Red Cell Distribution Width for Assessment of Activity of Inflammatory Bowel Disease

Single-Cell Analyses of Colon and Blood Reveal Distinct Immune Cell Signatures of Ulcerative Colitis and Crohn’s Disease

Cytokines serve a central function as key factors in the regulation of the intestinal immune response and mediation of tissue damage in inflammatory bowel disease (IBD). Abnormalities in the expression of immunoregulatory cytokines such as IL-2, IL-4, IL-10 and interferon-gamma (IFN-gamma) may indicate a dysregulation of intestinal immunity probably associated with pathogenic events. Therefore, cytokine mRNA concentrations were determined in the mucosa of patients with IBD at sites of active (n = 13) and inactive (n = 12) ulcerative colitis (UC), active (n = 11) and inactive (n = 11) Crohn's disease (CD) and in control patients (n = 14) using quantitative RT-PCR. IL-10 mRNA concentrations were significantly increased in patients with both active UC (P < 0.001) and active CD (P < 0.005) compared with control patients. IFN-gamma mRNA concentrations were also significantly increased both in patients with active UC (P < 0.02) and active CD (P < 0.05) compared with control patients, whereas IL-2 mRNA levels were significantly (P < 0.02) increased only in active CD. IL-4 mRNA expression in the intestinal mucosa was frequently below the detection limit. Our results demonstrate that chronic intestinal inflammation in patients with CD is characterized by an increase of Th1-like cytokines. Furthermore, the increased IL-10 mRNA expression at sites of active IBD suggests that IL-10 is an important regulatory component involved in the control of the inflammatory response in inflammatory bowel disease.

Fibroblast growth factor 21 (FGF21) is a stress-inducible hormone that regulates nutrient and metabolic homeostasis. Inflammatory state is one of the stimulators of FGF21 secretion. The aim of the study was to assess correlations between serum FGF21 level and inflammatory markers as well as nutritional status indicators in patients with inflammatory bowel disease (IBD). Fasting serum FGF21 level was measured using ELISA test in 105 IBD patients and 17 healthy controls. There were 31 subjects with active ulcerative colitis (UC), 16 with inactive UC, 36 with active Crohn's disease (CD), and 22 with inactive CD. Clinical and endoscopic activity of IBD was evaluated based on validated scales and indices. Fecal calprotectin, serum CRP, and selected parameters of nutritional status were tested in all patients. Serum FGF21 level was characterized by fluctuations depending on the IBD activity. FGF21 level was significantly higher in both active UC and CD compared to inactive phases of the diseases and to the controls. A correlation between FGF21 and fecal calprotectin levels was also found in UC and CD. Additionally, in CD, FGF21 level positively correlated with CRP level. In both UC and CD, a negative correlation was noted between FGF21 level and nutritional status parameters including cholesterol, protein, albumin levels, and BMI. The intensity of intestinal inflammation is related to FGF21 level, which correlates negatively with nutritional status indicators in IBD. The disturbances in FGF21 secretion may contribute to the multifactorial pathogenesis of malnutrition and weight loss in IBD patients.

Objective This study is designed to evaluate the role of fecal calprotectin as a marker for the diagnosis of ulcerative colitis (UC) and its correlation with disease activity and remission. Background Inflammatory bowel diseases (IBD) are lifelong intestinal inflammatory conditions of unknown etiology, characterized by remissions and exacerbations. The diagnosis and classification of IBD is usually established by a combination of tests (laboratory, endoscopic, and/or radiologic) in the presence of clinical symptoms. Fecal calprotectin serves as a noninvasive biomarker of intestinal inflammation, and has been found to be useful in the diagnosis of IBD, assessment of response to medical therapy, and in prediction of clinical relapse. Materials and methods This study was carried out on 40 patients. Twenty of these patients had clinical, laboratory, colonoscopic, and histopathological findings of UC. Group I was subdivided as follows: GIa: 20 patients with active UC and GIb: the same patients as in GIa while on remission. Group II included 20 patients as controls, matched for age and sex without clinical, laboratory, colonoscopic, and histopathological findings of UC. Complete stool analysis, erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) titer, complete blood count, colonoscopy and biopsy, histopathological examination of biopsy specimens, measurement of ulcerative colitis activity index (UCAI), and quantitative determination of calprotectin in stool sample were carried out for all patients. Results There was a highly significant increase in the mean value of fecal calprotectin in active UC patients in comparison with the inactive UC patients and controls. Also, there was a highly significant increase in the mean value of fecal calprotectin in the inactive UC patients in comparison with the controls. There was also a highly significant positive correlation between fecal calprotectin and UCAI, CRP, ESR, total leukocyte count, and platelets count. At the cut-off value of 131 μg/g, fecal calprotectin has 100% accuracy, sensitivity, specificity, positive predictive value, and negative predictive value in differentiating UC patients from other patients with lower gastrointestinal symptoms and at the cut-off value of 253 μg/g fecal calprotectin has 95% accuracy, sensitivity, specificity, positive predictive value, and negative predictive value in differentiating active from inactive UC patients. Conclusion Fecal calprotectin is a valuable, simple, easily performed, and cost-effective noninvasive marker for evaluation of patients with UC. It differentiated UC and other diseases causing colonic symptoms (cut-off value of 131 μg/g) and between active and inactive UC (cut-off value of 235 μg/g) with high accuracy, sensitivity, and specificity. It also correlates well with other markers for UC activity (UCAI, ESR, CRP, total leukocyte count, and platelets count) and could be a reliable surrogate marker for the severity of UC.

Oxidative stress is believed to play an important role in the pathogenesis of inflammatory bowel disease (IBD), specifically Crohn's disease (CD) and ulcerative colitis (UC). This meta-analysis aimed to identify and quantify the oxidative stress-related biomarkers in IBD and their associations with disease activity. We systematically searched Ovid MEDLINE, Ovid Embase, and Web of Science databases, identifying 54 studies for inclusion. Comparisons included: (i) active IBD versus healthy controls; (ii) inactive IBD versus healthy controls; (iii) active CD versus inactive CD; and (iv) active UC versus inactive UC. Our analysis revealed a significant accumulation of biomarkers of oxidative damage to biomacromolecules, coupled with reductions in various antioxidants, in both patients with active and inactive IBD compared to healthy controls. Additionally, we identified biomarkers that differentiate between active and inactive CD, including malondialdehyde, Paraoxonase 1, catalase, albumin, transferrin, and total antioxidant capacity. Similarly, levels of Paraoxonase 1, erythrocyte glutathione peroxidase, catalase, albumin, transferrin, and free thiols differed between active and inactive UC. Vitamins and carotenoids also emerged as potential disease activity biomarkers for CD and UC, but their intake should be monitored to obtain meaningful results. These findings emphasize the involvement of oxidative stress in the pathogenesis of IBD and highlight the potential of oxidative stress-related biomarkers as a minimally invasive and additional tool for monitoring the activity of IBD.

The innate immune system is currently seen as the probable initiator of events which culminate in the development of inflammatory bowel disease (IBD) with Toll-like receptors (TLRs) known to be involved in this disease process. Many regulators of TLRs have been described, and dysregulation of these may also be important in the pathogenesis of IBD. The aim of this study was to perform a co-ordinated analysis of the expression levels of both key intestinal TLRs and their inhibitory proteins in the same IBD cohorts, both ulcerative colitis (UC) and Crohn's disease (CD), in order to evaluate the potential roles of these proteins in the pathogenesis of IBD. Of the six TLRs (TLRs 1, 2, 4, 5, 6 and 9) examined, only TLR-4 was increased significantly in IBD, specifically in active UC. In contrast, differential alterations in expression of TLR inhibitory proteins were observed. A20 and suppressor of cytokine signalling 1 (SOCS1) were increased only in active UC while interleukin-1 receptor-associated kinase 1 (IRAK-m) and B cell lymphoma 3 protein (Bcl-3) were increased in both active UC and CD. In contrast, expression of both peroxisome proliferator-activated receptor gamma (PPARγ) and Toll interacting protein (Tollip) was decreased in both active and inactive UC and CD and at both mRNA and protein levels. In addition, expression of both PPARγ and A20 expression was increased by stimulation of a colonic epithelial cell line Caco-2 with both TLR ligands and commensal bacterial strains. These data suggest that IBD may be associated with distinctive changes in TLR-4 and TLR inhibitory proteins, implying that alterations in these may contribute to the pathogenesis of IBD.

Red blood cell distribution width is a parameter measured automatically in every complete blood count that actually reflects the degree of anisocytosis of the red blood cell population. It is a cost-effective tool used in everyday clinical practice along with other parameters to define and narrow the cause of anemia. In a series of pathologic entities, such as cardiovascular diseases, autoimmune diseases, malignancy, chronic renal diseases and chronic respiratory diseases, where inflammation and oxidative stress comprise the major pathophysiologic insults, red cell distribution width behaves as a significant and competent marker able to predict and assess disease activity and severity. A number of clinical studies based on these observations have aimed to evaluate its potential utility as an index of activity in inflammatory bowel disease. In this narrative review we present data from the international literature regarding its ability to express disease activity and we look into its relation with clinical, laboratory and endoscopic indices used to identify active disease. According to the results of published clinical trials, red cell distribution width is considerably correlated with disease activity and might serve as an index to differentiate Crohn’s disease from ulcerative colitis.

Background Individuals with inflammatory bowel disease (IBD) experience mental health disorders at a rate of 1.5 to 2 times higher than the general population. With the rising incidence of IBD, many people are affected by the disease during significant life stages such as pregnancy. Research on the impact of IBD on anxiety and depression in pregnant people is limited. Aims To assess the impact of active IBD on anxiety and depression during pregnancy. Methods This retrospective survey study analyzed data from patients seen at the ‘Preconception and Pregnancy in IBD’ clinic at Mount Sinai Hospital who conceived prior to January 1st, 2022. Standardized pre-visit surveys included validated measures of mental health (General Anxiety Disorder-7 for anxiety; Patient Health Questionnaire-9 for depression) and IBD disease activity (pMayo6 for ulcerative colitis (UC); mHBI for Crohn’s disease (CD)). Demographic data was also collected (age, marital status, occupational status, smoking habits, alcohol use, cannabis use, previous diagnosis of anxiety and/or depression, IBD type, IBD surgical history, and medication use). Linear regression models were used to assess associations between disease activity and anxiety/depression, adjusting for medication type and stratified by trimester and IBD subtype. Results A total of 94 patients were included in this study, 42 patients had UC (44.7%), and 52 patients had CD (55.3%). The median age for the UC group was 32 (IQR 31-35.8), and the median age for the CD group was 32.7 (IQR 30-35.2). Active CD in the second trimester was significantly associated with higher anxiety (p = 0.024) and depression scores (p = 0.021) compared to those in remission. For CD, 6 patients (35.3%) in trimester 1, 8 patients in trimester 2 (25.8%), and 8 patients (26.7%) in trimester 3 had a GAD-7 score of ≥ 5; 5 patients (29.4%) in trimester 1, 6 patients (19.4%) in trimester 2, and 7 patients (23.3%) in trimester 3 had a PHQ-9 score of ≥ 5. Active UC in the second trimester was associated with significantly higher anxiety (p = 0.034) and depression (p = 0.022) scores compared to inactive UC. In the third trimester, this pattern persisted, with active UC again associated with elevated anxiety (p = 0.030) and depression (p = 0.027) scores. For UC, 4 patients (33.3%) in trimester 1, 9 patients (30%) in trimester 2, and 9 patients (31.0%) in trimester 3 had a GAD-7 score of ≥ 5; 4 patients in trimester 1, 4 patients in trimester 2, and 7 patients in trimester 3 had a PHQ-9 score of ≥ 5. Conclusions Active IBD during pregnancy is significantly associated with increased anxiety and depression, therefore, a multidisciplinary approach including mental health support is imperative for pregnant people with IBD. Funding Agencies None

Soluble adhesion molecules are elevated in a number of inflammatory conditions. To investigate the correlation of soluble intercellular adhesion molecule-1 (sICAM-1) and sE-selectin with the activity of inflammatory bowel disease (IBD). sICAM-1 and sE-selectin were measured by an enzyme-linked immunosorbent assay (ELISA) in 53 patients with ulcerative colitis (UC) and 38 patients with Crohn's disease (CD). Patients with active UC and CD had significantly higher sICAM-1 than patients with inactive disease and controls. Patients with pancolitis had significantly higher levels than patients with distal colitis. There was a significant difference in sE-selectin levels between patients with active CD and control sICAM-1. sE-selectin did not correlate with the Harvey Bradshaw index (HBI). C-reactive protein (CRP) and microalbuminuria were better markers than sICAM-1 or sE-selectin which correlated with serum tumour necrosis factor (TNF)-alpha. sICAM-1 and sE-selectin are elevated in the serum of patients with IBD but CRP and microalbuminuria reflect clinical disease activity more accurately. This study does not support the routine use of soluble adhesion molecules as disease activity markers in IBD.

In this study, we investigated whether reticulated platelets (RP) would be useful markers in the evaluation of ulcerative colitis (UC) activity and also aimed to gain indirect information about the platelet kinetics. Complete blood count, C-reactive protein, erythrocyte sedimentation rate, and proportion of RP were measured in 16 active, 21 inactive UC patients, and 20 healthy blood donors. UC activity was assessed by Truelove-Witts criteria. Mean platelet count was increased in patients with active compared to inactive UC (p=0.008) or healthy donors (p=0.000). Mean platelet volume (MPV) was significantly decreased in patients with active compared to inactive (p=0.015) and healthy donors (p=0.001). RP values was significantly decreased in active and inactive UC groups compared to healthy donors (p=0.000, p=0.000, respectively), while there was no significant difference between active and inactive UC patients (p=0.980). Significant negative correlation between platelet count and MPV in patients with active UC (r=-0.542, p=0.030) was observed. RP values is reduced in active and inactive UC patients compared to healthy donors. To our knowledge, this is the first study about proportion of RP with UC in literature. However, the role of low RP values have not been determined clinically. Further studies are needed to evaluate the role of platelet abnormalities and changes in megakaryopoiesis caused by inflammatory state on low MPV and RP values during the course of UC.

To optimise treatment of ulcerative colitis (UC), patients need repeated assessment of mucosal inflammation. Current non-invasive biomarkers and clinical activity indices do not accurately reflect disease activity in all patients and cannot discriminate UC from non-UC colitis. Volatile organic compounds (VOCs) in exhaled air could be predictive of active disease or remission in Crohn's disease. To investigate whether VOCs are able to differentiate between active UC, UC in remission and non-UC colitis. UC patients participated in a 1-year study. Clinical activity index, blood, faecal and breath samples were collected at each out-patient visit. Patients with clear defined active faecal calprotectin >250 μg/g and inactive disease (Simple Clinical Colitis Activity Index <3, C-reactive protein <5 mg/L and faecal calprotectin <100 μg/g) were included for cross-sectional analysis. Non-UC colitis was confirmed by stool culture or radiological evaluation. Breath samples were analysed by gas chromatography time-of-flight mass spectrometry and kernel-based method to identify discriminating VOCs. In total, 72 UC (132 breath samples; 62 active; 70 remission) and 22 non-UC-colitis patients (22 samples) were included. Eleven VOCs predicted active vs. inactive UC in an independent internal validation set with 92% sensitivity and 77% specificity (AUC 0.94). Non-UC colitis patients could be clearly separated from active and inactive UC patients with principal component analysis. Volatile organic compounds can accurately distinguish active disease from remission in UC and profiles in UC are clearly different from profiles in non-UC colitis patients. VOCs have demonstrated potential as new non-invasive biomarker to monitor inflammation in UC.

The beta 2 integrin intercellular adhesion molecule (ICAM) adhesion pathway is likely pivotal in the immunopathogenesis of inflammatory bowel disease (IBD). We have undertaken a comprehensive study of peripheral blood lymphocyte (PBL) expression of all beta 2 integrins and ICAMs in patients with IBD using flow cytometry and assessed our data on the basis of IBD diagnosis, disease state of activity, and use of corticosteroids. Blood was collected from patients with Crohn's disease (N = 49), ulcerative colitis (N = 43), and normal control volunteers (N = 15). Mononuclear cells were separated using a Ficoll-Hypaque gradient and prepared for flow cytometry. The data were analyzed for percentage expression, mean fluorescent intensity (MFI) as well as for histogram patterns. The analysis was stratified for disease diagnosis, disease activity level, and for use of prednisone among patients with active disease. There was decreased percentage expression of CD11a, CD18, and ICAM-3 in Crohn's disease and ulcerative colitis compared with normal, but an increased MFI for these molecules among patients with Crohn's disease. Active Crohn's disease showed a greater change in this pattern compared with both inactive disease and active ulcerative colitis. CD11a and CD18 histograms typically had two peaks of expression. The predominance of one peak over the other varied with disease diagnosis and activity. CD11b and alpha d expression patterns were not different in IBD compared with normal. CD11c was not expressed by PBLs and, ICAM-2, typically an endothelial ligand, was expressed on PBLs. There were changes in the expression of beta 2 integrins in IBD, which were more evident in Crohn's disease than ulcerative colitis. We hypothesize that the decreased percentage expression and increased MFI of CD11a, CD18, and ICAM-3 may suggest that cells up-regulate these ligands following activation and are egressing into tissue.

Aims/Background Kawasaki disease is an acute inflammatory condition primarily affecting the young children. It can lead to coronary artery abnormalities, which can worsen the prognosis. Early diagnosis of coronary disease is crucial for the effective treatment and the prognosis evaluation. To explore the clinical significance of ultrasound examination characteristics, peripheral blood red cell distribution width, and changes in N-terminal pro-brain natriuretic peptide levels for the early detect coronary artery abnormality in children with Kawasaki disease. Methods The case-control study was conducted. 85 Kawasaki disease patients diagnosed in our hospital from January 2020 to December 2023 were selected as the Kawasaki disease group. 100 healthy children who received physical examination in the Department of Child Healthcare during the same period were selected as control group. The cardiac ultrasound indicators, erythrocyte sedimentation rate, C-reactive protein, white blood cell, neutrophil percentage, platelet count, D-dimer, red cell distribution width, N-terminal pro-brain natriuretic peptide of two groups were compared. The Kawasaki disease group was further divided into the coronary artery lesion group and the non-coronary artery lesion group based on whether coronary artery lesions occurred in the Kawasaki disease patients. The differences of above indicators were compared. Results The left main coronary artery, left anterior descending branch, and right coronary artery Z-scores of the Kawasaki disease group were all higher than those of the control group (p < 0.05). There was no significant difference in left ventricular ejection fraction between Kawasaki disease group and control group (p > 0.05). The erythrocyte sedimentation rate, C-reactive protein, neutrophil percentage, platelet count, D-dimer, red cell distribution width, and N-terminal pro-brain natriuretic peptide of Kawasaki disease group were all higher than those of control group (p < 0.05). The left main coronary artery, left anterior descending branch, and right coronary artery Z-scores of Kawasaki disease patients with coronary artery lesions were all higher than those of Kawasaki disease patients without coronary artery lesions (p < 0.05). The left ventricular ejection fraction of Kawasaki disease patients with coronary artery lesions was lower than that of Kawasaki disease patients without coronary artery lesions (p < 0.05). The erythrocyte sedimentation rate, C-reactive protein, white blood cell, neutrophil percentage, platelet count, D-dimer, red cell distribution width, and N-terminal pro-brain natriuretic peptide of Kawasaki disease patients with coronary artery lesions were all higher than those of Kawasaki disease patients without coronary artery lesions, and the differences were statistically significant (p < 0.05). After treatment, the left main coronary artery, left anterior descending branch, and right coronary artery Z-scores of Kawasaki disease patients with coronary artery lesions significantly decreased (p < 0.05), and the left ventricular ejection fraction significantly increased (p < 0.05). The erythrocyte sedimentation rate, C-reactive protein, white blood cell, neutrophil percentage, platelet count, D-dimer, red cell distribution width, and N-terminal pro-brain natriuretic peptide of Kawasaki disease patients with or without coronary artery lesions significantly decreased after treatment compared with before treatment in the same group (p < 0.05). Conclusion Kawasaki disease patients with coronary artery lesions exhibit significantly increased coronary artery vessel diameter, as well as elevated red cell distribution width and N-terminal pro-brain natriuretic peptide concentration. The combined use of ultrasound combined with red cell distribution width and N-terminal pro-brain natriuretic peptide examination can assist in determining whether Kawasaki disease patients have coronary artery lesions and assessing the clinical treatment effect.

Background: Inflammatory bowel disease (IBD) comprises primarily 2 disorders: ulcerative colitis (UC) and Crohn's disease (CD). The hallmark of IBD is chronic, uncontrolled inflammation of the intestinal mucosa. The diagnosis is based on a combination of disease history, colonscopy, inflammatory biomarkers, radiological and histological evaluation. Most biomarkers used are not reliable and not disease specific, but reflect generalized inflammation. Aim: The aim of the work is to identify plasma proteomic profiles of UC cases and correlating this profile with the other diagnostic markers and activity of the disease. Methods: We performed a study with 70 plasma samples collected from patients classified in two groups (37 UC, 33 healthy controls) according to accredited criteria. They were subjected to: complete history taking, thorough clinical examination, laboratory investigations (erythrocyte sedimentation rate (ESR), C- reactive protein (CRP), fecal calprotectin, perinuclear antineutrophil cytoplasmic antibodies (P-ANCA), ileocolonscopy, histopathology, imaging. Plasma proteomic pattern of UC patients and control subjects was determined using Matrix-Assisted Laser Desorption/Ionization (MALDI) Time of Flight (TOF) Mass Spectrometer (MS) analysis. All plasma samples were subjected to solid-phase extraction (SPE). We analyzed the spectra obtained from all the samples using ClinProTool software. Results: There was a statistically significant difference of the plasma proteome profiles of UC group in comparison to health volunteers. 64 signals were identified by the ClinProTool software and of these 16 peptide peaks were highly significant (sensitivity was 100%, specificity was 84.4%, positive predictive value was 100, and negative predictive value was 82). There was a statistically significant difference between active versus inactive UC group, 5 integration regions used for classification between active and inactive UC patients using Genetic Algorithm model (GA) which gave 88.5% cross validation and 100% recognition capability. Markers as ESR, CRP, fecal calprotectin, P-ANCA are statistically correlated to the plasma proteomics found in UC patients. Conclusion: Proteomic profile has the potential to improve diagnosis and evaluate UC activity, reducing the need for more invasive techniques. The pattern of these peptides holds the promise of distinguishing disease states and providing clinically important information such as prognosis, response to therapy or perhaps targets of therapy.

Laboratory data in children with newly diagnosed inflammatory bowel disease (IBD) have been reported from Europe and North America, but not Asia. The aim of this study was to clarify laboratory data in Japanese children with newly diagnosed IBD, and to compare them with those in Western reports. We retrospectively reviewed patients <16years old, newly diagnosed with ulcerative colitis (UC) or Crohn's disease (CD) at Kurume University Hospital between January 2008 and December 2015. UC and CD patients numbered 31 and 15, respectively. The percentages of patients with normal values for hemoglobin (Hb), platelet count (Plt), albumin (Alb), C-reactive protein (CRP), and erythrocyte sedimentation rate (ESR) in the UC and CD groups were 45% and 47%; 68% and 53%; 84% and 40%; 81% and 7%; and 35% and 0%, respectively. The frequency of normal results for these five tests were similar to Western findings except for the greater frequency of normal CRP in UC. Alb and ESR differed significantly between UC and CD in both mild and moderate-severe cases. Plt, Alb, CRP, and ESR differed significantly between diseases in late-onset IBD, whereas early onset IBD showed no differences. In UC, ESR correlated positively, while Hb and Alb correlated negatively, with disease activity. In CD, CRP and ESR correlated positively with activity. The proportion of Japanese children with IBD having normal values at diagnosis was mostly similar to that in Western reports. In early onset cases, UC parameters may be similar to CD. Of the five tests, ESR was particularly indicative of disease activity at diagnosis in both pediatric UC and CD.