoparticles on the Gut Microbiota and Liver Metabolism of Adult Zebrafish.

Metal-organic framework nanoparticles (MOF NPs) have received much attention for their potential use in nanopesticides. However, little is known about the potential health and environmental risks associated with these materials. In this study, the toxicological responses of zebrafish exposed to five MOF NPs for short and long periods of time were evaluated. The acute toxicity results showed that the toxicity of the five MOF NPs to zebrafish embryos and adult zebrafish was in the order of Cu-MOF > ZIF-90 > ZIF-8 > Fe-MOF > Zr-MOF. Histopathological analysis revealed that ZIF-8, ZIF-90, and Cu-MOF NPs caused liver swelling and vacuolization in zebrafish. The cellular ultrastructure showed that ZIF-8, ZIF-90, and Cu-MOF NPs severely damaged the mitochondrial structure in intestinal epithelial cells and liver cells. The 16S rDNA sequencing data showed that all five MOF NPs significantly altered the dominant microorganisms in the zebrafish intestine. The microbial markers of intestinal inflammation, Proteobacteria (Aeromonas, Plesiomonas, and Legionella), were significantly increased in the Fe-MOF, ZIF-8, Zr-MOF, and Cu-MOF treatment groups. Metabolomics results indicated that the levels of inflammatory promoting factors (Leukotriene E4, 20-hydroxyeicosatetraenoic acid) in arachidonic acid metabolism were decreased, and the levels of inflammatory suppressing factors (8,9-epoxyeicosatrienoic acid) were increased. Metabolites related to oxidative stress, such as glutamine, pyridoxamine, and l-glutamic acid in vitamin B6 metabolism and other signaling pathways, were significantly reduced. Overall, these results suggest that the different MOF NPs had widely varying toxicity to zebrafish, and further attention should be paid to the toxicity of MOF NPs in the real environment.

Impacts of Whole-Grain Soft Red, Whole-Grain Soft White, and Refined Soft White Wheat Flour Crackers on Gastrointestinal Inflammation and the Gut Microbiota of Adult Humans.

Consumption of whole-grain wheat has been associated with positive health outcomes, but it remains unclear whether different types of wheat elicit varying effects on the gut microbiome and intestinal inflammation. The objectives of this research were to investigate the effect of two whole-grain wheat flours versus refined wheat flour on the diversity of the human gut microbiota, as well as on butyrate production capacity and gastrointestinal inflammation, using one-week dietary interventions. For this study, 28 participants were recruited, with ages ranging from 18 to 55 years and a mean BMI of 26.0 kg/m2. For four weeks, participants were provided 80 g daily servings of different wheat crackers: Week A was a run-in period of crackers made from soft white wheat flour, Week B crackers were whole-grain soft white wheat flour, Week C crackers were a wash-out period identical to Week A, and Week D crackers were whole-grain soft red wheat flour. At the end of each week, participants provided fecal samples that were analyzed for markers of intestinal inflammation, including lipocalin and calprotectin, using enzyme-linked immunosorbent assays and quantitative real-time PCR. The primary outcome, gut bacterial community alpha and beta diversity, was similar across timepoints. Three taxa significantly differed in abundance following both whole-grain wheat flour interventions: Escherichia/Shigella and Acidaminococcus were significantly depleted, and Lachnospiraceae NK4A136 group was enriched. Secondary outcomes determined that protein markers of intestinal inflammation and genes related to putative butyrate production capacity were similar throughout the study period, with no significant changes. Lipocalin concentrations ranged from 14.8 to 22.6 ng/mL while calprotectin ranged from 33.2 to 62.5 ng/mL across all 4 weeks. The addition of wheat crackers to the adult human subjects' usual diet had a minimal impact on their gastrointestinal inflammation or the gut microbiota.

Gut Microbiota Profiling as a Promising Tool to Detect Equine Inflammatory Bowel Disease (IBD).

Gastrointestinal disorders are common and debilitating in horses, but their diagnosis is often difficult and invasive. Fecal samples offer a non-invasive alternative to assessing the gastrointestinal health of horses by providing information about the gut microbiota and inflammation. In this study, we used 16S sequencing to compare the fecal bacterial diversity and composition of 27 healthy horses and 49 horses diagnosed with inflammatory bowel disease (IBD). We also measured fecal calprotectin concentration, a marker of intestinal inflammation, in healthy horses and horses with IBD. We found that microbiota composition differed between healthy horses and horses with IBD, although less than five percent of the variation in microbiota composition was explained by individual health status and age. Several differentially abundant bacterial taxa associated with IBD, age, or body condition were depleted from the most dominant Firmicutes phylum and enriched with the Bacteroidota phylum. An artificial neural network model predicted the probability of IBD among the test samples with 100% accuracy. Our study is the first to demonstrate the association between gut microbiota composition and chronic forms of IBD in horses and highlights the potential of using fecal samples as a non-invasive source of biomarkers for equine IBD.

Gas Chromatography-Sensor System Aids Diagnosis of Inflammatory Bowel Disease, and Separates Crohn's from Ulcerative Colitis, in Children.

The diagnosis of inflammatory bowel disease (IBD) in children and the need to distinguish between subtypes (Crohn's disease (CD) and ulcerative colitis (UC)) requires lengthy investigative and invasive procedures. Non-invasive, rapid, and cost-effective tests to support these diagnoses are needed. Faecal volatile organic compounds (VOCs) are distinctive in IBD. VOC profiles can be rapidly determined using a gas chromatography-sensor device (OdoReader©). In an inception-cohort of children presenting with suspected IBD, we directly compared the diagnostic fidelity of faecal calprotectin (FCP, a non-specific protein marker of intestinal inflammation) with OdoReader© VOC profiles of children subsequently diagnosed with IBD with matched controls diagnosed with other gastrointestinal conditions. The OdoReader© was 82% (95% confidence interval 75-89%) sensitive and 71% (61-80%) specific but did not outperform FCP (sensitivity 93% (77-99%) and specificity 86% (67-96%); 250 µg/g FCP cut off) in the diagnosis of IBD from other gastrointestinal conditions when validated in a separate sample from the same cohort. However, unlike FCP and better than other similar technologies, the OdoReader© could distinguish paediatric CD from UC (up to 88% (82-93%) sensitivity and 80% (71-89%) specificity in the validation set) and justifies further validation in larger studies. A non-invasive test based on VOCs could help streamline and limit invasive investigations in children.

Long-term evaluation of faecal calprotectin levels in a European cohort of children with cystic fibrosis

ObjectiveIntestinal inflammation with contradictory data on faecal calprotectin (fCP) levels is documented in patients with cystic fibrosis (CF). The aim of this study was to longitudinally evaluate fCP in a...

Persistent dysbiosis of duodenal microbiota in patients with controlled pediatric Crohn’s disease after resolution of inflammation

Crohn’s disease is an inflammatory condition of the intestine characterized by largely unknown etiology and a relapse remission cycle of disease control. While possible triggers have been identified, research is inconsistent on the precise cause of these relapses, especially in the under-researched pediatric population. We hypothesized that patients in remission would have persistent microbial and inflammatory changes in small intestinal tissue that might trigger relapse. To this end, we analyzed intestinal biopsy samples from six patients with pediatric Crohn’s disease in remission and a control group of 16 pediatric patients with no evident pathogenic abnormality. We identified compositional microbiota differences, including decreases in the genera Streptococcus and Actinobacillus as well as increases in Oribacterium and Prevotella in patients with controlled Crohn’s disease compared to controls. Further, a histologic analysis found that patients with controlled Crohn’s disease had increased epithelial integrity, and decreased intraepithelial lymphocytes compared with controls. Additionally, we observed increased peripheral CD4+ T cells in patients with pediatric Crohn’s disease. These results indicate that markers of intestinal inflammation are responsive to Crohn’s disease treatment, however the interventions may not resolve the underlying dysbiosis. These findings suggest that persistent dysbiosis may increase vulnerability to relapse of pediatric Crohn’s disease. This study used a nested cohort of patients from the Bangladesh Environmental Enteric Dysfunction (BEED) study (ClinicalTrials.gov ID: NCT02812615 Date of first registration: 24/06/2016).

Rosmarinic acid mitigates intestinal inflammation and oxidative stress in bullfrogs (Lithobates catesbeiana) fed high soybean meal diets

High proportions of soybean meal in aquafeed have been confirmed to induce various intestinal pathologies. This study aims to investigate the regulatory effects of rosmarinic acid (RA), an antioxidant with anti-inflammatory and antimicrobial properties, when added to high soybean meal feeds in different doses, (0, 0.5, 1, and 4 g/kg). During the 56-day feeding trial, results indicated that, compared to the control group without RA (0 g/kg), the 1 g/kg and 4 g/kg RA groups increased bullfrog survival rates and total weight gain while reducing feed coefficient. Additionally, these doses markedly suppressed the expression of key intestinal inflammatory markers (tlr5, myd88, tnfα, il1β, cxcl8, cxcl12) and the activity and content of intestinal antioxidants (CAT, MDA, GSH, GPX). Concurrently, RA significantly downregulated the transcription levels of antioxidant-related genes (cat, gpx5, cyba, cybb, mgst, gclc, gsta, gstp), suggesting RA's potential to alleviate intestinal inflammation and oxidative stress induced by high soybean meal and to help downregulate and restore normal expression of antioxidant enzyme genes. However, the 0.5 g/kg RA group did not show a significant improvement in survival rates; instead, it upregulated the transcription of some antioxidant genes (cat, gpx5, cyba, cybb), revealing the complexity and dose-dependency of RA's antioxidant action. Furthermore, RA supplementation significantly reshaped the intestinal microbial community structure and relative abundance in bullfrogs, particularly affecting the genera Hafnia, Phascolarctobacterium, and Lactococcus. Notably, high doses of RA (1 g/kg, 4 g/kg) were able to downregulate pathways associated with the enrichment of gut microbiota in diseases such as Parkinson's, Staphylococcus aureus infection, and Systemic lupus erythematosus, suggesting its potential in anti-inflammatory action and health maintenance to prevent potential diseases.

PSIII-21 Impact of high zinc oxide supplementation or lignocellulose supplement on chronic markers of intestinal inflammation and growth of weaned piglets

Abstract The objective of this project was to study the effects of high zinc oxide (ZO) supplementation and fiber supplement from lignocellulose (LIGCEL) on chronic markers of intestinal inflammation and growth of weaned piglets. At weaning (21 d), 150 piglets (body weight = 6.39 ± 0.276 kg) were transferred from a farrowing farm to a nursery farm where they were divided into 30 pens of 5 piglets per pen according to their weanling weight. Each pen was assigned to one of the following three treatments: Control (CON, 150 mg/kg of zinc), ZO (2,500 mg/kg of zinc), and LIGCEL (CON+3% lignocellulose). The ZO and LIGCEL supplements were added to a phase 1 diet. The experimental diets (phase 1) were distributed for 14 d. Subsequently, all piglets received the same diets from phases 2 and 3 for two periods of 14 d. During the experiment, piglets were weighed upon arrival and after phases 1, 2, and 3. The feed was distributed every day and the total quantities calculated for each of the feeding phases. The average daily gain (ADG), average daily feed intake (ADFI) and feed conversion (FC) were subsequently calculated (Table 1). On d 6 and 13, the blood of two piglets per pen was collected to determine the content of D-lactate and diamine oxidase (DAO). Fecal samples from these two piglets were also taken to evaluate the calprotectin and neopterin contents. During phase 1, ADG was greater for ZO piglets compared with the CON animals with an intermediate value for the LIGCEL group (P < 0.001). The ADFI was greater for the ZO compared with the CON and LIGCEL treatments (P < 0.003). Piglets in the ZO and LIGNOCELL treatments had less FC than those in the CON group (P < 0.001). During phase 2, only the FC was greater for the ZO and LIGCEL treatments compared with CON animals (P < 0.013). During phase 3, growth performances were not affected by the dietary treatments. For the entire experiment, ZO piglets had greater ADG than those in the CON group with an intermediate value for LIGCEL treatment (P < 0.019). D-lactate decreased from d 6 to 13 while DAO increased (P < 0.001). The DAO was also greater for ZO and LIGCELL piglets compared with CON animals (P < 0.001). Neopterin was greater for ZO treatment but only on d 13 (Trt × day, P < 0.010). Finally, calprotectin decreased from d 6 to 13 (P < 0.001) but was not affected by dietary treatments. In conclusion, ZO and LIGCEL fiber act positively on growth by modulating markers of intestinal inflammation. LIGCEL fiber could, therefore, be an alternative to ZO in the diet of weaned piglets.

Fecal calprotectin levels in patients with non-celiac wheat sensitivity: a proof of concept

Some data suggest the existence of intestinal inflammation in patients with non-celiac wheat sensitivity (NCWS). We aimed to verify whether fecal calprotectin (FCP), a marker of intestinal inflammation, could be used to confirm this inflammatory status and to test its diagnostic performance in differentiating NCWS from irritable bowel syndrome/functional dyspepsia (IBS/FD). We conducted a multicenter study, comparing NCWS patients, diagnosed by a double-blind placebo-controlled wheat challenge, with IBS/FD subjects. In the retrospective phase, FCP values were analyzed to define the prevalence of its positivity and its role as a NCWS diagnostic biomarker. In the prospective phase we tested the effects of a strict 6-month wheat-free diet (WFD) on FCP values. 31.3% (n = 63/201) of NCWS patients had above normal FCP values (NCWS FCP +), whereas all IBS/FD patients proved negative (P = 0.0001). FCP using a cut-off value > 41 µg/g showed a 58.6% sensitivity and a 98.0% specificity (AUC 0.755, 95% C.I. 0.702–0.837) in distinguishing NCWS from IBS/FD patients. Of the 63 NCWS FCP+, 65.1% had negative FCP values after ≥ 6 months of WFD, with a significant reduction in FCP values (P < 0.0001). All NCWS FCP- subjects still preserved negative FCP values after ≥ 6 months of WFD. Our study showed that FCP can be a useful but supplementary diagnostic marker for differentiating between NCWS and IBS/FD. Strict WFD adherence reduced FCP values, normalizing them in 65.1% of NCWS FCP + subjects. These data suggest the existence of two NCWS subgroups: NCWS FCP + characterized by a probable predominantly inflammatory/immunologic pattern and NCWS FCP− featuring non-immuno-mediated etiopathogenetic mechanisms. (Registration number NCT01762579).

Association of intestinal inflammation and permeability markers with clinical manifestations of Parkinson's disease

IntroductionIntestinal inflammation and gut microbiota dysbiosis can stimulate degeneration of dopaminergic neurons and development of Parkinson's disease (PD) via the gut-brain axis in certain patients. MethodsIn a case-control study, fecal markers of intestinal inflammation and permeability were measured using the ELISA method in PD patients and healthy controls. Motor and nonmotor symptoms were assessed using the Movement Disorder Society (MDS) Unified PD Rating Scale, Hoehn & Yahr scale, MDS Non-Motor Symptom Scale, Scales for Outcomes in PD - Autonomic Dysfunction, PD Sleep Scale - 2, Montreal Cognitive Assessment, Beck Anxiety Inventory, and Beck Depression Inventory-II. A correlation was established between the intestinal inflammation and permeability markers and PD symptoms. ResultsHigher levels of beta-defensin 2, zonulin and lactoferrin were recorded in PD patients compared to controls. Calprotectin and secretory immunoglobulin A showed no significant differences. Regression analysis indicated the roles of beta-defensin 2 and lactoferrin in predicting PD likelihood. Calprotectin yielded positive correlations with disease duration, depression, motor fluctuations, and gastrointestinal symptoms; beta defensin 2 with thermoregulation; and secretory immunoglobulin A with depression. Secretory immunoglobulin A showed negative correlation with age and age at disease onset, while zonulin showed negative correlation with the MDS Unified PD Rating Scale total score. ConclusionsFecal markers differed in PD patients compared to controls and correlated with age, disease duration, and some nonmotor symptoms. Future studies should identify the subgroups of PD patients that are likely to develop intestinal inflammation.

Replacing fishmeal with salmon hydrolysate reduces the expression of intestinal inflammatory markers and modulates the gut microbiota in Atlantic salmon (Salmo salar)

The feed legislation allows the use of fish protein hydrolysates in feed for the same species in which it came from, since enzymatic hydrolysis degrades the proteins and eliminates potential prions, which have caused disease in mammals, but not in fish. In this trial, we investigated the effects of partially replacing dietary fishmeal (FM) with salmon protein hydrolysate (FPH) on the intestinal gene expression and microbiota. Atlantic salmon post smolts were either fed a control diet containing 30% fishmeal (FM), a 20% FM diet with 9% salmon hydrolysate (FPH-09) or a 10% FM diet with 18% salmon hydrolysate (FPH-18), until doubling of weight. Gene expression analysis by RNA sequencing of pyloric caeca (PC), midgut (MG) and hindgut (HG) revealed a downregulation of immunological genes involved in inflammation in the intestine of FPH-18 fed salmon compared to salmon fed the FM control. The gene expression of paralogous peptide transporters (PepT) was analyzed by real time quantitative PCR in PC, anterior midgut (AMG), posterior midgut (PMG) and HG of salmon fed all the three diets. The PepT1b paralog had highest relative expression levels in PC and AMG, suggesting that PepT1b is most important for peptide uptake in the anterior intestine. PepT1a was also mainly expressed in the PC and AMG, but at lower levels than PepT1b and PepT2b in the AMG. The PepT2b paralog had high levels of expression in AMG, PMG and HG indicating that it contributed significantly to peptide uptake in the posterior part of the gastrointestinal tract. The gut microbiota in the mucosa and digesta of the MG and HG, were dominated by the phyla Cyanobacteria and Proteobacteria, but also Firmicutes were present. The only dietary effect on the microbiota was the higher prevalence of the phyla Spirochaetes in the mucosa of FPH-18 fed salmon compared to the FM fed salmon. In conclusion, replacing FM with salmon hydrolysate reduced the expression of inflammatory markers in the Atlantic salmon intestine suggesting improved health benefits. The reduced inflammation may be related to the reduced FM content, potentially bioactive peptides in the hydrolysate and/or the altered gut microbial composition.

A5 DEVELOPMENT OF A PRE-CLINICAL MOUSE MODEL TO INVESTIGATE ADVERSE FOOD REACTIONS IN INTESTINAL INFLAMMATION

Abstract Background Patients with chronic intestinal conditions, including inflammatory bowel disease (IBD), experience diverse food-related adverse reactions. However,, the precise mechanisms of food intolerances in IBD remains unclear. Emerging evidence suggests that altered diet-microbiota interactions can contribute to the development of IBD. Previously we shown that intestinal microbiota plays an important role in the development of food intolerances. We hypothesize that microbial alterations in IBD patients facilitate adverse reactions to foods. Aims To establish a preclinical mouse model to study adverse food reactions in intestinal inflammation. Methods Eight-week-old C57BL/6 mice were treated with 3% dextran sodium sulfate (DSS) for 5 days, then divided into three groups. The first group was sensitized to dairy protein (3% casein and 3% whey), using 4 oral administrations of cholera toxin over 2 weeks, without any further diet intervention. The second group was also sensitized to casein and whey, then after 1 week of recovery, the mice were subsequently place on a chow containing casein and whey. Finally, the third group was subjected to diet intervention without sensitization. The same experiment was repeated in a different subset of C57BL/6 mice which included a second cycle of DSS (1.5%) during the dietary interventions. Markers of intestinal inflammation (disease activity index (DAI), histological damage, cell infiltration and pro-inflammatory genes expression (NanoString)) and intestinal microbiota were analysed after 1 week of the diet intervention. Results Mice previously sensitized on the dairy diet exhibited an enhanced activation of immune cells after intestinal inflammation. While the histological damage score did not reveal significant differences among experimental groups, the sensitized group under dairy intervention exhibited an elevated level of immune cell infiltration, increasing the polymorphonuclear, CD3+ and mast cells. Gene expression also revealed that dairy induced the expression of inflammatory genes, such as C6, Arg1, Tnf, Il6, Cxcl9 and Cxcl10. When animals are subjected to a second DSS-cycle, dairy worsen colitis in mice previously sensitized, as shown by higher DAI compared to group with control diet and dairy diet without sensitization. Conclusions In the context of intestinal inflammation, dairy protein leads to immune activation characterized by an increase in polymorphonuclear cells, mast cells and lymphocytes, as well as worsening colitis. Future studies using this model will provide valuable insights into the role of intestinal microbiota in food intolerances associated with intestinal inflammation. Funding Agencies CCC

Gut microbiota and inflammatory mediators differentiate IgE mediated and non-IgE mediated cases of cow's milk protein at diagnosis.

Analyze fecal and blood samples at point of diagnosis in IgE mediated cow's milk protein allergy (CMPA) and non-IgE mediated (NIM)-CMPA patients to look for potential new biomarkers. Fourteen patients with IgE mediated CMPA and 13 with NIM-CMPA were recruited in three hospitals in the north of Spain, and were compared with 25 infants from a control group of the same age range. To characterize intestinal microbiota, 16S rDNA gene and internal transcribed spaceramplicons of bifidobacteria were sequenced with Illumina technology. Fatty acids were analyzed by gas chromatography, meanwhile intestinal inflammation markers were quantified by enzyme-linked immunosorbent assayand a multiplex system. Immunological analysis of blood was performed by flow cytometry. The fecal results obtained in the NIM-CMPA group stand out. Among them, a significant reduction in the abundance of Bifidobacteriaceae and Bifidobacterium sequences with respect to controls was observed. Bifidobacterial species were also different, highlighting the lower abundance of Bifidobacterium breve sequences. Fecal calprotectin levels were found to be significantly elevated in relation to IgE mediated patients. Also, a higher excretion of IL-10 and a lower excretion of IL-1ra and platelet derived growth factor-BB was found in NIM-CMPA patients. The differential fecal parameters found in NIM-CMPA patients could be useful in the diagnosis of NIM food allergy to CM proteins.

P535 Effectiveness of upadacitinib in patients with Ulcerative Colitis: a real-life, multicenter, Italian report

Abstract Background Upadacitinib (UPA) is an oral, selective JAK inhibitor, that has recently received approval in Italy for the treatment of Ulcerative colitis (UC). To our knowledge, only few data of real-life studies are available. The aim of our observational data collection is to report the first real-life experiences with UPA in UC in terms of effectiveness and safety. Methods These data come from four Italian IBD centers. Adult patients with a diagnosis of UC, who have been prescribed UPA for active disease, after failure of at least one anti-TNF therapy, were enrolled. After an induction period of 8 weeks with a 45 mg daily dosage, treatment efficacy were assessed using the partial Mayo score (pMS) and the fecal calprotectin levels (CP). Treatment-related adverse events were also recorded. Results Among 20 patients who started UPA, 12 have completed the induction and were therefore enrolled and evaluated before starting UPA and at the end of the 8 weeks induction period. 83% (10 patients) had previously failed two or more biological therapies, including tofacitinib (3 patients, 25%). 3 patients (25%) were tapering systemic corticosteroids and 3 patients (25%) were receiving topical steroids when they started UPA. In the pre-treatment evaluation, the mean pMs was 5,25 (SD 2,26) and the median CP was 1270 ug/g (411- 3152). At the end of induction (week 8), 10 patients (83%) achieved clinical remission and one patient showed clinical response with a decrease of 6 points of pMS. One patient underwent colectomy after 4 weeks of ineffective therapy. The mean pMS at week 8 was 0 (SD 0,92) (Fig. 1), with a median CP of 88 ug/g (5-4639). Reported adverse events, that however did not cause drug discontinuation, were one case of leukopenia and one case of acne. To the date of the last evaluation, all patients maintained the achieved results, with a mean follow-up period of 92 days. A further analysis excluding the physician global assessment from pMS has been performed, to obtain a sort of patient-reported outcome 2 (PRO-2): the results, in terms of percentage reduction of the scores, were in line with the presented data. Despite the limited number of patients who completed the induction (11), we observed statistical significance in the difference between pretreatment and post-induction values of all the measured variables (Tab. 1, Fig. 1). Conclusion With this first Italian report in a real-life setting, UPA showed promising effects in rapidly inducing clinical remission and in normalizing a sensitive marker of intestinal inflammation. Moreover, it seems to be well tolerated. Long-term real-life studies, including endoscopic and histologic evaluations, are needed to confirm efficacy and safety of UPA in maintaining stable remission in UC patients.