Stool Calprotectin Levels Research Articles

Uncovering the Secret Clues: Using Stool Calprotectin to Improve Low Anterior Resection Syndrome after Surgery

Abstract Background: A majority of patients who undergo low rectal resection experience a frequent complication of low anterior resection syndrome (LARS). It is characterized by distressing bowel dysfunction such as urgency, frequency, and incontinence. Despite the high prevalence of LARS, its management remains challenging due to the lack of reliable diagnostic and monitoring tools. Objectives: This study investigates the relationship between stool calprotectin levels and LARS severity and evaluates the effectiveness of mesalamine treatment. Design: This prospective study involved 48 patients with low rectal cancer who had undergone resection with temporary ileostomy. The patients subsequently presented with LARS for over four weeks after ileostomy closure. Patient and Methods: LARS diagnosis was based on the LARS score, and stool calprotectin levels were measured at baseline and one month and six months post-ileostomy closure. Patients with elevated calprotectin levels received mesalamine treatment and dietary modifications. Statistical analyses were performed to determine the correlation between calprotectin levels and LARS scores using Pearson correlation analysis, linear regression analysis, paired t-test, and ANOVA. Main Outcome Measure: The predictive capacity for stool calprotectin for LARS was the main outcome variable. Sample Size: Forty-eight patients who underwent low rectal cancer resection were enrolled. Results: Among the study group, 28 were males and 20 were females. The mean age was 50 (9.998) years, 95% confidence interval CI = [17–78]) in both groups. The predictive value of calprotectin levels for LARS severity was high with β = 0.72, P-value < .001, and R² 0.55. The mean (SD) stool calprotectin level was 360 (6.282) mcg/g, which correlated with severe LARS symptoms (mean [SD] LARS score = 35 [2.449]). After six months, 60% of patients showed significant improvement (LARS scores <20), 30% had minor improvement (LARS scores 29–30), and 10% showed no improvement. Correlation analysis revealed a strong positive correlation between calprotectin levels and LARS scores (r = 0.75, P < .001). Conclusion: Stool calprotectin is demonstrated to be a promising biomarker for diagnosing and monitoring LARS. The findings highlight the association between LARS and high levels of stool calprotectin. However, the study had limitations of being a single-center study, comprising a small sample size, and not using a randomized clinical trial to evaluate the effectiveness of mesalamine. Conflicts of Interest: None.

Elevated type-17 cytokines are present in axial spondyloarthritis stool.

Axial spondyloarthritis (axSpA) is characterized by type-17 immune-driven joint inflammation, and intestinal inflammation is present in around 70% of patients. In this study, we asked whether axSpA stool contained Th17-associated cytokines and whether this related to systemic Th17 activation. We measured stool cytokine and calprotectin levels by ELISA and found that patients with axSpA have increased stool IL-17A, IL-23, GM-CSF, and calprotectin. We further identified increased levels of circulating IL-17A+ and IL-17F+ T-helper cell lymphocytes in patients with axSpA compared to healthy donors. We finally assessed stool metabolites by unbiased nuclear magnetic resonance spectroscopy and found that multiple stool amino acids were negatively correlated with stool IL-23 concentrations. These data provide evidence of type-17 immunity in the intestinal lumen, and suggest its association with microbial metabolism in the intestine.

1349. Unveiling the Link between COVID-19 and the Gut Microbiome in an Outpatient Cohort with Mild to Moderate Illness

Abstract Background As the acute phase of the COVID-19 pandemic subsides, an increasing number of individuals are recognized as having long COVID, affecting over 65 million people worldwide. Recent studies have suggested a potential link between long COVID and alterations in the gut microbiome. In this study, we examined the gut microbiome of a large outpatient cohort during acute phase of infection, with the ultimate goal of determining predictors of long-term COVID-19 health outcomes. Methods We conducted a study of 394 COVID-19 patients with positive SARS-CoV-2 testing from November 2020 to September 2021 at a multi-site healthcare system. Long COVID was defined as the persistence of symptoms for four weeks or more after illness onset. DNA extraction was performed using the MoBio PowerSoil Pro DNA isolation kit. Samples were sequenced using shotgun metagenomics. Stool SARS-CoV-2 RNA was tested using qRT-PCR of the E gene, and stool ELISA was performed for calprotectin levels. Standard biostatistical tools in R (v 4.2.3) were used for statistical analysis. Results Of the 394 COVID-19 patients, 84 (21%) had long COVID, with fatigue being the most common persistent symptom (Table 1). No significant differences were seen in age, BMI, and antibiotic use. Those with long COVID were more likely to be female, had higher Charlson's comorbidity scores, more severe index illnesses. However, the two groups had no difference in SARS-CoV-2 stool PCR positivity or stool calprotectin levels (Table 2). Principal coordinate analysis of Bray Curtis dissimilarities revealed that patients with long COVID had a distinct microbial composition compared with those without long COVID during the acute phase of infection (ANOSIM R2= 0.005, P= 0.01). Categorizing patients by number of long COVID symptoms showed significant differences among groups (ANOSIM R2=0.1, P=0.007), mainly between patients with >3 symptoms and those with 0-1 symptoms (Bonferroni, P= 0.018). Alpha diversity was lower in patients with > 3 symptoms, although this was not statistically significant (Shannon diversity, two-way-ANOVA, P=0.051). Figure 1 Conclusion Long COVID patients show distinct gut microbial composition during the acute phase of infection, suggesting a potential role in predicting long-term COVID health outcomes. Disclosures John C. O'Horo, Sr., MD, MPH, Janssen Pharmaceuticals.: Grant/Research Support|nference: Grant/Research Support Purna C. Kashyap, MD, Intrinsic Medicine: Advisor/Consultant|Novome Biotechnologies: Advisor/Consultant|Pendulum Therapeutics: Advisor/Consultant

A229 GUT MICROBIOTA PROFILES, DIET AND SHORT-CHAIN FATTY ACIDS AS PREDICTORS OF GENERALIZED ANXIETY DISORDER

Abstract Background Generalized anxiety disorder (GAD) is a debilitating chronic condition with a lifetime prevalence of 4–7% worldwide. Both diet and gut microbiota have been previously associated with anxiety. Aims To investigate whether bacterial taxa and/or nutrients associate with GAD, and whether they differ from those of healthy controls (HC). Methods Patients with GAD (n=82) and matched HC (n=97) were assessed by validated questionnaires for anxiety (DASS-21), gastrointestinal (GI) symptoms (Rome III, Short-Form Leeds Dyspepsia), and dietary profiles by the Dietary Questionnaire for Epidemiological Studies. We quantified several blood and stool biomarkers, including inflammatory and neuroactive metabolites, as well as short-chain fatty acids. Stool microbiota profiles were assessed by16S rRNA gene sequencing through Illumina. The data was then analyzed following the pipelines of dada2 and by multiple factor analysis (MFA), mean comparisons, correlation, LEfSe and XGBoost using R software (v.1.2.1335). Multiple comparison results were corrected allowing 5% of FDR. Results Using MFA to analyze all variables, we identified 3 clusters: one mainly composed of HC (n=99, 91% HC, GI symptoms in 25% of subjects), a second mixed cluster (n=30, 80% GAD, GI symptoms in 80%) and a third cluster mainly composed of GAD patients (n=50, 98% GAD, GI symptoms in 86%). When focusing only on the HCs of cluster 1 (n=90) and GADs of cluster 3 (n=49), we found higher GI symptoms, body mass index, serum C-reactive protein and stool calprotectin levels (adj. p=1.3x10-9, 0.001, 0.017 and 0.017, respectively) and lower concentrations of propionate, butyrate and acetate in GAD compared to HC. GADs also reported overall lower caloric intake (kJ/day; adj. p=1.7x10-4) in the food frequency questionnaire. Fibre (g/day) was the macronutrient most negatively associated with anxiety scores (R=-0.44; adj. p=4.2x10-5). Bacteroides was the only bacterial taxon significantly associated with GAD, as well as with anxiety scores (R=0.31, adj. p=0.003). Interestingly, Bacteroides/fiber ratio was strongly correlated to anxiety scores (R=0.58, adj. p=2.7x10-09). Furthermore, demographic, biomarkers and bacterial taxa data were predictive of the patients’ disease state with 92.8% accuracy. The features that aid the model to predict disease state were Bacteroides/fiber ratio, GI symptoms and stool acetate levels. Conclusions Our results suggest that most GAD patients differ in dietary and microbiota profiles from HCs, and that the Bacteroides/fiber ratio, stool acetate and GI symptoms might be good predictors of disease state. Furthermore, these data strongly support the role of microbiota-gut-brain axis in genesis of psychiatric diseases, and they will inform mechanistic studies in gnotobiotic mouse models. Funding Agencies NIH

Predictors of surgical intervention in patients with inflammatory bowel disease (two-center study)

BackgroundSixty percent of Crohn’s disease (CD) patients require intestinal resection, and 20% of ulcerative colitis (UC) patients undergo proctocolectomy for medically refractory disease. Scarcity of literature about predictors for surgical intervention in inflammatory bowel disease (IBD) encouraged the conduction of this study to assess risk factors for surgical intervention in IBD patients.ResultsThis cohort study included 80 Egyptian inflammatory bowel disease patients recruited from two medical centers. Patients were classified into two groups, 40 patients each, according to their need for surgical intervention to control inflammatory bowel disease. The two groups were compared regarding age of onset, type and location of disease, smoking, extraintestinal manifestations, perianal disease, granuloma, severity scores, stool calprotectin, complete blood count, erythrocyte sedimentation rate, C-reactive protein, and serum albumin at diagnosis for Crohn’s disease patients.Twelve ulcerative colitis and 28 Crohn’s disease patients required surgical intervention in the form of total colectomy (30%), fistulectomy (32.5%), resection anastomosis (17.5%) or abscess drainage (20%). Perianal disease, smoking, and disease severity scores showed high significant differences (P value < 0.001); disease type and presence of granuloma showed statistically significant difference (P value < 0.05) between both groups. But, patient age at onset, location of the disease or extraintestinal manifestation had no statistical significance (P value > 0.5). Surgical interventions were more likely to be needed in patients with higher stool calprotectin level, C-reactive protein, erythrocyte sedimentation rate, and lower serum albumin for Crohn’s disease patients (P value < 0.001 for each).ConclusionSmoking, perianal disease, higher severity scores, stool calprotectin, C-reactive protein, and erythrocyte sedimentation rate levels are predictors of surgical treatment.

Evaluation and comparison of stool calprotectin level in necrotizing enterocolitis infected and noninfected neonates of &lt;1500 g

Background: Necrotizing enterocolitis (NEC) is one of the most prevalent digestive emergencies in neonates, leading to mortality and morbidity in premature neonates. Since there is no specific test for NEC diagnosis, only clinical symptoms and radiological findings are used for diagnosis. Therefore, if a reliable biomarker found to detect NEC, it can help to reduce the mortality and morbidity; hence, the present study evaluated the stool calprotectin levels in the infected and noninfected neonates weighing 176 μg/g, sensitivity = 97.14%, Specificity = 100%, P

Atypical Presentation of a Rare Disease: Idiopathic Enterocolic Lymphocytic Phlebitis Mimicking Ulcerative Colitis

Enterocolic lymphocytic phlebitis (ELP) is a rare cause of ischemic bowel injury. It is characterized by phlebitis of the bowel wall and mesenteric veins, without arterial involvement. The etiology and pathogenesis of this condition remains poorly understood. A 33-year-old Caucasian male, presented with intermittent abdominal pain, and blood in stools for six months. He was reportedly diagnosed with ulcerative colitis and was started on mesalamine. He denied sick contacts, recent travel, tobacco and alcohol use. His physical examination revealed left lower quadrant abdominal tenderness, without guarding or rigidity. Bright red blood and tenderness to palpation were noted on rectal examination. His initial laboratory studies were unremarkable, except for hemoglobin level of 12.6 g/dl, and an elevated C-reactive protein level of 3.3 mg/dl. CT scan of the abdomen showed normal bowel wall thickness. A comprehensive laboratory work-up was negative for Anti-Nuclear Antibody (Ab), Anti-myeloperoxidase Ab, Anti-proteinase 3 Ab, Anti-mitochondrial Ab, Anti-smooth muscle Ab, Saccharomyces cerevisiae IgA/IgG Ab, celiac serology, anti- CCP Ab and rheumatoid factor. Complement and immunoglobin levels were within normal range. Stool infectious panel and stool calprotectin levels were negative. Colonoscopy revealed, moderate to severe proctosigmoiditis from the anorectal junction. Histopathologic examination showed regenerative mucosal changes without evidence of cryptitis, crypt abscesses, or granulomas. He underwent laparoscopic low anterior resection with diverting ileostomy, after failing treatment with steroids, azathioprine and infliximab for three months. Surgical pathology showed discrete ulcers that extended into submucosa, and lymphocytic cuffing of small submucosal and mesenteric veins suggestive of ELP. Flexible sigmoidoscopy during one month follow up showed, diffuse areas of severely congested, eroded, friable, dusky, and necrotic appearing mucosa. He underwent total proctocolectomy with end ileostomy for recurrence of ELP involving the entire colon. He has been asymptomatic for one year post-proctocolectomy. Discussion: ELP commonly presents as acute abdomen with hematochezia. Histological features can range from granulomatous phlebitis, necrotizing phlebitis, to myointimal and endothelial hyperplasia. Endoscopic appearance may lead to misdiagnosis, as deep submucosal biopsies are needed to diagnose ELP. Surgical resection of the affected bowel is curative.

Elevated fecal calprotectin levels during necrotizing enterocolitis are associated with activated neutrophils extruding neutrophil extracellular traps.

BACKGROUNDNeonates with necrotizing enterocolitis (NEC) have higher calprotectin levels in stool than do healthy neonates. However, it is not known whether high stool calprotectin at the onset of bowel symptoms identifies neonates who truly have NEC vs. other bowel disorders.STUDY DESIGNNeonates were eligible for this study when an x-ray was ordered to “rule-out NEC”. Stool calprotectin was quantified at that time and in a follow-up stool. Each episode was later categorized as NEC or not NEC. The location of calprotectin in the bowel was determined by immunohistochemistry.RESULTSNeonates with NEC had higher initial and follow-up stool calprotectin levels than did neonates without NEC. Calprotectin in bowel from neonates with NEC was within neutrophil extracellular traps (NETs).CONCLUSIONAt the onset of signs concerning for NEC, fecal calprotectin is likely to be higher in neonates with NEC. Calprotectin in their stools is exported from neutrophils via NETs.

Avoid Endoscopy in Children With Suspected Inflammatory Bowel Disease Who Have Normal Calprotectin Levels.

In children with suspected inflammatory bowel disease, adding calprotectin stool testing to the screening strategy has been recommended to distinguish organic from nonorganic disease. In this cohort study with historical controls, we could not confirm that screening with stool calprotectin improves the diagnostic yield (ratio inflammatory bowel disease-positive endoscopies and total number of endoscopies); however, in patients with normal fecal calprotectin levels (<50 μg/g) endoscopic and histological abnormalities were not seen. We propose to refrain from endoscopy when stool calprotectin levels are normal.

VAlidation of an 8-item-questionnaire predictive for a positive caLprotectin tEst and Real-life implemenTation in primary care to reduce diagnostic delay in inflammatory bowel disease (ALERT): protocol for a prospective diagnostic study

IntroductionDiagnosis of inflammatory bowel disease (IBD) in primary healthcare is challenging and often associated with a considerable diagnostic delay. This delay is associated with worse disease progression and outcomes. Although...

PP73 FROM CHILDHOOD TO ADOLESCENCE: THE COELIAC DISEASE EXPERIENCE FROM THE POINT OF VIEW OF THOSE WHO ARE GROWING UP AND THOSE WHO ARE TAKING CARE OF THEM

Stool calprotectin is useful in monitoring rejection in small bowel transplantation. We report about our initial experience in Children’s Hospital “Bambino Gesu” in Rome on monitoring gut rejection by using stool calprotectin together with intestinal biopsies. Case 1: a 12 year-old boy underwent at the age of 8 SBTx for short gut syndrome; he experienced one ACR, recovered on steroid bolus and he did well for 6 months. Sirolimus was started for initial renal damage, but it had to be discontinued for severe thrombotic micro-angiopathy. A second severe ACR occurred 1 year after transplant and again medical therapy was effective in recovery. After 3 years of well being, with no previous evidence of rejection at histology, he was admitted in Rome for acute diarrhea following Rotavirus infection. Hystology revealed ACR, initially treated by steroid without any effects so repeated steroid bolus, infliximab and mesenchymal cell infusion were administered, but severe exfoliative rejection led to re-transplant, performed in Rome on TAC, steroid, MMF and basiliximab; at + 3 months, CMV enteritis occurred, successfully treated by ganciclovir. Before and after re-Tx, stool calprotectin levels were strict monitoring: normal levels ( 1000 mcg/g) if severe ACR, while in CMV infection levels were always below 800 mcg/g. Case 2: C.S. is now a 10 yr-old boy affected by CIPO who underwent SBTx at the age of 7 for recurrence of severe dehydration and hemorrhagic events; surgical procedure was uneventful, on basiliximab, TAC and steroid therapy. Six months after transplant he experienced ACR and severe CMV enteritis, with complete recovery. The follow-up was continued in Rome, where he underwent protocol biopsies without any evidence of rejection so far. Stool calprotectin levels were always within normal ranges (<250 mcg/g), according to histology. In our experience stool calprotectin levels appear to correlate to gut inflammation, and high levels seem to be more associated to rejection. This could be a useful tool to suspect graft dysfunction.

Hot topics in postsmall bowel transplantation: noninvasive graft monitoring including stool calprotectin and plasma citrulline

Long-term success in intestinal transplantation could be significantly improved by the development of a reliable noninvasive marker of rejection, allowing early diagnosis and treatment without the need for repeated biopsy procedures. At present, no single test appears sufficiently reliable to replace the need for invasive testing. Falling plasma citrulline levels may correlate with mucosal damage already done, but do not appear to be specific for rejection or provide clinically useful prediction of an event in advance of its occurrence. Similarly, rising stool calprotectin levels are correlated with pathologic diagnosis of rejection, but do not appear to have sufficient predictive value. Both tests appear to be best described as exclusionary, in that if their values are 'normal', rejection is less likely, but each is hampered by wide variability within and between patients. These approaches and others are reviewed in an outline of the current progress towards developing a clinically useful noninvasive tool for graft monitoring.

Exogenous alkaline phosphatase for the treatment of patients with moderate to severe ulcerative colitis

Increased activity of intestinal alkaline phosphatase (AP) occurs locally in patients with ulcerative colitis (UC), aimed at repairing inflammatory tissue damage. We evaluated the safety and preliminary efficacy of exogenous AP administered to patients with UC in an open-label, first-in-patient exploratory trial, conducted in the Internal Medicine and Gastroenterology hospital departments in the Czech Republic and Italy. Twenty-one patients were enrolled (13 females), age 23-54 years, with steroid- and/or immunosuppressant-refractory, moderate/severe UC (Mayo score 6-11). Oral AP enzyme 30,000 U was administered daily for 7 days, intraduodenally. Efficacy outcomes were changes in Mayo score at Day 21 posttreatment; changes in Modified Truelove-Witts Severity index (MTWSI) at Days 21, 63; C-reactive protein and stool calprotectin levels at Days 7, 21, 63. Safety evaluations were adverse events and laboratory abnormalities reported up to Day 63 posttreatment. No clinically relevant adverse events causing withdrawal or considered serious, or laboratory abnormalities or antibody formation against AP were observed. Mayo scores were significantly decreased at Day 21, and MTWSI at Days 21 and 63. C-reactive protein and stool calprotectin levels were decreased at Days 21 and 63. Clinical response on the Mayo score after a single 7-day AP course was 48% at Day 21. In this uncontrolled trial, administration of exogenous AP enzyme daily over a 7-day course to patients with UC was associated with short-term improvement in disease activity scores, with clinical effects being observed within 21 days and associated with reductions in C-reactive protein and stool calprotectin. AP enzyme treatment was well tolerated and nonimmunogenic.

Stool calprotectin levels before and after moderate alcohol consumption in inactive inflammatory bowel disease

Stool calprotectin levels before and after moderate alcohol consumption in inactive inflammatory bowel disease

Stool calprotectin levels before and after moderate alcohol consumption in inactive inflammatory bowel disease

Stool calprotectin levels before and after moderate alcohol consumption in inactive inflammatory bowel disease

Calprotectin

To identify a noninvasive screening test for intestinal allograft monitoring. Intestinal allograft rejection is difficult to distinguish from other causes of diarrhea and can rapidly lead to severe exfoliation or death. Protocol biopsies are standard for allograft monitoring but may cause serious complications. No noninvasive test has shown clinical utility for monitoring of the intestinal allograft. Calprotectin levels (n = 68) were measured in this pilot study from ileostomy effluent in patients with histologic evidence of acute rejection (n = 12), viral enteritis (n = 5), and nonspecific inflammation (n = 16) and compared with those with normal allograft histology (n = 35). Median stool calprotectin levels from patients with rejection were significantly higher than those from patients with viral enteritis or normal biopsies [198 mg/kg compared with 7 and 19 mg/kg, respectively (P = 0.0002)]. Receiver operator characteristics suggest the optimal cut-off level to distinguish rejection from other diagnoses is 92 mg/kg with specificity of 77% and sensitivity of 83%. Although false-positive results occurred in 26% of patients with normal biopsies and 30% with nonspecific changes, no treated episode of acute rejection was below the cutoff. In addition, in 2 patients with serial levels, elevations in the calprotectin levels preceded histologic changes by 6 to 18 days. Low stool calprotectin levels correlate well with a low risk for intestinal allograft rejection. If confirmed, biopsies may be reserved in the future for confirmation of rejection, eliminating protocol biopsies, and immunosuppressive changes could potentially be made before allograft injury.

Small bowel transplantation: current status and new developments in allograft monitoring

Purpose of review This review aims to summarize the current practices for intestinal allograft monitoring, including the newest developments in the past year. The gold standard has been histologic examination of endoscopy-directed allograft biopsy. Recent findings Plasma citrulline levels, immunohistochemical staining of biopsy specimens, and stool calprotectin levels have all been examined as alternative methods of allograft monitoring. The aims of each diagnostic test are variable and include improvement in the accuracy of biopsy interpretation, early diagnosis of rejection, or a noninvasive method of monitoring the intestinal allograft that might avoid or decrease the risks associated with endoscopic biopsy. Summary Although the results are extremely limited for each of the tests currently being studied in humans, there is good reason to believe that significant progress over the next few years will satisfy all these aims. The development of accurate alternative methods of allograft monitoring will undoubtedly improve the results of intestinal transplantation by allowing lowering of immunosuppression levels in patients not at risk for rejection and limiting exposure to toxicities associated with these medications.