Blood calprotectin as a biomarker for infection and sepsis - the prospective CASCADE trial.

BACKGROUND: Early in the host-response to infection, neutrophils release calprotectin, triggering several immune signalling cascades. In acute infection management, identifying infected patients and stratifying these by risk of deterioration into sepsis, are crucial tasks. Recruiting a heterogenous population of patients with suspected infections from the emergency department, early in the care-path, the CASCADE trial aimed to evaluate the accuracy of blood calprotectin for detecting bacterial infections, estimating disease severity, and predicting clinical deterioration. METHODS: In a prospective, observational trial from February 2021 to August 2022, 395 patients (n = 194 clinically suspected infection; n = 201 controls) were enrolled. Blood samples were collected at enrolment. The accuracy of calprotectin to identify bacterial infections, and to predict and identify sepsis and mortality was analysed. These endpoints were determined by a panel of experts. RESULTS: The Area Under the Receiver Operating Characteristic (AUROC) of calprotectin for detecting bacterial infections was 0.90. For sepsis within 72 h, calprotectin's AUROC was 0.83. For 30-day mortality it was 0.78. In patients with diabetes, calprotectin had an AUROC of 0.94 for identifying bacterial infection. CONCLUSIONS: Calprotectin showed notable accuracy for all endpoints. Using calprotectin in the emergency department could improve diagnosis and management of severe infections, in combination with current biomarkers. CLINICAL TRIAL REGISTRATION NUMBER: DRKS00020521.

Serum Levels of S100A8/A9 as a Biomarker of Disease Activity in Patients with IgA Vasculitis.

S100A8/A9 protein is a well-known marker of disease activity or severity in many autoimmune and autoinflammatory diseases, but there have not been many studies about the role of S100A8/A9 in IgA vasculitis (IgAV). The aim of our study was to evaluate S100A8/A9 as a possible biomarker of activity in IgAV. We measured the serum levels of S100A8/A9 in pediatric patients with IgA vasculitis at the onset of the disease, after three months, and after six months. We compared these levels between patients with active disease, remission, and a control group, and assessed their correlation with disease activity and other markers of inflammation. Patients with active disease had significantly higher levels of serum S100A8/A9 (median ± SD) than those in the control group at the beginning of the disease (5740 ± 3157 ng/mL vs. 1447 ± 858.3 ng/mL; p < 0.0001), but also three months and six months after disease onset (p < 0.001). There was a positive correlation between S100A8/A9 serum levels and disease activity (p = 0.0003). Patients with active disease had significantly higher levels of S100A8/A9 than those in remission three months after disease onset (p = 0.0260). There was a correlation between S100A8/A9 and C-reactive protein, the C3 component of complement, ferritin, and fibrinogen. Serum levels of S100A8/A9 were also higher in patients with greater skin areas covered with rash. We demonstrated that serum levels of S100A8/A9 correlated well with disease activity and other biomarkers of inflammation in children with IgAV. According to our results, serum S100A8/A9 may be a good indicator of active disease in IgAV.

Combination of leucine-rich alpha-2 glycoprotein and fecal markers detect Crohn's disease activity confirmed by balloon-assisted enteroscopy.

BACKGROUND/AIMS: Endoscopic activity confirmed by enteroscopy is associated with poor clinical outcome in Crohn's disease (CD). We investigated which of the existing biomarkers best reflects endoscopic activity in CD patients including the small bowel, and whether their combined use can improve accuracy. METHODS: One hundred and four consecutive patients with ileal and ileocolonic type CD who underwent balloon-assisted enteroscopy (BAE) from October 2021 to August 2022 were enrolled, with clinical and laboratory data prospectively collected and analyzed. RESULTS: Hemoglobin, platelet count, C-reactive protein, leucine-rich alpha-2 glycoprotein (LRG), fecal calprotectin, and fecal hemoglobin all showed significant difference in those with ulcers found on BAE. LRG and fecal calprotectin showed the highest areas under the curve (0.841 and 0.853) for detecting ulcers. LRG showed a sensitivity of 78% and specificity of 80% at a cutoff value of 13 µg/mL, whereas fecal calprotectin showed a sensitivity of 91% and specificity of 67% at a cutoff value of 151 µg/g. Dual positivity for LRG and fecal calprotectin, as well as LRG and fecal hemoglobin, both predicted ulcers with an improved specificity of 92% and 100%. A positive LRG or fecal calprotectin/hemoglobin showed an improved sensitivity of 96% and 91%. Positivity for LRG and either of the fecal biomarkers was associated with increased risk of hospitalization, surgery, and relapse. CONCLUSIONS: The biomarkers LRG, fecal calprotectin, and fecal hemoglobin can serve as noninvasive and accurate tools for assessing activity in CD patients confirmed by BAE, especially when used in combination.

Evaluation of bowel wall flow by color Doppler ultrasound in the assessment of inflammatory bowel disease activity in pediatric patients / Avaliação do fluxo parietal pela ultrassonografia com Doppler em cores no diagnóstico de atividade na doença inflamatória intestinal em pacientes pediátricos

Abstract Objective: To assess inflammatory bowel disease (IBD) activity with Doppler ultrasound in pediatric patients, comparing the accuracy of the ultrasound findings with that of the concentrations of fecal calprotectin (FC). Materials and Methods: In a consecutive series, we evaluated 53 examinations of 44 pediatric patients seen between 2014 and 2020: 28 with Crohn's disease, 15 with ulcerative colitis, and one with IBD unclassified. The diagnosis of IBD was made in accordance with the Porto criteria. The alteration studied in the greatest detail was bowel wall flow, which was classified by the lead investigator and two pediatric radiologists, all of whom were blinded to the FC concentrations and the other ultrasound findings. Bowel wall flow was categorized as low if there were up to 2 Doppler ultrasound signals/cm2, moderate if there were 3-5 signals/cm2, and high if there were more than 5 signals/cm2. Results: The agreement among the radiologists was substantial (kappa = 0.73). In cases in which ultrasound showed low bowel wall flow, the median FC concentration was 92 µg/g (interquartile range, 33-661 µg/g), whereas it was 2,286 µg/g (interquartile range, 1,728-5,612 µg/g) in those in which ultrasound showed high bowel wall flow. In the sample as a whole, the sensitivity and specificity of ultrasound was 89.7% and 92.0%, respectively, for the detection of inflammatory activity; 95.5% and 90.9%, respectively, for the detection of Crohn's disease; and 81.3% and 100.0%, respectively, for the detection of ulcerative colitis. Conclusion: Ultrasound of the bowel wall showed a strong correlation with FC concentrations in the assessment of inflammatory activity in pediatric patients with IBD.

Resumo Objetivo: Avaliar a atividade da doença inflamatória intestinal (DII) por ultrassonografia (US) com Doppler em cores, comparada à concentração de calprotectina fecal (CF) em pacientes pediátricos. Materiais e Métodos: Em uma série consecutiva, no período entre 2014 e 2020, foram avaliados 53 exames de 44 pacientes pediátricos: 28 casos de doença de Crohn, 15 de colite ulcerativa e um de colite indeterminada. O diagnóstico da DII foi feito pelos critérios de Porto. O fluxo parietal foi a alteração estudada mais detalhadamente e classificada pelo pesquisador principal e por dois radiologistas pediátricos cegados aos valores de CF e de US Doppler. Baixo fluxo parietal foi definido pela captação de até 2 sinais de US Doppler/cm2, fluxo moderado entre 3 e 5 sinais/cm2 e alto fluxo mais de 5 sinais/cm2. Resultados: Houve concordância substancial entre os radiologistas (kappa = 0,73). Nos exames com baixo fluxo parietal a CF média foi 92 μg/g (intervalo interquartil: 33-661 μg/g) e nos exames com alto fluxo a CF média foi 2.286 μg/g (intervalo interquartil: 1.728-5.612 μg/g). Na amostra total, a US demonstrou sensibilidade de 89,7% e especificidade de 92,0% para detecção da atividade inflamatória, 95,5% e 90,9% na doença de Crohn e 81,3% e 100,0% na colite ulcerativa, respectivamente. Conclusão: Houve forte correlação entre a US da parede intestinal e os valores da concentração de CF na avaliação da atividade inflamatória na DII de pacientes pediátricos.

Narrow band imaging as a noninvasive tool for evaluation of the severity of inflammation in patients with ulcerative colitis in Kafr-El-Sheikh university, Egypt.

Objectives: To analyse the effectiveness of narrow-band imaging in determining the severity of inflammation in ulcerative colitis patients in relation to histological activity. Method: The cross-sectional study was conducted at Kafrelsheikh University Hospital, Egypt, from June 2021 to May 2022, and comprised patientsregardless of age and gender who had ulcerative colitis and visited the endoscopy unit. After taking detailed history, the patients were subjected to physical examination and laboratory tests which included complete blood count, international normalised ratio, erythrocyte sedimentation rate and faecal calprotectin. Colonoscopy with narrow-band imaging, biopsy and histopathological assessment were done as part of clinical evaluation. Data was analysed using SPSS 20. RESULTS: Of the 100 patients, 55(55%) were male and 45(45%) were females. The overall mean age was 33.72±10.29 years (range: 11-56 years). There were 73(73%) patients who were married, 12(12%) had positive family history and 19(19%) were smokers. Besides, 18(18%) patientsreceived biological therapy. There wassignificant positive correlation between histopathological and endoscopic scores(p<0.05). Also, there was a significant positive correlation between histopathologicalscore and findings of narrow-band imaging (p<0.05). There were 10(10%) patients who were found to have dysplasia that was not diagnosed with colonoscopy. CONCLUSIONS: Narrow-band imaging was found to have a significant correlation with the histologically determined degree of inflammation.

Calprotectin in chronic rhinosinusitis eosinophil extracellular traps.

BACKGROUND: Calprotectin is an antimicrobial peptide primarily secreted by neutrophils. Furthermore, calprotectin secretion increases in patients with chronic rhinosinusitis (CRS) with polyps (CRSwNP) and positively correlates with neutrophil markers. However, CRSwNP is known to be associated with type 2 inflammation related to tissue eosinophilia. Therefore, the authors investigated calprotectin expression in eosinophils and eosinophil extracellular traps (EETs) and explored the associations between tissue calprotectin and the clinical findings of patients with CRS. METHODS: A total of 63 patients participated, and patients diagnosed with CRS were classified based on the Japanese Epidemiological Survey of Refractory Eosinophilic Chronic Rhinosinusitis (JESREC) score. The authors performed hematoxylin and eosin staining, immunohistochemistry, immunofluorescence with calprotectin, myeloperoxidase (MPO), major basic protein (MBP), and citrullinated histone H3 with the participant's tissues. Finally, correlations between calprotectin and the clinical data were examined. RESULTS: Calprotectin-positive cells are co-localized not only in MPO-positive cells but also in MBP-positive cells in human tissues. Calprotectin was also involved in EETs and neutrophil extracellular traps. The number of calprotectin-positive cells in the tissue was positively correlated with the number of tissue and blood eosinophils. In addition, calprotectin in the tissue is associated with the olfactory function, Lund-Mackay computed tomography score, and JESREC score. CONCLUSIONS: Calprotectin, known to be secreted by neutrophils, in CRS was also expressed in eosinophils. In addition, calprotectin, which functions as an antimicrobial peptide, may play an important role in the innate immune response based on its EET involvement. Therefore, calprotectin expression could reflect as a disease severity biomarker for CRS.

Evaluation of bowel wall flow by color Doppler ultrasound in the assessment of inflammatory bowel disease activity in pediatric patients.

Objective: To assess inflammatory bowel disease (IBD) activity with Doppler ultrasound in pediatric patients, comparing the accuracy of the ultrasound findings with that of the concentrations of fecal calprotectin (FC). Materials and Methods: In a consecutive series, we evaluated 53 examinations of 44 pediatric patients seen between 2014 and 2020: 28 with Crohn's disease, 15 with ulcerative colitis, and one with IBD unclassified. The diagnosis of IBD was made in accordance with the Porto criteria. The alteration studied in the greatest detail was bowel wall flow, which was classified by the lead investigator and two pediatric radiologists, all of whom were blinded to the FC concentrations and the other ultrasound findings. Bowel wall flow was categorized as low if there were up to 2 Doppler ultrasound signals/cm2, moderate if there were 3-5 signals/cm2, and high if there were more than 5 signals/cm2. Results: The agreement among the radiologists was substantial (kappa = 0.73). In cases in which ultrasound showed low bowel wall flow, the median FC concentration was 92 µg/g (interquartile range, 33-661 µg/g), whereas it was 2,286 µg/g (interquartile range, 1,728-5,612 µg/g) in those in which ultrasound showed high bowel wall flow. In the sample as a whole, the sensitivity and specificity of ultrasound was 89.7% and 92.0%, respectively, for the detection of inflammatory activity; 95.5% and 90.9%, respectively, for the detection of Crohn's disease; and 81.3% and 100.0%, respectively, for the detection of ulcerative colitis. Conclusion: Ultrasound of the bowel wall showed a strong correlation with FC concentrations in the assessment of inflammatory activity in pediatric patients with IBD.

Objetivo: Avaliar a atividade da doença inflamatória intestinal (DII) por ultrassonografia (US) com Doppler em cores, comparada à concentração de calprotectina fecal (CF) em pacientes pediátricos. Materiais e Métodos: Em uma série consecutiva, no período entre 2014 e 2020, foram avaliados 53 exames de 44 pacientes pediátricos: 28 casos de doença de Crohn, 15 de colite ulcerativa e um de colite indeterminada. O diagnóstico da DII foi feito pelos critérios de Porto. O fluxo parietal foi a alteração estudada mais detalhadamente e classificada pelo pesquisador principal e por dois radiologistas pediátricos cegados aos valores de CF e de US Doppler. Baixo fluxo parietal foi definido pela captação de até 2 sinais de US Doppler/cm2, fluxo moderado entre 3 e 5 sinais/cm2 e alto fluxo mais de 5 sinais/cm2. Resultados: Houve concordância substancial entre os radiologistas (kappa = 0,73). Nos exames com baixo fluxo parietal a CF média foi 92 µg/g (intervalo interquartil: 33-661 µg/g) e nos exames com alto fluxo a CF média foi 2.286 µg/g (intervalo interquartil: 1.728-5.612 µg/g). Na amostra total, a US demonstrou sensibilidade de 89,7% e especificidade de 92,0% para detecção da atividade inflamatória, 95,5% e 90,9% na doença de Crohn e 81,3% e 100,0% na colite ulcerativa, respectivamente. Conclusão: Houve forte correlação entre a US da parede intestinal e os valores da concentração de CF na avaliação da atividade inflamatória na DII de pacientes pediátricos.

Fecal Calprotectin Assay at an Early Stage of Treatment Can Be Used as a Surrogate Marker to Predict Clinical Remission and Mucosal Healing in Pediatric Crohn's Disease.

Purpose: This study evaluated the predictive role of fecal calprotectin (FC) measured at an early stage of treatment for monitoring clinical remission (CR) after six months and endoscopic remission (ER) after one year of treatment in pediatric Crohn's disease (CD). Methods: This retrospective study included 45 patients who simultaneously underwent ileocolonoscopy and FC testing during follow-up. FC levels were measured before and after six weeks of treatment. CR was assessed after six months of treatment using Pediatric Crohn' s Disease Activity Index and acute-phase reactants. ER was assessed after one year using the Simple Endoscopic Score for Crohn's Disease. Results: Twenty-nine (64.4%) patients used oral prednisolone for remission induction and 16 (35.6%) patients used anti-tumor necrosis factor-alpha. Thirty (66.7%) patients achieved CR, while 24 (53.3%) achieved ER. The FC level measured after six weeks of treatment could predict CR (χ2=9.15, p=0.0025) and ER (χ2=12.31, p=0.0004). The δFC could predict CR (χ2=7.91, p=0.0049), but not ER (χ2=1.85, p=0.1738). With a threshold of ≤950.4 µg/g, FC at week six could predict CR with 76.7% sensitivity and 73.3% specificity. The area under the curve (AUC) was 0.769 (standard error 0.0773, p=0.0005). The same threshold predicted ER with 87.5% sensitivity and 71.4% specificity. The AUC was 0.774 (standard error 0.074, p=0.0002). Conclusion: FC assay at an early stage of treatment can be used as a surrogate marker to predict CR and mucosal healing in pediatric CD.

Plasma calprotectin as a biomarker of ultrasound synovitis in rheumatoid arthritis patients receiving IL-6 antagonists or JAK inhibitors.

Objectives: To analyse the accuracy of plasma calprotectin in patients with rheumatoid arthritis (RA) receiving monoclonal antibodies against IL-6 receptors (anti-rIL-6) or JAK inhibitors (JAKis) in detecting ultrasound (US) synovitis and compare it with acute phase reactants [high-sensitivity C-reactive protein (hs-CRP) and ESR]. Methods: An observational cross-sectional study of RA patients receiving anti-rIL-6 (tocilizumab or sarilumab) or JAKi, (baricitinib or tofacitinib) was made. Plasma calprotectin for the diagnosis of US synovitis [synovial hypertrophy grade (SH) ⩾ 2 plus power Doppler signal (PD) ⩾ 1] was analysed using receiver operating characteristic curves (ROCs). The performance of ESR and hs-CRP was also studied. The three ROC curves were compared to determine which had the highest discriminatory power. Associations between plasma calprotectin and US scores were made using correlation analysis. Results: Sixty-three RA patients were included. Mean plasma calprotectin levels were significantly higher in patients with US synovitis than in those without (0.89 ± 0.85 vs 0.30 ± 0.12 µg/ml; p = 0.0003). A moderate correlation between calprotectin and all US scores (HS score Rho = 0.479; PD score Rho = 0.492; and global score Rho = 0.495) was found. The discriminatory capacity of plasma calprotectin showed an AUC of 0.795 (95% CI: 0.687-0.904). The AUC of hs-CRP and ESR was 0.721 and 0.564, respectively. hs-CRP serum levels showed a low positive correlation with the three US scores (Rho < 0.40). After analysis according to the drugs administered, the correlation disappeared in patients receiving anti-rIL-6. Conclusion: Plasma calprotectin may be a sensitive biomarker of synovial inflammation in RA patients treated with anti-rIL-6 or JAKi.

Associations between Gut Microbiota and Intestinal Inflammation, Permeability and Damage in Young Malawian Children.

BACKGROUND: Environmental enteric dysfunction (EED) is common in low- and middle-income countries and associated with childhood undernutrition. The composition of gut microbiota has been implicated in the pathogenesis of EED. Our aim was to assess the associations between gut microbiota and EED biomarkers in rural Malawian children. We hypothesized that there would be an inverse association between microbiota maturity and diversity and fecal concentrations of EED biomarkers. METHODS: We used data from fecal samples collected at 6, 18 and 30 months from 611 children who were followed up during a nutrition intervention trial. The primary time point for analysis was 18 months. Microbiota data were obtained through 16S rRNA sequencing and variables included microbiota maturity and diversity, phylogenetic dissimilarity and relative abundances of individual taxa. EED biomarkers included calprotectin (marker of inflammation), alpha-1 antitrypsin (intestinal permeability) and REG1B (intestinal damage). RESULTS: There was an inverse association between microbiota maturity and diversity and fecal concentrations of all 3 EED biomarkers at 18 months (p≤0.001). The results were similar at 30 months, while at 6 months inverse associations were found only with calprotectin and alpha-1 antitrypsin concentrations. At 18 months, EED biomarkers were not associated with phylogenetic dissimilarity, but at 6 and 30 months several associations were observed. Individual taxa predicting EED biomarker concentrations at 18 months included several Bifidobacterium and Enterobacteriaceae taxa as well as potentially displaced oral taxa. CONCLUSIONS: Our findings support the hypothesis of an inverse association between microbiota maturity and diversity and EED in rural Malawian children.

Chronic childhood undernutrition is an important public health concern that affects about 150 million children, mostly in low- and middle-income countries. Undernutrition is caused by insufficient nutrient intake and frequent infections, but there are also other underlying factors. One of these is a condition called environmental enteric dysfunction (EED), which is characterized by intestinal inflammation and damage without apparent clinical symptoms. EED is thought to be caused by the ingestion of pathogenic bacteria that leads to changes in the intestine such as increased permeability and decreased absorptive capacity. This might make the intestinal wall vulnerable to bacterial invasion and reduce the absorption of nutrients. Besides potentially pathogenic bacteria, there are many commensal bacteria in the gastrointestinal tract that have beneficial functions and that interact with the immune system. The aim of our study was to assess the associations between all these bacteria, that is the intestinal microbiota and biomarkers of EED. We used data from fecal samples collected from young children participating in a nutrition intervention trial in rural Malawi. Our findings support an inverse association between the diversity and maturity of the intestinal microbiota and biomarkers of EED. Additionally, we identified the differences at the level of individual bacterial taxa (groups of bacteria defined by genetic similarity) between participants with different levels of EED biomarkers. Due to the type of study, we cannot determine whether the observed associations represent a causal relationship between the intestinal microbiota and EED. This as well as the exact mechanisms behind these associations should be assessed in further studies.

Fecal calprotectin levels used as a noninvasive method for screening for chronic gastritis in pediatric patients. A descriptive study

ABSTRACT BACKGROUND: Gastritis consists of inflammation of the gastric mucosa and is one of the main causes of dyspeptic symptoms in children. OBJECTIVE: To investigate the presence of inflammation by evaluating fecal calprotectin (FC) in children diagnosed with chronic gastritis. DESIGN AND SETTING: Descriptive study in Pediatric Gastroenterology Department of Ondokuz Mayis University Hospital in Turkey. METHODS: Between January 2016 and July 2018, FC levels were compared retrospectively in children with chronic gastritis (histopathology-based diagnosis), patients with inflammatory bowel disease (IBD) and healthy children. RESULTS: A total of 67 chronic gastritis patients (61.2% girls) with a mean age of 13.09 ± 3.5 years were evaluated. The mean FC levels were 153.4 μg/g in the chronic gastritis group, 589.7 μg/g in the IBD group and 43.8 μg/g in the healthy group. These levels were higher in chronic gastritis patients than in healthy individuals (P = 0.001) and higher in IBD patients than in the other two groups (P < 0.001). The FC level in the patients with chronic active gastritis (156.3 μg/g) was higher than in those with chronic inactive gastritis (150.95 μg/g) (P = 0.011). Among the patients with chronic active gastritis, the FC level was significantly higher in Helicobacter pylori-positive individuals than in negative individuals (P = 0.031). CONCLUSION: We confirmed the association between increased FC and chronic gastritis. Elevated FC levels may be seen in patients with chronic active gastritis. In order to be able to use FC as a screening tool for chronic gastritis, further studies in a larger study group are needed.

Plasma calprotectin in the emergency department: a potential clinical biomarker for patients with infectious diseases.

Increased levels of plasma calprotectin are reported in patients with infectious diseases. However, the clinical usefulness of calprotectin as a biomarker to identify patients with infectious diseases in the emergency department (ED) setting has not been investigated. To study the ability of calprotectin to discriminate patients with acute infectious diseases and dyspnea from patients with other causes of acute dyspnea in the ED setting. Patients aged ≥18 years seeking ED during daytime on weekdays between March 2013 and July 2018, with acute dyspnea, were included. Participants (n = 1287) were triaged according to Medical Emergency Triage and Treatment System-Adult score (METTS-A) or Rapid Emergency Triage and Treatment System (RETTS), and blood samples were collected. The association between calprotectin and other markers of infectious diseases, i.e. biomarkers (CRP, leucocytes) and body temperature, was studied. The predictive value of calprotectin for the outcome of acute infection was evaluated with receiver operating characteristic (ROC) analysis. Univariate cross-sectional regression showed significant associations between calprotectin and leucocytes, CRP and body temperature. Patients with severe infections including pneumonia (n = 119) had significantly higher concentrations of calprotectin compared to patients with heart failure (n = 162) or chronic obstructive pulmonary disease (n = 183). When tested for the outcome of acute infection (n = 109), the area under the ROC curve (AUROC) was for CRP 0.83 and for calprotectin 0.78. Plasma calprotectin identifies infectious diseases in ED patients with acute dyspnea, and the clinical usefulness of Calprotectin in the ED has to be further studied.

The comparison of the three assays for determination of fecal calprotectin in inflammatory bowel disease.

INTRODUCTION: Fecal calprotectin is a biomarker for monitoring inflammatory bowel disease (IBD) activity. Our aim, therefore, was to evaluate two new assays, the point of care test Quantum Blue and the Liaison Calprotectin with respect to the Calprest, commonly used assay, and to determine their performance for IBD diagnosis. MATERIALS AND METHODS: We included 73 prospective patients with IBD. Fecal calprotectin was measured and analysed with the routine Calprest assay and two recently introduced assays, the Quantum Blue and the Liaison Calprotectin. Furthermore, we compared the results by Bland and Altman analysis, and Passing-Bablok regression. RESULTS: We observed no difference in median calprotectin values obtained by the Calprest (94.6 µg/g, 95%CI 66.5 to 166.1) and Liaison assay (101.0 µg/g, 95%CI 48.1 to 180.1) whereas significantly higher concentrations were obtained with the Quantum Blue assay (240.0 µg/g, 95%CI 119.9 to 353.2). The mean absolute and relative difference between the Calprest and Quantum Blue methods was statistically significant (- 162.3 µg/g and- 143.1%). Mean absolute difference between the Calprest and Liaison calprotectin methods was positive (2.2 µg/g). The agreement between assays revealed that Quantum Blue and Calprest have fair agreement with Kappa coefficient of 0.38 (95%CI 0.26 to 0.51). Liaison Calprotectin and Calprest revealed moderate agreement with a weak Kappa coefficient of 0.47 (95%CI 0.32 to 0.62). CONCLUSION: Clinicians should be aware of these differences between the assays and avoid comparison of their respective results.

Fecal Calprotectin Is Increased in Stroke.

BACKGROUND: While there have been major advances in unveiling the mechanisms comprising the ischemic cascade of CNS, stroke continues to be a significant burden. There is a need to extend the focus toward peripheral changes, and the brain-gut axis has recently gained much attention. Our study aimed to evaluate gut inflammation and its association with blood variables in stroke using fecal calprotectin (FC). METHODS: Fecal samples were obtained from 27 stroke patients and 27 control subjects. FC was quantitatively measured using a commercial ELISA. Laboratory data on the fecal sample collection were also collected, including CBC, ESR, glucose, creatinine, total protein, albumin, transaminases, and CRP. RESULTS: There was a significant increase in FC levels in stroke patients compared to the controls. Furthermore, FC in stroke patients was negatively correlated with the Glasgow Coma Scale. Moreover, FC in stroke patients was positively correlated with CRP and negatively correlated with lymphocyte count and albumin. CONCLUSIONS: Our findings show that increased FC is associated with consciousness and systemic response in stroke and warrants further studies to elucidate the usefulness of FC in the management of stroke.

The feasibility of measuring calprotectin from a throat swab as a marker of infections caused by group A streptococcus: a case-control feasibility study.

BACKGROUND: Most people with sore throat do not benefit from antibiotic treatment, but nearly three-quarters of those presenting in primary care are prescribed antibiotics. A test that is predictive of bacterial infection could help guide antibiotic prescribing. Calprotectin is a biomarker of neutrophilic inflammation, and may be a useful marker of bacterial throat infections. AIM: To assess the feasibility of measuring calprotectin from throat swabs, and assess whether individuals with sore throats likely to be caused by streptococcal infections have apparently higher throat calprotectin levels than other individuals with sore throat and healthy volunteers. DESIGN & SETTING: A proof of concept case-control study was undertaken, which compared primary care patients with sore throats and healthy volunteers. METHOD: Baseline characteristics and throat swabs were collected from 30 primary care patients with suspected streptococcal sore throat, and throat swabs were taken from 10 volunteers without sore throat. Calprotectin level determination and rapid antigen streptococcal testing were conducted on the throat swab eluents. Calprotectin levels in the following groups were compared: volunteers without a sore throat; all patients with a sore throat; patients with a sore throat testing either negative or positive for streptococcal antigen; and those with lower and higher scores on clinical prediction rules for streptococcal sore throat. RESULTS: Calprotectin was detected in all throat swab samples. Mean calprotectin levels were numerically higher in patients with sore throat compared with healthy volunteers, and sore throat patients who had group A streptococci antigen detected compared with those who did not. CONCLUSION: Calprotectin can be measured from throat swab samples and levels are consistent with the hypothesis that streptococcal infection leads to higher throat calprotectin levels. This hypothesis will be tested in a larger study.

A novel prognostic biomarker for recurrent aphthous stomatitis: calprotectin.

OBJECTIVE: This study aims to investigate whether there is a correlation between serum calprotectin concentration and oral ulcer activity in patients with active recurrent aphthous stomatitis. METHOD: This prospective, cross-sectional, historical cohort study included 52 patients (26 with active lesions and 26 with inactive lesions) with recurrent aphthous stomatitis and 27 healthy volunteers. The relationship between the groups was statistically evaluated in terms of serum calprotectin level and other inflammatory parameters. RESULTS: The median calprotectin value of the active lesion group (80.5 ng/ml) was significantly higher than that of the control group (55.4 ng/ml, p = 0.002). Furthermore, the median calprotectin value of the active lesion group was significantly higher than that of the inactive lesion group (72.5 ng/ml, p = 0.017). CONCLUSION: In patients with recurrent aphthous stomatitis and active ulcer lesions, the serum calprotectin value can be used as a reliable and robust prognostic marker for inflammation.

Nivel de calprotectina fecal en niños sanos menores de 4 años

Introducción: la calprotectina es una proteína del citoplasma de neutrófilos, con propiedades bacteriostáticas. Aumenta en materia fecal en procesos inflamatorios de la mucosa intestinal, siendo un marcador de inflamación. Su cuantificación es un método no invasivo y estable en su procesamiento. Presenta una alta sensibilidad para diferenciar entre enfermedad orgánica y funcional y tiene alto valor predictivo positivo para hallazgo de lesiones a nivel endoscópico. Presenta baja especificidad para diferenciar la etiología de la inflamación. El nivel de calprotectina fecal (CF) es variable de acuerdo a la edad. En niños menores de 4 años se han encontrado concentraciones mayores que en otras edades. Esto se puede explicar por factores como: mayor migración de neutrófilos en la mucosa durante el desarrollo de tolerancia oral, regulación de la microbiota intestinal, inmadurez de la barrera epitelial y el ambiente que rodea al niño con permanente estímulo para su sistema inmunológico. Por lo que no se ha podido establecer el cut off en esta franja etaria. Objetivo: conocer los valores de la CF en niños sanos menores de 4 años para determinar valor normal o cut off. Determinarlo según grupos etarios y por sexo. Método: estudio descriptivo con componente analítico. Se obtiene la muestra de los niños que realizan el coproparasitario para el ingreso escolar. Catalogados como sanos: sin antecedentes personales de enfermedad crónica, sin síntomas por lo menos cuatro semanas anteriores a la toma de la muestra. Buen crecimiento según las curvas OMS. Distribuidos en diferentes regiones geográficas de la zona metropolitana de Montevideo. Se utilizó el kit RIDASCREEN®. Resultados: fueron analizadas 155 muestras; 73 de sexo femenino y 82 sexo masculino. Se distribuyeron en grupos etarios: menor de 1 año (n=11); 1 a 2 años (n=36); 2 a 3 años (n=45); 3 a 4 años (n=63). La mediana fue de 152,2 mg/kg (percentil 50). Rango de 4,1 a 1944 mg/kg. Nuestro cut off fue de 954 mg/kg (percentil 95). Se aplica Wilcoxon test p=0,68 y test Kruskal Wallis p=0,06, sin diferencias significativas entre sexo y grupos etarios respectivamente. Conclusiones: es el primer estudio de CF en una población de niños sanos menores de 4 años en Latinoamérica. La mediana encontrada es mayor que la habitual en niños mayores de 4 años y adultos. Obtuvimos un límite normal de 954 mg/kg. No se encontró diferencia en la concentración según sexo o rango etario. Limitaciones de nuestro estudio: el tamaño y la obtención de la muestra corresponde solamente al departamento de Montevideo.

Summary: Introduction: calprotectin is a protein with bacteriostatic properties, found in cytoplasmic neutrophils. Concentration in fecal matter increases in inflammatory processes of the intestinal mucous membrane, being it an inflammation marker. Quantification involves a non-invasive method, stable during processing. Fecal calprotectin concentration is highly sensitive to differentiate between organic and functional disease and has a high positive predictive value for the finding of endoscopic lesions. It evidences low specificity to distinguish the inflammation's etiology. Fecal calprotectin concentration (FCC) varies according to age. Higher concentrations have been found in children younger than 4 years old, what may be explained by the following factors: greater migration of neutrophils in the mucous during the development of oral tolerance, regulation of gut microbiota, non-mature epithelial tissue and the context the child grows in as a permanent stimulation for his immune system. Thus, cut off has not been established yet for this age group. Objective: to learn about FCC in healthy children younger than 4 years old to determine normal values or cut off. To determine values according to age and sex. Method: descriptive study with an analytic component. A sample of the children who undergo a coproparsitary test upon entering school is taken. A healthy child is regarded to: have no personal history of chronic disease, no symptoms at least in the 4 weeks prior to the sample being taken. Good growth is defined as per the WHO curves. Samples included came from different regions in the Montevideo metropolitan area. The RIDASCREEN® kit was used. Results: 155 samples were analysed, 73 of them corresponding to boys and 82 to girls. As to age: younger than 1 year old (n= 11), from 1 to 2 years old (n= 36), from 2 to 3 years old (n= 45), from 3 to 4 years old (n= 63). Median concentration was 152.2 mg/kg (Percentile 50). Range 4.1 to 1944 mg/kg. Our cut off was 954 mg/kg (percentile 95). Wolcoxon test was applied (p= 0.68) and Kruskal Wallis test (p=0.06), no significant differences were found between sex and age groups respectively. We obtained a normal limit of 954 mg/Kg. No difference was found in the concentration for different sex or age. Limitations of our study result from the size of the sample and the fact that all samples correspond to Montevideo.

Introdução: a calprotectina é uma proteína neutrofílica citoplasmática com propriedades bacteriostáticas. Ela aumenta na matéria fecal durante os processos inflamatórios da mucosa intestinal, e é um marcador de inflamação. Pode ser quantificada utilizando um método não invasivo, estável em seu processamento. Apresenta alta sensibilidade para diferenciar entre doença orgânica e funcional e tem alto valor preditivo positivo para encontrar lesões no nível endoscópico. Apresenta baixa especificidade para diferenciar a etiologia da inflamação. O nível de calprotectina fecal (CF) varia de acordo com a idade, e para o caso de crianças com menos de 4 anos ela teve concentrações mais elevadas, o que pode ser explicado por fatores tais como o aumento da migração de neutrófilos na mucosa durante o desenvolvimento de tolerância oral, regulação da microbiota intestinal, imaturidade da barreira epitelial e ambiente que envolve a criança como estímulo permanente do sistema imunológico. Portanto, não foi possível estabelecer o corte nesta faixa etária. Objetivo: conhecer os valores da FC em crianças saudáveis menores de 4 anos para determinar o valor normal ou o de corte de acordo com as faixas etárias e sexo. Método: estudo descritivo com componente analítico. A amostra foi obtida das crianças que realizaram o estudo coproparasitológico para a admissão escolar. Crianças catalogadas como saudáveis: sem histórico de doença crônica, sem sintomas durante pelo menos 4 semanas antes de experimentar a amostra. Bom crescimento: de acordo com as curvas da OMS. Distribuído em diferentes regiões geográficas da área metropolitana de Montevidéu. Se utilizou o kit RIDASCREEN®. Resultados: foram analisadas 155 amostras: 73 de mulheres e 82 de homens distribuídos em grupos etários: com menos de 1 ano (n = 11), 1 a 2 anos (n = 36), 2 a 3 anos (n = 45), 3 a 4 anos (n = 63). A mediana foi de 152,2 mg / kg (percentil 50). Faixa de 4,1 a 1944 mg / kg. Nosso corte foi 954mg / Kg (percentil 95). Aplicou-se o teste de Wilcoxon p = 0,68 e o teste de Kruskal Wallis p = 0,06, e não observamos diferenças significativas entre os grupos por sexo ou faixa etária, respectivamente. Conclusões: este foi o primeiro estudo de FC numa população de crianças saudáveis com menos de 4 anos de idade na América Latina. A mediana encontrada é maior que a usual em crianças com mais de 4 anos e em adultos. Obtivemos um limite normal de 954mg / Kg. Nenhuma diferença foi encontrada na concentração dependendo do sexo ou faixa etária. As limitações do nosso estudo são o tamanho da amostra e o fato de que a amostra foi obtida apenas de Montevidéu.

A Study of Fecal Calprotectin in Obese Children and Adults.

BACKGROUND: Obesity is a complex, medical condition causally contributing to many chronic diseases and a number of efforts have been made to find the associated markers for novel prevention and treatment of obesity. Our study was to evaluate the relationship between gut immune response and obesity and overweight with use of fecal calprotectin (FC) both in adult and children groups. METHODS: Fecal samples were obtained from 74 subjects: 14 non-obese and overweight children (PN), 13 obese and overweight children (PO), 20 non-obese and overweight adults (AN), and 27 obese and overweight adults (AO). FC was measured using a commercial Legend Max quantitative enzyme-linked immunosorbent assay (BioLegend). Mann-Whitney U-test was used for statistical analysis. RESULTS: Median FC concentration was 7.9 µg/g (range, 1.9-28.9 µg/g) for PN, 5.0 µg/g (range, 2.6-29.6 µg/g) for PO, 9.5 µg/g (range, 0.8-28.9 µg/g) for AN, and 10.0 µg/g (range, 1.6-25.6 µg/g) for AO, respectively. In both adults and children age groups, the FC showed no statistically significant difference between AO and AN or PO and PN. However, FC showed statistically significant difference (P<0.05) between AO and PO while not significant between AN and PN. CONCLUSION: FC level in AO was significantly higher than that in PO, suggestive of different pathophysiologic mechanism between children obesity and adults obesity.

Calprotectin (S100A8/A9) should preferably be measured in EDTA-plasma; results from a longitudinal study of patients with rheumatoid arthritis.

Calprotectin (S100A8/A9), a protein expressed in neutrophils and monocytes/macrophages in circulation and inflamed tissue, is associated with measures of disease activity in rheumatoid arthritis (RA) patients both when measured in ethylenediaminetetraacetic acid (EDTA)-plasma and in serum. We wanted to explore if EDTA-plasma or serum should be preferred for calprotectin as a marker of disease activity. Calprotectin was analysed in EDTA-plasma and serum by enzyme-linked immunosorbent assay (ELISA) at baseline in 141 RA patients, starting biologic disease-modifying anti-rheumatic drugs (bDMARDs), and after three months. Differences between plasma and serum levels of calprotectin were assessed by Wilcoxon signed rank test. Variability was assessed by quartile coefficient of dispersion. Spearman's test explored correlations between calprotectin in plasma and serum and between calprotectin (plasma or serum) and clinical/ultrasound (US) measures of disease activity. Bland Altman plots were used for method comparisons. Conventional inflammatory markers were evaluated for comparison. Calprotectin had similar variability when measured in plasma and serum, but there was a significant difference in concentrations between plasma and serum (p < .001). The correlation coefficients at baseline between calprotectin measured in plasma/serum and measures of disease activity were rs = 0.62/0.46 for sum power Doppler score (PD), rs = 0.60/0.48 for assessor's global visual analogue scale (VAS), rs = 0.59/0.43 for sum grey scale (GS) score and rs = 0.47/0.37 for swollen joint count of 32, all p < .001. Similar differences were found after three months. Calprotectin measured in plasma showed the strongest associations with assessments of disease activity, and EDTA-plasma should preferably be used when evaluating disease activity in RA patients.